United State s Enviro nmental Protection A gency

_________________________________________________________________________ 

# Manual for t he C ertification of Laborat ori es Ana l yz i ng Dri nki ng Water Criteriaand Procedures Qua lityAssu ran ce Fi ft h E di t i on

## EPA 815-R-05-004 J anuar y 2005 

# Manual for the C ertification of Laborat ori es Ana l yz i ng Dri nki ng Water Criteriaand Procedures Qua lityAssu ran ce

Fi fth E diti on Supersede s EPA/570/9-90/008, April 1990

EPA 814-B -92-002, September 1992 EPA 570/9-90-008A, October 1991 EPA 815-B -97-001, Ma

rch 1997

US Environmental Protection Agency

Offic e of Water Offic e of Ground Water a nd DrinkingWater Technical Support Center Cincinnati, Ohio 45268 t ap e o n rec ycl e l o go

## ii DI S CL AI M ER T h e U
. S. Env i r on men t alPr ot ec t i on Ag en cy ’ s ( E P A ’ s) O f f i ce of G r ou n d W at era n d D r i n k i n g W at er , i n th e O f f i ce of W a te r, a n d the O ff

ic e o f R e se a rc h a n d D e v e lo p m e n t o f th e U .S . E n v iro n m e n ta l P ro te c tio n A g e n c y p re p a re d this m a n u a l. T h o se O ff

ic e s a s w e ll a s E P A ’s te n R e g io n a l O ff ic e s h a v e r e v ie w e d th is m a n u a l. E P A inte n d s to us e this m a n u a l f o r its o wn use i n cert i f

y i ng l ab o rat o ri es f

o r anal y si s o f

dri nki ng wat er contaminant s. I n ord er t o ass um e p ri m aryenf o rcem ent res p o nsib i l i tyf

o r the d ri nki ng wat er r egul at i o ns, a St at e m ustei ther have avai l ab l e l ab o rat o ry f aci l i t i es, cert i f

i ed by the A d m inistra to r, c a p a b le o f co n d u c ting a n a lyti c a l m e a su re m e n ts o f d rin k ing w a te r c o n ta m ina n ts, o r e sta b lish a n d m a inta in i t s ow n pr og r am f

orc ert i f i ca t i on of

l abor at or i es . St at es wi s h i n g t o a daptth e pr oce du r es a n d c r i t er i a of th i s ma n u alf

or th e i row n c e r t i f

i c a t i onpr og r a m s h ou l d r e v i s e i tt o a c c u r a t e l yr e f l e c ta c c u r a t e l yth e i rS t a t e c e rt i f i c a t i onpr og r a m. Th i s i s a g u i d a n c e m a n u a l a n d n o t a r e g u l a t i o n . I td o e s n o tc h a n g e o rs u b s t i tu t e f o ra n yl e g a l r e q u i r e m e n t
. W h i l e EPA has mad e everyef

f o rtt o ensur e the accuracy of the m anual ’ s d i scus si o n, the o b l i gat i o ns o f the regul at ed co m m uni tyare d e te rm ine d b y the r e le v a n t sta tute s, r e g u la tio n s o r o the r le g a lly b ind ing re q u ire m e n ts. T h e m a n u a l o b v io u sly c a n o n ly r ef l ectthe r egu l at i ons i n pl ace atthi s t i me of i t s prepar at i on. Co ns equen t l y , f or a nydef

i ni t i ve des cr i pt i on of

cu rr entl ega l o b li gat i o ns, the p ub l i c shoul d no t rel y o nl y o n the d i scus si o n i n the m anual
. T hi s m anual i s no t a rul e, i s n o t l e g a lly enf o rceab l e, and do es notco nf er l egal ri ght s o r i m p o se l egal req ui rem ent s up o n a nym em b er o f the pub l i c, Stat es or any o the r F e d e ra l a g e n c y. In the e v e n t o f a c o n fli c t b e tw e e n the d isc u ssio n s in th is m a n u a l an d a n y sta tute o r re g u la tio n , th is d o c u m e n t i s n o t c o n tro lli n g . T h e w o rd “sh o u ld ” in th is m a n u a l d o e s n o t c o n n o te a re q u ire m e n t b u t d o e s ind ic a te E P A ’s stronglypref

erred ap p roac h t o ensur e the q ual i tyof l ab o rat o ry res ul t s. E P A m ay d eci d e t o revi se thi s m anual wi tho ut p u b lic n o tice to re flec t ch a n g e s to it s a p p ro a c h o r to c la rify an d up d a te the tex t. T h e men t i on of

c ommer ci alpr odu ct s i n th i s ma n u aldoe s n otc on s t i tu t e ane n dor s eme n tof th e u s e of

th atpr odu ctby EPA.

## iii A ckno w l ed g m en ts T h i s e di t i on of

th e manu alw as pr epa r ed th r ou g h th e ef f ort s of m an yi n di v i du al s , i n clu di n g r epr es en t at i v es f r om th e U
. S. E nvi ronm ent al P rotect i o n A gency’ s O f f i ce o f G round W at er and D ri nki ng W at er (O G W D W ), O f f i ce o f R esearch and D e v e lo p m e n t (O R D ), R e g io n a l O ff ic e s a n d the S ta te s. It ha s a s its f o u n d a tio n p re v io u s e d iti o n s o f th e m a n u a l. Contr i bu t or s t o th e pr ev i ou s e di t i on s of the m anu ala r e l i s t ed i n EP A docu men t s EP A 815- B
- 97- 001 M ar ch1997, E P A /5 7 0 /9 -9 0 /0 0 8 A p ril 1 9 9 0 , E P A -5 7 0 /9 -8 2 -0 0 2 O c to b e r 1 9 8 2 a n d E P A 6 0 0 /8 -7 8 -0 0 8 M a y 1 9 7 8 . C o n trib u to rs to thi s edi t i on ar e l i s t ed bel ow. M anu al R evi s i on C om mi t t ee: Pau l Be r g e r , OGW DW Ed w a r d Gl i c k , OGW DW Pa tr i c i a Hu rr , OGW DW C a r o l i n e M a d d i n g , OGW DW M i crobi ol og y Su bcom mi t t ee: P a u l S . B e rg e r, C h a ir K r i s t e n Br e n n e r , ORD T e rryC. Cov e r t , ORD M a ryA n n F e i g e , OGW DW Na n c yH Ha l l , UI HL St eph an i e H arr i s , Reg i on 10 M a r g o Hu n t , OEI Ki m b e r l yK L a u r i e , I N DOH Te d Pa s s I I , KY DEP G e n e Ri c e , ORD M a rkRod g e r s , ORD P eggy R y ker, C o nsul t ant L o i s S h a d i x , OGW DW J i m S i n c l a i r , OGW DW Joh n St an dr i dg e, W I SLH Ph i l l i p R Z i l l i n g e r , I N DOH R adi ochemi s t ry Sub comm i t t ee: Jo hn D ehart , D y nco rp J oh n Gr i g g s , OAR Ri c h a r d S h e i b l e y , PA DEP O th er M a n u a lR ev i ew er s: An n M a r i e A l l e n , M A DEP Da l e A l l e y , R I DOH C h e ry l Ba k e r , M E DHS R u s tyBa l d w i n , A R DOH Br e n d a Ba rn e tt , W V DOH A im e e B e n n e tt, W A Ev e Be rryA R DOH D a v e B ing h a m , W A Ge o r g e Bo w m a n , W I L OH El l e n Br a u n - Ho w l a n d , NYS DOH Da v i d Br i e r l e y , M A DEP D o ugl as B rune, R egi o n 7 Jo hn B o urbo n, R egi o n 2 Don a l d Bu c k l e y , M A DPH J o a n Bu rk a rt , L A DOHH Dw a yn e Bu rk h o l d e r , PA DEP I s a Cha mber l ai n , Reg i on 10 Fr e d Ch os k e , CA EL AP A rtC l ark, R egi o n 1 T e rryCov e rt , ORD C h r i s De a n , NM DOH V e r on i c a DeBoe r , AK M aryD i l l on , N JDH SS R o n D o nal y , ID Ch a r l e s Dy e r , NH DES S t a n F a l k , A R DOH C h a r l e s F e l d m a n n , OGW DW R ay Flores, R egi o n 6 Pau l Gr a y , NM DOH St eph an i e H arr i s , Reg i on 10 Ga rryHa w o rth , NH DES Pau l Hi c k e y , NYS DOH M y r o n Ho n d a , HI DOH J a n e J e n s e n , CA EL AP C harl es Jones, Jr. , R egi o n 3 C e c y l i a Ka r c h , VT DOH Ew a Ki n g , R I DOH M i c h a e l Ki tt o , NYS DOH R J Ki y o k a n e , HI DOH S h a r o n Kl u e n d e r , W DNR M ar g ar etK n i g h t , Reg i on 10 G e or g e Ku l a s i n g a m, CA EL AP D on La Fa r a, N V B LC M a ryBe th L a w h o rn , I L EPA A nd rew Li nco f

f , R egi o n 9 Al a n L ov e , AK Ni c h o l a s M a c e l l e t i i , C T DHS De L o i s M a n o r , A R DOH T e d M a rt i n , ORD G eral d M cK enna, R egi o n 2 Jos ephM i erzw i ck i , N JDH SS W i l l i a m Ge o r g e M i l l s , VT DOH C a th yM o o r e , A R DOH J e a n M u n c h , ORD Da v i d M u n c h , OGW DW Th o m a s O’ Co n n e l l , M A DPH

## iv

J i m O’ D e l l , ORD Al i ci a P. O r don a, V A D CLS Pa v i n P a r e k h , NYS DOH Vi r g i n a P a l o m o , OR Os c a rPa n c o rb o , M A DEP B ah manPar s a, N JDH SS G aryP errym an, R egi o n 8 B arryP haroa h, ID M a x Ph i l l i p s , TCEQ Sa nd ra R ad wi n, ID Sa r a Ra i r i c k , AK RobertRi ec k , Reg i on 10 J
. J a n e R o l l , A R DOH I r e n e R o n n i n g , OR P HL Ja ckRuc k man , N V B LC D avi d R uss el l , R egi o n 3 Ei l ee n Sa n der s , V A D CLS T i m Sand ers , R egi o n 6 Ph i l i p S c h l o s s b e r g , CT DPH Ri c h a r d S h e i b l e y , PA DEP Th o m a s S e m k o w , NYS DOH J od yS h oe ma k e r , O RD Sa nd ra Sp ence, R egi o n 8 Ma ryE
. T
. S t a n c a v a g e , MD H MH D avi d Stockt o n, R egi o n 6 M i guel R
. Suarez, Re gi o n 5 M ari l y n T ho rnt o n, R egi o n 4 L i s a Tou e t , M A DEP Lau r a T r a a s , W D O T CP Ti m o th yTr o u p , A R DOH W ayne T urnbull , R egi o n 4 De b r a W a l l e r , NJ DEP Te d W i tt e n , OK DEQ B ru ce W oods , Reg i on 10 Ke n n e th W u n s c h e l , R I DOH Yu e Z h a n g , TDH Edi t i ng Pa tr i c i a R. L ou i s , F SM D , ORD

## v P reface Si nce 197 8, the U
. S. E nv i r onment al P r ot ect i on Agenc y( E P A ) ha s i mp l ement ed a c ert i f i cat i on progr am f

or l aborat ori es p e rfo rm ing d rin k ing w a te r a n a lyse s fo r c o m p lian c e w ith re g u la tio n s issu e d p u rs u a n t t o the S a fe D rin k ing W a te r A c t. T hes e l aborat ori es i nclude EP A Regi onal l aborat ori es , ce rt ai n Feder al l aborat ori es , T r i bal N at i on l aborat ori es , pri ncipal St at e l aborat ori es i n pr i macySt at es , an d dri nk i ng wat erl aborat ori es i n n on- pri macySt at es . T hi s manu al des cr i bes cr i t er i a a nd procedur es thatE P A us es i n e valuat i ng l aborat ori es f or c ert i f i cat i on. E P A has co ncluded thatl aborat ori es th a ta d o p tth e a p p r o a c h e s d i s c u s s e d i n th i s m a n u a l w i l l g e n e r a t e r e l i a b l e a n a l y t i c a l d a t a . C o n s e q u e n t l y , EPA re c o m m e n d s tha t S ta te s fo llo w the se p ro c e d u re s a n d c rite ria in the ir c e rtifi c a tio n d e c isio n s. T hi s d ocumenti s th e f i f th e di t i onof

th e ma n u a l , de s c r i bi n g th e pr og r a m' s i mp l e me n t a t i onpr oc e du r e s a n d t e c h n i c al cr i t er i a. I ts u per s ede s th e M anu alf orth e Cert i f

i ca t i on of

Labor at or i es Ana l y zi n g D r i n k i n g W at er , EP A- 815- B
- 97- 001 (M a rc h 1 9 9 7 ). T he manua l was rev i s ed t o addres s : 1 ) the r ece nt l yp r om ul gat ed dri nk i ng wat erreg ul at i ons and methods; and 2) Agen cy pol i cy( O f

f i ce of G r ou n d W at eran d D r i n k i n g wat erm emo, O ct ober1, 2002)( s ee A ppen di x F) th at , a tth e di s cr et i on of e a c h s t a t e ’ s C e r t i f

i c a t i o n A u th o r i ty( CA ) , a l l o w s N a t i o n a l En v i r o n m e n t a l Lab o r a t o ryAc c r e d i t a t i o n Pr o g r a m ( NEL AP ) accred i t at i o n t o be accep t ed in l i eu o f dri nki ng wat er cert i f

i cat i o n i n t erm s o f

pro d uci ng da t a f

o r com p l i ance m o ni t o ri ng p urp o ses. A co m m i tt ee chair ed by the E P A ' s O f f i ce o f G round W at er and D ri nki ng W at er ( O G W D W ) wi th pa rt i ci p at i o n of the N at i o nal E xp o sur e R esearch L ab o rat o ri es i n C i nci nnat i (N E R L-Ci) a nd rep res ent at i ves f rom the E P A R egi o ns and the St a t e s pr e pa r e d th i s do c u me n t
. I t s g oa l i s t o i mp r ov e i mp l e me n t a t i onof th e S D W A i n l i g h tof n e w l ya pp r ov ed me th od ol og ya n d a dd i t i on a l e x pe r i e n c e w i th th e pr og r a m. Like previ o us ed i t i o ns, thi s ed i t i o n i s i n l o o se- l eaf f o rma t whi ch wi l l al l o w the E P A t o upd at e i tm o re easi l y . H o l d ers o f th is m a n u a l sho u ld c h e c kw ith the E P A R e g io n o r th e S ta te C e rtifi c a tio n O ff ic e rs to m a k e su re the ir m a n u a l i s c u rr e n t. A d d i t i o n a l c o p i e s o f

th i s m a n u a l m a yb e o b t a i n e d f r o m th e E PA , OGW DW , 2 6 W M
. L
. Ki n g Dr
. , C i n c i n n a t i , OH, 4 5 2 6 8 , f ax num b er 5 1 3 5 6 9
- 7 1 9 1 , o r b y call i ng the S afe D rinking W ater H o t l i ne at 8 0 0 4 2 6
- 4 7 9 1
. T he m an ua l i s also p o ste d o n the In te rn e t at w w w .e p a .go v /sa few a te r/c e rtla b /lab ind e x .htm l. T o ensur e uni f

o rmityi n i t s p rogram i n al l the R egi o ns, E P A uses the cert i f i cat i o n cri t eri a i n thi s m anual f o r eval uat i ng a ll d rin k ing w a te r la b o ra to rie s th a t it c e r ti fi e s. T h e A g e n c y a lso us e s this m a n u a l t o d e te rm ine the a d e q u a c y o f S ta te c e rtifi c a tio n p ro g ra m s fo r d rin k ing w a te r la b o ra to rie s. S ta te s a re e n c o u ra g e d to us e the c rite ria in this m a n u a l t o e v a lua te a ll la b o ra to rie s tha t th e y c e rtify. G ene r al l ythe t erm "m us t " i n thi s manu al r ef er s t o el ement s thatar e r equi r ed by the N at i onal P r i maryD r i nk i ng W at er R egul at i on s or th e appr ov ed dr i n k i n g wa t er meth ods
. T h i s manu al u s es th e t erm "s h ou l d” t o des cr i be cr i t er i a and p roce d u re s tha t i n O W ’ s judge m entare necess aryf o r l ab o rat o ri es t o pro d uce d at a thatare s ci ent i f i cal l yval i d and d e fen sib le , an d a re o f kn o w n a n d a c c e p ta b le p re c isio n a n d a c c u ra c y.

## vi

## vii Tabl e of Contents

P age C H A P T E R I: IN T R O D U C T IO N I-1 C H A P T E R II: R E S P O N S IB IL IT IE S II -1 C H A P T E R III : IM P L E M E N T A T IO N II I-1 E v a lua tio n o f C e rtifi c a tio nP ro g ra m s II I-1 R e q u ire m e n ts fo r C e rtifi c a tio n o f L a b o ra to rie s II I-1 In d ivid u a l(s) R e sp o n sib le fo r th e C e rtifi c a tio n P ro g ra m II I-1 O n -S ite L a b o ra to ry A u d it T e a m II I-2 P la n s fo r C e rtifi c a tio n o f L a b o ra to rie s II I-2 P rin c ip a l S ta te L a b o ra to rie s II I-2 C e rtifi c a tio n P ro c e ss II I-2 T yp e s o f C e rtifi c a tio n II I-3 D rin k ing W a te r L a b o ra to rie s II I-3 O the r C o n sid e ra tio n s fo r L a b o ra to ry C e rtifi c a tio n II I-4 L a b o ra to ry Q u a lity A ssu ra n c e P la n II I-4 C h a in o f C u sto d y P ro c e d u re s II I-6 R e q u ire m e n ts fo r M a inta inin g C e rtifi c a tio n S ta tus II I-6 C rite ria a n d P ro c e d u re s fo r D o w n g ra d ing /R e v o k ing C e rtifi c a tio n S ta tus II I-7 R e c o rd K e e p ing II I-9 R e c ip ro c ity II I-9 T ra inin g II I-9 A lter n a te T e st P ro c e d u re s (A T P s) II I-9 C H A P T E R IV : C R IT IC A L E L E M E N T S F O R C H E M IS T R Y IV -1 P e rs o n n e l IV -1 L a b o ra to ry F a c ili ties IV -1 L a b o ra to ry E q u ip m e n t an d In stru m e n ta tio n IV -2 G e n e ra l L a b o ra to ry P ra c tice s IV -2 A n a lyti c a l M e tho d s IV -3 S a m p le C o llec tio n , H a n d ling a n d P re se rv a tio n IV -3 Q u a lity C o n tro l IV -4 R e c o rd s a n d D a ta R e p o rting IV -9 Ac t i on i n Res pon s e t o N on compl i an tLabor at oryRes u l t s I V
- 10 T abl es I V
- 11- 37 Forms f

orO n - s i t e Eva l u at i on of Labor at or i es I V
- 39- 56 C H A P T E R V : C R IT IC A L E L E M E N T S F O R M IC R O B IO L O G Y V -1 P e rs o n n e l V -1 L a b o ra to ry F a c ili ties V -1 L a b o ra to ry E q u ip m e n t an d S u p p lies V -2 G e n e ra l L a b o ra to ry P ra c tice s V -6 A n a lyti c a l M e tho d o lo g y V -8P age



## vii i Sa mpl e Col l ec t i on , H an dl i n g , a n d Pr es erv at i on V
- 29 Q u al i tyAs s u r an ce V
- 30 Rec or ds a n d D at a Report i n g V
- 31 Ac t i on Res pon s e t o Labor at oryRes u l t s V
- 32 Forms f

orO n - s i t e Eva l u at i on of Labor at or i es V
- 34- 77 C H A P T E R V I: C R IT IC A L E L E M E N T S F O R R A D IO C H E M IS T R Y V I-1 P e rs o n n e l V I-1 L a b o ra to ry F a c ili ties V I-2 L a b o ra to ry E q u ip m e n t an d In stru m e n ta tio n V I-2 G e n e ra l L a b o ra to ry P ra c tice s V I-6 A n a lyti c a l M e tho d s V I-6 S a m p le C o llec tio n , H a n d ling , an d P re se rv a tio n V I-7 Q u a lity A ssu ra n c e V I-7 Rec or ds a n d D at a Report i n g V I
- 11 Ac t i on Res pon s e t o Labor at oryRes u l t s V I
- 13 T abl es V I
- 14- 16 S ampl e Forms f

orO n - s i t e Eva l u at i on of Labor at or i es V I
- 17- 29 AP PENDI CE S A p p e n d ix A : C h a in o f C u sto d y E v a lua tio n s A -1 A p p e n d ix B : R e c o m m e n d e d P ro to c o l f o r O n -S ite L a b o ra to ry E v a lua tio n s B -1 A p p e n d ix C : D e finiti o n s a n d A b b re v ia tio n s C -1 A p p e n d ix D : M e m o ra n d u m R e g a rd ing T h ird P a rty A u d ito rs D -1 A p p e n d ix E : R e q u ire d A n a lyti c a l C a p a b ili ty f o r P rin c ip a l S ta te L a b o ra to ry S yste m s E -1 A p p e n d ix F : M e m o ra n d u m R e g a rd ing N a tio n a l E n v iro n m e n ta l L a b o ra to ry A c c re d itatio n F -1 A p p e n d ix G : A n a lyti c a l M e tho d s fo r M ic ro b io lo g y G -1 TABLE S C hem i stry T abl e I V
- 1 Gl as s wa r e Cl ea n i n g Pr oce du r es I V
- 11 T abl e I V
- 2 Appr ov ed M eth ods f orPr i maryI n or g an i c Che mi ca l s I V
- 13 T abl e I V
- 3 Appr ov ed M eth ods f orPr i maryO r g an i c Che mi ca l s I V
- 17 T abl e I V
- 4 Appr ov ed M eth ods f orD i s i n f ec t an tRes i du al s I V
- 20 T abl e I V
- 5 Re comme n ded M eth ods f orSe con daryD r i n k i n g W at erCont ami n an t s I V
- 21 T abl e I V
- 6 S ampl e Cont ai n er s , Pre s erv at i on a n d H ol di n g T i mes f orReg u l at ed Par ame t er s I V
- 23 T abl e I V
- 7 M CLc an d D et ec t i on Li mi tRequ i r eme n t s t o Compos i t e I V
- 27 T abl e I V
- 8 VO C M CLs a n d M D Ls I V
- 28 T abl e I V
- 9 SO C M CLs a n d D et ec t i on Requ i r eme n t s t o Redu ce M on i t or i n g ( mg / L) I V
- 29 T abl e I V
- 10 M CL a n d Pr of i ci en cyPer f

orman ce Eva l u at i on Sa mpl e Acc ept an ce Cr i t er i a I V
- 30 T abl e I V
- 11 Promu l g at ed O r g an i c D r i n k i n g W at erM eth ods I V
- 35 R ad i o chem i stry T abl e V I
- 1 M eth ods f orRadi on u cl i de Ana l y s i s CFR 141. 25 V I
- 14 T abl e V I
- 2 S ampl e H an dl i n g , Pr es erv at i on , a n d I n s tru men t at i on V I
- 16

## I-1 Chapter I Introduction

P u b lic w a te r sys te m s se rv ing a t l e a st 2 5 p e rs o n s o r h a v ing a t l e a st 1 5 se rv ic e c o n n e c tio n s m u s t c o m p ly with the S a fe D r i n k i n g W at erAct( SD W A) a n d th e r equ i r eme n t s of th e N at i on al Pri maryD r i n k i n g W at erReg u l at i on s ( N PD W R) ( 40 C F R 1 4 1 ). S e c tio n 1 4 0 1 (1 )( D ) o f th e A c t d e fine s a N a tio n a l P rim a ry D rin k ing W a te r R e g u la tio n to inc lud e " c rite ria and p roce d ures . . . [ f

or ] qu al i tyc on tr ol a n d t es t i n g pr oce du r es t o i n s u r e compl i an ce . . . " EP A r eg u l at i on s r equ i re co m m uni tywat er s y st em s t o co nd u c t m o n itori ng f o r com p l i ance with the d ri nki ng wat er s t and ards. I n ad d i t i o n, reg ulati o ns p ro vid e that an alyses m ust b e co nd uc t ed b y l ab o rato ries ce rti f i ed b y E P A o r the S t ates. T he reg ulati o ns also e sta b lishe d re q u ire m e n ts fo r la b o ra to ry c e rtifi c a tio n .. T h e r e g u la tio n s g o v e rn ing p rim a c y a t 4 0 C F R 1 4 2 .1 0 (b )( 4 ) re q u ire , as a c o n d iti o n o f p rim a ry e n fo rc e m e n t re sp o n sib ili ty ( pri macy ) , thata s t at e h ave l aborat oryf aci l i t i es av ai l abl e ( the P r i ncipal St at e Labora t ory ) c erti f i ed b ythe r egi onal adm i ni s tr at or. I n addi t i on, the r egu l at i ons governi ng cert i f i cat i on ( 40 C FR 14 1. 28 ) r equi r e thatal l t es t i ng f or c om pl i anc e p u rp o se s b e p e rfo rm e d b y c e rtifi e d lab o ra to rie s e x c e p t th a t tu rb id ity, fre e c h lo rin e re sid u a l, te m p e ra tur e , p H , alk a linity, c a lc ium , co n d u c tivity, o rth o p h o sp h a te , T O C , S U V A , d a ily c h lo rite , a n d silica m a y b e p e rfo rm e d b y a n yo n e a c c e p ta b le t o th e St at e. T h i s manu al i s i n t en ded t o as s i s t EP A i n i mpl eme n t i n g 40 CFR 142. 10( b) ( 4) by s pec i f y i n g cr i t er i a and p ro ce d ure s E P A uses in ev alua t i ng p rincip al S t ate lab o rato ries f o r ce rti f i ca t i o n. S t ates w i th p rima cy m ay also ch o o se t o use e q uiva l en tcrit eria a nd p ro ce d ure s si m i l ar to tho se in this m an ua l f o r the i r o w n c erti f

i ca t i o n p ro gra m s. T o o b t ain an d m aintain p rima cy, a S t ate m ust co m p l y with 4 0 C F R 1 4 2 .10 , w hich i nc l ud es the fo l l o w i ng p ro visi o ns: T he est ab l i shment and m ai nt enance o f a St at e p rogram f

o r the cert i f

i cat i o n o f

l ab o rat o ri es cond uct i ng a n a lyti c a l m e a su re m e n ts o f d rin k ing w a te r c o n ta m ina n ts p u rs u a n t t o the re q u ire m e n ts o f th e S ta te pr i marydr i n k i n g w at err eg u l at i on s i n clu di n g th e des i g n at i on by th e St at e of a l abor at oryof

f i ce r , or of f i ce r s , ce r t i f

i ed by th e Admi n i s tr at or , as th e of f i ci al ( s ) r es pon s i bl e f

or th e St at e' s cert i f

i ca t i on p rogram . T he requir em ent s o f thi s p aragraph m ay b e wai ved by the A d m i ni strat o r f o r anyStat e where al l an al y t i calmeas ur ement s r equi r ed by the St at e' s pri marydri nk i ng wat err egu l at i ons ar e c onduct ed atl ab o rat o ri es op erat ed by the Stat e and cert i f i ed by the A gency. (40 C FR 14 2
. 1 0 (b)(3)(i )) A ss urance o f

the avai l ab i l i tyt o the Stat e o f l ab o rat o ryf

aci l i t i es c e rti f

i ed by the A d m i ni strat o r and cap ab l e o f

pe rf o rming anal y t i cal m easurements of all c o nt am i nant s specif i ed i n the Stat e p ri m ary d rin k ing w a te r re g u la tio n s . . . (4 0 C F R 1 4 2 .1 0 (b )( 4 ))
.
N O T E : Ref

er enc e t o the A dm i ni s tr at or of E P A al s o re f

er s t o hi s or herdes i gn ee. T he req uirem en tf

o r a l ab o rato ry ce rti f

i ca t i o n pro gra m ex t en d s t o E P A R eg i o na l l ab o rato ries, T riba l N ati o n lab o rato ries, pri ncipal St at e l aborat ori es i n pr i macy St at es , and l aborat ori es that perf orm ana l y s es un der the Saf e D r i nk i ng W at erAct i n States w i tho ut p rima cy. If all req uired an alyses are no tp erfo rm ed i n p rincip al S t ate lab o rato ries, p rima cy S t ates m ust h a v e a c e rtifi c a tio n p ro g ra m fo r c e rtifyi n g o the r d rin k ing w a te r la b o ra to rie s (4 0 C F R 1 4 2 .1 0 (b )( 3 )( i)). E P A ' s T ech ni calSuppo r tCent er( T SC ) i n Ci nci nn at i , O hi o, wi th the as s i s t anc e of the N at i onal E xp osu r e Res ear ch Labo r at ory i n C i nci nn at i , O hi o (N E R L- C i ) , i s r es po ns i bl e f or det ermi ni ng the cert i f i cat i on s t atus f

or EP A ’ s Regi onal l ab o rat or i es i n mi cr obi ol og y , ch emi s try , an d r adi och emi s try . Reg i on al ce rt i f i ca t i on of

f i ce r s ar e r es pon s i bl e f

or the cert i f i cat i on of

the p r i ncipal St at e l aborat oryi n e ach pri macySt at e, an d ar e a l s o re s po ns i bl e f or c ert i f y i ng al l T r i bal N a tio n lab o ra to rie s a n d la b o ra to rie s in no n -p rim a c y S ta te s. P rim a c y S ta te s w ith c e rtifi c a tio n p ro g ra m s a re re sp o n sib le fo r c e rtifyi n g the o the r d rin k ing w a te r la b o ra to rie s in th e ir S ta te , (i.e ., la b o ra to rie s o the r th a n the p rin c ip a l S ta te L a b o ra to ry)
. R e g io n a l L a b o ra to rie s m u st su c c e ssfu lly a n a lyz e a se t o f p r o fi c ie n c y tes ting sa m p le s (P T s) a t l e a st a n n u a lly fo r a ll r eg u l at ed c on t ami n an t s f orwh i chth eywi s h t o be ce rt i f i ed, by ea chm eth od f

orwh i chth eyw i s h t o be c ert i f i ed ( 40 CFR

## I-2

14 1. 23 an d 14 1. 24 ) , an d pass anon- s i t e e valuat i o n at l eas tev erythr ee y ear s . An on- s i t e e valuat i on det ermi nes con f orman ce w i th th e cr i t er i a s pec i f i ed i n th i s ma n u al
. Pr i n cipalSt at e l abor at or i es mu s t( 40 CFR 141. 23 an d 141. 24) suc ce ssf ull y ana l yze a c o m p l ete set o f p ro f i cien cy t esti ng (P T ) sam p l es from a so urc e a cc ep t ab l e to the R eg i o n a tl ea st a n n u a lly fo r th e c o n ta m ina n ts a n d m e tho d s inc lud e d in th e re g u la tio n s w h ic h the S ta te h a s a d o p te d , a n d p a ss an o n -sit e eval uat i o n everyth re e y ears. E P A aud i t o rs us e the cri t eri a i n thi s m anual f o r the o n- si t e aud i t s o f the R egi o nal and p rincip al S t ate lab o rato ries. C hapt erI I des cr i bes the r es po ns i bi l i t i es of eac h of the part i es i nv ol ved i n the cert i f i cat i on progr am. Chapt erI I I des cr i bes h ow th e pr og r am oper at es . Ch apt er s I V , V , a n d V I c ov erth e t ec h n i ca l c r i t er i a t o be u s ed du r i n g th e on - s i t e ev alu at i on of a l aborat oryf

or c hemi s try , mi cr ob i ol ogy , an d ra di ochemi s try , r es pect i vel y . O pt i onal au di tf orms ar e a l s o i ncluded i n C hap t ers IV , V , and V I. T he ap p end i ces i ncl ud e the f o l l o wi ng: a r eco m m end ed p rotoco l a nd f o rma tf o r cond uct i ng o n- si t e l ab o rat o ryeval uat i o ns whi ch m ay b e used b y the l ab o rat o ryaud i t o rs ; f reque nt l yus ed ab b revi at i o ns and def i n i t i on s ; EP A' s pol i cyon th i r d- partya u di t or s ; EP A’ s pol i cy o n N at i on alEnv i r on men t alLabor at oryAcc r edi t at i on P ro g ra m (N E L A P ) a c c re d itatio n a s a n a lter n a te fo r d rin k ing w a te r ce rtifi c a tio n ; a list o f c o n ta m ina n ts a p rin c ip a l S ta te l ab o rato ry m ust(pri m acy ci t at i o n 14 2
. 1 0 (b)(4)) have the cap ab i l i tyt o anal y ze, anal y t i cal m etho d s f o r mi crob i o l o gy a n a lyse s; a n d re c o m m e n d e d c h a in-o f- c u sto d y p ro c e d u re s to b e us e d w h e n ne c e ssa ry.

## II-1 Chapt e rII Responsib ilities

T h e s u cc es s of

th e l abor at oryc ert i f

i ca t i on pr og r am depe n ds upon coope r at i on amon g th e or g an i za t i on s r es pon s i bl e f or i t s i m p l em ent at i o n. W i thi n the A gency, res p o nsib i l i t i es f o r l ab o rat o rycert i f

i cat i o n are s hared by the O f f i ce o f G round W ater a nd D rinking W ater (O G W D W ), the O f f i ce o f R ese arc h a nd D ev elo p m en t(O R D ), an d the R eg i o na l O f f i ce s. 

## Offic eof Ground W ate rand D rinkin g W ate r(O GWDW)and O fficeof Re se arc h and Develop m e nt (ORD) O G W D W , w i th a s s i s t an ce f r om O RD , h as th e r es pon s i bi l i ty f or dev el opi n g an d i mpl eme n t i n g th e n at i on alc ert i f i ca t i on p ro gra m f

o r l ab o rato ries tha tan alyze d rinking w ater sam p l es an d fo r i m p l em en t i ng the S afe D rinking W ater A ct. T he se res p o nsib i l i t i es i ncl ud e the f o l l o wi ng:

• P ro p o se a nd p ro m ulga t e re gu l ati o ns;
• A sse ss n a tio n a l l a b o ra to ry c a p a c ity a n d c a p a b ili ty;

• R e v ie w the E P A R e g io n a l c e rtifi c a tio n p ro g ra m s a n n u a lly a n d e v a lua te th e re so u rc e s a n d p e rs o n n e l av a ilab le i n e a c h EPA Re g i ont o c a rryou tth e c e rt i f i c a t i onpr og r a m;
• D e v e lo p gu id a n c e a n d re sp o n d to q u e stio n s a n d c o m m e n ts;

• D ev elo p t ec hn i ca l an d ad m i nistrati ve ce rti f i ca t i o n c rit eria to sup p o rt f uture reg ulati o ns;
• R e v ise this m a n u a l wh e n ne c e ssa ry;
• C o nd ucttr i enni al o n- si t e aud i t s o f e ach R egi o nal l ab o rat o ryf o r chemis try , m i crob i o l o gy , and radi o chem i stry
• M ai nt ai n adat abas e of l aborat oryI D Co des

• D e v e l op a n d p a r t i c i pa t e i n tr a i n i n g c ou r s e s t o s u pp ortth e c e rt i f i c a t i onpr og r a m;
• P ro v id e tec h n ic a l as sista n c e to E P A a n d the S ta te s;
• D ev elo p an d ev alua t e m etho d s f o r the an alysi s o f

d rinking w ater c o ntam i na nts. 

## EPA Re gio ns T h e Reg i on s ov er s ee th e ce r t i f i ca t i on pr og r ams i n th e St at es a n d T r i balN at i on s . T h e Reg i on s ' r es pon s i bi l i t i es a r e t o:

• D et ermi ne the cert i f i cat i on s t atus f

or the pri ncipal St at e/ T r i bal l aborat orysy s t em i n e ach pri macySt at e/ T r i be;

• P erf o rm an annual r evi ew o f Stat e/ T rib al cert i f

i cat i o n pro gram s and pro f i ci ency t est i ng resul t s and m o ni t o r the ad eq uacy o f

Stat e/ T rib al pro gram s f o r cert i f y i ng l ab o rat o ri es, as d escrib ed i n C hap t er I II ;
• Sp o nsor annual m eet i ngs f o r the st at e C ert i f

i cat i o n O f

f i cers (C O s) ;
• P ro v id e tec h n ic a l as sista n c e to the S ta te s' E P A -ce rtifi e d d rin k ing w a te r la b o ra to rie s, a s n e e d e d ;
• M a n a g e the c e rtifi c a tio n p ro g ra m fo r d rin k ing w a te r la b o ra to rie s in n o n -p rim a c y S ta te s u sin g th e c rite ria in this m a n u a l.

• M a n a g e th e c e rtifi c a tio n p ro g ra m fo r d rin k ing w a te r la b o ra to rie s o n T rib a l N a tio n la n d s u sin g th e c rite ria in this m a n u a l. T hi s l astdutym ay b e p erf o rme d by the Stat e, but the R e g io n ret ai ns res p o nsib i l i tyf o r the o n- si t e eval uat i o n of the des i gn at ed T r i bal N at i on pri ncipal l aborat ory . D r i nk i ng wat erl aborat ori es mayb e evaluat ed by the R egi on, or un der a R egi o n- ap p roved pro gram carri ed out b ya de si gnat ed Stat e p rogram . I n e i ther ca se, thi s m anual i s the b asi s f o r the on- s i t e audi t s , co nduct ed by E P A , o f p r i ncipal S t at e l aborat ori es , l aborat ori es o n T r i bal N at i on l ands , an d dri nk i ng wat er l ab o rato ries i n no n-p rima cy S t ates. T h e R e g io n a l l a b o ra to ry sh o u ld m a inta in ce rtifi c a tio n fo r as m a n y re g u la te d c o n ta m ina n ts a s its re so u rc e s p e rm it. T h is enhances b o th E P A ' s t echni cal ass i st ance capa b i l i t i es and cred i b i l i ty wi th tho se i t cert i f i es. Ital so ensur es the l ab o rat o ry capabi l i tyt o ana l y ze s amp l es f or pos s i bl e e nf

orce mentac t i ons an d f or St at es wh i ch do notha ve pri macy . Reci procal ag r ee men t s w i th oth err eg i on s t o s h ar e s ca r ce r es ou r ce s a r e r ec ommen ded.

## II-2

P rim acy State s P rima cySt at es , i n w hi ch al l dri nk i ng wat ercomp l i anc e s amp l es ar e not anal y zed atStat e o p erat ed l ab o rat o ri es, are requir ed t o est ab l i sh and m ai nt ai n a St at e p rogram f o r the cert i f

i cat i o n o f l ab o rat o ri es cond uct i ng anal y ses of d ri nki ng waterco m p l i ance sam p l es [ see 40 C FR 14 2
. 1 0 (b)(3)(i )]
. E P A enco urages the Stat es t o ba se cert i f i cat i o n of d ri nki ng wat erl aborat ori es ei therupo n cr i t er i a c ont ai ned i n thi s manu al or upon s t at e- devel op ed equi val ent s thatar e a tl eas tas s tr i ng entas thi s manu al
. Pri macyS t at es m us test abl i s h and m ai nt ai n a st at e p r ogr am f or the cert i f

i cat i on of

l aborat ori es co nd uct i ng anal y t i cal m easurements of d ri nki ng w at er co nt am i nant s p urs uantto the requir em ent s o f the Stat e p ri m ary dr i n k i n g w at err eg u l at i on s . T h e St at es m u s tdes i g n at e a “ l abor at oryof f i ce ror of

f i ce r s , c ert i f

i ed byth e Admi n i s tr at or o r desi gnee as the o f

f i ci al (s ) r espo nsib l e f

o r the cert i f

i cat i o n pro gram . ” (40 C FR §1 4 2
. 1 0 (a) (3)(i )) Stat es ar e res p o nsib l e f o r the cert i f

i cat i o n o f the p ub l i c and pri vat e l ab o rat o ri es i n thei r St at e. T hi s i ncl ud es audi t i ng the l ab o rato ries and review i ng the P T d ata. S t ates sho uld also p ro vid e t ec hn i ca l assis t an ce t o l ab o rato ries. T he y m ay also c h o o se to c e rtify la b o ra to rie s o u tsid e the ir S ta te e ithe r b y a n o n -sit e e v a lua tio n o r re c ip ro c ity.

## III-1 Chapte rIII Implem entation
1 . E v alu ati o no f

Ce rti f i cati o nPro g ra ms O G W D W and the R egi o ns m o ni t o r the cert i f i cat i o n pro gram s und er thei r purvi ew annuall y . T hese of f i ces as ses s the ad eq ua cy o f

p ro gra m s f

o r ce rti f yi ng l ab o rato ries b y eva l ua t i ng ea ch p ro gra m ' s sco p e, staff i ng , reso urc es, p o l i cies, pr oce du r es , a n d e f

f ec t i v en es s . T h i s s h ou l d be don e i n pe r s on du r i n g a n on - s i t e au di tw h enpos s i bl e, a n d a tl ea s tby m e a n s o f a q u e stio n n a ire in the o the r ye a rs . T h e a d e q u a c y o f th e se e sse n tial p ro g ra m e le m e n ts is ev a lua te d b y:

• R evi ewing the p rogram p l an, respo nsib i l i t i es, o rgani zat i o nal structure, st af f (i ncl ud i ng ed ucat i o nal ba ckground an d ex p erien ce ), sco p e and d esc ripti o n of the ce rti f i ca t i o n pro ce ss, d o w ng rad i ng crit eria a nd p ro ce sses, and use o f P T sam p l es;
• R eq ue st i ng an an nu al p ro gra m rep o rt tha ti nc l ud es p ro gra m highli gh t s, training, co nti nu i ng ed uc ati o n e f f o rts , num b er o f o n- si t e eval uat i o ns p erf o rme d , l i st i ng of l ab o rat o ri es cert i f i ed by di scip l i ne o r cont am i nant , and any ce rti f i ca t i o n d o w ng rad i ng o r up gra d i ng ac t i o ns alo ng with re aso ns f o r tho se a cti o ns;
• O b serv i ng the st ate c erti f i ca t i o n o f

f i ce rs o n-si t e a ud i t s o f

d rinking w ater lab o rato ries;

• E ncour agi ng St at e and Regi onal l aborat ory audi t ors t o obs erve on- s i t e audi t s of thei rown a nd otherl aborat ori es a s o n -the -jo b tra inin g ;
• Spo ns ori ng an nu al meet i ng s of ce r t i f i cat i on of

f i cer s t o di s cus s program i s s ues , pol i ci es , an d probl ems. K ey R egi onal , N E R L, O G W D W , an d St at e per s onnels houl d be i nv i t ed t o part i ci pat e. 

## 2. Requirem ents for Certification of Laboratories In o rd e r to b e e ligib le to a n a lyz e c o m p lian c e s a m p le s u n d e r the S a fe D rin k ing W a te r A c t, R e g io n a l an d P rin c ip a l S ta te l ab o rato ries sho uld m ee tthe m i nim um crit eria s p ec i f i ed i n thi s m an ua l , w hich i nc l ud es p assing a n o n-si t e a ud i tat l ea st o n c e e v e ry th re e ye a rs , an d sa tisf a c to rily an a lyz ing a se t o f P T sa m p le s a n n u a lly. T he O f f i ce o f G round W at er and D ri nki ng W at er (O G W D W ) wi l l accep tN E LA P accred i t at i o n as eq ui val entt o D ri nki ng W a te r c e rtifi c a tio n , i f all re q u ire m e n ts o f th e d rin k ing w a te r p ro g ra m a re m e t. 

## 3. Ind ividu al(s) Re spon sibl e for the Ce rtification Program T he T ec hn i ca l S up p o rt C en t er, with the a ssi st an ce f ro m N E R L
- C i i s resp o nsib l e fo r ce rti f yi ng the reg i o na l l ab o rato ries; the R egi o ns are respo nsib l e f o r cert i f y i ng thei r S t at es’ p ri nci p al labo rat o rysy st em s and T rib al N at i o ns’ labo rat o ri es; and the S t ates ar e re sp o nsib l e fo r ce rti f yi ng p rivate, m un i cip al, no n-p rincip al st ate, and go ve rnm en t s l ab o rato ries. T h e ce r t i f

i ca t i on pr og r am per s on n eli n e ac h R eg i on s h ou l d con s i s tof a c ert i f i ca t i on a u th or i ty ( s ) ( CA), th e ce rt i f i ca t i on progr am manag er , and a cert i f i cat i on t eam com pri s ed of cert i f i cat i on of

f i cer s ( C O s ) and t ech ni calex pert s . Ad di t i onal third p arty au d i t o rs an d ex p erts m ay b e u sed
. H o w ev er, third p arti es ha s hav e n o au tho rity f o r ce rti f i ca t i o n d ec i si o ns. T h i r d pa r tya u di t i n g i s di s cu s s ed i n Se ct i on 4. 2. T he Ce r t i f

i c a t i o n Au t h o r i t y ( CA) i s the p ers o n w ho has si gnature autho ri tyf o r al l cert i f

i cat i o n de ci si o ns. T hi s i s the C h ie f o f th e T e c h n ic a l S u p p o rt C e n te r a n d the R e g io n a l A d m inistra to r in the R e g io n s. T h e R A m a y d e le g a te this a u tho rity to a lo w e r le v e l. T he C ert i f

i cati on P rogra m M ana ger (CP M ) i s the p ers o n res p o nsib l e f o r managing th e dri nki ng wat er l ab o rat o ry c e rtifi c a tio n a c tivities in the R e g io n . T he C e r tifi ca tion O fficer s (C O s) a r e th e r eg i on ala n d s t at e per s on n elw h o h av e th e r es pon s i bi l i tyof c ert i f y i ng l ab o rat o ri es under thei r p urvi ew. 4 0 C FR 1 4 2
. 1 0 (b)(3)(i ) requir es P ri m acy Stat es t o d esi gnat e a p ers o n cert i f i ed b ythe Admi n i s tr at ora s th e of

f i ci alr es pon s i bl e f

orth e St at e' s ce rt i f

i ca t i on prog r am. T h i s per s on w ou l d be th e St at e ce rt i f i ca t i on a u tho rity.

## III-2

4. On- SiteLabo r atory Audit 4. 1 O n-Site Labor ator y Audit Team T he cert i f

i cat i o n p rogram m anager shoul d est ab l i sh o ne o r m o re t eam s o f cert i f i cat i o n o f

f i cers and aud ito rs to a u d it l aborat ori es . I ti s the r es po ns i bi l i tyof thes e t eams t o perf

orm the on- s i t e l aborat oryaudi t s , r evi ew the l aborat oryP T dat a, a n d m a k e re c o m m e n d a tio n s to the C A c o n c e rn ing the c e rtifi c a tio n sta tus o f th e lab o ra to rie s. T eam memb er s s houl d be exper i enc ed prof es s i onal s , hol d atl eas ta bachel or' s degr ee or e qui val enteduc at i on/ exper i enc e i n the d i scip l i ne (chem i stry , radi o chem i stry , m i crob i o l o gyor a rel at ed f i el d ) f o r whi ch they ce rtify, a nd have recent l aborat oryex peri enc e. T eam m em b ers sho ul d a l so have exp eri ence i n l ab o rat o ryeval uat i o n and qua l i tyass urance, be f am i l i ar wi th the d ri nki ng wat err egu l at i ons an d dat a r educt i on an d re po r t i ng t ech ni ques , b e t ech ni cal l yconv er s antwi th the ana l y t i calt ech ni ques b e ing e v a lua te d , an d b e a b le to c o m m u n ic a te e ff e c tive ly, b o th o ra lly a n d in w riting
. T h e o n -sit e tea m sh o u ld inc lud e a t le a st o n e p e rs o n k n o w le d g e a b le in e a c h a re a b e ing a u d ited (e .g., ino rg a n ic a n d o rg a n ic chem i stry , radi o chem i stry , and m i crob i o l o gy ). C O s need t o s uccessf ul l yco m p l et e the ap p rop ri at e E P A l ab o ratory c e rtifi c a tio n c o u rs e . In a d d iti o n , the re sh o u ld b e a m e c h a n ism fo r C O s to re c e ive p e rio d ic tra inin g re g a rd in g n e w ly p r o m u l g a t e d r e g u l a t i o n s , n e w l ya d o p t e d c e r t i f i c a t i o n c r i t e r i a , a n d n e w m e th o d s . Th i s c o u l d b e d o n e b y a u d i t i n g th e EPA C O T raining C o urse an d / o r att en d i ng the req uired an nu al R eg i o na l an d S t ate C O M ee t i ng s. 4 .2 T h irdP a rt

y A u dito rs C ert i f i cat i o n pro gram s m ay em p l o ythi rd p artyaud i t o rs who m eetal l of the q ual i f i cat i o ns l i st ed ab o ve. I n areas where exp eri ence d o es no t exi st wi thi n the cert i f i cat i o n t eam (e. g. , asbest o s) , o ut si d e exp ert ass i st ance may b e o b t ai ned i n the need ed area s to a ssi stthe o n- si t e t eam . O ut si d e exp ert s who have no tatt end ed the E P A cert i f i cat i o n off

i cer trai ni ng sh o u ld b e a c c o m p a n ie d b y a c e rtifi c a tio n o ff ic e r. A ltho u g h the se third p a rtie s m a y b e us e d to a ssist E P A o r S ta te ce r t i f i ca t i on of

f i ce r s , th eyh av e no a ut ho r i tyf orc ert i f i ca t i on de ci s i on s a n d th eyma yn otma k e f i n alc ert i f

i ca t i on deci s i ons . T hes e deci s i ons r es twi th the E P A or the St at e. W h enu s i n g th i r d pa r tye x pert s , i ti s cr i t i ca l t o av oi d con f l i ct s of i n t er es t
. A th i r d pa rtya u di t orw h o i n a n yw ays t an ds t o be n ef i tbyth e ce r t i f

i ca t i on s t atu s of

th e l abor at orya u di t ed mayn otbe u s ed. 

## 5. Plans f

or Ce rtification of

Laboratories T h e c e rtifi c a tio n p ro g ra m m a n a g e r sh o u ld d e v e lo p p la n s fo r c e rtifyi n g d rin k ing w a te r la b o ra to rie s u n d e r h e r/h is autho ri ty . W ri tt en p l ans s ho ul d i ncl ud e the f o l l o wi ng:

• D ocument at i on of

ce r t i f i cat i on au thor i tyan d cert i f i cat i on of

f i cer s an d thei reduc at i on/ exper i enc e;
• Sc h edu l es of

l abor at or i es t o be a u di t ed;

• Spe ci f

i c types of

a n al y s es t o be e x ami n ed;

• Prot ocolt o be f

ol l owe d;

• S tra te g y fo r a sse ssin g lab o ra to ry p e rfo rm a n c e (e .g., P T s, d a ta a u d its, e tc .);

• P l ans f

o r providi ng t echni cal ass i st ance t o l ab o rat o ri es whi ch need upgrad i ng. 

## 6. P rincipal S tateLaboratories T o r ec ei v e an d r et ai n pr i mac y , the S t at e mu s t( 40 CFR 142. 10( b) ( 4) ) h av e av ai l abl e l abor at oryf ac i l i t i es c apabl e of p erf o rming anal y t i cal m easurements f o r al l the f

ed eral l y m and at ed co nt am i nant s specif i ed i n the Stat e P ri m aryD ri nki ng W at erReg u l at i on s . T h i s l abor at ory or l abor at or i es ar e con s i der ed th e Pri n cipalSt at e La bor at orySy s t em an d mu s tbe c e r t i f i e d b yEPA. 

## 7. Ce rtific ationP r oc e ss T h e ce r t i f

i ca t i on pr oce s s be g i n s w h enth e l abor at orydi r ec t orma k es a f ormalr equ es ti n w r i t i n g t o th e ce rt i f i ca t i on autho ri tyt o be cert i f

i ed
. T hi s ap p l i cat i o n m ay b e o ne o f the f o l l o wi ng:

## III-3 • A re q u e st fo r first-t im e c e rtifi c a tio n fo r m ic ro b io lo g y, ch e m istry, o r ra d io c h e m istry;

• A re q u e st fo r c e rtifi c a tio n to a n a lyz e a d d iti o n a l o r n e w ly re g u la te d c o n ta m ina n ts;

• A requestt o reapp l yf o r cert i f

i cat i o n af

t er correct i o n of d ef i ci enci es whi ch res u lted in the d o w ng rad i ng /rev o ca t i o n o f ce rti f i ca t i o n status. T he r es po ns e t o a f

ormal appl i cat i on f

or a nyof

the above req ue s t s s houl d be gi venwi thi n 30 day s . Atthi s t i me a m u tua lly a g re e a b le d a te a n d ti m e sh o u ld b e se t f o r th e o n -sit e lab o ra to ry a u d it. S u b se q u e n t au d its m a y b e initiate d b y the C A o r th e lab o ra to ry. A re c o m m e n d e d p ro to c o l f o r c o n d u c ting the se a u d its i s g ive n in A p p e n d ix B
. D rinking w ater lab o rato ries sho uld sp ec i f i ca l l y stat e tha tthe y plan t o an alyze d rinking w ater sa m p l es w he n they req ue st cert i f i cat i on. I f

a l aborat oryha s not been ana l y zi ng dri nk i ng wat ers amp l es an d do es notpl an t o ana l y ze dri nk i ng wat er sa m p le s, O G W D W , th e R e g io n o r S ta te m a y c h o o se no t t o e x p e n d the re so u rc e s to re n e w the ir c e rtifi c a tio n . 

## 8. Types of

Certification A fterre v ie w o f P T sa m p le re su lts an d a n o n -sit e visit, the c e rtifi c a tio n a u tho rity sh o u ld p ro v id e a w ritten re p o rt w ithin 4 5 da y s and cl ass i f

ythe l ab o rat o ryf

o r each contaminantor group of co nta m ina nt s acco rding t o the f o l l o wi ng rat i ng s che me: 8 .1 Certi

f ied
- a l aborat ory that meet s the r egu l at ory perf ormanc e cr i t er i a as expl ai ned i n thi s manu al an d al l other a p p lica b le re g u la to ry re q u ire m e n ts. 8. 2 P r ovis iona ll

y Cer ti f ied
- a l aborat ory that has def i ci enc i es but dem ons tr at es i t s abi l i ty t o c ons i s t ent l yp ro d uce v al i d da t a wi th i n th e ac ce pt an ce l i mi t s s pec i f i ed i n th e N PD W R, a n d w i th i n th e pol i cyr equ i r ed byth ei rc ert i f i ca t i on a u tho rity. A p ro v isio n a lly c e rtifi e d la b o ra to ry m a y a n a lyz e d rin k ing w a te r sa m p le s fo r c o m p lian c e p u rp o se s, if the sa id cl i ent s are no t i f

i ed o f it s d o wngrad ed st atus i n w ri t i ng, o n a nyrep o rt
. P rovis i o nal c ert i f i cat i o n m ay no tb e gi ven i f the eval uat i o n t eam be l i eves thatthe l ab o rat o rycanno tpe rf o rm an analysis w ithin the acc ep t ance l i m i t s specif i ed i n the reg ulati o ns. 8. 3 NotCerti

f ied
- a l abor at oryth atpos s es s es de f i ci en ci es a n d, i n th e opi n i on of th e Cert i f

i ca t i on Au th or i ty , c an n ot cons i s t ent l yproduce val i d dat a. 8. 4 I nterim Certi

f ication - i n t er i m c ert i f

i ca t i on ma ybe g r an t ed i n cert ai n ci rcum st ance s w h eni ti s i mpos s i bl e or un nec es s aryt o perf

orm an on- s i t e a udi t
. I nt er i m cert i f i cat i on s t atus maybe gra nt ed i f , f or exampl e, the C A det ermi nes thatthe l aborat oryha s the appropri at e i ns trument at i on, i s us i ng the approved m ethods, has ad equat el ytr ai ned per s onnel t o pe rf o rm the anal y ses , and has sat i sf act o ri l yanal y zed P T s am p l es, i f avai l ab l e, f

o r the co nt am i nant s i n que st i o n. T he C O s ho ul d p erf

o rm an o n- si t e aud i ta s soo n a s p o ss i b l e b utno l at er than three y ears. A n exam p l e o f a s i tuat i o n where thi s type of

ce r t i f i cat i on i s warr ant ed woul d be a l aborat orythatha s r eques t ed cert i f i cat i on f or the ana l y s i s of

addi t i onal a n a lytes th a t i n v o lve a m e tho d f o r w h ic h ita lre a d y ha s c e rtifi c a tio n . T h e C O sh o u ld re v ie w the lab o ra to ry' s q u a lity co ntrol d at a b ef

o re gr ant i ng thi s typ e o f cert i f i cat i o n. 

## 9. Drinkin g WaterLaboratories Forth e pu rpos e of

c ert i f i ca t i on , a n yl abor at oryw h i cha n al y ze s dr i n k i n g w at erc ompl i an ce s ampl es i s c on s i der ed a dri nk i ng wat erl aborat ory . T hi s i ncludes Feder al l aborat ori es thatan al y ze com pl i anc e s amp l es an d anyl aborat ori es that a n a lyz e c o m p lian c e sa m p le s fo r F e d e ra l f a c ili ties. A lls uchl aborat ori es mus t(40 C FR 1 41
. 21 , . 23 , . 24 , . 25 ) be ce rt i f i ed b ythe Stat e o r E P A
. I f request ed by the Stat e, a R egi o n m ay cert i f yFed eral l ab o rat o ri es i n i t s R egi o n.
T he R eg i o n c erti f

i es in d iv id u a l l ab o rato ries o n T riba l N ati o n land s, wh en req ue st ed b y the triba l ch airp erso n. T he se l a bo r a t or i e s mu s tme e tth e c r i t e r i a f orc e r t i f i c a t i ona s s pe c i f

i e d i n th e NP D W R.

## III-4

T he R egi o ns use the cri t eri a, p roce d ures , and m echanis m as specif i ed i n the N P D W R and thi s m anual i n thei r decis i o ns t o ce rti f y m un i cip al o r p rivate d rinking w ater lab o rato ries i n no n-p rima cy st ates. 

## 10. Other Considerations for Lab oratory Certification 10 . 1 L ab oratoryP ers onn el T he l aborat orys houl d hav e su f f i ci entsuperv i s oryand otherp er s onnel , wi th the nec es s aryeduc at i on, tr ai ni ng , t ech ni cal kn o w l ed ge , an d ex p erien ce f o r the i r assigne d f

un cti o ns. 10. 2 Labor ator

y D ir ec tor

/ Manage rorTec hnical D ir ec tor T he l abor at ory di r ec t or/man ag er s h ou l d be a qu al i f i ed pr of es s i on al wi th th e t ec h n i ca l edu ca t i on an d ex per i en ce, and m a n a g e ria l ca p a b ili ty c o m m e n su ra te w ith the siz e /typ e o f the la b o ra to ry. T h e la b o ra to ry d ire c to r/m a n a g e r is u lti m a te ly r es po ns i bl e f

or e ns ur i ng thatal l l aborat ory pers onnel hav e dem ons tr at ed prof i ci enc y f or thei ras s i gn ed f

un ct i ons an d that all d ata re p o rted b y the l ab o rato ry m ee tthe req uired q ua l i ty assuran ce (Q A ) crit eria a nd reg ulato ry req uirem en t s. 10 . 3 Q ualit

y Ass ur ance Manager T he Q A m anager shoul d be i nd ep end entf rom the l ab o rat o rym anagem ent , i f po ss i b l e, and have d i rectacce ss t o the hi gh es tl eve l of

manag ement
. T he Q A manag ers houl d hav e abach el or' s degr ee i n s ci enc e, tr ai ni ng i n qua l i tyas s ur anc e p rin c ip le s c o m m e n su ra te w ith the siz e a n d so p h istica tio n o f th e lab o ra to ry, a n d a t l e a st o n e ye a r o f ex p e rie n c e in q u a lity as s u r an ce . T h e Q A man ag ers h ou l d h av e atl eas ta w ork i n g kn owl edg e of th e s t at i s t i cs i n v ol v ed i n qu al i tyc on tr ol of l ab o rato ry ana l ysi s and a b asic un d erstan d i ng o f the m etho d s wh i ch the l ab o rato ry em p l o ys. 

## 11. Laboratory Quality Assurance Plan A ll la b o ra to rie s a n a lyz ing d rin k ing w a te r c o m p lian c e s a m p le s m u st a d h e re to a n y r e q u ire d Q C p ro c e d u re s sp e c ifi e d in th e meth ods
. T h i s i s t o e n s u r e th atr ou t i n el yg en er at ed an al y t i ca l da t a ar e s ci en t i f i ca l l yv al i d a n d de f

en s i bl e, a n d a r e of k n o w n a n d a c c e p ta b le p re c isio n a n d a c c u ra c y. T o a c c o m p lish th e se go a ls, ea c h lab o ra to ry sh o u ld (E P A O rd e r 5 3 6 0 .1 A 2 ) prep are a wri tt en d escrip t i o n of i t s Q A act i vi t i es ( a Q A plan)
. I ti s the res p o nsib i l i ty o f the Q A m anager t o keep the Q A pl anup t o da t e. Al l l abor at orype r s on n eln ee d t o be f ami l i arw i th th e con t en t s of th e Q A pl an . T h i s pl ans h ou l d be su b m itt e d to the a u d ito rs fo r re v ie w p rio r to the o n -sit e visit o r sh o u ld b e re v ie w e d a s p a rt o f th e o n -sit e visit. T he l aborat oryQ A pl an s houl d be a s epar at el ypre pare d t ext
. H owever , d ocument at i on f or m anyo f

the l i s t ed Q A pl an i t em s m ay b e m ad e b y ref ere nc e to a p p ro p riat e sec t i o ns o f this m an ua l , the lab o rato ry' s stand ard o p era t i ng p ro ce d ure s, (S O P s) o r o the r li t era ture (e.g., p ro m ulga t ed m etho d s, Stan da rd Metho ds f or the Exam i na t i on of

W at erandW

astewa ter , et c. ). T he Q A P l an s ho ul d be upd at ed atl eastannual l y(E P A O rder 5 3 6 0
. 1 A 2 ). A ta m i ni m um , the f

o l l o wi ng i t em s shoul d be ad d res sed i n each Q A plan: 1 1 .1 L a b o ra to ry o rg a n iza tio n a n dre sp o n sib ility
• i nc l ud e a ch art o r tab l e sho w i ng the l ab o rato ry org an i za t i o n a nd l i ne s o f resp o nsib i l i ty, i nc l ud i ng Q A m an ag ers;
• l i stthe key i nd i vi d ual s who a re respo nsib l e f o r ensuri ng the p rod uct i o n o f va l i d m easurements a nd the rout i ne ass ess m entof m easurement s y st em s f o r preci si o n and accuracy ( e. g. , who i s res p o nsib l e f o r i nt ernal aud i t s and revi ews of

the i m p l em ent at i o n of the p l an and i t s requir em ent s) ;
• ref erence the job de scr i p t i o ns o f the p ers o nnel a nd de scr i b e trai ni ng t o keep pe rs o nnel upd at ed on regul at i o ns and methodo l ogy , an d do cumen tthatl aborat oryp er s onnelhave d emo ns tr at ed prof i ci enc yf or the methods they pe r f orm. 1 1 .2 P ro ce ss use dto iden ti fy c lien t s' D a ta Q u a lit

y O b jec tive s 11. 3 SO Ps with datesof las t rev is ion 

### C T he l aborat orys houl d m ai nt ai n SO P s thatac cur at el yr ef l ectal l phas es of

cu rr entl aborat oryac t i vi t i es 

### C k e e p a l i s to f

S OPs


### C en sure tha tcu rren tco p i es o f S O P s are i n the lab o rato ry and i n the Q A M an ag ers fi l es; 

### C ens ur e thatSO P s ar e r evi ewed an nu al l yan d re vi s ed as ch ang es ar e made;


### C en s u r e th atSO Ps h av e s i g n atu r e pag es a n d r ev i s i on s da t ed.

## III-5

11 . 4 Field s ampling p r ocedures

• des cr i be the proces s us ed t o i dent i f ys amp l e c ol l ect ors , s amp l i ng procedur es a nd l ocat i ons , r equi r ed pr es er v at i on , pr operc on t ai n er s , c orr ec ts ampl e con t ai n erc l ea n i n g pr oce du r es , s ampl e h ol di n g t i mes f r om co l l ec t i o n to an alysi s, an d sam p l e ship p i ng an d st o rag e c o nd i t i o ns;
• ensur e thatap p rop ri at e f o rms are l egi b l yf

i l l ed out i n i nd el i b l e ink o r har d co p i es of el ectronic d at a are av ai l abl e. Se e Chapt er s I V , V , a n d V I f ors pec i f i c i t ems t o be i n clu ded;

• des cr i be how s amp l es ar e c hec ked whe n theyarr i ve f or pr op ercont ai ner s an d t emp er atur e a nd how s amp l es are ch ec ke d f

o r p ro p er p rese rva t i o n (e .g. , p H , ch l o rine re si d ua l ) b efo re a na l ysi s;
• en sure tha tsam p l i ng p ro t o co l i s writt en an d av ail ab l e to sam p l ers. 11 . 5 L ab oratorys ample rece i ptandhan dling p r ocedures

• b o und l ab o rat o rynote bo o ks, i f us ed , s ho ul d be f

i l l ed out i n i nk; entri es dat ed and si gned (A secure, pa ss wo rd p rotect ed , el ectronic d at a b ase i s accep t ab l e);

• st o re u np ro ce ssed an d p ro ce sse d s a m p l es a tthe p ro p er tem p era ture , i so l ated f ro m l ab o rato ry co ntam i na nts, s t an dar ds a n d h i g h l yc on t ami n at ed s ampl es a n d, s omet i mes , e ac h oth er ; h ol di n g t i mes ma yn otbe e x ce ede d;

• m a inta in in te g rity o f all sa m p le s, (e .g., b y tr a c k ing sa m p le s fro m re c e ip t b y lab o ra to ry th ro u g h a n a lysis to d i spo sal );

• re q u ire C h a in-o f- C u sto d y p ro c e d u re s fo r sam p le s like ly to b e the b a sis fo r a n e n fo rc e m e n t ac tio n (se e A p p e n d ix A );

• s pec i f

yc r i t er i a f

orr ej ec t i on of

s ampl es w h i chdo n otme ets h i ppi n g , h ol di n g ti m e a n d/ orpr es erv at i on req uirem en t s and p ro ce d ure s f o r no t i f i ca t i o n o f

sam p l e o rigi na t o rs. 1 1 . 6 I n s tr u m e n t

c a lib r a tio n p r o c e d u r e s ( m a y re f e r e n c e S OP)

• sp e c ify typ e o f ca lib ra tio n us e d fo r e a c h m e tho d a n d fre q u e n c y o f use ;
• d escrib e cal i b rat i o n s t and ards' s o urce, age, s t o rage, l ab el i ng;
• p erfo rm d ata c o m p ara b i l i ty che ck s;
• use co ntro l ch arts and f o r rad i o ch em i stry, rep o rt co un t i ng erro rs with their co nfi d en ce l ev els. 1 1 . 7An a l

y tic a l p r o c e d u r e s (

m a y re f e r e n c e S OP)

• c ite c o m p le te m e tho d m a n u a l;

• des cr i be qu al i tyc on tr ol pr oce du r es r equ i r ed byth e meth ods th atn ee d t o be f ol l owe d. 1 1 . 8 Da ta re d u c tio n , v a lida tio n , re p o rtin ga n dv e r i

f ic a tio n (

m a y re f e r e n c e S OP)

• des cr i be dat a r edu ct i on pr oce s s : me th od of c on v er s i on of

r aw da t a t o mg / L, pi coc u r i es / L, c ol i f orms / 100 mL, e tc .;
• d esc ribe d ata va l i d ati o n p ro ce ss;
• d e sc rib e re p o rting p ro c e d u re s, inc lud e p ro c e d u re s a n d fo rm a t;

• d esc ribe d ata ve rif i ca t i o n p ro ce ss;
• f o r rad i o ch em i stry, d esc ribe rep o rti ng o f co un t i ng unc ertainti es a nd co nfi d en ce l ev els;

• d esc ribe p ro ce d ure f o r d ata c o rrec t i o ns. 11 . 9 T ype of qua lit

y c ontr ol

( Q C ) checks and the f

r equency of theirus e ( s ee Chap ters I V

, V a nd V

I ) . ( m ay r ef erence S OP) P ar amet er s f

or c hemi s tryan d ra di ochemi s trys houl d i nclude orr ef er enc e:
• i nstrum en tp erfo rm an ce ch ec k stand ard s;
• f req ue nc y and ac ce p t ab i l i ty of m etho d d etec t i o n li m i t(M D L ) ca l cu l ati o ns;
• fre q u e n c y a n d a c c e p ta b ili ty o f d e m o n stra tio n o f l o w lev e l ca p a b ili ty;

• ca l i b rati o n, i ntern al and surro ga t e stand ard s;
• l ab o rat o ryreagentblank, f i el d reagentblank and trip blank;

• f i eld an d l ab o rato ry m atri x re p l i ca t es;
• q ua l i ty co ntro l an d p ro f i cien cy t esti ng sam p l es;
• l ab o rato ry f

o rti f i ed b l an k a nd l ab o rato ry f o rti f i ed sam p l e m atri x re p l i ca t es;
• initial d e m o n stra tio n o f m e tho d c a p a b ili ty

## III-6 • use o f

co ntro l ch arts;

• q ua l i t ati ve i d en t i f

i ca t i o n/co nfi rm ati o n o f co ntam i na nts. P ar amet er s f

or mi cr ob i ol ogys houl d i nclude orr ef er enc e:
• p o si t i ve an d ne ga t i ve cu l ture co ntro l s;
• co nfi rm ati o n/ve rif i ca t i o n o f

p resu m p t i ve t o t al co l i f o rm p o si t i ve sam p l es;
• st eril i ty co ntro l s;
• p ro f i cien cy t esti ng an d q ua l i ty co ntro l sam p l es. 11. 10 Lis t s ch edu les of internal and external s ys tem and data qualit

y audit s and inter labor ator

y compar is ons (

m ay r e f e r e n c e S OP) 11 . 11 P r eve ntive m aintenance procedur es ands chedules
• des cr i be l ocat i on of i ns trument manua l s and s che dul es and document at i on of r out i ne equi pm entmai nt ena nce ;
• d e sc rib e a v a ilab ili ty o f i n stru m e n t sp a re p a rts in the lab o ra to ry;
• l i s tan ymai nt ena nce contr act s i n pl ace . 11 . 12 Co rr ective action con tingencies
• d esc ribe resp o nse t o o b t aining un ac ce p t ab l e re sult s f ro m an alysi s o f P T sam p l es and f ro m i ntern al Q C ch ec ks;

• na m e p erso ns resp o nsib l e fo r the va riou s co rrec t i ve ac t i o ns;
• des cr i be h ow corr ec t i v e ac t i on s t ak ena r e docu men t ed; 11 . 13 Record keeping p r ocedures

• d e sc rib e p ro c e d u re s a n d d o c u m e n ta tio n o f th o se p ro c e d u re s;
• l i stl ength of st o rage, m ed i a typ e (el ectronic o r har d co p y );

• d esc ribe sec urity po l i cy o f elec tro nic d atab ase s;
• al l e l ec tr on i c dat a s h ou l d h av e s of twa r e s u ppor ts o i tma ybe r eg en er at ed. I f a part i cul ari t em i s notr el eva nt , the Q A p l an s houl d s t at e thi s an d provi de a br i ef ex pl ana t i on. A l aborat oryQ A pl an sh o u ld b e re sp o n siv e to the a b o v e it e m s w h il e r e m a inin g b rie f an d e a sy to fo llo w . M inim iz ing p a p e rw o rk , w h ile i m p ro ving d ep en d ab i l i ty and q ua l i ty of d ata, are the i nten d ed go als. 

## 12. Chain-of

-Custody P roce dures C ert i f i ed l aborat ori es , wh en r eques t ed t o p ro ce ss a sam pl e f or pos s i bl e l ega l ac t i on ag ai ns ta s uppl i er , s houl d us e a n ade qu at e ch ai n - of

- cu s t odypr oce du r e. An ex ampl e of s u ch a pr oce du r e i s f ou n d i n Appen di x A. T h e St at e orReg i on shoul d s eek i np utf

rom i t s att o rney general ' s o f f i ce t o ensur e thatthe l ab o rat o ry ' s p roce d ures are adeq uat e. T he p roce d ure u s ed s h ou l d be doc u men t ed. 

## 13 . Re quir e m e nt s forM aintainingCe rtific ationSt atus 13 . 1 P r oficiency Tes ting (PT ) Samples A t le a st a n n u a lly d rin k ing w a te r lab o ra to rie s c e rtifi e d fo r c h e m ic a l co n ta m ina n ts m u st sa tisf a c to rily an a lyz e a P T sa m p le t o ma i n t ai n c ert i f

i ca t i on ( 40CFR 141. 23( k ) ( 3) ( i ) , 141. 24( h ) ( 17) ( i ) ( A) a n d 141. 89( a) (1)( i ) )
. PT s ampl es s h ou l d be a n a lyz e d in the sa m e m a n n e r a s ro u tine sa m p le s. L a b o ra to rie s m u st a c q u ire the P T sa m p le fro m a su p p liera c c e p ta b le to the a p p ro p ria te c e rtifi c a tio n a u tho rity. I f th e ce r t i f

i ed l abor at orydoe s n ota n al y ze th e PT s ampl e wi th i n th e ac ce pt an ce l i mi t s s pec i f i ed i n th e r eg u l at i on s , or w ithin p o licy d e sc rib e d b y th e ir c e rtifyi n g a u tho rity, th e c e rtifyi n g a u tho rity sh o u ld fo llo w the p ro c e d u re d isc u sse d in th e s ec t i on e n t i t l ed, "Cr i t er i a an d Pr oce du r es f orD own g r adi n g /Rev ok i n g Cert i f i ca t i on St atu s . " I f a l abor at oryw i s h es t o be c ert i f i ed f ora c on t ami n an tbymor e th anon e meth od, i tm u s t( 40CFR 14 1. 23 ( k) ( 3)( i i ) , 14 1. 24 ( h) ( 17 ) ( i ) ( A ) an d 14 1. 89 ( a) ( 1)( i ) ) an al y ze the P T s amp l es byea ch method f or w hi ch i twi s hes t o b e c e r t i f

i e d
. The m e th o d s l i s t e d o n th e l a b o r a t o ry ' s c e rt i f i c a t i o n c e rt i f

i c a t e m u s tb e th e m e th o d s b yw h i c h th e PT s ampl es w er e an al y ze d.

## III-7

T he l ab o rat o rys ho ul d b e ab l e t o pro vi d e d o cum ent at i o n t o the cert i f i cat i o n autho ri tythatthe p ers o n(s ) anal y zi ng any P T sam p l e is a l ab o rato ry em p l o yee wh o ro uti ne l y ana l yze s d rinking w ater c o m p l i an ce sam p l es. 13. 2 Met

hodology La b o rat o ri es mustus e the m etho d s specif i ed i n the d ri nki ng water regul at i o ns at 4 0 C FR pa rt14 1 T hese method s are l i s t e d i n C h a p t e r s I V, V, VI , a n d A p p e n d i x G. 13. 3 O n-Site Evaluation T h e C A s h o u ld b e s a tisf ie d th a t a la b o ra to ry is m a inta inin g the re q u ire d sta n d a rd o f q u a lity fo r c e rtifi c a tio n . N o rm a lly, this s ho uld b e b ase d o n a rec o m m en d ati o n f ro m a tri en nial o n-si t e ev alua t i o n. H o w ev er, if the lab o rato ry und erg o es a majorch ang e orr epeat edl yf ai l s a PT s amp l e, the C A s houl d cons i der conduct i ng anev al uat i on bef ore the us ualthr ee y ea rpe r i od h as e x pi r ed. 13 . 4 Noti

f ication o f Certi f ying A uthorit

y ( C A) of Major Chan ges Ce r t i f i ed l a bo r a t or i e s s h ou l d n ot i f yth e a pp r op r i a t e CA ( Re g i on a l Ad mi n i s tr a t ororde s i g n e e orth e Ch i e f , T SCO G W D W ) i n w ri t i ng , wi thi n 30 da y s o f m ajo r changes i n pe rs o nnel , eq ui p m ent , o r l ab o rat o ry l o cat i o n. A m ajo r change i n pe rs o nnel i s d ef

i ned as the l o ss o r re p la c e m e n tof the l ab o rat o rys up ervi sor or a s i tuat i o n i n whi ch a trai ned and e x p e r i e n c e d a n a l y s ti s n o l o n g e ra v a i l a b l e t o a n a l y z e a p a rt i c u l a rp a r a m e t e rf o rw h i c h c e rt i f

i c a t i o n h a s b e e n g r an t ed. T he C A sho uld d i scu ss the si tua t i o n w i th the lab o rato ry sup erv i so r a nd e st ab l i sh a sch ed ule f o r the lab o rato ry t o ad d ress m ajo r changes. I f

the C A de t erm i n e s thatthe l ab o rat o rycan no l o nger pro d uce val i d da t a, the C A s ho ul d f o l l o w the p roce d ure f

o r r evo cat i o n of cert i f

i cat i o n. 

## 14 . Crite ria and P r oc e dur e s forDowngr ading/Revo kingCe rtific ationSt atus 14. 1 Cr iteria f

orD owngr ading Certi

f ication Status A l abor at orys h ou l d be down g r ade d t o "prov i s i on al l yce rt i f i ed" s t atu s f

ora c on t ami n an torg r ou p of c on t ami n an t s f

or an y of the f

o l l o w i ng rea so ns:

• Fa i l ure t o anal y ze a P T s am p l e atl eastannual l ywi thi n the accep t ance l i m i t s specif i ed i n the regul at i o ns, or, if the re a re no re q u ire m e n ts sp e c ifi e d in the r e g u la tio n s, w ithin p o licy d e sc rib e d b y the ir c e rtifyi n g a u tho rity;

• F a ilure o f a c e rtifi e d lab o ra to ry to no tif y the C A w ithin 3 0 d a ys o f m a jo r ch a n g e s (e .g., in p e rs o n n e l, e q u ip m e n t, o r l ab o rat o ryl o cat i o n);

• F a i l u r e t o s a t i s f

yth e CA th a tth e l a b o r a t o ryi s m a i n t a i n i n g th e r e q u i r e d s t a n d a r d o f q u a l i ty , b a s e d up o n a EP A o n- si t e eval uat i o n;

• F a ilure to re p o rt c o m p lian c e d a ta to the p u b lic w a te r sys te m o r th e S ta te d rin k ing w a te r p ro g r a m i n a ti m e ly m a n n e r, the re b y p re v e n ting c o m p lian c e w ith F e d e ra l o r S ta te re g u la tio n s a n d e n d a n g e rin g p u b lic h e a lth. D a ta w h ic h m a y c a u se the syste m to e x c e e d a n M C L sh o u ld b e re p o rte d a s so o n a s p o ssib le
. 14. 2 Pr oce duresf

orD owngr ading to "Pr ovis ionall y Certi f ied"Status If a l ab o rat o ryi s subjectt o do wngrad i ng on the b asi s o f the ab o ve i nd i cat ed cri t eri a, the C A s ho ul d noti f ythe l ab o rat o ry d i rect o r or ow ner ( b yregi st ered or cert i f i ed m ai l ) of

i t s i nt entt o do wngrad e wi thi n 14 da y s f rom be co m i ng aware o f the s i tu at i on w arr an t i n g dow n g r adi n g . T h e l abor at orydi r ec t o r should revi ew th e pr obl ems c i t ed, a n d w i th i n 30 da y s of recei p tof th e l ett er , s en d a l ett er t o th e CA s pec i f y i n g wh at i mmedi at e corr ec t i v e ac t i on s ar e bei n g t ak en and any p ro p o se d a c tio n s tha t ne e d the c o n c u rr e n c e o f the C A
. T h e C A sh o u ld c o n sid e r th e a d e q u a c y o f the re sp o n se a n d no tif y the lab o ra to ry in w riting (b y re g ister e d o r c e rtifi e d m a il) o f i ts c e rtifi c a tio n sta tus w ithin 1 4 d a ys o f re c e ip t o f i ts res p o nse. T he C A s ho ul d f o l l o w up t o ensur e thatco rr ect i ve act i o ns have b een t aken. I f a l abor at oryf

ai l s t o a n al y ze a PT oroth eru n k n owns ampl e wi th i n th e ac ce pt an ce l i mi t s , th e CA s h ou l d n otdow n g r ade cert i f i cat i on i f the l aborat oryi dent i f

i es and corr ect s the probl em t o the C A ' s s at i s f act i on wi thi n 30 day s of bei ng not i f

i ed of th e f ai l u r e. I f

, a f t era r ev i ew of

th e s u bmi tt ed i n f

ormat i on , th e CA det ermi n es th atth e l abor at ory ne ed no tbe do wng r aded, thenwi thi n 30 day s o f this deci s i on, the C A s houl d not i f ythe l aborat orythati ti s r equi r ed t o ana l y ze

## III-8

a n o th e r PT. I f th e l a b o r a t o ry a n a l y z e s th i s s e c o n d u n k n o w n s a m p l e w i th i n th e a c c e p t a n c e l i m i t s e s t a b l i s h e d b y th e EPA o r S ta te , th e lab o ra to ry sh o u ld n o t b e d o w n g ra d e d . If the lab o ra to ry fa ils t o a n a lyz e this se c o n d un k n o w n sa m p le w ithin the esta b lishe d li m i t s, the C A s ho ul d do wngrad e the l ab o rat o ryt o "p rovis i o nal l ycert i f i ed " s t atus and noti f

ythe la b o ra to ry w ithin 1 4 d a ys (b y re g ister e d o r c e rtifi e d m a il). L a b o r a to r ie s s h o u ld b e d o w n g ra d e d o n ly f o r th e a n a lyte f ai l ed, e x ce ptw h er e EP A/ St at e ce r t i f i es a g r ou p of

r el at ed an al y t es ba s ed on a l i mi t ed n u mberof an al y t es i n th e g r ou p. (S e e C h a p te r 4 , S e c tio n 7 .2 .1 fo r a d d iti o n a l i n fo rm a tio n .) D u rin g a n y p h a se o f th is p ro c e d u re , a la b o ra to ry m a y re q u e st tha t th e E P A o r S ta te p ro v id e tec h n ic a l as sista n c e to he lp i de n t i f ya n d r e s ol v e a n ypr ob l e m. Af t erth e CA not i f

i es a l abor at ory , i n w r i t i n g , th ati th as be endow n g r aded t o " p rovis i o nal l yc ert i f i ed" s t atu s f

or procedur al , admi nistrati ve, equipm entor pers onneldef i ci enc y , the l aborat orys houl d corr ecti t s probl em wi thi n thr ee m o n ths . If the la b o ra to ry wa s d o w n g ra d e d to " p ro v isio n a lly c e rtifi e d " sta tus b e c a u se o f a failu re to a n a lyz e a P T sa m p le (o r o the r u n k n o w n tes t sa m p le ) w ithin the a c c e p ta n c e li m its sp e c ifi e d in the re g u la tio n s, o r w ithin p o licy re q u ire d b y the ir cert i f y i ng author i ty , the l aborat orys houl d corr ecti t s probl ems and s at i s f act ori l y ana l y ze anotherP T s amp l e ( or other un kn own sa mpl e ) w i th i n on e mo n th of r e c e i ptof

th e s e c on d PT s a mp l e . A pr ov i s i on a l l yc e rt i f i e d l a bo r a t oryma y con t i n u e t o a n al y ze s ampl es f orc ompl i an ce pu rpos es , bu t s h ou l d n ot i f y i t s cl i en t s of i t s down g r ade d s t atu s a n d pr ov i de tha t i n fo rm a tio n , i n w riting , o n a n y re p o rt. 14. 3 Cr iteria f

orRev oking Certi

f ication Status A l abor at orys h ou l d be dow n g r ade d f r om c ert i f

i ed, pr ov i s i on al l yc ert i f i ed or i n t er i m ce rt i f

i ed s t atu s t o "n otc ert i f i ed" f o r a p arti cu l ar c o ntam i na nt ana l ysi s f o r the f o l l o w i ng rea so ns:

• R e p o rting P T d a ta fro m a n o the r la b o ra to ry a s its o w n ;
• F alsi f

i ca t i o n o f

d ata o r o the r d ec ep t i ve p rac t i ce s;
• F ail ure t o use the an alyt i ca l m etho d o l o gy sp ec i f i ed i n the re gu l ati o ns;
• F o r p ro visi o na l l y ce rti f i ed l ab o rato ries, f

ail ure t o suc ce ssf

ull y an alyze a P T sam p l e o r an y o the r un kn o w n test sa m p le fo r a p a rtic u la r c o n ta m ina n t with in th e a c c e p ta n c e li m its sp e c ifi e d ;
• Fo r p rovis i o nal l y cert i f i ed l ab o rat o ri es, f

ai l ure t o sat i sf

y the C A that the l ab o rat o ry has co rr ect ed d e v ia tions i d en t i f i ed d uring a n o n-si t e e va l ua t i o ns;
• For pr ovi s i onal l yce r t i f i ed l aborat ori es , pers i s t entf ai l ur e t o re po rtcomp l i anc e dat a t o the publ i c wa t ers y s t em o r th e S ta te d rin k ing w a te r p ro g ra m in a tim e ly m a n n e r th e re b y p re v e n ting c o m p lian c e w ith F e d e ra l an d /o r S ta te r eg u l at i on s a n d e n dan ger i n g publ i c h ea l th
. D at a wh i chma yc au s e th e s y s t em t o e x ce ed anM CL s h ou l d be r eport ed as s oo n a s poss i bl e.
• Re f u s a l t o p a r t i c i pa t e i n a n on - s i t e e v a l u a t i onc on du c t e d b yth e CA 14. 4 Pr oce duresf

orRev ocation T he C A shoul d no t i f

y the l ab o rat o ry , i n wri t i ng (by regi st ered o r cert i f i ed m ai l ) of

the i nt entt o revoke cert i f i cat i o n. I f th e l abor at oryw i s h es t o c h al l en g e th i s de ci s i on , a n ot i ce of a ppea l s h ou l d be s u bmi tt ed i n wr i t i n g t o th e CA wi th i n 30 day s of r ec eiptof

th e n ot i ce of

i n t en tt o r ev ok e ce rt i f i ca t i on . I f

n o n ot i ce of

a ppea l i s f

i l ed , c ert i f

i ca t i on s h ou l d be r ev ok ed. T h e n ot i ce of

appe als h ou l d be s u pport ed wi th an ex pl an at i on of th e r ea s on s f

orth e ch al l en g e an d s h ou l d be s i g n ed by a res p o nsib l e o f

f i ci al f rom the l ab o rat o rys uch as the p res i d ent/ o wner f o r a com m erci al l ab o rat o ry , or the lab o rat o ry su p e rv iso r in the c a se o f a m u n ic ip a l l a b o ra to ry o r th e lab o ra to ry d ire c to r fo r a S ta te o r R e g io n a l l a b o ra to ry. W i th i n 30 da y s of

r ec eiptof

th e appe al , th e CA s h ou l d ma k e a dec i s i on a n d n ot i f yth e l abor at oryi n w r i t i n g ( by regi st ered or cert i f

i ed m ai l ). D eni al of the ap p eal s ho ul d res ul ti n i m m ed i at e revoc at i o n of the l ab o rat o ry ' s cert i f i cat i o n. O n ce c ert i f

i ca t i on i s re v ok ed, a l abor at orym aynotan al y ze dri n k i n g wat ers ampl es f orc ompl i an ce u n t i l i t s c ert i f i ca t i on h as be enr ei n s t at ed. If the ap p eal i s d et erm i ned t o b e val i d , the C A sho ul d take ap p rop ri at e m easures t o reeval uat e the f aci l i tyand noti f

ythe l ab o rat o ry , i n wri t i ng (by regi st ered or cert i f i ed m ai l ), of i t s d eci si o n wi thi n 30 da y s o f the reeval uat i o n.

## III-9

14. 5 U pgr ading orReins tatem en t of Certi f ication T hrough a wri tt en reque st , a labo rat o rym ay seek up grading o r rei nst at em ento f c ert i f i cat i o n, w hen and if

the l ab o rat o ry ca n de mon s tr at e t o th e CA' s s at i s f ac t i on th atth e def

i ci en ci es w h i chpr odu ce d pr ov i s i on al l yc ert i f i ed s t atu s orr ev oca t i on hav e been corr ect ed. T hi s may i nclude an on- s i t e eva luat i on, s ucc es s f ul ana l y s i s of un kn own s amp l es or a ny o t her meas ur e the C A deems appropri at e. 

## 15 . Re c or d Ke eping T h e ce r t i f

i ca t i on pr og r am man ag ers h ou l d e n s u r e th atr ec or ds f o r o n- si t e l abor at orya s s es s men t s a n d c ert i f i ca t i on p rogram revi ews b e m ai nt ai ned i n an easi l y accessib l e central l o cat i o n f o r a p eri o d o f 6 y ears t o i ncl ud e the l asttwo onsi t e aud i t s, or l o nger i f requir ed b y specif i c Stat e regul at i o ns. T hi s i ncl ud es r eco rds/ co rr espo nd ence used t o de t erm i ne co m p l i an ce with the re q uirem en t s i n this m an ua l
. R ec o rd s m ay i nc l ud e che ck l i st s, co rrec t i ve ac t i o n rep o rts , f i na l rep o rts , c e rtifi c a te s, P T stud y re su lts an d re la te d d o c u m e n ts 

## 16. Reciprocity R eciproci ty( mutual l y acc ept abl e cert i f i cat i on amo ng R egi ons and/ or pri macy St at es ) i s s tr ongl y endors ed by E P A as a h i gh l y de si rab l e e l em en ti n the c erti f i ca t i o n p ro gra m f

o r d rinking w ater lab o rato ries. S ta te s a re e n c o u ra g e d to a d o p t p ro v isio n s in th e ir law s a n d re g u la tio n s to p e rm it r e c ip r o c ity. E v e n tho u g h ultim a te res p o nsib i l i tyf

o r r eci p roca l cert i f

i cat i o n res i d es wi th the R egi o ns and pri m acy Stat es, the St at es may as k f o r the a ssista n c e o f E P A in c a se s inv o lvin g c la rifica tio n o f w h a t sh o u ld b e c o n sid e re d in a re c ip ro c a l ag re e m e n t. S u c h re q u e sts shoul d be s ub m i tt ed t o the R egi o n or O G W D W through the R egi o n. 

## 17 . Tr ain ing T r ai n i n g i s a n int egral partof th e l abor at oryc ert i f

i ca t i on pr oce s s f

orpe r s on n elc on du ct i n g on - s i t e ev alu at i on s of l aborat ori es on beha l f

of ei therthe R egi onal O f f i ce or apri macySt at e. EP A pol i cyre qu i r es th atal l R eg i on alC ert i f i ca t i on O f

f i ce r s i n i t i al l ypas s th e a ppr opr i at e EP A l abor at oryc ert i f i ca t i on tr ai n i n g c ou r s es f

orth e di s cipl i n e f

orw h i ch th eyc ert i f

y( ch emi s tryormi cr obi ol og y )
. Al l l abor at orya u di t or s s h ou l d be ex per i en ce d pr of

es s i on al s , a n d h av e at l ea s t a bac h el or ' s de g r ee ore qu i v al en te du ca t i on / ex per i en ce i n th e di s cipl i n e f or whi ch they cert i f

ya nd rece ntlabo rat o ry exp eri ence i n the f i el d for w hi ch they aud i tlabo rat o ri es. T hi rd p artyaud i t o rs (s ee A p p end i x D ) al so need t o pa ss the E P A cert i f i cat i o n trai ni ng co urs e. O ut si d e exp ert s, ret ai ned f o r thei r knowledge i n al i mi t ed ar ea ( e. g. , as bes t os) are notrequi r ed t o pas s the l aborat orycert i f i cat i on co ur s e i f

theyar e u s ed as partof an o n -sit e a u d it te a m w h ic h inc lud e s a c e rtifi c a tio n o ff ic e r. P e rio d ic tra inin g fo r b o th la b o ra to ry a u d ito rs a n d a n a lysts shoul d be pro vi d ed by the R egi o ns. C ert i f i cat i o n off

i cers shoul d att end ref

res her trai ni ng p rogram s atleas te veryf i ve y ear s t o kee p thei rkn owl edge of the methods and the dri nk i ng wat er progr am curr ent
. I ti s hi gh l yr ecomm ended that c e rtifi c a tio n o ff

ic e rs h a v e re c e n t b e n c h e x p e rie n c e in th e m e tho d s fo r wh ic h the y c e rtify. O G W D W w il l n o tif y ce rti f i ca t i o n o f

f i ce rs o f m aj o r up d ates/cha ng e s to E P A ' s ce rti f i ca t i o n p ro gra m . Iti s rec o m m en d ed tha tthe S t ates use the se sa m e c rite ria in the ir c e rtifi c a tio n p ro g ra m s. 

## 18 . Alte r nateTe stP r oc e dur e s (AT P s) E P A promul gat es an al y t i calmethods f or a l l r egu l at ed d rinki ng wat ercont ami nan t s . A r egu l at i on f or apart i cul ar co nt am i nantwi l l i ncl ud e o ne o r mo re m e tho d s thatm ustbe us ed t o de t erm i ne thatco nt am i nant
. Sub sequentl y , the A ge n c y m ay ap p ro ve ad d i t i o na l m etho d s o r m o d i f i ca t i o ns o f

E P A ap p ro ve d m etho d s i n a no the r rule. E P A m ay also author i ze the us e of

al t ernat e a nal y t i calmethods as provi ded i n 40 C FR 141
. 27 , "W i th the wr i tt en permi s s i on of the St at e, co ncurred b ythe A d m i ni strat o r of the E P A , an al t ernat e anal y t i cal t echni q ue m ay b e em p l o y ed
. A n al t ernat e t echni q ue maybe ac cept ed onl y i f i ti s s ubs t ant i al l yequi val entt o the pres cr ibed t es ti n both pr eci s i on an d accu r acyas i tr el at es to the d e te rm ina tio n o f co m p lian c e w ith a n y M C L ." Any on e ca n r equ es tth atEP A appr ov e a n ew me th od ormodi f i ca t i on of

a m et ho d a l r ea dya ppr ov ed byEP A, by s u bmi tt i n g EP A- s pec i f i ed da t a a n d oth eri n f

ormat i on t o th e D i r ec t or , A n al y t i ca l M eth ods St af f , ( M S 4303T ) O f

f i ce of S c ie n c e a n d T e c h n o lo g y, O ff ic e o f W a te r, E P A , 1 2 0 0 P e n n s y lv a n ia A v e ., N W , W a sh ing to n D C 2 0 4 6 0 . E P A w ill e v a lua te the m a te ria l t o d e te rm ine w h e the r th e m e tho d o r m e tho d m o d ifi c a tio n m e e ts E P A c rite ria
.

## III-10

In the c a se o f “ac c e p ta b le ve rs io n s” o f m e tho d s, (m ino r m o d ifi c a tio n s to a p p ro v e d m e tho d s) , a le tter o f a p p ro v a l will be i s s ued byO W
. A l i s tof thes e app r oved mi normo di f i cat i ons ca n be f

ound on the O W webs i t e a t http : //w w w .ep a.go v/O G W D W /m etho d s.

## IV- 1 Chapt e rIV Critic al Elem ents for Chemistry
1. P ersonnel 1. 1 Labor ator

y Supe r vis or T he l ab o rato ry s up erv i so r sho uld ha ve at l ea st a b ac he l o r' s d eg ree w i th a m aj o r i n che m i stry o r eq uiva l en t , an d at l ea st o ne y ear of exp eri ence i n the anal y si s o f dri nki ng w a te r. T h e labo rat o rys up ervi sor s ho ul d have atl easta wo rki ng kno wl ed ge o f

qua l i tyass urance p ri nci p l es. T he l ab o rat o rys up ervi sor has the res p o nsib i l i tyt o ensur e thatal l l ab o rat o ry p e rs o n n e l ha v e d e m o n stra te d the ir ab ili ty t o sa tisf a c to rily p e rfo rm the a n a lyse s to w h ic h the y a re a ssig n e d a n d tha t all dat a r eport ed by the l aborat orymeetthe r equi r ed qual i tyas s ur anc e a nd r egu l at orycr i t er i a. 1 .2 L a b o ra to ry A n a l

yst T he l ab o rat o ryanal y sts ho ul d have atl easta ba chel o r' s d egree wi th a m ajo r i n chem i stryor eq ui val ent , and atl eastone y earof ex peri enc e i n the an alysi s of dri nk i ng wat er . I f

the ana l y s ti s r es po ns i bl e f or the op er at i on of

an al y t i cal i ns trument at i on, he or s he s houl d hav e c om pl et ed s peci al i zed tr ai ni ng o f f er ed by the manuf

actur eror anotherqual i f i ed tra inin g fac ili ty o r se rv e d a p e rio d o f a p p re n tice sh ip u n d e r a n e x p e rie n c e d a n a lyst. T h e d u ra tio n o f th is a p p re n tice sh ip sh o u ld b e p ro p o rtio n a l t o the so p h istica tio n o f th e instru m e n t. D a ta p ro d u c e d b y a n a lysts a n d instru m e n t o p e ra to rs w h ile in th e p ro c e ss o f o b ta inin g the re q u ire d tra inin g o r ex p e rie n c e a re a c c e p ta b le o n ly wh e n re v ie w e d a n d va lid a te d b y a fully q ual i f i ed anal y stor the l ab o rat o rys up ervi sor. B ef ore begi nn i ng the ana l y s i s of comp l i anc e sa mp l es , the an al y s tm us tad her e t o anyrequi r ed Q C p r ocedur es s peci f i ed in the m e tho d s fo r b la n k s, p re c isio n , ac c u ra c y, se n sitivity, sp e c ifi c ity a n d sa tisf a c to ry a n a lysis o n un k n o w n sa m p le s. T h is shoul d be do cum ent ed acco rding t o the l ab o rat o ry ’ s Q A P l an. 1. 3 Tec hnician T he l aborat oryt ech ni ci an s houl d hav e a t l eas t a hi gh s chool di pl om a or equi val ent , com pl et e amethod tr ai ni ng progr am u n d e r a n e x p e rie n c e d a n a lyst an d ha v e six m o n ths b e n c h e x p e rie n c e in the a n a lysis o f d rin k ing w a te r sa m p le s. B ef ore begi nn i ng the a na l ys i s of comp l i anc e s amp l es , the t ech ni ci an mus tadhe r e t o anyr equi r ed Q C procedur es s peci f i ed i n the methods f

or bl ank s , pre ci s i on, ac cur acy , s ens i t i vi ty , s peci f i ci tyan d s at i sf

act oryan al y s i s on u nknown samp l es. T hi s shoul d be do cum ent ed acco rding t o the l ab o rat o ry ’ s Q A P l an. 1. 4 Sam pling P ers onn el P er s onnelwh o col l ects amp l es s houl d be tr ai ned i n the proper col l ect i on t ech ni que f or al l types of

s amp l es wh i ch they c o llec t. T h e ir tec h n iq u e sh o u ld b e re v ie w e d b y e x p e rie n c e d sa m p ling o r la b o ra to ry p e rs o n n e l. 1. 5 Waive rof

Acade m ic Tr aining Requ ir em en t T h e c e rtifi c a tio n o ff

ic e r m a y w a ive the ne e d fo r sp e c ifi e d a c a d e m ic t ra inin g , o n a c a se -b y-ca se b a sis, fo r h igh ly ex p erien ce d an alyst s. 1. 6 Tr aining Rec or ds T rai ni ng record s shoul d b e m a i nt ai ned f o r al l pe rs o nnel
. T hese s ho ul d i ncl ud e al l job

- rel at ed f o rma l ed ucat i o n and tr ai ni ng t ake n by the ana l y s twh i ch pert ai ns t o anyas pectof hi s /herr es po ns i bi l i t i es , i ncludi ng butnotl i mi t ed t o ana l y t i cal methodo l ogy , l aborat orys af ety , s amp l i ng , qual i tyas s ur anc e, dat a a nal y s i s , et c. 

## 2. Laboratory F acilities T he ana l y s i s of

comp l i anc e s amp l es i s t o be conduct ed i n al aborat orywher e the s ecu r i tyand i nt egr i tyo f the s amp l es a n d th e d a ta can b e m ai nt ai ned
. T he l ab o rat o ryf aci l i t i es s ho ul d be cl ean, have ad eq uat e t em p erature and hum id ity co ntro l , ha ve ad eq ua t e li gh t i ng at the b en ch t o p an d sho uld m ee tap p l i ca b l e O S H A st an d ar d s. T h e l ab o rato ry m ust adher e t o anyr equi r ed Q C p r ocedur es s peci f i ed i n the methods by ha vi ng provi s i ons f or the proper s t orag e a nd di s po s al of c h emi ca l w as t es ; s ec on daryc on t ai n men tf orh az ar dou s w as t e st o rage is recommen ded. T h e appr opr i at e type of exh aus thood i s r equi r ed whe r e app l i cabl e.

## IV- 2

T here s ho ul d be s uf

f i ci entbe nch space f

o r proc ess i ng s am p l es. W o rkbenc h s p ace should b e co nveni entto si nk, wat er, ga s, va cu um an d elec trical sou rce s f ree f ro m surge s. Instrum en t s sho uld b e p ro p erly gro un d ed
. F o r safetyrea so ns, i n or g an i c an d or g an i c f ac i l i t i es s h ou l d be i n s epa r at e r ooms ; or g an i c an al y s i s a n d s ampl e ex tr ac t i on s h ou l d a l s o be s epar at ed t o prev entcr oss cont ami nat i on. T he ana l y t i cal and s amp l e s t orag e ar eas s houl d be i s ol at ed f r om al l pot ent i al sources o f c o n ta m ina ti o n. T here s ho ul d be s uf f i ci ents t o rage s p ace f

o r the saf

e st o rage of chem i cal s, gl ass ware and p o rta b le e q u ip m e n t, su ff ic ie n t f lo o r a n d b e n c h sp a c e fo r sta tio n a ry e q u ip m e n t an d a re a s fo r c le a n ing m a te ria ls. 

## 3. Laboratory Equipm ent and In strum entation T h e l abor at oryi s t o ha v e th e i n s tru men t s a n d e qu i pmen tn ee ded t o pe r f orm th e appr ov ed meth ods f orw h i chc ert i f

i ca t i on has been req uest ed
. T he checkli sto n p a g e s 4 4 t o 50 of thi s chap t er pro vi d es mo re i nf o rma t i o n on the necessary equ i pmen t
. Al l i n s tru men t s a r e t o be pr oper l yma i n t ai n ed an d c al i br at ed. 

## 4. Ge neral Laboratory P ractices 4. 1 G en eral 4 .1.1 C h em i ca l s/ re a g en ts: C he m i ca l s and rea ge nts used m ust m ee tan y req uirem en t s sp ec i f i ed i n the m etho d s. I f n ots pec i f

i ed, th en"An al y t i ca l r ea g en tg r ade " ( AR) orAmer i ca n Che mi ca l Soc i ety( ACS) g r ad e chem i ca l s or bett ers h ou l d be us ed f

oran al y s es i n c ert i f

i ed l abor at or i es . Con s u l tth e curr en t l ypr omu l g at ed edi t i on s of S ta n da rd Metho dsf

or the Exam i na t i on of

W at erandW

astewa ter , p art 1 0 7 0 f

o r m o re d etail ed i nfo rm ati o n o n re ag en tgra d es. 4 .2 In o rg a n ic Co n ta m in a n ts 4. 2. 1 R eagent w ater : T h e lab o ra to ry m u st h a v e a so u rc e o f re a g e n t w a te r ha v ing a re sista n c e v a lue o f at lea st 0 .5 megohms ( conduct i vi tyl es s than2. 0 m i cr om hos / cm) at25 C wh en r equi r ed by the method. H i gh qual i t y w at er o m ee t i ng such sp ec ific a tio ns m ay b e p urc ha sed f ro m co m m erc i al sup p l i ers. Q ua l i ty of rea ge nt wa t er is be st m ai nt ai ned by s eal i ng i tf rom the atm o sphere. Q ual i tychecks t o m eets p eci f i cat i o ns ab o v e s h o u ld b e m ad e and d o c u m e n te d a t p la n n e d inte rv a ls b a se d o n u se . Ind ivid u a l an a lyti c a l m e tho d s m a y sp e c ify a d d iti o n a l re q u ire m e n ts f orthe r ea g en tw at ert o be u s ed. I n or g an i c meth ods r equ i r e di s t i l l ed orde i on i ze d w at erf r ee of th e an al y t e( s ) of i nt erestand trace m et al s m etho d s requir e A ST M T yp e 1 wat er. 4. 2. 2 Gl as s w are pr ep arat i on: G l ass ware cl eani ng requir em ent s specif i ed i n the m etho d s m ustbe f o l l o wed
. I f no s peci f

i cat i ons are l i s t ed, thengl as s war e sh oul d be was hed i n a warm det er gen ts ol ut i on an d thor ough l yr i ns ed f i rstwith tap wa t er a nd the n w i th re ag en tw ater. T his c l ea ning p ro ce d ure is suff i cien tfo r ge ne ral ana l yt i ca l ne ed s. Iti s ad vant ageo us t o m ai nt ai n s ep arat e set s o f s ui t ab l yprep ared gl ass ware f o r the ni trat e and m ercuryanal y ses due t o the po t ent i al f

or c ont ami nat i on f

r om the l aborat ory env i r onment
. T abl e I V
- 1 s ummari zes the cl ean i ng procedur es sp ec i f i ed i n the E P A m etho d s. 4 .3 O rg a n ic Co n ta m in a n ts 4. 3. 1 R eagent w ater : R ea ge nt w ater f o r o rga nic an alysi s m ust ad he re t o an y req uired Q C sp ec i f i ed i n the m etho d s. M os tme th ods s pec i f

yth e r ea g en tw at ern otc on t ai n a n aly t es of int eresta b o ve th ei rr es pec t i v e meth od de t ec t i on l eve l s ( M D Ls )
. I tmaybe ne ces s aryt o tr eat wat er wi th act i vat ed carbo n t o el i mi nat e a l l i nt er f er enc es . Reage ntwat er r equ i r eme n t s of

i n di v i du alme th ods mu s tbe f ol l owe d. 4. 3. 2 Gl as s w are pr ep arat i on: G l as s wa r e cl ea n i n g r equ i rem ent s s pec i f i ed i n th e meth ods mu s tbe f ol l owe d. T ab l e IV
- 1 sum m arize s the clea ning p ro ce d ure s sp ec i f i ed i n the E P A m etho d s. 4 .4 L a b o ra to ry S a

fe ty W h i l e s af

ety cr i t er i a ar e n ot an as pec t of l abor at ory ce rt i f i ca t i on , l abor at ory per s on n el s h ou l d appl y g ener al and cus t om arys af

etypra ct i ces as a p arto f goo d l aborat oryp r act i ces . E ach l aborat oryi s en cour age d t o hav e as af etypl an a s p a rt o f th e ir sta n d a rd o p e ra ting p r o c e d u r e w h ic h inc lud e s p e rs o n n e l sa fety, tra inin g a n d p ro te c tio n . W h e re sa fety p rac t i ce s a r e r equi r ed i n a n app r oved method (i
. e. , 515
. 1), theymus tbe f ol l owed. See S ta n da rd Me th o ds f o rthe Exam i na t i on of

W at erandW

astewa ter , p a rt 1 0 9 0 fo r a d isc u ssio n o f l a b o ra to ry sa fety. 4. 5 Q ualit

y Ass ur ance Labo r at ori es s houl d m ai nt ai n c urr entQ ual i tyAss ur anc e Pl ans as des cr i bed i n Chapt er3. Al l l aborat oryac t i vi t i es

## IV- 3

i n clu di ng, but no tl i m i t ed t o , s am p l i ng, t estm etho d s, i nstrume ntop erat i o n, da t a generat i o n, da t a val i d at i o n and co rr ect i ve a c tio n p ro c e d u re s sh o u ld b e d e sc rib e d in the P la n . P la n s n e e d to b e re a d b y a ll p e rs o n n e l. 

## 5.Analytic al M eth ods 5. 1 G en eral A l i s tof

pr omu l g at ed meth ods f ori n or g an i c an d or g an i c co nt am i nant s c anbe f ou n d i n T abl es I V
- 2 a n d I V
- 3, r es pec t i v el y . M eth ods ma n u al s s h ou l d be a v ai l abl e t o a ppl i cabl e per s on n el
. O th erme th ods c an n otbe u s ed f or c o m p lian c e sa m p le s u n le ss a p p ro v a l ha s b e e n gr a n te d b y the A g e n c y b y o b ta inin g a n A lter n a te T e st P ro c e d u re a p p ro v a l. A l low edm o d ifi c a tio n to the m e tho d s m u st b e d o c u m e n te d . C o n ta c t th e a p p ro p ria te c e rtifyi n g a u tho rity fo r th e a lter n a te t es tpr oce du r e pr oce s s ( s ee Chapt er3, p 10)
. T abl e I V
- 4 l i s t s th e meth ods w h i chmu s tbe u s ed f orth e an al y si s of d isinfe c ta n t re sid u a ls. R e c o m m e n d e d m e tho d s fo r S e c o n d a ry c o n ta m ina n ts a re li ste d in T a b le IV -5
. 5. 2 An al

ys es Ap p ro v e db y th e S ta te M easurements f

o r turbidi ty , pH , t em p erature, dis i nf ect antres i d ual , cal ci um , ortho p ho sphate, s i l i ca, al kal i ni ty , and conduct i vi tyne ed notbe made i n cert i f i ed l aborat ori es , butmaybe per f ormed by an yper s ons ac cept abl e t o the St at e. H owever , approved methodo l ogymus t ) be us ed ( T abl es I V
- 2 t o IV
- 5). T he St at e sh oul d i ns t i tut e aqual i tyas s ur anc e p ro gra m t o assure va l i d i ty of d ata fro m the se m ea sure m en t s. 5 .2 .1 T u rb i d i ty sta n d a rd s: Se al ed l i qu i d s ec on darytu rbi di tys t an dar ds pu r ch as ed f r om th e i n s tru ment m anuf acturer o r other s o urces shoul d be cal ib ra ted agai nstpro p erl yprep ared and dilut ed f o rma zi n or sty rene di v i n y l ben ze n e pol y merpr i marys t an dar ds a n d r ev i s ed v alu es as s i g n ed a tl ea s te v eryf ou rmon th s i n or dert o mon i t or f ora n yde t er i or at i on . T h i s c al i br at i on s h ou l d be docu men t ed. T h es e s t an dar ds s h ou l d be r epl ac ed wh enth eydo n otf al l w i th i n 15% of

th e i n i t i ala s s i g n ed con ce n tr at i on of th e s t an dar d. Sol i d turb i d i tys t an dar ds c ompos ed of pl as t i c, g l as s , oroth erma t er i al s a r e n otr el i abl e an d s h ou l d n otbe u s ed. 5 .2.2 R esid u a lch l o rin e sta n d a rd s: I f vi s ualcomp ar i s on devi ces s uchas col or w hee l s or s eal ed ampul es ar e u s ed f o r d eterm i ning free ch l o rine re si d ua l , the st an d ard s i nc o rp o rated i nto such d ev i ce s sho uld b e c ali b r a t ed at l ea st everys i x m o nths. T hese cal i b rat i o ns need to b e d o cum ent ed
. D i rect i o ns f o r prep ari ng t em p o raryand pe rma nent type v i s u als t an dar ds c anbe f ou n d i n M eth od 4500- Cl

- G , of th e curr en t l ypr omu l g at ed edi t i on s of Standard Methods f or the Exam i na t i on of

W at erandW

astewa ter . B yc ompar i n g s t an dar ds a n d pl ott i n g s u cha c ompar i s on on g r aph p ap er, a c o rrec t i o n facto r ca n b e d erive d an d ap p l i ed t o f uture result s o b t aine d o n the n o w ca l i b rated ap p ara tus. 

## 6. Sample Collection, Hand ling, an d Preservation T he manne ri n w hi ch s amp l es ar e c ol l ect ed an d han dl ed i s cr i t i cal t o obt ai ni ng va l i d dat a. I ti s i mp ort antthata w r i tt en s ampl i n g pr ot ocolw i th s pec i f i c s ampl i n g i n s tru ct i on s be a v ai l abl e t o and u s ed bys ampl e col l ec t or s a n d a v ai l abl e f or i nspecti o n by the cert i f i cat i o n off

i cer. ( A p p end i x A , C hai n- o f

- C ust o d y ). 6. 1 R ejection o

f Samples T he l ab o rat o ry ’ s reject i o n cri t eri a shoul d be do cum ent ed i n wri t i ng i n the l ab o rat o ry ’ s Q A P l an o r i n an SO P
. T he l abor at orys h ou l d r ej ec t an y s ampl e t ak en f orc ompl i an ce pu rpos es w h i chdoe s n otme etth e cr i t er i a i n 6. 2 th r ou g h 6. 6. T he l ab o rat o rym ust(1 4 1
. 2 3 (a)( 4 )( i )) no t i f ythe autho ri tyrequest i ng the anal y ses and ask f o r a r esam p l e. If

res am p l i ng i s no tpo ss i b l e and the samp l e i s anal y zed , the samp l e d a ta s h o u ld b e cl earl yi d ent i f i ed i n the d at a p ackage as be i ng un us abl e f

or i t s i nt ended purpo s e. I n a dd i t i on, the i nadmi s s i bi l i ty of thes e s amp l e dat a n eed t o be cl ear l ycomm un i cat ed to a ll e n d d a ta us e rs . 6. 2 Sample Containe rs and Pr es ervation T he typ e o f

s am p l e co nt ai ner and the requir ed preservat i ve f o r each i no rgani c and organi c chem i cal co nt am ina ntare l i s t ed i n T abl e I V
- 6. T he l aborat orymus tmeas ur e a nd r ecor d the t emp er atur e o f the s amp l e wh en i tarr i ves wh en t empe r atu r e pr es erv at i on i s re qu i r ed byth e meth od. T h e u s e of "bl u e i ce " i s di s cour ag ed be cau s e i tg en er al l ydoe s n ot mai n t ai n th e t emper at ure o f the sam p l e at4° C ±2° C orl es s . I f bl u e i ce i s u s ed, i ts h ou l d be f r oze n a tth e t i me of samp l i ng, the samp l e shoul d be chi l l ed be f o re pac ki ng, and s p eci al noti ce t aken ats am p l e recei p tt o be c e rta in the r equ i r ed t empe r atu r e ( 4° C) h as be enma i n t ai n ed.

## IV- 4

6. 3 Max im um Holding Tim es Sa mpl es mu s tbe a n al y ze d w i th i n th e max i mu m h ol di n g t i mes r equ i r ed by th e meth od. T h es e ar e l i s t ed i n T abl e I V
- 6. 6.4 S a mp le Co llec tio n a n dTra n sp o rt T her e mus tbe s tr i ctadhe r enc e t o corr ectsampl i ng p r ocedur es , sampl e han dl i ng , co mp l et e i dent i f i cat i on o f

the s amp l e, a n d p ro m p t tra n sfe r o f th e s a m p le to th e la b o ra to ry w h e n re q u ire d b y the m e tho d . W h e n the lab o ra to ry is n o t re sp o n sib le f o r s am p l e co l l ect i o n a nd tr anspo rt , itm ustve ri f ythatthe p ap erwo rk, p res ervat i ves, c o nt ai ners a nd ho l d i ng ti m es ar e corr ectas r equi r ed by the methods or re j ectthe s amp l e. T he r ej ect i on cri t er i a sh oul d ( E P A O r der 53 60
. 1) be document ed i n wri t i ng. 6. 5 Sample Collec tor T he sam p l e c o l l ec t o r sho uld b e trained i n sam p l i ng p ro ce d ure s and ha ve co m p l ete w ritt en sam p l i ng i nstructi o ns (S O P s) f or e ach type of

s amp l e t o be col l ect ed. T he s amp l er s ar e t o be abl e t o dem ons tr at e pr op ers amp l i ng t ech ni que. 6.6 S a mp le R ep o rt

Fo rm T h e s a m p l e c o l l e c t i o n r e p o r tf o rm s h o u l d c o n t a i n , a t a m i n i m u m , th e I D , l o c a t i o n , d a t e a n d t i m e o f c o l l e c t i o n , c o l l e c t o r 's name, p rese rva t i ve added an d s hipp i ng r equi r ement s , cont ai neran d vol ume, s amp l e type, an al y s i s , an d anys peci al re m a rk s c o n c e rn ing the sa m p le . Ind e lib le ink sh o u ld b e us e d
. 6. 7 Sample Compos iting I f s ampl es a r e compos i t ed, th e compos i t i n g mu s t( 40 CFR 141. 23, 24) be don e i n th e l abor at ory . Sa mpl es ma yon l ybe co m p o si t ed i f

the l ab o rat o ryde t ect i o n l i m i ti s ad eq uat e f o r the num b er o f

sam p l es b ei ng c o m p o si t ed (up to a m axi m um o f f i ve). Fo r examp l e, f

o r i no rgani c samp l es, co m p o si t e samp l es f rom a m axi m um of f

i v e s a m p le s are al l o wed i f the d e te c tio n li m it o f th e m e tho d us e d fo r a n a lysis is l e ss th a n o n e -f ifth t h e M C L . If the c o n c e n tra tio n o f an y ino rg a n ic c h e m ic a l i n the c o m p o site is g re a te r th a n o r e q u a l t o o n e -f ifth o f th e M C L , th e n a fo llo w u p s a m p le m u st b e tak e n w ithin 14 day s atea ch s amp l i ng poi nti ncluded i n the com po s i t e. T hes e sa mp l es m us tb e anal y zed f or the cont ami nan t s wh i ch ex ce ede d one

- f

i f th th e M CL i n th e compos i t e s ampl e. [ CFR 144. 23( a) ( 4) ] Compos i t i n g of V O Cs i s n otr ec ommen ded. 

## 7. Quality Control 7 .1 G e n e ra l R e q u ire m e n ts 7 .1.1 A v a i l a b i l i ty o f Q A D o cu m en ts: T he l aborat ory ' s Q A pl an and app r op r i at e St andar d O pera t i ng P r ocedur es ( SO Ps ) s h ou l d be r ea di l ya v ai l abl e t o th e an al y s t s a n d f o r i n s pec t i on bya u di t or s . ( s ee Chapt erI I I ' s di s cu s s i on of Q ual i tyA ss urance). 7. 1. 2 A vai l abi l i t y of

Q C I nf ormat i on: Al l qua l i tycon tr oli n f ormat i on s h ou l d be re adi l ya v ai l abl e f ori n s pec t i on b y aud i t o rs. 7 . 1 . 3 B a l a nc es an d W ei g hts: B al an ce r an g e s h ou l d be a ppr opr i at e f orth e appl i ca t i on f

orw h i chi ti s t o be u s ed. D r i n k i n g w at erc h emi s tryl abor at or i es s h ou l d u s e bal an ce s th atw ei g h t o a tl ea s t0. 0001 g
. T h e bal an ce s s h ou l d be c a l i b r a t e d a tl e a s ta n n u a l l yw i th AST M T y pe I , C l as s 1 o r2 w e i g h t s . ( AS TM , 1 9 1 6 Ra c e S t
. , Ph i l a d e l p h i a , PA 1 9 1 0 3 ) T h is m a y b e d o n e b y lab o ra to ry p e rs o n n e l o r u n d e r c o n tra c t b y a m a n u fa c tu r e r ' s r e p re se n ta tive . W e s tr on g l yr ec ommen d th atl abor at or i es have a con tr ac tt o c al i br at e bal an ce s du e t o th e ex pen s e of th e ca l i br at i on w eigh t s, an d t o serv e as an o utsi d e Q C ch ec k o f the we i gh t s and b alan ce s. W eigh t s m ee t i ng A S T M T yp e I, C l ass 1 or 2 sp eci f

i cat i o ns shoul d b e recert i f i ed a tleas te veryfi ve y ears o r i f ther e i s reason t o b el i eve d am age (co rr o si o n, n ic k s) ha s o c c u rr e d
. E a c h d a y the m e c h a n ic a l o r d igita l b a la n c e is u se d , a v e rifica tio n sh o u ld b e p e rfo rm e d
. T h e ve rifica tio n c o n sists of a c h ec kof

a r ef er en ce ma s s a ta ppr ox i mat el yth e s ame n omi n al mas s t o be de t ermi n ed. V er i f i ca t i on s s h ou l d be d o ne each we i ghi ng s ess i o n unl ess i tcan b e shown thatfluctuat i o ns i n the envir o nm entd o no ta f f ectthe cal i b rat i o n. W ei ght s m eet i ng A ST M T yp e 1 s p eci f i cat i o ns m ay b e used. T hese s ho ul d be cal i b rat ed annual l yagai nstthe ref erence weight s att i m e o f ba l ance calib rat i o n. T he checks and thei r f req u e n cy shoul d be as p res cri b ed i n the la b o ra to ry' s Q A P la n . A re c o rd o f all ch e c k s sh o u ld b e ke p t an d b e a v a ilab le fo r in sp e c tio n .

## IV- 5 7 .1.4 C o l o r S ta n d a rd s: W av el en g th s ett i n g s on s pec tr oph ot omet er s s h ou l d be v er i f i ed atl ea s ta n n u al l yw i th c ol or sta n d a rd s. T h e sp e c ifi c c h e c k s a n d the ir fre q u e n c y sh o u ld b e a s p re sc rib e d in th e lab o ra to ry’s Q A d o c u m e n ts. A r ec or d of

th ese che cks sh ou l d be k epta s pr es cr i bed i n th e l abor at ory ’ s Q A docu men t s a n d be a v ai l abl e f or ins p e c tio n . 7 .1.5 T em p er a tu re M ea su rin g D ev i ce s Li q ui d be ari ng therm o m et ers s uch as mercuryor al co ho l therm o m et ers nee d t o be tr ace able to N IS T c al i brat i on an d ver i f i ed atl eas tan nu al l yan d when eve rthe thermo met erha s been exp o sed t o t em p erature extrem es. T he co rr ect i o n fact o r should b e i nd i cat ed on the therm o m et er and the d at e the the rm o m e te r wa s c a lib ra te d a n d the c a lib ra tio n fac to r sho u ld b e k e p t a s p re sc rib e d in the lab o ra to ry’s Q A d o c u m e n ts and be a vai l abl e f

or i ns pect i on. T he N I ST thermo met ers houl d be re cal i brat ed atl eas tev eryf i ve y ear s or whe nev er the the rm o m eter ha s b ee n e xp o sed t o t em p era ture ex trem es. D i g i t alth ermomet er s , th ermocoupl es a n d oth ers i m i l are l ec tr on i c t empe r atu r e mea s u r i n g de v i ce s s h ou l d be cal i brat ed atl eas tquart er l y . T he dat e the thermo met erwas ca l i brat ed an d the cal i brat i on f act or s houl d be keptas p res cri b ed i n the l ab o rat o ry ’ s Q A do cum ent s and be avai l ab l e f o r i nspecti o n. W henani nf

r ar ed det ect i on devi ce i s us ed t o meas ur e the t emp er atur e of s amp l es , the devi ce s houl d be ver i f i ed at l eas tev ery s i x mo nths us i ng a N I ST ce r t i f i ed thermo met eroverthe f ul l t emp er atur e r ang e thatthe I R thermo met er w illb e us e d . T h is w o u ld inc lud e a m b ie n t (2 0 -3 0 °C ), ic e d (4 °C ) a n d fro z e n ( 0 t o -5 °C ). E a c h d a y o f use a sing le che ck of the I R s houl d be m ade bych eck i ng the t emp er atur e of a bo tt l e o f

wat eratthe t emp er atur e o f i nt er es tthat con t ai n s a c a l i b r a t e d th e rm o m e t e r . Ag r e e m e n t b e tw e e n th e tw o s h o u l d b e w i th i n 0
. 5 ° C, o r th e d e v i c e s h ou l d be r ec al i br at ed. 7. 1. 6 Trac eab i l i t y of

C al i br at i on: Cal i brat i ons of al l meas urem en td evi ces ne ed t o be tr ace abl e t o nat i onal s t andar ds whe nev erappl i cabl e. 7 .2 S p ec i fic R eq u irem en t

s: T h e f ol l owi n g a r e r equ i r ed f ore ac h a n al yt e f

orw h i cha l abor at oryi s c ert i f i ed: 7 .2.1 P ro fici en cy T estin g (P T ) S a m p l es: I n ord er t o recei ve and m ai n ta in full cert i f i cat i o n f o r an anal y t e, the l aborat orymus t( 40 C FR 141
. 23 ( k) ( 3)( i ) , 14 1. 24 ( h) ( 17 ) ( i ) ( A ) an d 14 1. 89 ( a) ( 1)( i ) ) an al y ze P T s amp l es (i f av ai l abl e) a c c e p ta b le to the C e rtifyi n g A u tho rity a t le a st o n c e e v e ry 1 2 m o n ths fo r e a c h a n a lyte a n d b y e a c h m e tho d us e d to a n a lyz e c o m p lian c e sa m p le s. R e su lts fro m a n a lysis o f the P T sa m p le m u st b e w ithin the a c c e p ta b le li m its e sta b lishe d b y U .S . E P A . T h e se a c c e p ta n c e lim its ar e li ste d in T a b le IV -1 0 , “M C L a n d P ro fec ie n c y T e sting S a m p le A ccep t ance C ri t eri a in the C FR , P ri m aryand Sec o nd aryD ri nki ng W at er R egul at i o ns [§1 4 1
. 2 3 (k) (3)(i i ) and 1 4 1 .2 4 (f)( 1 7 ) a n d (1 9 )] .” T h e lab o ra to ry sh o u ld d o c u m e n t th e c o rr e c tive a c tio n s ta k e n w h e n a P T sa m p le is anal y zed uns uccessf

ul l y . A co p y o f thi s d o cum ent at i o n s ho ul d be avai l ab l e f o r r evi ew b ythe cert i f

i cat i o n off

i cer. A m a k e up P T sa m p le m u st b e su c c e ssfu lly a n a lyz e d . If p ro b le m s a rise , th e a p p ro p ria te a c tio n t o b e tak e n is sp e c ifi e d in C h ap te r II I, Im p le m e n ta tio n o f C e rtifi c a tio n P ro g ra m . Exc l u di n g v i n y l c h l or i de, th e l abor at ory may be ce rt i f i ed f or al l V O Cs i f th eys u cc es s f u l l ya n al y ze a tl ea s t80% of th e r e g u l a t e d VOCs ( 1 4 1
. 2 4 ( 1 7 ) ( f ) ( i ) ( B)
. Th e i n t e n t i o n o f th i s r e g u l a t i o n i s t o a l l o w s o m e f l e x i b i l i tyf

or r andom mi s s es becau s e the V O C m ethods i nclude 2 0 r egu l at ed ana l y t es . A l aborat orysh oul d notbe ce rt i f i ed fo r a n a n a lyte w h ic h i t fa il s r e p e a te d ly. T h is “8 0 % ru le ” fo r V O C s h a s re c e n tly b e e n m a d e m o re d iff ic u lt to i n t erpr ets i n ce s ome PT pr ov i der s ar e i n clu di n g T H M s i n th e s ame v i ala s th e V O Cs
. T h e 80% Rul e does n ot ap p l y t o the T H M s. T he Stage 1 D i sinf

ect i o n B yP rod uct s (D B P ) R ul e, whi ch b ecam e ef f ect i ve i n January20 0 2 , regul at es the sum o f f ive ha lo a c e tic a c id s (H A A 5 ): m o n o c h lo ro a c e tic a c id , d ic h lo ro a c e tic a c id , tric h lo ro a c e tic a c id , mo nobromo ace t i c a ci d and di bromo ace t i c a ci d. Labora t ori es ar e c ert i f i ed f or H A A 5, but s ucc es s f

ulan al y s es o f the H A A P T s am p l es ar e b ased o n the res ul t s f o r the i nd i vi d ual co m p o und s. T he 8 0 % R ul e ap p l i es t o the H AA5s , s o i f

f ou rof th e 5 H AA5s ar e s u c c e s s f u l l yan a l y z e d, th e l a bo r a t oryma ybe ce rt i f i e d f orH AA5. As b e fo re , a la b o ra to ry sh o u ld no t b e c e rtifi e d if the sa m e a n a lyte is faile d re p e a te d ly.

## IV- 6 T he D B P Rul e a l s o chang ed the waythe tr i hal om ethan es ( T H M s ) : ch l orof orm, di chl orobromo methan e, chl o rod i b rom o m ethane and b rom o f o rm P T s are eval uat ed
. Lab o rat o ri es ar e st i l l cert i f i ed f o r t o t al T H M s b ut un der the D B P Rul e, ea ch T H M conc entr at i on mus t( 14 1. 13 1(b) ( 2) be r eport ed, ev al uat ed an d pass ed i n di v i du al l yt o pa s s th e PT s ampl e. T h e D B P R u l e al s o s t at es th ati f a l abor at oryf

ai l s on e T H M , i tc an n otbe cert i f i ed f o r T T H M s, but m ust (14 1
. 1 3 1 (b)(2)anal y ze ano ther P T samp l e and p ass al l f o ur of the T H M s i n a P T sam p l e to b e c erti f

i ed t o an alyze c o m p l i an ce m o nit o ring sam p l es for to t al trihalo m etha ne s. T he f ol l owi ng t abl e s ummari zes the 80 % Rul e.

A nalyt e( s ) PT Suc ce s s R eq ui re men t V i n y l Chl or i de 100% 2 0 V O C s 8 0 % 1 4 T H M s 100% 5 H AA5s 80% 1 A l ab sho uld no tm aintain ce rti f i ca t i o n for a na l yt e(s) w hich i trep ea t ed l y f ail s. 1 7 .2.2 Q u a l i ty C o n tro lS a m p l es: Atl ea s tonc e e ac h qua rt er , th e l abor at orys h ou l d a n al y ze a qu al i tyc on tr ol s amp l e f or the ana l y t es they ar e det ermi ni ng i n that quart er . T he s amp l e s houl d be prepar ed f r om a s our ce other th anth atf

r om wh i chth ei rw ork i n g s t an dar ds a r e pr epa r ed. T h e s ampl e s h ou l d be i n th e s ame c on ce n tr at i on r an g e as th e dr i n k i n g w at erc al i br at i on c u rv e. I f err or s ex ce ed l i mi t s r equ i r ed i n th e meth ods , c orr ec t i v e ac t i on m u st b e tak e n a n d d o c u m e n te d , an d a fo llo w -up q u a lity c o n tro l sa m p le a n a lyz e d a s so o n a s p o ssib le to d e m o n stra te the p ro b le m ha s b e e n c o rr e c te d
. 7. 2. 3 C al i brati on C urve: C a lib r a tio n r e q u ir e m e n ts in th e m e th o d s m u s t b e fo llo w e d . If there are no cal i brat i on r equi r ement s i n the method, the f ol l owi ng ar e g ui del i nes t o be us ed. Atthe begi nn i ng of ea ch day thats amp l es ar e t o be ana l y zed, a c al i brat i on cu rve cover i ng the s amp l e c oncen tr at i on r ang e a nd al l t ar get anal y t es s ho ul d be generat ed acco rding t o the ap p roved S O P
. D ep end i ng o n c o ncentrat i o n ranges , the curve s h ou l d be c ompos ed of th r ee ormor e poi n t s . Fi el d mea s u r eme n t s ( e. g . pH an d ch l or i n e r es i du al ) n ee d t o be m ad e o n ins trume nt s whi ch have b een p rop erl yc al i b rat ed a s specif i ed i n the m etho d or i nstrume ntm anual and che cke d eachdayof

us e. T he l es s p r eci s e the meas ur ement , the gr eat erthe nu mb ero f co nce ntr at i ons wh i ch sh o u ld b e inc lud e d in the c a lib ra tio n c u rv e . 7 . 2 . 4 C a l i br a ti o n C he ck: T he cal i brat i on f or s om e methods i s s o t i me- cons umi ng that7. 2. 3 i s i mp r act i cal on a da i l yba s i s . W h er e th e det ermi n at i v e t i me i s ex t en s i v e s u cha s M eth ods 508/ 508. 1, 515. 1, 524. 2, 525. 2, et c. a n d th e i n s tru men t i s v ery s t abl e, th e ca l i br at i on curv e s h ou l d be i n i t i al l yde v el oped as s pec i f i ed i n 7. 2. 3. T hereaf t er, e ach d ay anal y ses a re p erf o rme d , thi s curve sho ul d b e veri f i ed b ya nal y si s o f

a tleas to ne st and ard f o r eac h o f

the t argetanal y t es atthe exp ect ed co ncentrat i o n range. T hi s veri f i cat i o n s ho ul d be do ne atbo th the b egi nni ng and end of the anal y ses . A l l checks mustbe wi thi n the co ntrol l i m i t s requir ed i n the m etho d o r the s y s t em i s t o be re cal i brat ed as s peci f i ed i n 7. 2. 3. T he conce ntr at i on of the che ck s t andar d s houl d varyf r om day to d a y a c ro ss th e ra n g e o f an a lyte c o n c e n tra tio n s b e ing m e a su re d
. Fo r s o m e m etho d s an i ni t i al co nd i t i o ni ng i nje c tio n is t o be m ad e t o de act i vat e act i ve si t es thatm ay have d ev elo p ed o ve rnight. D ep en d i ng o n the m etho d , the b l an k m ay b e a p p ro p riat e fo r this. S p e c ifi c c a lib ra tio n re q u ire m e n ts in the m e tho d s m u st b e fo llo w e d if d iff e re n t th a n the a b o v e . Iti s reco m m end ed thata cal i b rat i o n s t and ard o f one co m p o nent o f a m ul t i co m p o nent anal y t e (P C B s, t o xap hene orc h l or dan e) a l s o be an al y ze d ea chda yorw orks h i f t
. B yr ot at i n g th e an al yt e ch os en , c on t i n u i n g c al i br at i on dat a ca n be obt ai n ed on a l l th e mu l t i compon en ta n al y t es ov era pe r i od of on e t o two we ek s . I f

a pos i t i v e f

or a mu l t compon en ta n al y t e i s f ou n d i n a s ampl e, a c al i br at i on c h ec kf orth ata n al y t e s h ou l d be pe r f ormed as s oon a s p o ssib le
.

## IV- 7 7 .2.5 B l a n ks: R eq ui re men t s i n t he met hods mus t be f ol l ow ed . A l aborat oryr eag entbl anks houl d be carr i ed through the f

ul l anal y t i cal p roce d ure with e verysam p l e b at ch. In general , res ul t s f rom l ab o rat o ryreagentblanks s h ou l d n ote x ce ed th e l abor at ory ' s M i n i mu m Report i n g Li mi t , th e l owe s tc on ce n tr at i on of s t an dar d u s ed f

or q uant i t at i o n. (M R L). 7 .2.6 L a b o ra to ry F o rtifi ed B l a n ks: R eq ui re men t s i n t he me t hods mus t be f ol l ow ed . LFB s sh ou l d be ana l y zed atthe l eve l speci f i ed i n the method. S om e m ethods r equi r e thata l aborat oryf orti f i ed bl ankatt en t i mes the M D L o r a m id l e v e l co n c e n tra tio n b e a n a lyz e d w ith e a c h b a tc h o f sam p le s. P re c isio n a n d a c c u ra c y d a ta shoul d b e d o cum ent ed f o r thi s d et erm i nat i o n. In ad d i t i o n, the anal y st shoul d routi nel y veri fy th e m i ni m um re p o rting li m it fo r e a c h a n a lyte b y a n a lyz ing a lab o ra to ry fo rtifi e d b la n ka t th e m inim u m re p o rting lev e l. 7 . 2 . 7 L a bo ra tory F o r tif i ed Sa m ple M a tri x: Laborat ory fort i f i ed s ampl e mat ri x r eq ui re men t s i n t he m etho ds m us t b e m e t. If there are no l ab o rat o ryf o rt i f i ed s am p l e m atri x requi rem ent s i n the m etho d , the f o l l o wi ng are gui d el i nes t o b e used. T he l ab o rat o rys ho ul d ad d a know n qua nt i tyof anal y t es t o a pe rcent age (t o be de scr i b ed i n the ap p roved SO P ) of the ro u tine s a m p le s t o de t erm i ne samp l e m atri x i nt erf erence. T he f o rti f i ed co nc en trati o n sho uld no tb e less tha n the co nc en trati o n of the sam p l e sel ec t ed f o r f o rti f i ca t i o n un l ess sp e c ifi e d b y the m e tho d
. If the sa m p le c o n c e n tra tio n is un k n o w n o r le ss th a n d e te c ta b le , th e a n a lyst sho u ld c h o o se a n a p p ro p ria te c o n c e n tra tio n (e .g., a p e rc e n ta g e o f th e M C L o r m id p o int in the c a lib ra tio n ra n g e ). O ver t i me, s am pl es from a l lro uti ne samp l e sources shoul d be fort i f i ed. The procedure s hou l d be des cri bed i n t h e S O P. I f

an yof thes e c hec ks ar e n otwi thi n the cr i t er i a s peci f i ed i n the method or contr ol l i mi t s s peci f i ed i n 7.2.7, and the l ab o rat o rype rf o rma nce i s i n co ntrol, the res ul t f o r thats am p l e shoul d be f l agged t o i nf

o rm the d a ta us e r th a t th e re su lts ar e su sp e c t d u e to m a trix e ff e c ts. 7 .2.8 C o n tro lC h a rts: C o ntrol chart s f o r accuracyand precis i o n, generat ed f rom l ab o rat o ryf

o rt i f i ed blanks (L F B s) sh o u ld b e m a inta ine d a n d u se d b y the la b o ra to ry. U n tilsu ff ic ie n t d a ta a re a v a ilab le fro m the lab o ra to ry, u su a lly a m inim u m o f 2 0 to 3 0 te st re su lts o n a sp e c ifi c a n a lysis, th e lab o ra to ry sh o u ld us e the c o n tro l l im its sp e c ifi e d in the m e tho d s. I f th e re a re n o c o n tro l lim its sp e c ifi e d in the m e tho d , th e li m its m a y b e sta tisti c a lly cal i brat ed us i ng the procedur e bel ow. W hen suf

f i ci entda t a b eco m e avai l ab l e, the l abor at o rys ho ul d de vel o p LF B co ntrol chart s f rom the m ean p ercent re c o v e ry ( 

### 0 ) an d the s t andard d evi at i on ( S) of the perc entr ecove ryf or the Q C ch eck s s peci f i ed above ( s ee Stan da rd Metho ds f

or the Exam i na t i on of

W at erandW

astewa ter , p a rt 1 0 2 0 B , o r sim ilarQ C re fer e n c e tex ts f o r f urthe r i nfo rm ati o n). T he se d ata a re u sed t o estab l i sh up p er a nd l o w er c o ntro l l i m i t s as f o l l o w s:
u pp e rc on tr oll i mi t= 

### 0 + 3S ( u pperw arn i n g l i mi t+ 2S) l ow e rc on tr oll i mi t= 

### 0 - 3S ( l owe rw arn i n g l i mi t- 2S) A f t er each f i ve t o ten new reco verym easurements , new co ntrol l i m i t s shoul d be cal cul at ed us i ng the m o strecent 2 0
- 3 0 da t a p o i nt s. T hese cal cul at ed co ntrol l i m i t s shoul d not exceed tho se es t ab l i shed i n the m etho d
. I f any of thes e c ontr ol l i mi t s ar e t i gh t erthanthe method s peci f i cat i ons , the l aborat orys houl d us e the t i gh t ercr i t er i a. 7. 2. 9 I ni t i alD emon s t rat i on of C apabi l i t y: R eq ui re men t s i n t he m ethods mus t be f ol l ow ed . B ef

o re begi nn i ng the ana l y s i s of co mp l i anc e sa mp l es , an i ni t i al demo ns tr at i on of capabi l i ty( I D C ) mus tbe per f ormed f ore ac h me th od a s r equ i r ed i n th e meth od. T h e I D C i n clu des a de mon s tr at i on of th e abi l i tyt o ac h i ev e a l ow b a c k g ro u n d , th e p rec isio n a n d a c c u ra c y re q u ire d b y the m e tho d , an d d e te rm ina tio n o f th e m e tho d d e te c tio n li m it ( M D L) ( s ee be l ow)
. An I D C s h ou l d be per f ormed f orea chi n s tru men t
. I ti s a l s o r ec ommen ded th ata n I D C be perf ormed by ea ch ana l y s t
. In ad di t i on, i ti s recomm ended thatthe I D C al s o addres s the va r i abi l i tyi ntr od uce d i f m or e th a n o n e s a m p l e p r e p a r a t i o n t e c h n i c i a n i s u s e d
. Pr e c i s i o n , a c c u r a c ya n d M DL s h o u l d b e s i m i l a rf or e a c h tec h n ic ia n . T h e a n a lyst sho u ld r e c a lc u la te ID C s w h e n a c h a n g e in the m e tho d , an a lyst o r in stru m e n t i s m a d e w h ic h c o u ld a ff

e c t th e p re c isio n o r a c c u r a c y o r s e n sitivity. M ino r c h a n g e s sh o u ld p ro m p t a c h e c kto as ce rt ai n th atth e pr ec i s i on , a cc u r ac ya n d s en s i t i v i tyh av e bee n ma i n t ai n ed.

## IV- 8 7. 2. 10 Q uan t i t ati on o f M ul t i comp onent O rgan i c A nal ytes (t o x a p h e n e , chlord ane and P C B s) T he q u a n tit a tio n o f m u lti c o m p o n e n t an a ly te s r e q u ire s p ro fes sio n a l ju d g m e n t o n the p a rt o f th e a n a lyst. T h is is requir ed due t o the c o m p le x n a ture of the chrom at o graphy i nvo l ved , s am p l e weather i ng, de grada t i o n and i nt er f er enc es thatmaybe pre s enti n the s amp l es . T he patt ern of peak s f ound i n the s amp l e s houl d be exa mi ned car ef ul l yan d com pare d t o a s t andar d. T he peaks i n the s amp l e thatm at ch the peak r at i os i n the s t andar d can b e used i n qua nt i ta t i o n. P eaks thathave o b vi o us i nt erf erences ( such as pe st i ci d es or phthal at es or pea ks exhi b i t i ng po o r peak shap e) or ap p ear t o have b een d egrad ed o r weather ed shoul d no t b e used f o r quant i t at i o n. A rep rese ntati ve num b er (5

- 9 ) o f p ea ks i s su gg ested . P ea k are a sho uld b e use d f o r q ua nti t ati o n a nd the an alyst s h ou l d e n s u r e th a t th e s a m p l e s a n d s t a n d a r d s h a v e b e e n i n t e g r a t e d i n th e s a m e m a n n e r . Qu a n t i t a t i o n c an be d o ne b yus i ng the t o t al pe a k a re a o r hei ght(com p ari ng the area of the 5

- 9 pe aks us ed f o r quant i t at i o n of the samp l e t o the area o f the st and ard) o r b y cal cul at i ng each peak s ep arat el y (us i ng area) and t aki ng the average c o n c e n tra tio n o f th e 5 -9 p e a k s. B e c a u se o f f a c to rs s u c h a s p e a k s h a p e a n d b a se line rise , th e m o st a c c u ra te q u a n tit a tio n is o b ta ine d w h e n the c o n c e n tra tio n o f th e sa m p le c lo se ly m a tc h e s tha t o f th e sta n d a rd (e .g., w ithin 20 % o f the s t andar d). See E P A M ethod 80 81 , O r gan ochl ori ne P es t i ci des an d P C B s as Arocl ors byG as C h ro m a to g ra p h y: C a p ill a ry C o lum n T e c h n iq u e , (E P A S W 8 4 6 T e st M e th o d s fo r E v a lua ting S o lid W a ste P hy si cal / C hem i cal M etho d s, T hi rd E d i t i o n) f o r a m o re d et ai l ed d i scus si o n o f q uant i t at i o n o f

m ul t i co m p o nent an alyt es. N ote: P C B s ar e qua l i t at i vel yi dent i f i ed as Arocl ors an d m eas ur ed f or c om pl i anc e purpo s es as d e c a c h lo ro b ip h e n yl. C h lo rd a n e is re g u la te d a s te c h n ic a l c h lo rd a n e , a m ixtu re o f at lea st 1 1 m a jo r c o m p o n e n ts an d 3 0 m i no r o ne s. 7. 2. 11 M D L C al cu l at i on: R eq ui re men t s i n t he me t hods mus t be f ol l ow ed . M ost methods r equi r e i ni t i al M D L c a lc u la tio n s fo r a ll a n a ly te s a n d c e rtifi c a tio n o ff ic e rs sh o u ld re q u ire the lab o ra to rie s to c a lc u la te the ir d e te c tio n li m its f

o r a ll re g u la te d c o n ta m ina n ts. If the re is no p ro c e d u re to d e te rm in e t h e d e te c ti o n li m its i n the m etho d , its ho ul d be de t erm i ned i n acco rdanc e wi th the p roce d ure gi ven i n 40 C FR 13 6 , A p p end i x B
. T he C F R , at1 4 1 .2 4 ( f)(1 7 )( ii)(C ) re q u ire s a n M D L o f 0 .0 0 0 5 m g /L b e a ttain e d fo r V O C s, a n d 1 4 1 .8 9 (a )( 1 )( iii ) re q u ire s a n M D L o f 0 .0 0 1 m g /L b e a ttain e d fo r le a d if the lab w illb e p r o c e s s in g s o u rc e w a te r c o m p o site s ampl es . Fori n or g an i cs a n d SO Cs , a me th od de t ec t i on l i mi tof 1/ 5 of th e M CL mu s tbe a tt ai n ed f or c o m p o s i t i n g [ C F R 1 4 1
. 2 3 ( a ) ( 4 ) ] a n d [ C F R 1 4 1
. 2 4 ( f ) ( 1 0 ) ]
. VOC s s h ou l d n otb e c o m p o s i t e d
. Th e S OC d e te c tio n lim its liste d a t C F R (1 4 1 .2 4 (h )( 1 8 ) a re re q u ire d to re d u c e m o n ito rin g (C F R 1 4 1 .2 4 (f)( 1 1 )( iv) . T a b le IV -8 li sts the M C L s, M C L G s a n d M D L s, fo r V O C s w h ic h a re in the d rin k ing w a te r re g u la tio n s. T a b le IV -9 l i st s the S O C M C L s, M C L G s and M o nit o ring T rigge rs. S a m p le p re p a ra tio n a n d a n a lyse s fo r th e M D L c a lc u la tio n sh o u ld b e m a d e o v e r a p e rio d o f at lea st thr e e d a ys to inc lud e d a y-to -d a y va ria tio n a s a n a d d iti o n a l so u rc e o f e rr o r. T h e a n a lyst sho u ld d e te rm ine M D L s initia lly, wh ena n yc h an g e i s ma de wh i chc ou l d a f f ec tth e M D Ls , ormor e f r equ en t l yi f

req u i r ed byth e meth od. ( I n or g an i c meth ods ma yr equ i r e M D Ls t o be de t ermi n ed di f f er en t l y , a n d ina l l c as es th e meth ods mu s tbe f o l l o w e d
. ) I n a d d i t i o n , th e a n a l y s tmu s tde m o n s tr a t e l o w l e v e l c a p a b i l i tyona n on g o i n g b a s i s th r o u g h a n M DL d e te rm ina tio n o r re p e a te d lo w lev e l an a lyse s (M R L ). T he cal cul at i on of

M D Ls bythe C FR procedur e maynotbe adequat e f or t oxaphen e a nd ch l ordane becau s e they re q u ire p a tter n o r p e a kp ro fil e re c o g n iti o n fo r id e n tif ic a tio n . P re se n tly, no sta n d a rd p ro c e d u re e x ists, so itis r ecomm ended thatthe M D L be def i ned a s the l owes tconc entr at i on f or whi ch patt ern r ecogn i t i on i s poss i bl e. P att ern r ecogn i t i on i s us ed f or qua l i t at i ve i dent i f

i cat i on of

P C B s as Arocl ors . Q uan t i t at i on o f P C B s i s ac hi eve d b y c o n v e rs io n o f P C B s to d e c a c h lo ro b ip h e n yl (D C B ). 7. 2. 12 Low Leve lQ uant i t at i on : T he l ab o rat o ry ' s m i ni m um rep o rt i ng li m i t s (M R L) shoul d b e repo rt ed to the cl i en ta l on g w i th th e dat a. T h e r eport i n g l i mi tmu s tb e b el o w th e M CL. Labor at or i es s h ou l d ru n a LFB at t he i r M R L e ver y analys i s day and s houl d notreportco nt ami nan t s atl eve l s l es s thanthe l eve l atwh i ch they ro u tine ly an a lyz e the ir lo w e st sta n d a rd
. W h ile th is is a sc ie n tif ic a lly so u n d p ra c tice , w h e the r it i s a n a c c e p ta b le p ract i ce wi l l de p end o n S t at e and F ed eral rep o rt i ng req ui rem ent s. Iti s i m p o rt antf o r us ers of d at a t o unde rs t and the st at i st i cal and qua l i t at i ve si gni f i cance o f

the d at a. La b o rat o ri es m ay b e requir ed by the Stat es t o achi eve a s pe c i f i c M D L orqu a n t i t a t i onl i mi tmo r e s tr i n g e n tth a n th a tr e qu i r e d b yEPA.

## IV- 9

8.Re c or ds a nd D at a Reporting 8. 1 L egalD ef

ens ibilit y: C o m p liance mo ni t o ri ng da t a shoul d be m ad e l egal l yde f ensib l e b ykeep i ng tho rough and ac cur at e r ec or ds
. T h e Q A pl ana n d/ orSO Ps n ee d t o ( EP A O r der5360. 1) des cr i be th e pol i ci es an d pr oce du r es u s ed by the fac ili ty f

o r re c o rd inte g rity, re te n tio n a n d sto ra g e . If sa m p le s a re e x p e c te d to b e c o m e p a rt o f a le g a l ac tio n , ch a in o f cust o d ypro ced ures s ho ul d be us ed (See A p p end i x A ). 8 .2 Ma in ten a n ce o

f R ec o rds: Pu b l i c W a t e r S ys t e ms a r e r equ i r ed t o ma i n t ai n r ec or ds of c h emi ca l a n al y s es of co m p l i ance sam p l es f

o r 10 y ears (40 C FR 1 4 1
. 3 3 ) and lead and c o p p er f o r 1 2 y ears (40 C FR 14 1
. 9 1 ). T he l ab o rat o ry s houl d m ai nt ai n eas i l yac ces s i bl e r ecor ds f or f i ve y ear s or un t i l the nex tce rt i f i cat i on dat a a udi ti s comp l et e, wh i che ver i s l o nger. C hanges i n ow ners hi p , m ergers , or cl o sur es of l ab o rat o ri es d o no t el i m i nat e these requir em ent s. T he cl i ent wat ers y s t em s houl d be n ot i f i ed bef ore di s po s i ng of

r ecor ds s o theymayr eques tcopi es i f ne eded. T hi s i ncludes al l r aw dat a, ca l cul at i ons , an d qual i tycontr ol dat a. T hes e dat a f i l es maybe ei therha r d cop y , mi cr of i che or el ectr oni c. E le c tro n ic d a ta sh o u ld a lw a ys b e b a c k e d up b y p ro te c te d tap e o r d isko r h a rd c o p y. If the lab o ra to ry c h a n g e s its co m p ut er hardware o r s o f tware, i tsho ul d m ake p rovis i o ns f o r trans f erri ng o l d d at a t o the new sy st em s o thati tremains r etr i evabl e wi thi n the t i me f r ames s peci f

i ed above. D at a wh i ch i s ex pect ed t o bec om e part of a l ega l ac t i on mayne ed to b e m a inta ine d fo r a lo n g e r p e rio d o f t im e . C h e c kw ith yo u r le g a l co u n se l. 8 . 3 Sa mp ling R eco rds: D at a shoul d b e reco rded i n i nk wi th any changes l i ned through s uch thatthe o ri gi nal entry i s v i s i bl e. D at a maya l s o be k epte l ec tr on i ca l l y . Ch an g es need to b e i ni t i al ed an d da t ed. T h e f ol l owi n g i n f

ormat i on s houl d be re adi l yav ai l abl e: 8 .3 .1 D at e, l o cat i o n (i ncl ud i ng nam e o f ut i l i ty and P W SS ID # ), si t e wi thi n the sy st em , t i m e o f s am p l i ng, n ame , or g an i za t i on a n d ph on e n u mberof th e s ampl er , a n d a n al y s es r equ i r ed; 8 .3 .2 I dent i f

i cat i on o f

the s amp l e a s t o whetheri ti s a r out i ne di s tr i but i on s y s t em s amp l e, ch ecks amp l e, r aw or f i ni s hed wa t ers amp l e, r epeator conf i rmat i on s amp l e orothers peci al purpo s e s amp l e; 8 .3 .3 D at e of

r ece i ptof

the s amp l e; 8 .3 .4 S a m p le vo lum e /w e igh t, c o n ta ine r typ e , p re se rv a tio n a n d ho ld ing ti m e a n d c o n d iti o n o n re c e ip t; 8 .3 .5 p H and dis i nf ect antres i d ual att i m e o f s am p l i ng (i f

requi red) (f

rom plantrecord s) ; 8 .3 .6 T rans p o rt at i o n and de l i veryof the samp l e (person/carri er, co nd i t i o ns)
. 8. 4 Anal

ytical Rec or ds D a ta s h o u ld b e r e c o rd e d in ink w ith a n y c h a n g e s line d thr o u g h su c h tha t o rig ina l en try is vi s i bl e. Chang es ne ed t o be i ni t i al ed an d dat ed T he f ol l owi ng i nf

ormat i on s houl d be re adi l yav ai l abl e: 8 .4 .1 L ab o rato ry and p erso ns resp o nsib l e fo r p erfo rm i ng an alysi s; 8 .4 .2 Ana l y t i ca l t ec h n i qu es /meth ods u s ed; 8 .4 .3 D ate a nd t i m e o f an alysi s; 8 .4 .4 R esu l t s o f

sam p l e a nd q ua l i ty co ntro l an alyses; 8 .4 .5 C al ib rat i o n and s t and ards i nf o rma t i o n. 8 .4 .6 A naly s tan d t ech ni ci an I ni t i al D emo ns tr at i on of C apabi l i tydocument at i on s houl d be kepton f i l e a s wellas res ul t s o f

pro f i ci ency t est i ng. 8. 5 Rec ons tr uc tion of D ata: A d eq ua t e i nfo rm at i on s houl d be ava i l abl e t o al l ow the audi t or t o re cons tructthe f i nal resu l t s f o r co m p l i an ce sam p l es a nd P T sam p l es.

## IV- 10

8 . 6 Co mp uterP rog ram s: C o m p ut er pro gram s shoul d be veri f i ed i ni t i al l y and pe ri o d i cal l yby ma nual cal cul at i o ns and the cal cul at i ons s houl d be ava i l abl e f or i ns pect i on. A cce s s t o c om put er progr ams and el ectr oni c dat a nee d t o be l i mi t ed to a p p ro p ria te p e rs o n n e l. 

## 9. Action inResponse to Noncomplian t Laboratory Results W hena l aborat oryi s r es po ns i bl e, ei therbycontr actor St at e pol i cy , t o re po rts amp l e r es ul t s which w o uld indi cat e a sy st em i s o utof co m p l i ance, the l ab o rat o rym ust(14 1
. 2 3 (a)( 4 )( i )) pro m p t l ynoti f ythe p rop er autho ri tys o tha tthe autho ri tycan requestthe wat er ut i l i tyt o res am p l e f rom the same sam p l i ng l o cat i o n(s ) i m m ed i at el y . See C hap t er I II
.

## IV- 11

T abl e I V - 1 G l ass w are C l eani ng P rocedures - c o n su lt the m e tho d fo r c o m p le te d e ta ils; d o n o t o v e r-h e a t vo lum e tric gl ass ware
M et hod W as hi ng D ryi ng 50 2. 2/ 50 4/ 50 4. 1/ 52 4. 2 D et er gen twas h, r i ns e wi th t ap and dis t i l l ed wat er 1 0 5 °C f o r 1 ho ur
50 5 D et er gen twas h, r i ns e wi th t ap and reagentwat er 4 0 0 °C fo r 1 ho u r o r rin se w ith acetone 5 0 6 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and det er gen t , r i ns e wi th t ap and r eag entwat er 4 0 0 °C fo r 1 ho u r o r rin se w ith ace t one

5 0 7 / 5 0 8 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and de t ergent , ri nse wi th t ap and r eag entwat er 4 0 0 °C fo r 1 ho u r o r rin se w ith acetone

50 8. 1 D et er gen twas h, r i ns e wi th t ap an d rea ge nt wa t er o r so l ve nt ri nse 4 0 0 °C f o r 2 ho urs
5 0 8 A N o s p eci f i cat i o ns, s uggestthe sa m e a s 5 1 5 .1 /5 1 5 .2 no s p eci f i cat i o n, s uggestthe sa m e a s 5 1 5 .1 /5 1 5 .2 5 1 5 .1/5 1 5 .2 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er a n d d e te rg e n t, rin se w ith d ilute aci d, t ap an d re age ntwat er 4 0 0 °C fo r 1 ho u r o r rin se w ith acetone

5 1 5 .3 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and de t ergent , ri nse wi th t ap and r eag entwat er 4 0 0 °C fo r 1 ho u r o r rin se w ith acetone

5 1 5
. 4 W ash wi th t ap wat er and d et ergent , ri nse wi th t ap and rea ge nt wa t er. A so l ve nt ri nse m a y b e ne c e ssa ry. In p la c e o f so lve n t rins e , m u ff le a t 4 0 0 °C fo r 2 ho u rs . H eatvol umetr i cs a t12 0° C. 524. 2 N otde s cr i bed i n me th od N otde s cr i bed i n me th od 52 5. 2 D et er gen twas h, r i ns e wi th t ap and dis t i l l ed wat er or s o l vent rinse air d ry or m uff l e(n o sp ec s) (s uggest40 0 °C f

o r 1 ho ur)
5 3 1 .1/6 6 1 0 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and de t ergent , ri nse wi th t ap and r eag entwat er 4 5 0 °C fo r 1 ho u r o r rin se w ith acetoneM et hod W as hi ng D ryi ng



## IV- 12 5 4 7 / 5 4 8 .1 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and de t ergent , ri nse wi th t ap and r eag entwat er 4 0 0 °C f o r seve ral ho urs o r ri nse wi th me th an ol

5 4 9 .2 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and de t ergent , ri nse wi th t ap and r eag entwat er 1 3 0 °C f o r seve ral ho urs o r ri nse wi th me th an ol

5 5 0 / 5 5 0 .1 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and de t ergent , ri nse wi th t ap and r eag entwat er 4 0 0 °C f o r 1 5
- 3 0 m i nu t es o r ri nse w ith a c e to n e o r p e sticid e q u a lity hexane
5 5 1 .1 W a sh w ith w a te r a n d d e te rg e n t, ri nse wi th t ap and reagentwat er. R i nse caps i n acet o ne V i al s : 400 ° C f or 30 mi nu t es C ap s: 80 °C f

o r 1 ho ur
5 5 2 .1, 5 5 2 .2, 5 5 5 R i nse i m m ed i ately w i th last s ol ven tus ed, was h w i th h otwat er and de t ergent , ri nse wi th t ap and r eag entwat er 40 0°C f or 1 h ouror r i ns e 3 t i mes wi th acet o ne

1 6 1 3 R ins e w ith so lve n t, so n ic a te w ith d eterg en tf

o r 3 0 m i nu t es, ri nse s equ en t i al l yw i th me th an ol , h ot t ap wat er, m ethano l , acet o ne and meth y l en e ch l or i de A i r dry

M e ta ls W a sh w ith d e te rg e n t, rin se w ith t ap wa t er , s oak4 h ou r s i n 20% (V /V ) n itric a c id o r d ilute ni tr i c( 

### - 8 % )/ hy drochl ori c( 

### - 1 7 % ), rin se w ith r eag entwat er A i r dry

In o rg a n ic s W a sh w ith d e te rg e n t, rin se w ith t ap and rea ge nt wa t er (u se ph os ph at e f

r ee de t er g en tf

or o - p ho sp ha t e a na l ysi s) A i r dry

## IV- 13 * ICPwillnot b eanac cepta bl epro ce dureaf

ter 200 6 T abl e I V - 2 A pprov ed M et hod s for P ri ma ry I norg ani c C hemi cal s , P aram et ers i n t he Lea d an d C opp er R ul e, So dium , an d T ur bidi ty [§1 4 1 . 2 3 (k) (1)]
C ontam i nan t M et hod ol og y E P A A ST M SM O t her 1 2 A n tim o n y IC P -M S 2 0 0 .8 3

H y dr i de- AA D 3697- 92 A A -P la tfo rm 2 0 0 .9 3 AA- Fu rn ac e 3113B Ar s en i c I CP * 200. 7 3120B 3 IC P -M S 2 0 0 .8 3 A A -P la tfo rm 2 0 0 .9 3 AA- Fu rn ac e D 2972- 93C 3113B H y dr i de- AA D 2972- 93B 3114B A sb e sto s T E M 1 0 0 .1 4

T E M 1 0 0 .2 5
Ba r i u m I C P 2 0 0
. 7 3 1 2 0 B 3 IC P -M S 2 0 0 .8 3 AA- D i r ec t 3111D AA- Fu rn ac e 3113B B ery l l i u m I CP 200. 7 3120B 3 IC P -M S 2 0 0 .8 3 A A -P la tfo rm 2 0 0 .9 3 AA- Fu rn ac e D 3645- 93B 3113B B ro m a te IC 3 0 0 .1 6 C ad m i um IC P 2 0 0 .7 3

IC P -M S 2 0 0 .8 3 A A -P la tfo rm 2 0 0 .9 3 A A
- F u rn a c e 3 1 1 3 B C h lo rite IC 3 0 0 .0 7

IC 3 0 0 .1 6C ontam i nan t M et hod ol og y E P A A ST M SM O t her 1 2



## IV- 14 C hrom i um I C P 2 0 0
. 7 3 1 2 0 B 3 IC P -M S 2 0 0 .8 3 A A -P la tfo rm 2 0 0 .9 3 AA- Fu rn ac e 3113B Cy an i de M an . D i s t i l l at i on

fo llo w e d b y: D 2 0 3 6 -9 8 A 4 5 0 0 -C N -C
S p e c ., A m e n a b le D 2 0 3 6 -9 8 B 4 5 0 0 -C N -G S pec . M anu al D 2036- 98A 4500- CN
- E I
- 3300- 85 8 S e m i-au to 3 3 5 .4 7 Io n S e l. E le c .(IS E ) 4 5 0 0 C N -F La ch at K en da Fl u or i de I on Chr omat og r aph y 300. 0 D 4327- 91 4110B 7 M anual D i st i l l at i o n, S PA DNS 4 5 0 0 F -B ,D
M a n u a l IS E D 1 1 7 9 -9 3 B 4 5 0 0 F -C Aut omat ed I SE 380- 75W E 9 Aut o. Al i za r i n 4500F- E 129- 71W 9 M e rc u ry M anu alCol d V apor 245. 1 D 3223- 91 3112B 3 A u to . C o ld V a p o r 2 4 5 .2 10 IC P -M S 2 0 0 .8 3 N itra te I on Chr omat og r aph y 300. 0 D 4327- 97 4110B B
- 1011 7 1 1

3 A u to C d R e d u c tio n 3 5 3 .2 D 3 8 6 7 -9 0 A 4 5 0 0 -N O -F 7

3 I on Se l ec t i v e El ec . 4500- N O
- D 601 12

3 M a n C d R e d u c tio n D 3 8 6 7 -9 0 B 4 5 0 0 -N O -E N itrit e I on Chr omat og r aph y 300. 0 D 4327- 97 4110B B
- 1011 7 1 1

3 A u to C d R e d u c tio n 3 5 3 .2 D 3 8 6 7 -9 0 A 4 5 0 0 -N O -F 7

3 M a n C d R e d u c tio n D 3 8 6 7 -9 0 B 4 5 0 0 -N O -E

2 S p e c tro p h o to m e tric 4 5 0 0 -N O -BC ontam i nan t M et hod ol og y E P A A ST M SM O t her 1 2



## IV- 15 Se l en i u m H y dr i de- AA D 3859- 98A 3114B

IC P -M S 2 0 0 .8 3 A A -P la tfo rm 2 0 0 .9 3 AA- Fu rn ac e D 3859- 93B 3113B T h a lli u m IC P -M S 2 0 0 .8 3

A A -P la tfo rm 2 0 0 .9 3 Le ad AA- Fu rn ac e D 3559- 96D 3113B

IC P -M S 2 0 0 .8 3 A A -P la tfo rm 2 0 0 .9 3 Copper AA- Fu rn ac e D 1688- 90C 3113B

A A
- Di r e c t D1 6 8 8
- 9 0 A 3 1 1 1 B I CP 200. 7 3120B 3 IC P -M S 2 0 0 .8 3 A A -P la tfo rm 2 0 0 .9 3 p H E le c tro m e tric 1 5 0 .1 D 1 2 9 3 -8 4 4 5 0 0 -H -B 10 + 1 5 0 .2 10 Condu ct i v i ty Condu ct an ce D 1125- 91A 2510B
C al ci um E D T A ti tra tio n 3 5 0 0 -C a -B 2a

E D T A ti tra tio n D 5 1 1 -9 3 A 3 5 0 0 -C a -D 2a AA- D i r ec t D 511- 93B 3111B I CP 200. 7 3120B 3 Al k al i n i ty T i tr at i on D 1067- 92B 2320B

El ec . t i tr at i on I
- 1030- 85 8 O r th op h o sp h a te u n f i l t er ed, n o di g es t i on or h yd ro lysis Col or , a u t omat ed

a sc o rb ic a c id 3 6 5 .1 4 5 0 0 -P -F 7
C o lo r, a sc o rb ic a c id D 5 1 5 -8 8 A 4 5 0 0 -P -E Col or , ph os ph omol ybdat e I
- 1601- 85 8 Aut oSe g men t ed Fl ow I
- 2601- 90 8 Aut o di s cr et e I
- 2598- 85 8 I on Chr omat og r aph y 300. 0 D 4327- 97 4110 7C ontam i nan t M et hod ol og y E P A A ST M SM O t her 1 2



## IV- 16 Si l i ca Col or , mol ybdat e bl u e; I
- 1700- 85 8

a u t o s eg . f

l ow I
- 2700- 85 8 Col or D 859- 88 M o lyb d o silica te 4 5 0 0 -S i-D 2a H e te ro p o ly b lue 4 5 0 0 -S i-E 2a A ut o . m o l yb d at e react i ve s i l i ca 4 5 0 0 -S i F 2a

I CP 200. 7 3120B 3 T empe r atu r e T h ermometr i c 2550B S o d ium IC P 2 0 0 .7 3

AA- D i r ec t 3111B T u rbi di ty N eph el ometr i c 180. 1 2130B G LI M eth od 2 7 1 3 H ac h 10133 Foo tnot es Annual Book of ASTM Sta n dards , Vols . 11.01 and 11.02, A meric an Soc iety

f or Testingand Materials , 1916 Rac e Street, 1 Philade lphia, PA

19103. Sta nd ard Metho ds for th e Examination of Water and Wastewater, 18 , 19 or 20th E diti on, A meric an Public He alth Ass oci ation, 2 th th 1015 Fif te ent h Street NW, Washington, D. C. 20005. Except 3111B, 3111D, 3112B, 3113B, 3114B a renot approved in t

he 20 th edition. Only a pproved in 20 e diti on 2a th " Methodsf

or t he Determination of

Metals in Environmental Sam ple s - Supplement I," EPA-600/R-94- 111, May 1994. A vai la ble 3 at NTIS, PB 94-184942. Method 100. 1, " Analy

tic alMethod f

or D eter mination of

Asbe stos Fibersin Water," EPA-600/4- 83-043, EPA , Septem ber 1983. 4 Avai la

bleat NTIS, PB 83-260471. Method 100.2, " Determination of Asbe stos Str

uct ureOver 10- : m In L ength in DrinkingWater," EP A-600/R-94- 134, J une 1994. 5 Avai la

bleat NTIS, PB 94-201902. M eth o d sf

o r th eD etermin ati o no f

Org ani candIn o rg ani c C o m p ound s in DrinkingWater -V o lume1 , ”do cumentnum b er EPA 6 815-R-00- 014, A

ugust 2000. " Methodsf

or t he Determination of

Inorg ani c Substa

nce s in EnvironmentalSam ple s, " EPA-600/R-93- 100, A ugust 1993. 7 Avai la

bleat NTIS, PB94-121811. Avai la blef rom Books and Open-Fi leReports Se ction, U. S. Geol ogic al Survey, Fe deral Center, Box 25425, D enver, C O 80225- 8 0425. Industri alMethod No. 129-71W , "Fluoridein Water and Wastewater," Dec em ber 1972, and Method No. 380- 75WE, " Fluoride 9 in Water

and Wastewater," Februa ry 1976, Technic on Industri alSystems, T ar ry tow n, NY 10591. Methods150.1, 150.2 a

nd 245.2 a

reavai la ble f rom US EPA , NERL , Cincinnati, OH 45268. The identic al met hods were f

ormerly 10 in " Methodsf

or Chemic alA

nalysi s of Water and Waste s, " EPA-600/4- 79-020, March 1983. Method B -1011, " Waters T est Method f or Determination ofNitrite/Nitrate in Water U singSingleColum

n Ion Chromatogra

phy," 11 Mil lipore Cor

poration, Waters Chromatogra phy D ivis ion, 34 Ma

pleStreet, Mil f

ord, MA 01757. T echnic alBulletin 601 " Sta ndard Method of

T est f or Nitrate in DrinkingWater," July 1994, PN 221890-001, Ther mo Orion, 12 500 Cum

minsCenter, Beverly, MA 01915-9846. Thismet

hod isidentic alto Orion WeW WG/5880, w

hich isa pproved f or nitra te analysis. ATI Orionrepub lished the metho d in 19 94 , and renumbered itas 60 1, because th e 1 98 5 manual "Orion Guid e to Water and Wastewater

A nalysi s, " w

hich c ontaine d WeW

WG/5880, isno l ong er

avai la ble
. GLI M et hod 2, " Turbidity," Novem ber 2, 1992,GLI International, 9020 W D ea n Rd. Milwaukee , Wis cons in 53224. 13

## IV- 17

T a ble I V -3 A pp ro v ed M etho ds for P ri m a ry O rg a nic C he m i ca l s [§1 4 1 . 2 4 (e)]
C ont ami nant M et hod ( R evi s i on N umb er) 1 B en ze n e 502. 2( 2. 1) , 524. 2( 4. 1) Carbon t etr ac h l or i de 502. 2( 2. 1) , 524. 2( 4. 1) , 551. 1( 1. 0) Chl or oben ze n e 502. 2( 2. 1) , 524. 2( 4. 1) 1, 2- D i ch l or oben ze n e 502. 2( 2. 1) , 524. 2( 4. 1) 1, 4- D i ch l or oben ze n e 502. 2( 2. 1) , 524. 2( 4. 1) 1, 2- D i ch l or oeth an e 502. 2( 2. 1) , 524. 2( 4. 1) ci s - 1, 2- D i ch l or oeth y l en e 502. 2( 2. 1) , 524. 2( 4. 1) tr an s - 1, 2- D i ch l or oeth y l en e 502. 2( 2. 1) , 524. 2( 4. 1) D i ch l or ometh an e 502. 2( 2. 1) , 524. 2( 4. 1) 1, 2- D i ch l or opr opan e 502. 2( 2. 1) , 524. 2( 4. 1) Eth y l ben ze n e 502. 2( 2. 1) , 524. 2( 4. 1) Sty r en e 502. 2( 2. 1) , 524. 2( 4. 1) T etr ac h l or oeth y l en e 502. 2( 2. 1) , 524. 2( 4. 1) , 551. 1( 1. 0) 1, 1, 1- T r i ch l or oeth an e 502. 2( 2. 1) , 524. 2( 4. 1) , 551. 1( 1. 0) T r i ch l or oeth y l en e 502. 2( 2. 1) , 524. 2( 4. 1) , 551. 1( 1. 0) T ol u en e 502. 2( 2. 1) , 524. 2( 4. 1) 1, 2, 4- T r i ch l or oben ze n e 502. 2( 2. 1) , 524. 2( 4. 1) 1, 1- D i ch l or oeth y l en e 502. 2( 2. 1) , 524. 2( 4. 1) 1, 1, 2- T r i ch l or oeth an e 502. 2( 2. 1) , 524. 2( 4. 1) , 551. 1( 1. 0) V i n y l c h l or i de 502. 2( 2. 1) , 524. 2( 4. 1) X y l en es ( t ot al ) 502. 2( 2. 1) , 524. 2( 4. 1) 2, 3, 7, 8- T CD D ( di ox i n ) 1613 2, 4- D ( as a ci ds , s al t s a n d e s t er s ) 515. 1( 4. 0) , 515. 2( 1. 1) , 515. 3( 1. 0) , 555( 1. 0) , D 5317- 93, 515. 4( 1. 0) Al ac h l or 505( 2. 1) , 507( 2. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) 1 Atr az i n e 505( 2. 1) , 507( 2. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) 1 B e n z o (a )p yre n e 5 2 5 .2 (2 .0 ), 5 5 0 , 5 5 0 .1 Carbof u r an 531. 1( 3. 1) , 6610, 531. 2( 1. 0) Chl or dan e 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0)C ont ami nant M et hod ( R evi s i on N umb er) 1



## IV- 18 D al apon 515. 1( 4. 0) , 515. 3( 1. 0) , 552. 1( 1. 0) , 552. 2( 1. 0) , 515. 4( 1. 0) D i ( 2- eth y l h ex y l ) adipat e 506( 1. 1) , 525. 2( 2. 0) D i ( 2- eth y l h ex y l ) ph th al at e 506( 1. 1) , 525. 2( 2. 0) D i br omoch l or opr opan e ( D B CP ) 504. 1( 1. 1) , 551. 1( 1. 0) D i n os eb 515. 1( 4. 0) , 515. 2( 1. 1) , 515. 3( 1. 0) , 555( 1. 0) , 515. 4( 1. 0) D i qu at549. 2( 1. 0) Endoth al l 548. 1( 1. 0) Endr i n 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) Eth y l en e di br omi de ( ED B ) 504. 1( 1. 1) , 551. 1( 1. 0) G l yph os at e 547, 6651 H ept ac h l or 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) H ept ac h l orEpoxi de 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) H ex ac h l or oben ze n e 505( 2. 1) , 508( 3. 1) , 508. l ( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) H ex ac h l or ocy cl open t adi en e 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) Li n dan e 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) M eth ox y ch l or 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) O x amy l 531. 1( 3. 1) , 6610, 531. 2( 1. 0) P C B s (a s d e c a c h lo ro b ip h e n yl) 2 (as A ro clo rs) 508A( 1. 0) 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) Pen t ac h l or oph en ol 515. 1( 4. 0) , 515. 2( 1. 1) , 515. 3( 1. 0) , 525. 2( 2. 0) , 555( 1. 0) , D 5317- 93, 515. 4( 1. 0) Pi cl or am 515. 1( 4. 0) , 515. 2( 1. 1) , 515. 3( 1. 0) , 555( 1. 0) , D 5317- 93, 515. 4( 1. 0) Si maz i n e 505( 2. 1) , 507( 2. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) , 551. 1( 1. 0) 2, 4, 5- T P ( Si l v ex ) 515. 1( 4. 0) , 515. 2( 1. 1) , 515. 3( 1. 0) , 555( 1. 0) , D 5317- 93, 515. 4( 1. 0) T ox aph en e 505( 2. 1) , 508( 3. 1) , 508. 1( 2. 0) , 525. 2( 2. 0) H AA5 552. 1( 1. 0) , 552. 2( 1. 0) , SM 6251

4 T ot alT r i h al ometh an es 502. 2( 2. 1) , 524. 2( 4. 1) , 551. 1( 1. 0) Foo tnot es M e th od s 50 8A, a n d 5 15
. 1 a r e i n M e th od s f or th e Det e rmi n a t i onof Or g a n i c Com po u n ds i n Dr i n k i n g W a t e r , EPA- 1 600/4- 88- 039, D ec embe r1988, Rev i s ed, J u l y1991. M eth ods 547, 550, a n d 550. 1 a r e i n M eth ods f orth e D et ermi n at i on

## IV- 19

of O r g an i c Compoun ds i n D r i n k i n g W at er- Suppl eme n tI , EP A- 600- 4- 90- 020, Ju l y 1990. M eth ods 515. 2, 524. 2, 548. 1, 55 2. 1 a n d 5 55 a r e i n M e th od s f orth e Det e rmi n a t i onof Or g a n i c Com po u n ds i n Dr i n k i n g W a t e r- Su pp l e me n tI I , EPA6 0 0 /R -9 2 -1 2 9 . M e tho d s 5 0 2 .2 , 5 0 4 .1 , 5 0 5 , 5 0 6 , 5 0 7 , 5 0 8 , 5 0 8 .1 , 5 1 5 .1 , 5 1 5 .2 , 5 2 4 .2 , 5 2 5 .2 , 5 3 1 .1 , 5 5 1 .1 , 5 5 2 .2 a re in M eth ods f

orth e D et ermi n at i on of O r g an i c Compoun ds i n D r i n k i n g W at er- Suppl eme n tI I I , EP A 600/ R- 95/ 131. M eth ods 51 3. 3 a n d 5 49
. 2 a r e i n M e th od s f orth e Det e rmi n a t i onof Or g a n i c a n d I n or g a n i c Com po u n ds i n Dr i n k i n g W a t e r- Vol u me 1 , E P A 8 1 5 -R -0 0 -0 1 4 , A u g u st 2 0 0 0
. M e tho d 1 6 1 3 , T e tra -T h ro u g h O c ta - C h lo rin a te d D io x ins a n d F u ra n s b y Iso to p ic D i l ut i on H R G C /H R M S, E P A
- 81 /B
- 94
- 00 3, O ct ob er199 4 T hes e document s ar e a vai l abl e f r om the N at i onal T ech ni cal I n f ormat i on Se rv i ce , N T I S PB 91- 231480, PB 91- 146027, PB 92- 207703, PB 2000- 106981 a n d PB 95- 104774, U
. S. D epa r tmen tof

C ommer ce , 5285 PortRoy alRoad, Spr i n g f i el d, V i r g i n i a 22161. T h e t ol l

- f r ee n u mberi s 800- 553- 6847. M etho d 16 1 3 i s avai l ab l e f rom U SE P A O f

f i ce o f W at er R esource C ent er ( R C
- 4 1 0 0 ), 40 1 M
. Str eetS.W
. , W ashi ngt o n, D
. C
. 204 60
. T he phone n umber i s 2 0 2
- 2 6 0
- 7 7 86
. M ethod 66 51 an d 66 10 ar e c ont ai ned i n the curr ent l yapproved e d iti o n s o f Stan da rd Metho dsf or the Exam i na t i on of

W at erandW

astewa ter , A mer i ca n P u bl i c H ea l th As s oci at i on , 1015 Fi f t ee n th Str ee tN W , W as h i n g t on , D
. C. 20005. P C B s are qua l i t ative lyi d ent i f i ed as A roclors and m easured f o r com p l i ance p urpo ses as d ecachloro b i p henylus i ng 2 M e th od 50 8A. A ni tr ogen- phosphorus det ect or s houl d be s ubs t i tut ed f o r the e l ectr on captur e det ect or i n M ethod 50 5 (oran other 3 appr ov ed meth od s h ou l d be u s ed) t o de t ermi n e al ac h l or , a tr az i n e an d s i maz i n e, i f l owe rde t ec t i on l i mi t s a r e r equ i r ed. T h e to ta l o f m o n o c h lo ro a c e tic a c id , d ic h lo ro a c e tic a c id , tric h lo ro a c e tic a c id , m o n o b ro m o a c e tic a c id a n d d ib ro m o a c e tic 4 ac i d.

## IV- 20

Tabl e I V - 4 A ppr oved M et hods f or D i s i nfec t ant R es i dual s P ub l i c wat er s y st em s ne e d to m e a su re resi d ual dis i nf ect antco ncentrat i o ns wi th one of the anal y t i cal m etho d s i n the fo llo w ing ta b le
. T h e m e tho d s a re c o n ta ine d in th e 1 8 , 1 9 a n d 2 0 ed iti o n s o f Standard Methods f orthe Exam i nat i on th th th of W at erandW

astewa ter .

R es i dual M et hodol ogy SM

1 3 Fr ee Chl or i n e Amper ometr i c T i tr at i on

2 D P D F e rr o u s T itrim e tric D P D C o lo rim e tric Syr i n g al dah y de ( FACT S) 4 5 0 0 -C l D D 1253- 86 4 5 0 0 -C l F 4 5 0 0 -C l G 4 5 0 0 -C l H
C o m b i ned C hl o ri ne (C hlo ram i ne s) Amper ometr i c T i tr at i on D P D F e rr o u s T itrim e tric D P D C o lo rim e tric 4 5 0 0 -C l D D 1253- 86 4 5 0 0 -C l F 4 5 0 0 -C l G
T ot alChl or i n e Amper ometr i c T i tr at i on

2 Amper ometr i c T i tr at i on (lo w lev e l m e a su re m e n t) D P D F e rr o u s T itrim e tric D P D C o lo rim e tric I odometr i c El ec tr ode 4 5 0 0 -C l D D 1253- 86 4 5 0 0 -C l E 4 5 0 0 -C l F 4 5 0 0 -C l G 4 5 0 0 -C l I Chl or i n e D i ox i de Amper ometr i c T i tr at i on D PD M eth od Amper ometr i c T i tr at i on 2 4 5 0 0 -C lO C 4

2 4 5 0 0 -C lO D

2 4 5 0 0 -C lO E

3 O z o n e In d igo M e tho d 4 5 0 0 -O B Foo tnot es I f a ppr ov ed byth e St at e, r es i d ual d i si n f ec t an tc on ce n tr at i on s f orf r ee c h l or i n e an d c ombi n ed ch l or i n e al s o ma ybe 1 m e a su re d b y us ing D P D c o lo rim e tric tes t kits. F ree an d t o t al chlo rine re si d ua l s m ay b e m ea sure d co nti nu o uslyb y ad ap t i ng a s p ec i f i ed ch l o rine re si d ua l m etho d f o r use 2 wi th a co nt i nuo us m o ni t o ri ng i nstrume nt p rovided the chem i stry , accuracy, and p reci si o n of the m easurement r em ai n the same . I nstrume nt s used f o r cont i nuo us m o ni t o ri ng ne ed to b e cal i b rat ed wi th a grab sam p l e m easurement atl eastevery f i ve day s , or wi th pr ot ocol approved bythe St at e. Stan da rd Metho ds f

or the Exam i na t i on of

W at erandW

astewa ter , 1 8 , 1 9 o r 2 0 th E d iti o n , A m e ric a n P u b lic H e a lth 3 th th As s oci at i on , 1015 Fi f

t ee n th Str ee tN W , W as h i n g t on , D
. C. 20005. 2 M etho d 45 0 0
- C l 0 i s no ta p p ro v e d fo r d et erm i ni ng co m p l i ance at 1 4 1
. 1 3 1 (c) b ecause the o ther two m etho d s are 4 superi o r.

## IV- 21

T a ble I V -5 R eco m m en de d M etho ds for S eco nd a ry D ri nking W a ter C o nta m i na nts A nal y s es of

alumi nu m, ch l ori de, col or, f l uor i de, f

oam i ng ag ent s , i r on, mang ane s e, odo r , s i l ver , s ul f at e, t ot al di s s ol ved s ol i ds ( T D S) and z i nc t o det ermi ne comp l i anc e under§ 14 3. 3 mayb e conduct ed wi th the methods i n the f ol l owi ng t abl e. C ri t eri a f

o r anal y zi ng al um i num , i ron, m anganese, si l ver, and zi nc samp l es wi th d i gest i o n or d i rect l ywi tho utdiges t i o n, an d o the r m an d ato ry pro ce d ure s are co ntained i n S ec t i o n IV o f “T ec hn i ca l N o t es o n D rinking W ater M etho d s” E P A /6 0 0 /R -9 4 /1 7 3 , O c to b e r 1 9 9 4 . M e a su re m e n t o f p H m a y b e c o n d u c te d w ith o n e o f th e m e tho d s l is te d a b o v e in S e c tio n I u n d e r " M e tho d s fo r In o rg a n ic C h e m ic a ls."

C ontam i nan t E P A A ST M SM O t her 1 2 Al u mi n u m 200. 7 3120B 3 200. 8 3113B 3 200. 9 3111D 3 Chl or i de 300. 0 D 4327- 91 4110B 4 D 5 1 2 -8 9 B 4 5 0 0 -C l B ,-D – Col or 2120B Fl u or i de 300. 0 D 4327- 91 D 1179- 93 4110 B

4 5 0 0 -F B , C , D , E

- 3 8 0
- 7 5 WE

129- 71W 5 Foa mi n g Ag en t s 5540C I r on 200. 7 3120B 3 200. 9 3111B 3 3113B M an g an es e 200. 7 3120B 3 200. 8 3111B 3 200. 9 3113B 3 O dor 2150B Si l v er 200. 7 3120B I
- 3720- 85 3 6 200. 8 3111B 3 200. 9 3113B 3 Su l f at e 300. 0 D 4327- 91 4110B 4

4 3 7 5 .2 D 5 1 6 -9 0 4 5 0 0 -S O -E ,-F 4

4 4 5 0 0 -S O -C ,D T D S 2540C Zi n c 200. 7 3120B 3 200. 8 3111B 3

## IV- 22

Foo tnot es AnnualBook of

ASTM Standards , V o ls. 1 1 .0 1 a n d 1 1 .0 2 , A m e ric a n S o c ie ty fo r T e sting a n d M a te ria ls, 1 9 1 6 R a c e S tre e t, 1 Phi l ade l ph i a, PA 19103. Stan da rd Metho ds f

or the Exam i na t i on of

W at er an d W

astewa ter , 1 8 , 1 9 o r 2 0 th E d iti o n , A m e ric a n P u b lic H e a lth 2 th th As s oci at i on , 1015 Fi f

t ee n th Str ee t N W , W as h i n g t on , D
. C. 20005. Exc ept 3111B , 3111D , 3112B , 3113B , 3114B a r e n ot ap p roved i n the 2 0 ed i t i o n. th "M eth ods f

orth e D et ermi n at i on of M et al s i n Env i r on men t alSa mpl es - Suppl eme n tI , " EP A- 600/R- 94- 111, M ay1994. 3 Ava i l abl e atN T I S, PB 94- 184942. "M eth ods f

orth e D et ermi n at i on of I n or g an i c Subs t an ce s i n E n v i r on men t alSampl es , " E PA
- 600/R- 93- 100, A u g u s t1993. 4 Ava i l abl e atN T I S, PB 94- 121811. I n d u s tr i a l M e th o d No
. 1 2 9
- 7 1 W , " F l u o r i d e i n W a t e ra n d W a s t e w a t e r , " De c e m b e r1 9 7 2 , a n d M e th o d No
. 3 8 0
- 7 5 W E, 5 "Fl u or i de i n W at er an d W as t ew at er , " Fe bru ary 1976, B r an an d Lubbe, 1025 B u s ch Park wa y B u f f al o G r ov e, I L 60089. ( Former l yT ec h n i conI n du s tr i alSy s t ems , T arry t own , N Y 10591) A vai l ab l e f

rom B o o ks and O p en- Fil e R ep o rt s Se ct i o n, U
. S. G eo l o gi cal Survey, Fe d eral C ent er, B o x 2 5 4 2 5 , D enver, 6 CO 80225- 0425.

## IV- 23

Ta bl e I V - 6 Sam pl e C ontai ners , P res ervati on a nd H ol di ng T i mes f or R egul ated Pa ramete rs P aram et er/ M et hod Pr es er vat i ve S ampl e H ol di ng Ti me Ext rac t H ol di ng

Ti me and S t orage C ondi t i ons Sug ges t ed Sam pl e Si ze Type of

C ontai ner M e ta ls (excep tH g) 3

H N O p H < 2 6 m o nths 1 L P l asti c o r G l ass

3 M erc ury H N O p H < 2 2 8 d ays 1 0 0 m L P l asti c o r G l ass A l ka l i nity C o o l , 4 C 1 4 d ays 1 0 0 m L P l asti c o r G l ass A sb esto s C o o l , 4 C 4 8 ho urs 1 L P l asti c o r G l ass C hlo ride no ne 2 8 d ays 1 0 0 m L P l asti c o r G l ass R e sid u a l D i si nf ect ant no ne i m m ed i ately 2 0 0 m L P l asti c o r G l ass

C o l o r C o o l , 4 C 4 8 ho urs 1 0 0 m L P l asti c o r G l ass C o nd uc t i vity C o o l , 4 C 2 8 d ays 1 0 0 m L P l asti c o r G l ass Cy a n i de Coo l , 4C,

A sc o rb ic a c id (if chl o ri nat ed ), N aOH pH >12 1 4 d ays 1 L P l asti c o r G l ass

F l uo ride no ne 1 m o nth 1 0 0 m L P l asti c o r G l ass Fo am i ng A gent s C o o l , 4C 4 8 hours N itra te ( ch l or i n at ed) Cool , 4C

non- aci di f

i ed 1 4 d ays 1 0 0 m L P l asti c o r G l ass

N itra te ( n on c h l or i n at ed) C o o l, 4 C ,

non- aci di f

i ed 4 8 ho urs 1 0 0 m L P l asti c o r G l ass

N i trit e C o o l , 4 C 4 8 ho urs 1 0 0 m L P l asti c o r G l ass N i trate+ N i trit e H 2 S O 4 p H < 2 2 8 d ays 1 0 0 m L P l asti c o r G l ass O d o r C o o l , 4 C 2 4 ho urs 2 0 0 m L G l ass p H no ne i m m ed i ately 2 5 m L P l asti c o r G l ass

o - P ho sp ha t e C o o l , 4 C 4 8 ho urs 1 0 0 m L P l asti c o r G l assP aram et er/

M et hod Pr es er vat i ve S ampl e H ol di ng Ti me Ext rac t H ol di ng

Ti me and S t orage C ondi t i ons Sug ges t ed Sam pl e Si ze Type of

C ontai ner



## IV- 24 S ili c a C o o l, 4 C 2 8 d a ys 1 0 0 m L P la stic
S o l i d s (T D S ) C o o l , 4 C 7 d ays 1 0 0 m L P l asti c o r G l ass S ulf ate C o o l , 4 C 2 8 d ays 1 0 0 m L P l asti c o r G l ass T em p era ture no ne i m m ed i ately 1 L P l asti c o r G l ass T urb i d i ty C o o l , 4 C 4 8 ho urs 1 0 0 m L P l asti c o r G l ass 5 0 2
. 2 So d i um

T h io su lfate o r A sc o rb ic Ac i d, 4C, HCl pH<2 14 da y s 40- 120 mL G l as s w i th PT FE Lined S ep tum

5 0 4
. 1 So d i um

T h io su lfate Coo l , 4C, 14 da y s 4C, 24 h ou r s 40 mL G l as s w i th PT FE Lined S ep tum

5 0 5 So d i um

T h io su lfate Cool , 4C 1 4 d a ys

( 7 da y s f or H ep t achl o r) 4C, 24 h ou r s 40 mL G l as s w i th PT FE Lined S ep tum

5 0 6 So d i um

T h io su lfate C o o l , 4C , D ark 1 4 da y s 4 C , da rk 1 4 d a ys 1 L A m b e r G la ss w ith P T FE Li ned Cap

5 0 7 So d i um

T h io su lfate C o o l , 4C , D ark 14 day s ( s ee

meth od f or ex ce p t i o ns) 4 C , da rk 1 4 d a ys 1 L A m b er G l ass wi th P T FE Li ned Cap

5 0 8 So d i um

T h io su lfate C o o l , 4C , D ark 7 day s ( s ee

meth od f or ex ce p t i o ns) 4 C , da rk 1 4 d a ys 1 L G l as s w i th PT FE Li ned Cap

5 0 8 A C o o l , 4C 1 4 da y s 3 0 da y s 1 L A m b er G l ass wi th P T FE Li ned Cap 5 0 8 .1 S o d ium S u lfi te

H Cl pH<2 Cool , 4C 14 day s ( s ee

meth od f or ex ce p t i o ns) 3 0 d ays 1 L G l ass wi th PT FE Li ned Cap 5 1 5
. 1 So d i um

T h io su lfate C o o l , 4C , D ark 1 4 da y s 4 C , da rk 2 8 d a ys 1 L A m b er G l ass wi th P T FE Li ned CapP aram et er/

M et hod Pr es er vat i ve S ampl e H ol di ng Ti me Ext rac t H ol di ng

Ti me and S t orage C ondi t i ons Sug ges t ed Sam pl e Si ze Type of

C ontai ner



## IV- 25 5 1 5
. 2 So d i um

T h i os u l f at e or S o d ium S u lfi te H Cl pH<2 C o o l , 4C , D ark 1 4 d a ys 

### # 4 C , da rk 1 4 d a ys 1 L A m b e r G la ss w ith P T FE Li ned Cap

5 1 5
. 3 So d i um

T h io su lfate C o o l , 4C , D ark 1 4 d a ys 

### # 4 C , da rk 1 4 d a ys 5 0 m L A m b e r G la ss w ith P T FE Li ned Cap

51 5. 4 Sod i um Sul f i t e, d a rk , co o l 

### # 10C fro fi rs t 4 8 hr
. 

### # 6C ther eaf

t er 1 4 d a ys 

### # 0C 2 1 d ays 4 0 m L A m b e r g la ss w ith P T FE l i ned septum

5 2 4 .2 A sc o rb ic A c id

o r So d i um T h io su lfate H Cl pH <2, Cool 4C 14 da y s 40- 120 mL G l as s w i th PT FE Lined S ep tum

52 5. 2 Sod i um Sul f i t e, D a rk , Coo l , 4C, HCl pH<2 14 day s ( s ee

m etho d f o r ex ce p t i o ns) 

### # 4C

3 0 d a ys 1 L A m b er G l ass wi th P T FE Li ned Cap

5 3 1
. 1 , 66 1 0 So d i um

T hi osu l f at e, M onochl oroace t i c ac i d, pH <3, Cool , 4C Cool 4C 28 da y s 60 mL G l as s w i th PT FE Lined S ep tum

5 3 1
. 2 So d i um

T hi osu l f at e, P o t ass i um D i hy drogen C itra te b u ff

e r to p H 4, dark, 

### # 10C f orf i r s t48 h r, 

### # 6C ther eaf t er 28 da y s 40 mL G l as s w i th PT FE Lined S ep tum

5 4 7 So d i um

T h io su lfate Cool , 4C 14 da y s ( 18

m o . f roze n) 60 mL G l as s w i th PT FE Lined S ep tumP aram et er/

M et hod Pr es er vat i ve S ampl e H ol di ng Ti me Ext rac t H ol di ng

Ti me and S t orage C ondi t i ons Sug ges t ed Sam pl e Si ze Type of

C ontai ner



## IV- 26 5 4 8
. 1 So d i um

T h io su lfate (H C l p H 1 .5 -2 if hi gh bi ol ogi cal a c tivity) C o o l , 4C , D ark 7 d a ys 

### # 4C 1 4 d a ys 

### $ 25 0 m L A m b er G l ass wi th PT FE Lined S ep tum

5 4 9
. 2 So d i um

T hi osu l f at e, 2 4 (H S O p H < 2 if b io lo g ic a lly act i ve) C o o l , 4C , D ark 7 d a ys 2 1 d a ys 

### $ 2 5 0 m L H igh D e n sity AmberPl as t i c or Si l ani zed Am ber G l ass

5 5 0 , 55 0
. 1 So d i um

T h io su lfate Coo l , 4C, HCl pH<2 7 da y s 4 C , D ark 5 5 0 , 3 0 d a ys 5 5 0 .1 , 4 0 d a ys 1 L A m b e r G la ss w ith P T FE Li ned Cap

5 5 1 .1 S o d ium S u lfi te ,

A m m o ni um C hl ori de, p H 4 .5 -5 .0 w ith phospha t e buf

f er Cool , 4C 1 4 d a ys 

### $ 40 mL G l as s w i th PT FE Lined S ep tum

5 5 2
. 1 A m m o ni um

ch l or i de C o o l , 4C , D ark 2 8 d a ys 

### # 4 C , da rk 4 8 hours 2 5 0 m L A m b e r G la ss w ith P T FE Li ned Cap

5 5 2
. 2 A m m o ni um

ch l or i de C o o l , 4C , D ark 1 4 d a ys 

### # 4 C , d a rk 7 d a ys


### # -1 0 C 1 4 d a ys 5 0 m L A m b e r G la ss w ith P T FE Li ned Cap

5 5 5 S o d ium S u lfi te

H Cl , pH 

### # 2 D ark , Cool 4C 1 4 d a ys 

### $ 100 mL G l as s w i th PT FE Li ned c ap

1 6 1 3 So d i um

T h io su lfate Coo l , 0- 4C, D ark R eco m m end 4 0 d a ys 1 L A m b e r G la ss w ith P T FE Li ned Cap

## IV- 27

T a ble I V -7 M C L s a nd D etecti o n L i m i ts R eq uirem en ts i n th e C F R (mg / L ) t o C o m po si te

Ino rg a nics M C L * M C L G D etecti o n L i m i t R eq uired to C o m po si te [ § 1 4 1 . 2 3 (a)(4)] As bes t os 7 M FL 7 M FL 1. 4 M FL Br o m a t e 0
. 0 1 0 NA C h l o r i t e 1
. 0 NA Cy an i de 0. 2 0. 2 0. 04 F luo rid e 4 .0 0 .8 N i tr at e 10 10 2 N itrit e 1 1 0 .2

* T he m o ni t o ri ng tri gger f o r the i no rgani cs i s the M CL e x c e ptf

orbo th n i tr a t e a n d n i tr i t e , w h i c h a r e 1⁄2 th e M CL

M etals M C L * M C L G D etecti o n L i m i t R eq uired to C o m po si te [ § 1 4 1 . 2 3 (a)(4)] Ant i mon y 0. 006 0. 006 0. 001 Ar s en i c 0. 01 - 0. 01 B a riu m 2 2 0 .4 B ery l l i u m 0. 004 0. 004 0. 0008 Cadmi u m 0. 005 0. 005 0. 001 Chr omi u m 0. 1 0. 1 0. 02 Copper * * 1. 3 1. 3 0. 001 0. 02 ( f ordi r e c ta s pi r a t i onAA) Le ad* * 0. 015 ze r o 0. 001 M er cury 0. 002 0. 002 0. 0004 Se l en i u m 0. 05 0. 05 0. 01 T h al l i u m 0. 002 0. 0005 0. 0004 * T he mo ni t ori ng tr i gg erf or met al s i s the M C L un l es s comp osi t i ng , then1/ 5 M C L i s r equi r ed

* * A ct i on Lev el

## IV- 28

TA B LE I V - 8 V O C M C Ls and D et ec t i on Li mi t R eq ui re men t s i n t he C FR ( mg/ L) f or C ompl i anc e M oni t ori ng
Vo l a t i l e Or g a n i c s M CL M CL G Re q u i r e d M DL * THM s 0
. 0 8 NA NA HA A 5 0
. 0 6 NA NA B en ze n e 0. 005 ze r o 0. 0005 Carbon t etr ac h l or i de 0. 005 ze r o 0. 0005 Chl or oben ze n e 0. 1 0. 1 0. 0005
o- D i ch l or oben ze n e 0. 6 0. 6 0. 0005 p- D i ch l or oben ze n e 0. 075 0. 075 0. 0005 1. 2- D i ch l or oeth an e 0. 005 ze r o 0. 0005 1, 1- D i ch l or oeth y l en e 0. 007 0. 007 0. 0005
c- 1, 2- D i ch l or oeth y l en e 0. 07 0. 07 0. 0005 t - 1, 2- D i ch l or oeth y l en e 0. 1 0. 1 0. 0005 D i ch l or ometh an e 0. 005 ze r o 0. 0005 1, 2- D i ch l or opr opan e 0. 005 ze r o 0. 0005 Eth y l ben ze n e 0. 7 0. 7 0. 0005 Sty r en e 0. 1 0. 1 0. 0005 T etr ac h l or oeth y l en e 0. 005 ze r o 0. 0005 T ol u en e 1 1 0. 0005 1, 2, 4- T r i ch l or oben ze n e 0. 07 0. 07 0. 0005 1, 1, 1- T r i ch l or oeth an e 0. 2 0. 2 0. 0005 1, 1, 2- T r i ch l or oeth an e 0. 005 0. 003 0. 0005 T r i ch l or oeth y l en e 0. 005 ze r o 0. 0005 V i n y l c h l or i de 0. 002 ze r o 0. 0005 X y l en es 10 10 0. 0005 * A l aborat orymus tbe abl e t o ach i eve anM D L of 0. 00 05 mg/ L t o be cert i f i ed t o ana l y ze s amp l es f

or c om pl i anc e moni t ori ng [ §1 41
. 24 ( f ) ( 17 ) ( i ) ( E ) an d (i i ) ( C ) ]
. T hi s i s al s o the m o ni t o ri ng tri gger f

o r V O C s [§1 4 1
. 2 4 (f

)( 1 1 )]
.

## IV- 29

TAB LE I V - 9 S O C M C Ls an d D e t e c t i on Li mi t Req ui r e me nt s i n t he CFR t o Re du c e M on i t or i ng ( mg / L)
SO C s M C L M C L G M oni t ori ng T ri gg er* Al ac h l or 0. 002 ze r o 0. 0002 Atr az i n e 0. 003 0. 003 0. 0001 B en zo( a) py r en e 0. 0002 ze r o 0. 00002 Carbof u r an 0. 04 0. 04 0. 0009 Chl or dan e 0. 002 ze r o 0. 0002 2, 4- D 0. 07 0. 07 0. 0001 D i ( 2- eth y l h ex y l ) adipat e 0. 4 0. 4 0. 0006 D i ( 2- eth y l h ex y l ) ph th al at e 0. 006 ze r o 0. 0006 D i br omoch l or opr opan e ( D B CP ) 0. 0002 ze r o 0. 00002 D al apon 0. 2 0. 2 0. 001 D i n os eb 0. 007 0. 007 0. 0002 D i ox i n ( 2, 3, 7, 8- T CD D ) 3x 10 ze r o 5x 10 -8 -9 D i qu at 0. 02 0. 02 0. 0004 Endoth al l 0. 1 0. 1 0. 009 Endr i n 0. 002 0. 002 0. 00001 Eth y l en edibr omi de ( ED B ) 0. 00005 ze r o 0. 00001 G l yph os at e 0. 7 0. 7 0. 006 H ept ac h l or 0. 0004 ze r o 0. 00004 H ept ac h l orEpoxi de 0. 0002 ze r o 0. 00002 H ex ac h l or oben ze n e 0. 001 ze r o 0. 0001 H ex ac h l or ocy cl open t adi en e 0. 05 0. 05 0. 0001 Li n dan e 0. 0002 0. 0002 0. 00002 M eth ox y ch l or 0. 04 0. 04 0. 0001 O x amy l 0. 2 0. 2 0. 002 PC B s ( as de ca ch l or obi ph en y l ) 0. 0005 ze r o 0. 0001 Pen t ac h l or oph en ol 0. 001 ze r o 0. 00004 Pi cl or am 0. 5 0. 5 0. 0001 Si maz i n e 0. 004 0. 004 0. 00007 T ox aph en e 0. 003 ze r o 0. 001 2, 4, 5- T P ( Si l v ex ) 0. 05 0. 05 0. 0002 *T h e m o n ito rin g trigg e rs fo r S O C s listed in the re g u la tio n a re a lso re q u ire d fo r c o m p o siting b u t are not requir ed by r egul at i o n f o r cert i f i cat i o n [§1 4 1
. 2 4 (g) (7), (10 )( i ) and (18 )]
.Rec ommendedL evel 1 

## IV- 30 T a b l e I V- 1 0 M CL a n d P r o f i c i e n c y T e s t i n g S a m p l e Ac c e p t a n c e Cr i t e r i a i n t h e CF R P ri m a ry an d S eco nd a ry D ri nking W a ter R eg ulati o ns [§1 4 1 . 2 3 (k) (3)(i i ) an d 1 4 1 . 2 4 (f )( 1 7 ) an d (1 9 )]
R egul ated Pa ramete r M C L /[ SM C L ] M C L G A cce ptance
M ETALS Al u mi n u m [ 0. 05- 0. 2] - Ant i mon y 0. 006 0. 006 + 30% Ar s en i c 0. 01 - B ar i u m 2. 0 2. 0 + 15% B ery l l i u m 0. 004 0. 004 + 15% Cadmi u m 0. 005 0. 005 + 20% C al ci um - - Chr omi u m 0. 1 0. 1 + 15% Copper 1. 3/ 90% [ 1. 0] 1. 3 + 10%

Ir o n [0 .3 ] - Le ad 0. 015/ 90% ze r o + 30% M an g an es e [ 0. 05] - M er cury 0. 002 0. 002 + 30% Se l en i u m 0. 05 0. 05 + 20% S ili c a - - Si l v er [ 0. 10] Sodi u m 20 - 1
T h al l i u m 0. 002 0. 0005 + 30% Zi n c [ 5. 0] -R egul ated Pa ramete r M C L /[ SM C L ] M C L G A cce ptance



## IV- 31 I NORGANI CS A lka linity - - A s bes t os 7M F/ L> 10 u 7M F/ L> 10 u 2 St d D ev B r omat e 0. 010 95% conf interval around mean Chl or i de [ 250] - C h lo rite 1 .0 95% conf interval around mean R e sid u a l D isinfe c ta n t d e te c ta b le - C o l o r [15 cu] - C o n d u c tivity - - C orr osi vi ty [ non- corr osi ve] - Cy an i de 0. 2 0. 2 + 25% F luo rid e 4 .0 [ 2. 0] - + 10%

Foa mi n g Ag en t s [ 0. 5] - N i tr at e ( as N ) 10 10 + 10% N i tr i t e ( a s N ) 1 1 + 1 5 % N i tr at e/N i tr i t e ( as N ) 10 10 O d o r [3 t. o .n.] - p H 6 .5 -8 .5 [ 6. 5- 8. 5] -

o -P h o sp h a te - - Sol i ds ( T D S) [ 500] - Sul f at e def err ed [ 250] def err ed

T em p erature - -R egul ated Pa ramete r M C L /[ SM C L ] M C L G A cce ptance



## IV- 32 V O LATI LES T r i h al ometh an es ( T ot al ) 0. 080 95% conf interval around mean H A A 5 0. 06 0 95 % conf i nt erval ar ound mean B enzene 0
. 0 0 5 zero * C arbo n t etrachl o ri d e 0
. 0 0 5 zero * C h lo ro b e n z e n e 0 .1 0 .1 *
p- D i ch l or oben ze n e 0. 075 [ 0. 005] 0. 075 *

o -D ic h lo ro b e n z e n e 0 .6 0 .6 * 1 , 2
- D i chl o roethane 0
. 0 0 5 zero * 1, 1- D i ch l or oeth y l en e 0. 007 0. 007 *

c- 1, 2- D i ch l or oeth y l en e 0. 07 0. 07 * t-1 ,2 -D ic h lo ro e thyle n e 0 .1 0 .1 * D i chl o rom ethane 0
. 0 0 5 zero * 1, 2- D i ch l or opr opan e 0. 005 ze r o * E thylb e n z e n e 0 .7 0 .7 * S tyre n e 0 .1 0 .1 * T etrachl o roethy l ene 0
. 0 0 5 zero * T o l uene 1 1 * 1, 2, 4- T r i ch l or oben ze n e 0. 07 0. 07 * 1 ,1 ,1 -T ric h lo ro e tha n e 0 .2 0 .2 * 1, 1, 2- T r i ch l or oeth an e 0. 005 0. 003 * T ri chl o roethy l ene 0
. 0 0 5 zero * V i n y l c h l or i de 0. 002 ze r o ± 40% X y l en es ( T ot al ) 10 10 *R egul ated Pa ramete r M C L /[ SM C L ] M C L G A cce ptance



## IV- 33 S YNT HET I C ORGANI CS Al ac h l or 0. 002 ze r o + 45% A l di carb P ostpo ned P ostpo ned 2 St d D ev A l di carb Sul f

oxi de P ostpo ned P ostpo ned 2 St d D ev A l di carb Sul f

one P ostpo ned P ostpo ned 2 St d D ev Atr az i n e 0. 003 0. 003 + 45% Carbof u r an 0. 04 0. 04 + 45% Chl or dan e 0. 002 ze r o + 45% 2, 4- D 0. 07 0. 07 + 50% D al apon 0. 2 0. 2 2 St d D ev D i br omoch l or opr opan e( D B CP ) 0. 0002 ze r o + 40% D i nos eb 0. 00 7 0. 00 7 2 St d D ev D i quat 0. 02 0. 02 2 St d D ev E ndothal l 0. 1 0. 1 2 St d D ev Endr i n 0. 002 0. 002 + 30% Eth y l en edibr omi de( ED B ) 0. 00005 ze r o + 40% G l yphosa t e 0. 7 0. 7 2 St d D ev H ept ac h l or 0. 0004 ze r o + 45% H ept ac h l ore pox i de 0. 0002 ze r o + 45% Li n dan e 0. 0002 0. 0002 + 45% M eth ox y ch l or 0. 04 0. 04 + 45% O xamy l ( V y dat e) 0. 2 0. 2 2 St d D ev

Pen t ac h l or oph en ol 0. 001 ze r o + 50%R egul ated Pa ramete r M C L /[ SM C L ] M C L G A cce ptance



## IV- 34 P i cl oram 0. 5 0. 5 2 St d D ev Si mazi ne 0. 00 4 0. 00 4 2 St d D ev T ox aph en e 0. 003 ze r o + 45% 2, 4, 5- T P(S i l v ex ) 0. 05 0. 05 + 50% H exa chl orobenze ne 0. 00 1 zer o 2 St d D ev H exa chl orocy cl op ent adi ene 0. 05 0. 05 2 St d D ev B enz o(a) py r ene 0. 00 02 zer o 2 St d D ev PC Bs (a s d e c a c h lo ro b ip h e n yl) 0
. 0 0 0 5 z e r o 0
- 2 0 0 %
2, 3, 7, 8- T C D D ( D i oxi n) 3x10 zer o 2 St d D ev -8
A c ry l a m i d e Tr e a tm e n t z e r o NA Ep i c h l o r o h y d r i n Tr e a tm e n t z e r o NA D i ( 2- ethy l hex y l ) adi pat e 0. 4 0. 4 2 St d D ev D i ( 2- ethy l hex y l ) phthal at e 0. 00 6 zer o 2 St d D ev * th e a c c e p ta n c e li m its f o r V O C s a re ± 2 0 % a t 

### $ 0 .0 1 0 m g /L a n d ± 4 0 % a t < 0 .0 1 0 m g /L N A - N o t A p p lica b leTabl e I V - 11 Promul gat ed O rgan i c D ri nk i ng W at er M et hods (As of January 2 0 0 5 )

M et hod N umbe r M et hod Ti t l e Revi s i on



## IV- 35 502. 2 V ol at i l e O r g an i c Compoun ds i n W at erB yPur g e an d T r ap Capi l l ary 2. 1 a C o lum n G a s C h ro m a to g ra p h y w ith P h o to io n iz a tio a n d E le c tro lyti c

C o nd uc t i vity D etec t o rs i n S eries 5 0 4 .1 1 ,2 -D ib ro m o e tha n e (E D B ), 1 ,2 -D ib ro m o -3 -C h lo ro p ro p a n e (D B C P ), 1 .1 a and 1,2,3- T ri chl o ro- p rop ane (12 3 T C P ) i n W at er by M i croe xtract i o n and G as Chro m at o graphy 505 Ana l y s i s of O r g an oh al i de Pes t i ci des a n d Commer ci alPol y ch l or i n at ed 2. 1 a B i p henylP rod uct s i n W at er by M i cro-E xtract i o n and G as Chro m at o graphy 506 D et ermi n at i on of Phth al at e an d Adi pat e Es t er s i n D r i n k i n g W at erby 1. 1 a Li qu i d- Li qu i d Extr ac t i on orLi qu i d- Sol i d Extr ac t i on a n d

G as Chro m at o graphy wi th P ho t o i o ni zat i o n D et ect i o n 5 0 7 D e te rm ina tio n o f N itro g e n a n d P h o sp h o ru s-C o n ta inin g P e sticid e s 2 .1 a i n W at erbyG as Chr omat og r aph yw i th a N i tr og en - Phos ph oru s D et ec t or 508 D et ermi n at i on of Chl or i n at ed Pes t i ci des i n W at erbyG as 3. 1 a Chr omat og r aph yw i th An El ec tr on Captu r e D et ec t or 5 0 8 A S c re e n ing fo r P o lyc h lo rin a te d B ip h e n yls b y P e rc h lo rin a tio n a n d 1 .0 b G as Chro m at o graphy 5 0 8 .1 D e te rm ina tio n o f C h lo rin a te d P e sticid e s, H e rb ic id e s, a n d O rg a n o h a lid e s 2 .0 a b yLi q ui d

- So l i d E xtract i o n and E l ectron C ap ture G as Chro m at o graphy 5 1 5 .1 D e te rm ina tio n o f C h lo rin a te d A c id s in W a te r b y G a s C h ro m a to g ra p h y 4 .0 b wi th an E l ectron C ap ture D et ect o r 5 1 5 .2 D e te rm ina tio n o f C h lo rin a te d A c id s in W a te r U sin g L iq u id -S o lid 1 .1 a E xtract i o n and G as Chro m at o graphy W i th an E l ectron C ap ture D et ect o r 5 1 5 .3 D e te rm ina tio n o f C h lo rin a te d A c id s in D rin k ing W a te r b y L iq u id -L iq u id 1 .0 e E xtract i o n, D eri vat i zat i o n and G as Chro m at o graphy wi th E l ectron C ap ture

D et ec t i on 5 1 5 .4 D e te rm ina tio n o f C h lo rin a te d A c id s in D rin k ing W a te r b y L iq u id -L iq u id 1 .0 f M i croe xtract i o n, D eri vat i zat i o n and FastG as Chro m at o graphy wi th

El ec tr on Captu r e D et ec t i on 5 2 4 .2 M e a su re m e n t o f P u rg e a b le O rg a n ic C o m p o u n d s in W a te r b y C a p ill a ry 4 .1 a C o l um n G as Chro m at o graphy/M ass Sp ectrom etry 5 2 5 .2 D e te rm ina tio n o f O rg a n ic C o m p o u n d s in D rin k ing W a te r b y L iq u id -S o lid 2 .0 (a) E xtract i o n and C ap i l l aryC o l um n G as Chro m at o graphy/M ass Sp ectrom etry 531. 1 M ea s u r eme n tof N
- M eth y l ca rbamoy l ox i mes a n d N

- M eth y l ca rbama t es i n 3. 1 a W at erbyD i r ec tAqu eou s I n j ec t i on H PLC w i th Pos tCol u mn D er i v at i za t i onTabl e I V - 11 Promul gat ed O rgan i c D ri nk i ng W at er M et hods (As of January 2 0 0 5 )

M et hod N umbe r M et hod Ti t l e Revi s i on



## IV- 36 5 3 1 .2 M e a su re m e n t o f N -M e thylc a rb a m o ylo x im e s a n d N -M e thylc a rb a m a te s in 1 .0 f W at erbyD i r ec tAqu eou s I n j ec t i on H PLC w i th Pos tCol u mn D er i v at i za t i on 5 4 7 D eterm i na t i o n o f G l yp ho sate in D rinking W ater B y D i rec t - A q ue o us- c I n

j ec t i on H PLC , Pos t - Col u mn D er i v at i za t i on , a n d Fl u or es ce n ce D et ec t i on 548. 1 D et ermi n at i on of Endoth al l i n D r i n k i n g W at erbyI on Exc h an g e Extr ac t i on , 1. 0 d A ci di c M ethan ol M ethy l at i on G as Chromat ogr aphy/M as s Spec.

5 4 9 .2 D e te rm ina tio n o f D iq u a t an d P a ra q u a t i n D rin k ing W a te r b y L iq u id -S o lid 1 .0 e E xtract i o n and H i gh P erf o rma nce Liqui d C hrom at o graphy wi th U l tr av i ol etD et ec t i on 5 5 0 D et erm i nat i o n of P o l y cycl i c A rom at i c H y d ro- carb o ns i n D ri nki ng W at er c byLi qu i d- Li qu i d Extr ac t i on a n d H PLC w i th Coupl ed U l tr av i ol eta n d

Fl u or es ce n ce D et ec t i on 5 5 0
. 1 D et erm i nat i o n of P o l y cycl i c A rom at i c H y d ro- carb o ns i n D ri nki ng W at er c byLi qu i d- Sol i d Extr ac t i on a n d H PLC w i th Coupl ed U l tr av i ol eta n d

Fluorescence D et ect i o n
551. 1 D et ermi n at i on of Chl or i n at i on D i s i n f

ec t i on B ypr odu ct s , Chl or i n at ed Sol v en t s , 1. 0 a an d H al og en at ed Pes t i ci des /H erbi ci des i n D r i n k i n g W at erbyLi qu i d- Li qu i d

Extr ac t i on a n d G as Chr omat og r aph yw i th El ec tr on - Captu r e D et ec t i on 5 5 2 .1 D e te rm ina tio n o f H a lo a c e tic A c id s a n d D a lap o n in D rin k ing W a te r b y 1 .0 d Ion E xchange L i q ui d

- So l i d E xtract i o n and G as Chro m at o graphy W i th

El ec tr on Captu r e D et ec t i on 552. 2 D et ermi n at i on of H al oac et i c Aci ds a n d D al aponi n D r i n k i n g W at erby 1. 0 a Liqui d

- Liqui d E xtract i o n, D eri vat i zat i o n and G as Chro m at o graphy W i th

E l ectron C ap ture D et ect i o n 5 5 5 D e te rm ina tio n o f C h lo rin a te d A c id s in W a te r b y H igh P e rfo rm a n c e L iq u id 1 .0 d C hrom at o graphy wi th a P ho t o d i o d e A rr ay U l travi o l etD et ect o r

## IV- 37

Foo tnot es T hese method s are i n the m anual t i t l ed "M etho d s f o r the D et erm i nat i o n of O rgani c C o m p o und s i n D ri nki ng W at er - a Suppl ementI I I , " do cumen tnu mb erE P A / 60 0/ R
- 95 / 13 1, Augu s t199 5. T hi s manu al i s av ai l abl e f r om the N at i onal T ec h n i ca l I n f

ormat i on Se rv i ce ( N T I S* )
. Addr es s y ou rr equ es tt o N T I S a n d a s kf orth ei ror dern u mberPB 95- 261616, cos ti s $101. 00 T h e se m e tho d s a re in the m a n u a l t itl e d " M e tho d s fo r th e D e te rm ina tio n o f O rg a n ic C o m p o u n d s in D rin k ing W a te r," b d o cum entnum b er E P A / 6 0 0 /4
- 8 8 / 0 3 9 , D ecem b er 19 8 8 (Re vi sed July19 9 1 ). T hi s m anual i s avai l ab l e f rom the N at i onal T ech ni calI nf ormat i on Servi ce ( N T I S* )
. Ad dres s y ourr eques tt o N T I S and as k f or thei rordernu mb er PB 91- 231480; c os ti s $77. 50. T hese method s are i n the m anual t i t l ed "M etho d s f o r the D et erm i nat i o n of O rgani c C o m p o und s i n D ri nki ng W at er - c Suppl ementI , " do cumen tnu mb erE P A / 60 0/ 4- 90 / 02 0, Ju l y199 0. T hi s manu al i s av ai l abl e f r om the N at i onal T ec h n i ca l I n f

ormat i on Se rv i ce ( N T I S* )
. Addr es s y ou rr equ es tt o N T I S a n d a s kf orth ei ror dern u mberPB 91- 146027; cos ti s $58. 50. T hese method s are i n the m anual t i t l ed "M etho d s f o r the D et erm i nat i o n of O rgani c C o m p o und s i n D ri nki ng W at er d Suppl ementI I , " do cumen tnu mb erE P A / 60 0/ R
- 92 / 12 9, Augu s t199 2. T hi s manu al i s av ai l abl e f r om the N at i onal T ec h n i ca l I n f

ormat i on Se rv i ce ( N T I S* )
. Addr es s y ou rr equ es tt o N T I S a n d a s kf orth ei ror dern u mberPB 92- 207703; cos ti s $63. 00. T h e se m e tho d s a re in the m a n u a l t itl e d , “M e tho d s fo r th e D e te rm ina tio n o f O rg a n ic a n d In o rg a n ic C o m p o u n d s in e D ri nki ng W at er - V o l um e 1 , ” d o cum entnum b er E P A 81 5
- R
- 0 0
- 0 1 4 , A ugust20 0 0
. T hi s m anual i s avai l ab l e f rom the N at i onal T ech ni calI nf ormat i on Servi ce ( N T I S* )
. Ad dres s y ourr eques tt o N T I S and as k f or thei rordernu mb er PB 2000- 106981; c os ti s $71. 50. S ta n d a lo n e m e tho d s. T h e se m a n u a ls a re a v a ilab le a t th e E P A w e b site a t: f h ttp: //w w w . e pa . g ov / og w dw /me th od s / s ou r c a l t
. h tml

* N at i o nal T echni cal Inf o rma t i o n Servi ce (N T IS) 528 5 P ortRo y al Ro ad Spr i n g f i el d, V A 22161 Phon e n u mber : 800- 553- 6847 Fa x n u mber : 703- 0605- 6900 T her e i s a $5. 00 ha ndl i ng ch ar ge f or the t ot al pur cha s e.

* * M i crob i o l o gi cal and C hem i cal E xp o sur e A ss ess m entR esearch D i vi si o n, Ch emi ca l Expos u r e Res ea r chB r an ch , Room 564 U
. S. E nv i r onment al P r ot ect i on Agenc y 26 W estM art i n Luther K i ng D ri ve Ci n ci n n at i , O H 45268 Ph on e n u mber : 513- 569- 7586 F ax n u mber : 513- 569- 7757 E- mai l a ddr es s : dw meth ods

- h elp@ epa mai l
. epa . g ov

## IV- 38

## IV- 39



# E xampleCh e c kli st s forOn -S iteE valuati on of Laboratori es Analyzin g Drin king Water

## IV- 40



# Ge ne ral AuditIn fo rm ation
L a b o ra to ry

Str eet

C ity, S ta te
Z ip
T el eph on e N o. Fa x N o. A udi tT eam Leader
A ud i tT eam M em b ers

Audi tT ea m Af

f i l i at i on D a te

## IV- 41

Laborat ory ___________________________________________ Eval uat or

_________________________ Locat i on _____________________________________________ D at e

______________________________ PH Y SI C A L FAC I LI TY

Ite m A c c e p ta b le Ye s N o C o m m e n ts
E nvi ronm ent H e a ting /C o o ling /H u m id ity Li ght i ng V en t i l at i on / Exh au s th oods C l ea nli ne ss E l ec trical and wa t er se rvice s W orkSpace Separ at i on of

i ncomp at i bl e t es t i ng ar eas C o n tro lled a c c e ss w h e re a p p ro p ria te H o usekeep i ng U ne nc um b ere d ac ce ss Ad equat e work s pace Storage Chemi cal s proper l ys t ored an d dat ed St and ards pro p erl ys t o red, dat ed and l ab el ed wi th co ncentrat i o n, prep arer' s nam e and s o l vent , ori gi n, p u rity & tra c e a b ili ty C o m p ut ers & aut o m at ed eq ui p m ent Saf etyprocedur es

## IV- 42

Laborat ory ___________________________________________ Eval uat or

_________________________ Locat i on _____________________________________________ D at e

______________________________ PER SO N N EL (U se a d d iti o n a l p a p e r if ne c e ssa ry.) P osi t i on/Ti t l e N am e E ducati on L evel D egree/M ajor* Speci al i zed Trai ni ng Pre s en t

Sp eci a l ty E xperi ence
La b o rat o ry D i r ec t or
M ana ger Sup ervi sors
Ins trume nt O p erat o rs AA
TEM
H PLC
GC
I CP
G C /MS
I C
O the r a n a lysts

Y e s N o C o m m e n ts A n o rg a n iz a tio n c h a rt a v a ilab le Q A m a n a g e r h a s line a u tho rity P er s onnelj ob des cr i pt i ons an d re s umes a v a ilab le P er s onneltr ai ni ng document ed *If the m a

jo r is n o t i n c h e m istry, li st ho u rs o f co lleg e lev e l co u rs e s in c h e m istry.

## IV- 43

Laborat ory ___________________________________________ Eval uat or

_________________________ Locat i on _____________________________________________ D at e

______________________________ QUAL I T Y AS S URANCE AND DAT A RE PORT I NG
I t em C om ments Sati s factory Y es No Q A pl an O rgan i zat i on Sampl i ng SO P s av ai l abl e a nd us ed Pr es erv at i on C o nt ai ners H ol di ng t i mes Sampl er s tr ai ned Sampl e R ej ec t i on Laborat ory s ampl e handl i ng Lo g i n pro ced ure B o und l o g bo o k or secure c o m p u te r lo g in St o rage T racki ng A nalyt i calM et hods W ritten m e tho d s a v a ilab le Ap proved methods us ed SO P s av ai l abl e a nd us ed C al i br at i on T ype an d f

r equen cy Sou r ce of

s t an dar ds D a ta c o m p a ra b ili ty I nstrume nttuni ng B l a nks T rip F ie ld M eth od M etho d D etecti o n L i m i ts I ni t i al Fr equen cy A c c e p ta b ili ty P rec i s i on a nd A ccuracy I ni t i al Fr equen cy A c c e p ta b ili ty C o n tro l ch a rts Lab o rat o ryf

o rt i f i ed blanks M atr i x dupl i cat esI t em C om ments Sati s factory Y es No



## IV- 44 O ther Q C C he cks Perf ormanc e c hec k s amp l es I n t ern ala n d s u rr og at e s t an dar ds M atr i x s pi kes an d re pl i cat es Q ual i t at i ve I de nt i f

i cat i on/ C onf i rmat i on Per forman ce Eval uat i on Samp l es A nal yzed D at a R ed uc t i on and V al i dat i on C al cul at i o ns T r an s cr i pt i on Si gn i f i can tFi gu r es V al i dat i on P revent i ve M ai nte nan ce R ec ord s R et en t i on C orr ec t i ve A ct i on

## IV- 45

Laborat ory Eval uat or L o ca ti o n D a te
I t em N o. of U nit s M et hod M anu facture r M odel Sati s factory Ye s No A nal yti calB al ance 0 .1 m g re a d a b ili ty S t ab l e b ase A S T M typ e 1 o r 2 we i gh t s S e rv ic e c o n tra c ts M ag net i c St i rre r V ar i abl e s peed, T FE coat ed s t i rbar pH M et er A c c u ra c y ± 0 .1 un its L ine o r b a tter y U s abl e wi th s peci f

i c i on el ectr od es C ond uct i vi t y M et er Rea dabl e i n oh ms ormh os Ran g e of

2 oh ms t o 2 mh os Li ne o r batt ery H o t P l a te - t emp con tr ol C en t ri f uge T o 3 00 0 rpm , Opt i onof 4 x 50 mL C ol or St andar ds T o veri f ywavel engths p ho t o m et ers Sho ul d co ver 20 0
- 8 0 0 nm R ef ri ger at or / F ree zer St andar d l aborat ory , ex pl osi on proof f or or gan i cs Capabl e of

ma i n t ai n i n g n omi n alt empe r atu r e of 4CI t em N o. of U nit s M et hod M anu facture r M odel Sati s factory Ye s No



## IV- 46 D ryi ng O ven G r av i tyorc on v ec t i on Contr ol l ed f

r om r oom t emp t o 180° C or h i g h e r ( ±2° C) M uff l e Fu rnace T o 45 0 °C f

o r cl eani ng organi c gl ass ware T hermom et er M ercuryf

i l l ed C el si us 1 °C o r f i ne r sub d i visi o n to 1 8 0 °C N IS T C e rtifi e d o r tra c e a b le G l ass w are B o ro silica te V o l um etri cs sho uld b e C l ass A Spect ropho t om et er R ange 4 0 0 - 70 0 nm B and wi d th - < 20 nm U se se v e ra l size & sh a p e c e lls P ath l eng th 1 - 5 cm Cy an i de, Fl u or i de M ercury N itra te /N itrit e

F i l t er P hotom et er R ange 4 0 0 - 70 0 nm B and wi d th 10 - 7 0 nm U se se v e ra l size & sh a p e c e lls P athl eng th 1 - 5 cm same as ab o ve

A mpe rome t ri c Ti t rat or D isinfe c ta n ts Speci f i c I on M et er Ac c u r a c y± 1 mV Cy an i de Fl u or i de N itra teI t em N o. of U nit s M et hod M anu facture r M odel Sati s factory Ye s No



## IV- 47 I nd uc t i ve l y Cou pl e d Pl as ma (se q u e n tial, si m ult an eo us) Com put ercontr ol l ed Bac k g r ou n d c orr ec t i on Ra di o f

r equ en cyg en er at or A rg o n ga s su p p ly M as s Spectr om et er Ra n g e 5- 25 0 a mu Res ol ut i on 1 amu peak wi d th at5% pe ak hei ght 200. 7, 3120B

2 0 0 .8

W a ter B a th E l ectr i c ors t eam he at Con tr ol l a bl e w i th i n 5° C t o 1 00 ° C M ercury N itra te P es t i ci des I on C hr omat ograp hy Condu ct i v i tyde t ec t or , U V de t ec t or S u pp r e s s orc olu mn , S e pa r a t orc olu mn Fl u or i de, Chl or i de N itra te /N itrit e B ro m a te , C h lo rite
A t omi c A bs orp t i on Spect ropho t om et er Si ng l e c han nel , Si ng l e ordoubl e beam G r at i ng mo nochr om et er Phot o mu l t i pl i erde t ec t or A d jus t ab l e sl i t s, R ange 1 9 0
- 8 0 0 nm Re a do u ts y s t e m: Re s po n s e t i me c om pa t i bl e w i th AA Ab l e t o det ectposi t i ve i nt er f er enc e f or f urnac e C hartrecord er, C R T or hard co p ypri nt er M e ta ls

A i r/A cety l ene c ommer ci alg r ade B a r i u m, C op perI t em N o. of U nit s M et hod M anu facture r M odel Sati s factory Ye s No



## IV- 48 N i trous O xi d e - co m m . grade B ari um G r aphi t e Furnac e Arg on or N i tr ogen ( com merc i al gr ade) Reach r equi r ed t emp er atur es B a c k g ro u n d c o rr e c to r p ro v isio n fo r o ff line a n a lysis Pi pet s a n d t i ps mi cr ol i t erc apa ci tyw i th di s pos abl e t i ps 5- 1 0 0 m i croli t ers me t alf r ee t i ps A nt i mo ny , Lead A rs eni c, B ari um B ery l l i um C adm i um, N i cke l C hrom i um Se l eni um T hal l i um C op per A rs i ne G enerat o r A rs eni c, Seleni um H y dr i de G en er at or h y dr og en , c ommer ci alg r ade A nt i m o ny A rs eni c, Seleni um M erc ury A nal yzer Spectr op hot om et er Ded i c a t e d a n a l y z e rh a v i n g a me r c u ryl a mp ac c e p ta b le A d so rp tio n c e ll: 10 cm qu art z ce l l w i th qu art z en d w i n dows or 11
. 5 cm plexi gl ass cel l wi th 2.5 cm ID A i r p um p t o d eli ve r f

l o w o f at l ea st 1 L /m in A e ra tio ntub e w ith c o a rs e gla ss frit F lo w m e te r to m e a su re a ir fl o w o f 1 L /m in D ryin g un it: 6 i n. tub e wi th 20 grams m agnesi um p e rc h lo ra te o r h e a ting d e v ic e o r la m p to p re v e n t co n d e n sa tio n o n ce ll M ercuryI t em N o. of U nit s M et hod M anu facture r M odel Sati s factory Ye s No



## IV- 49 G l ass w are S e p a ra to ry fun n e ls K ud erna D ani sh (K

- D ) conce ntrat o rs SOCs

G as C hr omat ograp hy Spl i t/ s pl i t l es s i n

j ec t i on O ve n tem p . co ntro l ± 0 .2°C Rec or der , h ar d c opy O vent emp . progr amm er Sub

- am b i entaccessory V ar i abl e- con s t an tdi f

f er en t i alf l ow con tr ol O r gan i cs

El ec tr on Captu r e det ec t or Li nea r i zed R adi ol ogi calCheck 504. 1, 505 5 0 8 , 5 0 8 .1 5 0 8 A , 5 1 5 .1 5 1 5 .2 , 5 5 1 , 5 5 2 .1 E le c tro lyti c C o n d u c tivity/P h o to io n iz a tio n d e te c to r 5 0 2 .2 5 0 6 (P ID o n ly) N i tr og enPhos ph oru s de t ec t or 507 M ass s pect romete r ( qu adru pol e ori on tr ap) Al l gl as s en r i chmen tdevi ce A l l gl ass trans f

er l i ne E le c tro n io n iz a tio n a t 

### $ 70 eV Sca nni ng 35
- 2 6 0 am u 

### # 2 s ec I nt er f ace d dat a s y s t em 5 2 4 .2 , 5 2 5 .2 5 4 8 .1

P urge & Tra p s yst em Al l gl as s pur ger 5/ 25 mL s amp l e s i ze 5 0 2 .2 , 5 2 4 .2I t em N o. of U nit s M et hod M anu facture r M odel Sati s factory Ye s No



## IV- 50 H i gh Per forman ce Li qui d C hr omat ograp hy Cons t an tf

l ow C ap ab l e o f

i nject i ng 2 0- 50 0 μ L G r adi ents y s t em pos t - colu mn r ea ct or f l uo res cence d et ect o r A b sorp t i o n at34 0 nm and 30 8 nm
G r adi ents y s t em U V de t ect o r at25 4 nm f l u or es ce n ce de t ec t or exci t at i o n at28 0 nm de t ect i o n at> 38 9 nm I s ocra t i c s y s t em ph ot odi ode arr ayde t ec t or e x c i t a t i ona t25 7 n m de t ect i o n at> 3 0 8 nm I s ocra t i c s y s t em pos t - colu mn r ea ct or f l u or es ce n ce de t ec t or exci t at i o n at34 0 nm de t ect i o n at> 4 5 5 nm G r adi ents y s t em ph ot odi ode arr ay/U V de t ec t or 2 1 0
- 3 1 0 nm 531. 1, 6610

5 5 0 , 5 5 0 .1

5 4 9 .2

547

555I t em N o. of U nit s M et hod M anu facture r M odel Sati s factory Ye s No



## IV- 51 A uto A nal ysi s Syst em m u l t i

- c h a n n e l pu mp mani f ol d, col ori met er C ya n id e , N itra te /nitrite

Trans mi s s i on El ec t ron M i cr os cop e 80 kV 300- 100, 000X ma g n i f i ca t i on 1 n m r es ol u t i on cal i b rat e scr een S AE D a n d ED As bes t os

## IV- 52

L abo ratory Eval uat or Locat i on D a te M E T HO DOLOGY C o n ta m i n a n t M eth o d (s)

N ame /N u mbera n d r ev i s i on R efere nce C i t e source, y ear, pa ge Sam pl es / M o Sati s factory Ye s No A nt i m o ny A rs e n ic B ari um B ery l l i um C ad m i um C hrom i um C op per Lead M ercury Se l eni um T hal l i um As bes t os Cy an i de B ro m a te C h lo rite Fl u or i deC o n ta m i n a n t M eth o d (s) N ame /N u mbera n d r ev i s i on R efere nce C i t e source, y ear, pa ge Sam pl es / M o Sati s factory Ye s No



## IV- 53 N itra te N itrit e T o t a l T H Ms H AA5 V O Cs H erbi ci des P es t i ci des ED B /D B CP D io x in O th e rS O Cs PC Bs C arbam at es D i quat E n d o tha ll G lyp h o sa te

## IV- 54

L abo ratory Eval uat or Locat i on D a te
S A M P L E C OL L E C T I ON
I t em C om ments Sati s factory Y es N o T r ai n ed Sa mpl e Col l ec t or
R ep res ent at i ve samp l i ng
C o m p l et e samp l e f

o rm
A p p rop ri at e samp l i ng a n d pr es erv at i on

Sa m p l es exceed i ng hol di ng t i mes di s car ded V O C s & T H Ms H ermet i c s eal

## IV- 55

L abo ratory Eval uat or Locat i on D a te S A M P L E HA N D L I N G A N D P R E S E R V A T I ON
C ontam i nan t C ontai ner

mat er i al & s i ze Pr e s e r va t i ve s H ol di ng Ti me Samp l e Extr act Sati s factory Ye s No M ercury M e ta ls As bes t os Cy an i de Fl u or i de N itra te N itrit e B ro m a te C lo rite T o t a l T H Ms H AA5 V O Cs H erbi ci des P es t i ci des ED B /D B CPC ontam i nan t C ontai ner mat er i al & s i ze Pr e s e r va t i ve s H ol di ng Ti me Samp l e Extr act Sati s factory Ye s No



## IV- 56 D io x in O th e rS O Cs C arbam at es D i quat E n d o tha ll G lyp h o sa teV -1 

## Chapter V Critic al Elem ents for Microbiology N o t e 1 : T hi s chap t er us es the t erm "m ust " t o ref er t o cert i f

i cat io n c rite ri a thatare r eq ui red b ythe N at i o nal P ri m ary D rin k ing W a te r Re gul at i o ns. T he t erm "sho ul d " i s used f o r proc ed ures that , whi l e no ts p eci f i cal l yrequi red b ythe r egu l at i ons , ar e c ons i dere d good l aborat orypra ct i ces . T o as s ur e the val i d i t y o f the dat a, i ti s cr i t i calthatl aborat ori es obs erv e both th e r eg u l at ory an d n on - r eg u l at ory cr i t er i a. Cert i f i ca t i on O f

f i ce r s h av e th e pr er og at i v e t o r ef u s e ce rt i f

i ca t i on if the q u a lity c o n tro l d a ta a re ju d g e d un sa tisf a c to ry o r in su ff

ic ie n t. N ot e 2: Q ual i tycontr ol i t ems, des i gn at ed by a “ QC ,” ne c e ssi ta te w ritten re c o rd s. E a c h re c o rd sh o u ld inc lud e a n a lyst’ s initials a n d d a te (s ). N ot e 3: Ref

er en ce s t o Stan da rd Metho ds f orthe Ex ami na t i on of

Wate ran d Was te wat era r e to the 1 8 , 1 9 , an d 2 0 th th th ed i t i o ns (excep twhere s p eci f i cal l ynoted). 

## 1. P ersonnel 1. 1 Supe r vis or

/ Cons ultant T h e su p e rv iso r o f th e m ic ro b io lo g y lab o ra to ry sh o u ld ha v e a b a c h e lo r' s d e g re e in m ic ro b io lo g y, b io lo g y, o r e q u iva le n t. Sup ervi sors who have a d egree i n a subject o ther than m i crob i o l o gy shoul d have had atl eastone co l l ege- l evel m i crob i o l o gy l ab o rat o ry co urs e i n whi ch envi ronm ent al m i crob i o l o gy was co vered
. In ad d i t i o n, the supervi sor s ho ul d have a m i ni m um o f two weeks trai ni ng ata Fed eral agency, Stat e agency, or a c ad em i c i nst i tut i o n i n m i crob i o l o gi cal anal y si s o f dri nki ng wateror 8 0 ho urs o f

o n- the- job tr ai ni ng in waterm i crob i o l o gya ta c ert i f i ed labo rat o ry , o r other trai ni ng accep t ab l e t o the St a t e orEPA. I f

a s u pe rv i s ori s no tav a i l a bl e ( a n d a wa i v e rno tgr a n t e d pe rpa r a g r a ph 1. 3) , a c on s u l t a n th a v i n g th e s a me q ual i f i cat i o ns m ay b e subst i tut ed , as l o ng as the l ab o rat o ry can d o cum ent thatthe co nsul t anti s accep t ab l e t o the Stat e and i s p rese nt o n-si t e freq ue ntl y eno ug h to sati sf ac t o ril y pe rf

o rm a sup erv i so r' s d uti es. T h e lab o ra to ry su p e rv iso r h a s the re sp o n sib ili ty t o e n su re th a t a ll la b o ra to ry p e rs o n n e l h a v e d e m o n stra te d th e ir a b ili ty t o s at i s f act ori l yper f

orm the ana l y s es t o whi ch theyar e a s s i gn ed an d thatal l dat a r eport ed by the l aborat ory meet the r equi r ed qual i tyas s ur anc e a nd r egu l at orycr i t er i a. 1. 2 A nal

ys t ( or equi valent

job title) T he anal y st shoul d p erf o rm micr o b i o l o gi cal t est s wi th m i ni m al supervi si o n and have atl easta hi gh s cho o l ed ucat i o n. I n addi t i o n , th e a n a l y s t s h o u l d h a v e a m i n i m u m o f a t l e a s t th r e e m o n th s o f b e n c h e x p e r i e n c e i n w a t e r , m i l k , or f ood m ic ro b io lo g y. T h e a n a lyst sho u ld a lso ha v e tra inin g a c c e p ta b le to the S ta te (o r E P A fo r n o n -p rim a c y S ta te s) in m i crob i o l o gi cal anal y si s o f dri nki ng w at er and a m i ni m um o f 3 0 d ays o f

on-the- job trai ni ng i n dri nki ng wat er micr o b i o l o gy u n d e r a n e x p e rie n c e d a n a lyst. A n a lysts s h o u ld ta k e a d v a n ta g e o f wo rk sh o p s a n d tra inin g p ro g ra m s tha t m a y b e a v a ilab le f rom St at e r eg u l at ory ag en ci es , pr of es s i on al s oci et i es , an d manu f ac tu r er s . B ef

or e an al y zi n g compl i an ce s ampl es, the an alystsho uld d em o nstrate ac ce p t ab l e re sult s o n un kn o w n sam p l es. 1. 3 Waive rof

Acade m ic Tr aining T h e c e rtifi c a tio n a u tho rity m a y w a ive the ne e d fo r th e a b o v e sp e c ifi e d a c a d e m ic tra inin g , o n a c a se -b y-ca se b a sis, fo r h igh ly e x p e rie n c e d a n a lysts. T h e c e rtifi c a tio n a u tho rity m a y a lso w a ive the n e e d fo r th e a b o v e sp e c ifi e d tra inin g , o n a c a se -b ycas e bas i s , f

or s upervi s o rs o f

lab orat ori es as s oci at ed wi th dr i nk i ng wat ers y s t ems thatonl yan al y ze s amp l es f r om that s y s t em. I f

s uch a w aive r fo r s upervi s or tr ai ni ng i s gr ant ed, the cert i f i cat i on au thor i tywi l l pre pare a w r i tt en and s i gn ed j u s t i f i ca t i on f ors u cha w ai v era n d h av e i ta v ai l abl e f ori n s pec t i on . Labor at or i es s h ou l d a l s o k ee p a co p yof the wai ver. 1. 4 Pers onne l

Rec or ds P er s onnelrecor ds thati nclude academi c b ack gr ound, speci al i zed tr ai ni ng cour s es comp l et ed, an d types of mi cr ob i ol ogi cal an alyses co nd uc t ed sho uld b e m aintained o n lab o rato ry ana l yst s. 

## 2. Laboratory F acilities La b o rat o ryf

aci l i t i es shoul d b e cl ean, t em p erature and hum i d i ty co ntroll ed , and have ad eq uat e l i ght i ng atbe nch t o p s. T heV -2

l ab o rat o rys ho ul d m ai nt ai n ef f ect i ve separat i o n b etween areas where act i vi t i es ar e i nco m p at i b l e, m i ni m i ze traf f i c f l o w and e n su re tha t co n ta m ina tio n d o e s n o t ad v e rs e ly affe c t d a ta q u a lity. B e n c h to p s a n d fl o o rs sh o u ld b e o f a m a te ria l th a t i s e a sily cl ean ed an d di s i nf

ect ed. Labora t ory f

aci l i t i es s houl d h ave s uf f i ci entbenc h- t op ar ea f

or pr oces s i ng s amp l es ; s t orag e s pace f o r m ed i a, glasswa re, an d p o rtab l e eq uip m en t ; f l o o r sp ac e f

o r st ati o na ry eq uip m en t(i nc ub ato rs, wa t er b aths, ref rigera t o rs, et c. ) ; an d as s oci at ed ar ea( s ) f or cl ean i ng gl as s war e and s t er i l i zi ng mat er i al s . T hey s houl d a l s o h ave provi s i ons f or di s po s al o f m ic ro b io lo g ic a l wa ste
. 

## 3. Laboratory Equipm ent and S upp lies T he l aborat orymus tha ve the equi pm entand s uppl i es needed t o perf orm the approved methods f or w hi ch cert i f

i cat i on ha s bee n r equ es t ed. 3 .1 p HMeter 3. 1. 1 A cc ura cy an d sca l e g rad ua t i o ns sho uld b e w i thin ± 0 .1 u nit s. 3. 1. 2 pH buf

f eral i quot s s houl d be us ed onl yonce . 3. 1. 3 E l ec tro d es sho uld b e m aintained ac co rd i ng t o the m an ufac ture r' s rec o m m en d ati o ns. Q C 3. 1. 4 p H m eters sho uld b e st an d ard i ze d b efo re ea ch u se p erio d w i th p H 7 .0 a nd eithe r p H 4 .0 o r 1 0 .0 stand ard b uff ers, whi chever range cove rs the d esi red p H of the m ed i a o r r eagent. T he d at e and buff ers us ed s ho ul d be record ed i n a l o gb o o k, alo ng with a na l yst ’ s i nit i als. Q C 3 .1 .5 R eco rd p H m et er s l o p e m o nthl y , af t er cal i b rat i o n. 3 .1 .5 .1 If th e p H m e te r d o e s n o t ha v e a fea tur e to a u to m a tica lly c a lc u la te th e s lo p e , b u t ca n p ro v id e the p H in mi l l i vol t s ( mV ) , us e the f ol l owi ng f

ormul a t o cal cul at e the s l op e. Sl op e ( a s % ) = 

### * m V a tp H 7 - m V a tp H 4 

### * x 100/ 177 3 .1 .5 .2 I f

th e s l ope i s be l ow 95% ora bov e 105% , th e el ec tr ode orme t erma yn ee d m ai nt enance. Fol l ow m a n u fac tur e r’s ins tru c tio n s fo r e le c tro d e m a inte n a n c e a n d ge n e ra l cle a n ing
. Q C 3. 1. 6 Commer ci albu f f ers ol u t i on c on t ai n er s s h ou l d be da t ed u pon r ec eipta n d w h enope n ed. B uf f ers sho ul d be di s car ded b ythe expi r at i on dat e. 3 .2 Balanc e (

top loaderorpan) 3. 2. 1 B al an ce s s h ou l d h av e r ea dabi l i tyof 0. 1 g
. 3 .2 .2 B alan ce s sho uld p ro vid e a sensit i vity of at l ea st0 .1 g f o r a l o ad o f

1 5 0 g, and 1 m g for a l o ad o f 1 0 g o r l ess. Q C 3. 2. 3 B a la n c e s sh o u ld b e c a lib ra te d m o n thl y u s in g A S T M C la ss 1 , 2 , o r 3 w e igh ts (m inim u m o f th re e tra c e a b le wei gh t s wh i ch brac ketl aborat orywei gh i ng ne eds , wi th ar eadabi l i tyof 0
. 1 g. ) ( A ST M , 191 6 R ace St
. , P hi l adel phi a, P A 19 1 0 3 ). N o n- ref

erence weight s shoul d b e cal i b rat ed e verysi x m o nths wi th ref erence weight s. R eco rd calib rat i o ns i n a l og bookw i th th e i n i t i al s of th e i n di v i du al per f ormi n g th e ca l i br at i on . Cor r ec t i on v alu es s h ou l d be on f i l e an d u s ed. A r ef er en ce w ei g h ts h ou l d be r e- ce r t i f i ed ev eryf

i v e y ea r s . D ama g ed orc orr oded we i g h t s s h ou l d be r epl ac ed. Q C 3. 2. 4 Servi ce contr act s or i nt ernal mai nt ena nce prot ocol s an d m ai nt ena nce r ecor ds s houl d be ava i l abl e. M ai nt ena nce , cal i b rat i o n, and cl eani ng s ho ul d be co nd uct ed at l east annual l y b y a q ual i f i ed i nd ep end entt echni ci an. I n cases where a l aborat oryi s geogr aphi cal l y i s ol at ed s uch that an annualvi s i tf r om a t ech ni ci an i s i mp r act i cal , the cert i f i cat i on of

f i cer m a y m o d ify o r w a ive the ne e d fo r a tec h n ic ia n .V -3

3. 3 Temper atur e Monitoring D evice 3. 3. 1 G l as s , di al , or el ectr oni c thermo met er s mus tbe gr aduat ed i n 0. 5°C i ncr ement s (0. 2°C i ncr ement s f or t es t s wh i ch a r e i n c u ba t e d a t44
. 5° C) or l e s s , e x c e pt a s n ot e d f or h ota i rov e n s ( 3. 6. 1) a n d r e f r i g e r a t or s ( 3. 9. 1)
. The f l u i d c olu mn i n g l as s th ermomet er s s h ou l d n otbe s epa r at ed. D i alth ermomet er s th atc an n otbe c al i br at ed s h ou l d n otbe u s ed. Q C 3. 3. 2 T he cal i b rat i o n of gl ass and el ectro n ic th e rm o m et ers s ho ul d be checked annual l y , and dialtherm o m et ers quart er ly, at the t em p era ture used , ag ainst a N ati o na l Insti tute o f S t an d ard s and T ec hn o l o gy (N IS T )- trac ea b l e re f er enc e th ermomet eroron e th atme et s th e r equ i r eme n t s of N B S M on og r aphSP 250- 23. T h e c al i br at i on f ac t oran d dat e of cal i b rat i o n s ho ul d be i nd i cat ed on the therm o m et er. I n ad d i t i o n, the l ab o rat o ry shoul d reco rd i n a Q C record bo o k the f o l l o wi ng i nf

o rma t i o n:



### C s er i al nu mb erof

l aborat orythermo met er 

### C s eri al num b er of

N IST
- traceab l e therm o m et er ( o r other r ef erence therm o m et er)


### C t emp er atur e of

l aborat orythermo met er 

### C t em p erature of

N IST
- traceab l e therm o m et er ( o r other r ef erence therm o m et er)


### C c orr ec t i on ( orc al i br at i on ) f ac t or 

### C da t e o f check


### C a n a lyst’ s initia ls Q C 3 .3 .3 If

a thermo met erd i f

f er s b ym ore than1°C f

r om the r ef

er enc e thermo met er , i ts houl d be di s car ded. Ref er enc e th ermomet er s s h ou l d be r ec al i br at ed atl ea s te v eryf i v e y ea r s. Ref

er en ce th ermomet erc al i br at i on doc u men t at i on s h ou l d be ma i n t ai n ed. Q C 3. 3 . 4 C o nti nu o us rec o rd i ng d ev i ce s tha tare used t o m o nit o r i nc ub ato r t em p era ture sho uld b e re ca l i b rated at l ea st an n u al l y . A r ef

er en ce th ermomet erth atme et s th e s pec i f i ca t i on s de s cr i bed i n pa r ag r aph3
. 3
. 2 sh ou l d be u s ed f or cal i b rat i o n. 3 .4 In c u b a to r Un it 3. 4. 1 I n cubat orun i t s m u s tha v e a n i n t ern alt empe r atu r e mon i t or i n g de v i ce a n d ma i n t ai n th e t empe r atu r e s pec i f i ed by the method us ed, us ual l y 35 ° ±0
. 5°C a nd 44
. 5°±0. 2°C. For non- po rt abl e i ncubat ors , thermo met er s s houl d be pl ace d on the t o p a nd bott om s h el v es of th e u s e ar eaa n d i mmer s ed i n l i qu i d a s di r ec t ed byth e manu f ac tu r er( ex ce ptf

or e le c tro n ic the rm o m e te rs ). If an a lum inu m b lo c kinc u b a to r is u se d , cu lt u r e d is h e s a n d tub e s sh o u ld fi t snu g ly. L a b o ra to rie s w h ic h u se th e e n z ym e s u b stra te te sts w ith a ir-typ e inc u b a to rs sh o u ld no te the p ro d u c t i n c u b a tio n d e ta ils i n di ca t ed i n pa r ag r aph5. 3. 1. 5. Q C 3. 4. 2 C al i brat i on- corr ect ed t emp er atur e s houl d be re corded f or e ach thermo met er bei ng us ed atl eas ttwi ce per day du r i n g e ac h dayth e i n cubat ori s i n us e, wi th re adi n g s s epar at ed byatl ea s t4 h ou r s . D ocu men t at i on s h ou l d i n clu de the d ate a nd t i m e o f

rea d i ng , t em p era ture , an d t ec hn i cian ’ s i nit i als. 3 .4 .3 A n inc u b a tio n tem p e ra tur e o f 4 4 .5 °± 0 .2 °C c a n b e st b e m a inta ine d w ith a c irc u la ting w a te r b a th e q u ip p e d w ith a gab l e co ver. 3. 5 A utoclave 3. 5. 1 T he aut o cl ave shoul d have an i nt ernal he atso urce, a temp erature gauge w i th a sensor o n the exhaus t , a press ure gauge, and an op erat i o nal s af etyval ve. T he aut o cl ave shoul d m ai nt ai n a s t eri l i zat i o n t em p erature duri ng the st eri l i zi ng c yc le a n d c o m p le te a n e n tire c yc le (i.e ., t im e b e tw e e n sta rting a u to c la v e a n d re m o v ing it e m s fro m a u to c la v e ) w ithin 4 5 m in u te s w h e n a 1 2 -1 5 m inu te ste riliza tio n p e rio d is use d . T h e a u to c la v e sh o u ld d e p re ssu riz e slo w ly en o u g h to en sure tha tm ed i a w i l l no tb o i l o ve r an d b ub b l es w i l l no tf o rm i n inve rted tub es. 3. 5. 2 B e c a u se o f safe ty co n c e rn s a n d d iff ic u lti e s w ith o p e ra tio n a l co n tro l, p re ssu re c o o k e rs sh o u ld no t b e us e d
. Q C 3. 5. 3 T h e dat e, c on t en t s , s t er i l i za t i on t i me an d t empe r atu r e, t ot alt i me i n au t ocl av e, an d an al y s t ’ s i n i t i al s s h ou l d be reco rded each t i m e the aut o cl ave i s used. C o p i es of the servi ce co ntractor i nt ernal m ai nt enance p rotoco l and m ai nt enance reco rds s ho ul d be kep t
. M ai nt enance sho ul d be co nd uct ed atl eastannual l y . A record o f the mo strecent servi ce p erf

o rme d s ho ul d be on f

i l e, avai l ab l e f

o r i nspecti o n.V -4

Q C 3 .5 .4 A m axi m um - t em p erature- regi st eri ng therm o m et er, el ectronic t em p erature r ead o utde vi ce, or co nt i nuo us reco rding d evi ce shoul d be us ed d uri ng e ach aut o cl ave cycl e t o e nsur e thatthe p rop er t em p erature was reached , and the tem p e ra tur e re c o rd e d . O v e rc ro w d ing s h o u ld b e a v o id e d . S p o re strip s o r sp o re a m p u le s sh o u ld b e us e d m o n thly as bi o i nd i cat o rs t o co nf i rm ster ili z a tio n . ( Since chem i cal i nd i cat o rs wi l l res p o nd t o a wi d e range of t i m es and t em p eratures , i
. e. , a l o nger t i m e ata l o wer t em p erature, as wellas a short er t i m e ata hi gher t em p erature, a p o si t i ve res ul twi th the i nd i cat o r do es notnecess ari l ys ho w thats t eri l i zat i o n has o ccurred.) Q C 3. 5. 5 Au t om a t i c t i mi n g me c h a n i s ms s h ou l d be c h e c k e d qu a rt e r l y w i th a s t op w a t c h oroth e ra c c u r a t e t i me pi e c e ort i me s i g n al , a n d th e r es u l t s r ec or ded an d i n i t i al ed. 3 .5 .6 Au t ocl av e door s ea l s s h ou l d be c l ea n a n d f r ee of c ar ame l i ze d me di a. Al s o, a u t ocl av e dr ai n s cr ee n s s h ou l d be cl ea n ed f r equ en t l ya n d de br i s r emov ed. 3. 6 HotAirO ven 3. 6. 1 T he o ven shoul d m ai nt ai n a s t ab l e st eri l i zat i o n t em p erature of 17 0 °- 1 8 0 °C f

o r atleas ttwo ho urs . O vercrowd i ng sh o u ld b e a v o id e d . T h e o v e n the rm o m e te r sh o u ld b e g ra d u a te d in 1 0 °C inc re m e n ts o r l e ss, w ith the b u lb p la c e d in s and dur i ng us e. Q C 3 .6 .2 T h e d a te , co n te n ts, ster ili z a tio n ti m e a n d tem p e ra tur e , an d a n a lyst' s initia ls sh o u ld b e re c o rd e d
. Q C 3 .6 .3 Sp o re s trip s shoul d be us ed m o nthl yt o co nf i rm st eri l i zat i o n. A m p ul es ar e no trecom m end ed f o r ho tai r ovens b e c a u se the y m a y e x p lo d e o r m e lt. 3. 7 Colony Coun ter A d ark f i eld co l o ny co un t er sho uld b e u sed t o co un tH etero tro p hic P l ate C o un tco l o nies. 3. 8 Cond uctivit

y Meter 3 .8 .1 M et er s s houl d be s ui t able fo r c he ck i ng l aborat oryr eag ent - gr ade wat eran d re adabl e i n uni t s of ei ther m ic ro m h o s/c m o r m ic ro sie m e n s/c m . Q C 3 .8 .2 C a lib ra te the m e te r at le a st m o n thly, fo llo w ing the m a n u fac tur e r’s re c o m m e n d a tio n s a n d us ing a n a p p ro p ria te c e rtifi e d a n d tra c e a b le lo w -lev e l stan d a rd . If the m e te r c a n n o t b e c a lib ra te d w ith a c o m m e rc ia l stan d a rd , th e c e ll co nst a n t should b e d et erm i ned atm o nthl yi nt erval s, us i ng a m etho d i n Sec t i o n 25 1 0 , "C o nd uct i vi ty , " i n S ta n da rd Methods
. 3 .8 .3 I f an i n- l i ne uni tcanno tbe cal i b rat ed , i ts ho ul d not b e used t o check reagent

- grade wat er. 3. 9 Ref r i gerator 3. 9. 1 R efri ge rato rs sho uld m aintain a tem p era ture o f 1 °- 5 °C
. C ali b rated the rm o m eters sho uld b e g rad ua t ed i n a tl ea st 1° C i n cr eme n t s a n d th e th ermomet erbu l b i mmer s ed i n l i qu i d. Q C 3. 9. 2 O n day s th e r ef r i g er at or i s i n u s e, an d th e l abor at ory i s s t af f ed, th e ca l i br at ed- corr ec t ed t empe r atu r e s h ou l d be re c o rd e d a t l e a st o n c e p e r d a y. 3. 10 I noc ulating Equi

pm en t St er i l e met alordi s pos abl e pl as t i c l oops , w ood a ppl i ca t ors t i ck s , s t er i l e s wabs , ors t er i l e pl as t i c di s pos abl e pi pett i ps s h ou l d be u s ed. I f

wood appl i ca t ors t i ck s ar e u s ed, th eys h ou l d be s t er i l i ze d by dryhe at
. T h e met ali n ocu l at i n g l oops an d/ or n ee dl es s h ou l d be made of n i ck el al l oy or pl at i n u m. ( W h enpe r f ormi n g a n ox i das e t es t , do n otu s e n i ck ela l l oy l o o p s b ecause they m ay i nt erf ere wi th the t est ). 3. 11 Mem br ane Filtr ation Equi pm en t ( i f M F pr oce dur e is us ed) 3. 11. 1 M F un i t s m ust b e st ainless st ee l , glass, p o rce l ain, o r au t o clav ab l e p l asti c, no t scra t ch ed o r co rro d ed , an d m ust no tl eak.V -5

Q C 3 .1 1 .2 If gra d u a tio n m a rk s o n c le a r gla ss o r p la stic fu n n e ls a re us e d to m e a su re sa m p le vo lum e , th e ir a c c u ra c y sh o u ld be ch ec k ed wi th a C l as s B gr adu at ed c y l i n derorbe tt er( oroth erCl as s B g l as s wa r e) , a n d a r ec or d of th i s c al i br at i on ch ec kr et ai n ed. 3. 11. 3 A 1 0 X to 1 5 X s te re o m ic ro sc o p e w ith a flu o re sc e n t ligh t so u rc e m u st b e us e d to c o u n t th e tar g e t co lo n ie s (e .g., sheen coloni es on M
- E nd o or E nd o LE S m ed i a). 3. 11. 4 M emb r ane f

i l t er s mus tbe approved by the manuf actur er f or t ot al col i f

orm wat er ana l y s i s . Ap provali s bas ed on da t a f r om t es t s f

ort ox i ci ty , r ec ov ery , r et en t i on , a n d a bs en ce of g r owth

- pr omot i n g s u bs t an ce s . Fi l t er s mu s tbe gr i dm arked, 47 mm di amet er , and 0. 45 μm po r e s i ze, or al t ernat e po r e s i zes i f the manuf

actur erprovi des per f

ormanc e d at a eq ual t o or b ett er than the 0
. 4 5 μ m p o re si ze. T hey shoul d a l so b e whi t e, a nd o f c el l uo se es t er. M em b rane f i l t ers and pads mus tbe pur cha s ed pres t er i l i zed or aut ocl ave d f or 10 mi nu t es at121 ° C bef ore us e. Q C 3 .1 1 .5 T he l o tnum b er f o r mem b rane f

i l t ers and the d at e recei ved shoul d b e reco rded
. E nsur e thatm em b rane f i l t ers a re no t b rittl e o r d isto rte d , an d tha t m a n u fac tur e r’s sp e c ifi c a tio n /c e rtifi c a tio n sh e e t i s a v a ilab le
. 3. 11. 6 Fo rce p s used sho uld b e b l un tan d sm o o th-t i p p ed witho ut co rrug ati o ns o n the inne r si d es o f t i p s. 3 .12 Cu ltu re D ish es (lo o se o r ti g h t lids) 3 .1 2 .1 P res t eri l i zed p l ast i c o r st eri l i zab l e gl ass c ul ture d i shes sho ul d b e used. T o m ai nt ai n s t eri l i tyof gl ass cul ture d i shes , us e st ai nl ess s t eel or al um i num cani st ers , or a wrap of heavy al um i num f o i l or char- res i st antpa p er. 3. 12. 2 Lo o se- l i d p etri d i shes s ho ul d be i ncub at ed i n a t i ght

- f i tt i ng co nt ai ner, e. g. , plas t i c veget ab l e cri sper co nt ai ni ng a m o isten e d p a p e r to w e l t o p re v e n t d e h yd ra tio n o f m e m b ra n e fi ltera n d m e d ium
. 3. 12. 3 O p en ed p ac ks o f d i sp o sab l e c ulture d i she s sho uld b e re sea l ed b etwe en use p erio d s. 3 .1 2 .4 F o r m e m b ra n e fi lterm e tho d s, c u lture d ish e s sh o u ld b e o f an a p p ro p ria te siz e to a llo w fo r th e tra n sfe r o f a sin g le mem bran e perpl at e. 3 .1 3 P ip e ts 3 . 1 3 . 1 To s t e r i l i z e a n d m a i n t a i n s t e r i l i ty o f g l a s s p i p e t s , s t a i n l e s s s t e e l o r a l u m i n u m c a n i s t e r s s h o u l d b e u s e d, or ind ivid u a l p ip e ts sh o u ld b e w ra p p e d in c h a r-re sista n t p a p e r o r a lum inu m fo il. 3. 13. 2 Pi pet s s h ou l d h av e l eg i bl e mark i n g s a n d s h ou l d n otbe c h i pped ore t ch ed. 3. 13. 3 O p en ed p ac ks o f d i sp o sab l e steril e p i p ets sho uld b e re sea l ed b etwe en use p erio d s. 3 .1 3 .4 P ip e ts d e live rin g vo lum e s o f 1 0 m L o r le ss m u st b e a c c u ra te to w ithin a 2 .5 % t o le ra n c e . Q C 3 .1 3 .5 C al i b rat ed m i crop i p ett ers m ay b e used i f t i p s are s t eri l e. M i crop i p ett ers s ho ul d be cal i b rat ed an n u a lly and a d j u s t e d o rr e p l a c e d i f

th e p r e c i s i o n o ra c c u r a c yi s g r e a t e rth a n 2
. 5 %. 3. 14 G lass w are andP lasticwa r e 3 .1 4 .1 G l ass ware shoul d be bo rosi l i cat e gl ass o r o ther co rr o si o n- res i st ant gl ass and f ree o f chi p s and cracks. M arki ngs o n gra d ua t ed cyl i nd ers a nd p i p ets m ust be l eg i b l e. P l asti c it em s sho uld b e c l ea r an d no n-t o xic to m i cro o rga nism s. Q C 3 .1 4 .2 G r aduat ed cy l i nder s f or m eas ur emento f

sampl e vol umes m us tbe ac cur at e t o wi thi n a2. 5% t ol er anc e. I n l i eu of gr aduat ed cy l i nder s , prec al i brat ed cont ai ner s that hav e cl ear l y mark ed vol umes acc ur at e t o w i thi n a 2. 5% t ol er anc e maybe u s ed. 3 .1 4 .3 C ul ture tub es and co nt ai ners co nt ai ni ng f erm ent at i o n m ed i um s ho ul d be of suf f i ci ents i ze t o co nt ai n m ed i um p lus sa m p le w itho u t b e ing m o re tha n thr e e q u a rte rs full.V -6 3. 14. 4 T ub e cl o sur es shoul d b e st ai nl ess st eel , p l ast i c, al um i num , o r scr ew caps wi th no n- t o xi c l i ners . C o tt o n plugs a n d fo a m p lug s sh o u ld no t b e us e d
. 3 .15 S a mp le Co n ta in ers 3 .1 5 .1 Sa m p l e c o ntainers sho uld b e w i d e-m o uth p l asti c o r no n-co rro si ve glass b o tt l es w i th no n-l ea king g ro un d glass st o p p ers or cap s wi th non-t o xi c l i ners thats ho ul d wi thst and repea t ed s t eri l i zat i o n, or st eri l e p l ast i c b ags contai ni ng s odi u m th i os u l f

at e. O th era ppr opr i at e s ampl e c on t ai n er s ma ybe u s ed. T h e capac i tyof s ampl e con t ai n er s s h ou l d be a t l e a st 1 2 0 m L (4 o z .) to a llo w a t l e a st a 1 -inc h he a d sp a c e . 3 .1 5 .2 G l ass s t o p p ers m ustbe co vered wi th al um i num f o i l or char- res i st antpa p er f o r s t eri l i zat i o n. 3 .1 5 .3 G l as s an d pl as t i c bott l es that hav e not been pres t er i l i zed s houl d be s t er i l i zed byau t ocl avi ng . G l as s bot t l es may al so b e st eri l i zed by dry heat
. E m p tyco nt ai ners s ho ul d be m o i st ened wi th s everal dro p s o f wat er before aut o cl avi ng t o prev entan"ai rl ock" s t er i l i zat i on f ai l ur e.

2 2 3 3 .1 5 .4 If ch lo rin a te d w a te r is to b e a n a lyz e d , sufficie n t so d ium thio su lfate (N a S O ) m u st b e a d d e d to the sa m p le b o ttl e b e fo re ste riliza tio n to ne u tra lize a n y re sid u a l ch lo rin e in the w a te r sa m p le
. D e c h lo rin a tio n is ad d re sse d in Se ct i on 9060A of

Standard Methods
. 3. 16 Ultr aviolet

lamp ( i f u s ed) 3 .1 6 .1 A g e rm ic id a l u n it (2 5 4 -nm ) sho u ld b e d isc o n n e c te d m o n thly a n d the lam p c le a n e d b y w ip ing w ith a so ft c lo th m o i st ened wi th ethano l
. A l o ngwave uni t(36 5
- 3 6 6
- nm ), us ed f o r f l uo rom etri c t est s, s ho ul d al so b e kep tcl ean. Q C 3 .1 6 .2 A g ermi ci dalu n i ts h ou l d be t es t ed qu art er l ywi th a U V l i g h tm et eror ag ars pr ea d pl at e. T h e l amp s h ou l d be rep l ac ed i f

i tem i t s l ess than 7 0 % o f i t s i nit i al o utp ut o r i f

an ag ar sp rea d p l ate c o ntaini ng 2 0 0 t o 2 5 0 m i cro o rga nism s, ex p o sed t o the U V l i gh tf o r tw o m i nu t es, d o es no t sho w a co un t red uc t i o n of 9 9 %
. O the r m etho d s m ay b e u sed t o t est a lam p if d a ta d e m o n stra te tha t th e y a re a s e ff e c tive a s the tw o s u g g e ste d m e tho d s. (U V p ro te c tive e ye w e a r sh o u ld b e us e d w h e n c h e c k ing the o p e ra tio n o f a 2 5 4 -nm lam p .) 3. 17 Spectr op hotom eterorcolorim eter( i f us ed) 3. 17. 1 W a v e le n g ths sh o u ld b e in the visib le ra n g e
— S p e c tro n ic 2 0 (T h e rm o S p e c tro n ic ), o r e q u iva le n t, w ith c e ll ho l d er fo r 1⁄2 " d i am eter c uv ett es (M o d el # 4 0 1 5 ) o r 1 3 m m × 1 0 0 m m cu ve tt es. Q C 3. 17. 2 A ca l i br at i on s t an dar d a n d a me th od- s pec i f i c bl an k s h ou l d be an al y ze d e v eryda yth e i n s tru men ti s u s ed, pr i or t o s ampl e an al y s i s . T h e ca l i br at i on s t an dar d s h ou l d g i v e a r ea di n g i n th e des i r ed abs orbance ran g e an d s h ou l d be ob t ai ned f

r om anout s i de s our ce. 

## 4. Ge neral Laboratory P ractices Al th ou g h s af

etyc r i t er i a ar e n otc ov er ed i n th e l abor at oryc ert i f i ca t i on pr og r am, l abor at orype r s on n el s h ou l d be a wa r e of gen er al an d cus t om arysafet y p ract i ces f or l aborat ori es . Each l aborat oryi s en cour age d t o hav e as af etypl an ava i l abl e. A l s o, ea ch l aborat orys houl d kee p a copy , an d f ol l ow the pers onal p r ot ect i on gui del i nes , of an ymat er i al s af

etydat a s hee t a c c o m p a n ying the re c e ip t o f a to x ic m a te ria l. 4. 1 Steriliz ation P r ocedures 4 .1 .1 A ut o cl avi ng t i m es at12 1 °C are l i st ed be l o w. E xcep tf o r mem b rane f

i l t ers and pa d s and carbo hy d rat e- co nt ai ni ng m ed i a, i nd i cat ed t i m es are m i ni m um t i m es and m ay necessi t at e ad jus tm entde p end i ng upo n volumes, co nt ai ners , and l oads
. Carboh y dr at e- bas ed medi a s h ou l d n otbe ov er - s t er i l i ze d.V -7 It em T i m e (min) M embr an e f

i l t er s & pa ds 10 Carboh y dr at e con t ai n i n g me di a 12- 15 1 Cont ami n at ed t es tma t er i al s 30 2 M embr an e f

i l t era s s embl i es 15 Sa mpl e col l ec t i on bott l es 15 I n di v i du alg l as s wa r e 15 D i l u t i on w at erbl an k 15 Ri n s e wa t er( 0. 5 - 1 L) 15- 30 2

ex ceptwh en otherwi s e s peci f i ed by the manuf

actur er 1 t i me depends upon wat ervol ume percont ai neran d aut ocl ave l oad 2 4. 1. 2 A u to c la ved m em b rane f i l t ers and pa d s and al l m ed i a shoul d be remo ved i m m ed i at el yaf t er com p l et i o n of the s t er i l i zat i on cy cl e. 4. 1. 3 M embr an e f

i l t ere qu i pmen tmu s tbe a u t ocl av ed be f or e th e be g i n n i n g of

a f i l tr at i on s er i es . A f

i l tr at i on s er i es e n ds wh en30 mi n u t es orl on g ere l aps es a f t era s ampl e i s f

i l t er ed. 4. 1. 4 U ltra v io le t l igh t (2 5 4 nm ) m a y b e us e d to sa n iti z e e q u ip m e n t (a fter initial au to c la v ing fo r ste riliza tio n ), if a ll supp l i es ar e p res t eri l i zed
. U l travi o l etl i ghtm ay b e used to r e d u ce b act eri al carry - o ver between s am p l es duri ng a fil tra tio n se rie s. 4 .2 S a mp le Co n ta in ers QC A tl eas to ne sa m p l e cont ai ners houl d be s el ect ed atr andom f r om ea ch bat ch of

s t er i l e s amp l e bott l es or other c o n ta ine rs (o r lo t o f co m m e rc ia lly a v a ilab le sa m p le c o n ta ine rs ), a n d th e ste rility c o n firm e d b y a d d ing a p p ro x im a te ly 25 mL of

a s t er i l e n on - s el ect i ve b roth (e
. g . , trypt i c s oy , trypt i ca s e s oy , ortrypt on e br oth )
. T h e br oth s h ou l d be inc u b a te d a t 3 5 ° ± 0 .5 °C , an d c h e c k e d a fter2 4 a n d 4 8 ho u rs fo r g ro w th. R e c o rd re su lts. R e ste rilize e n tire b a tc h if g r owth i s de t ec t ed.
4. 3 R eagent- G r ade W ater 4. 3. 1 O nly sati sf

ac t o ril y t ested rea ge nt w ater f ro m st i l l s o r d eio niza t i o n un i t s m ay b e u sed t o p rep are m ed i a, rea ge nts, an d d i l uti o n/ri nse wa t er fo r p erfo rm i ng m i cro b i al ana l yses. Q C 4. 3. 2 T he qual i tyof the r eag entwat ers houl d be t es t ed an d s houl d m eetthe f ol l owi ng cr i t er i a: Parame t er Li mi t s Fre que nc y C onduct i vi ty >0
. 5 m egohms r es i s t anc e or<2 mi cr om hos / cm ( mi cr os i eme n s / cm) a t25 C ° M o n thly 4

P b , C d , C r, C u , N i, Z n N o t gr e a te r th a n 0 .0 5 m g /L p e r c o n ta m ina n t. C o llec tive ly, n o gr e a te r th a n 0 .1 m g /L A n n u a lly
T o ta l C h lo rin e R e sid u a l < 0 .1 m g /L M o n thly 1 H e te ro tro p h ic P la te C o u n t < 5 0 0 C F U /m L M o n thly 2 5Parame t er Li mi t s Fre que nc y

V -8 B a c te rio lo g ic a l Q u a lity o f R eag entW at er 3 R a tio o f gro w th r a te 0 .8 to 3 .0 A n n u a lly D PD M eth od s h ou l d be u s ed. N otr equ i r ed i f s ou r ce w at eri s n otc h l or i n at ed. 1 P ourP l at e M ethod. See Standard Methods 9 2 1 5 B. 2See Standard Methods (18 or 1 9 ed s. ), Sec t i o n 90 2 0 B , unde r Lab oratory Sup pl i es . T his ba cterio l o gica l q ua l i ty t est 3 th th i s no tneed ed f

o r T yp e II wat er or be tt er, as d ef i ned i n Standard Methods (1 8 a nd 1 9 e d s) , S ect i o n 10 8 0 C , or M ed i um th th q ual i tywat er or be tt er, as d ef i ned i n Standard Methods ( 20 e d. ) , Se ct i on 1080C. I f T ype I I orM ediu m qu al i tyw at eror th b e tteris no t av a ilab le , an d a gla ss still is us e d fo r re a g e n t wa te r, a sili c o n tes t th a t m e e ts the sp e c ifi c a tio n s o f S ta n da rd Me th o ds, Se ct i on 1080C ( 20 e d. ) s h ou l d a l s o be a cc ompl i s h ed. th M o nthl y , i f

m et er i s i n- l i ne o r has a res i st i vi tyi nd i cat o r l i ght ; otherwi se, wi th each new b at ch o f reagentwat er. 4C FU m eans colony- f

o rming uni t s (s am e as col o ni es, but i s a m o re preci se t erm ). 5 4. 4 D ilution/

R ins e W ater 4. 4. 1 Stock b uf

f er s o l ut i o n or p ep t o ne watersho ul d b e p repa red, as s p eci f i ed i n Standard Methods , S e c t i on90 50 C. 4. 4. 2 Stock b uf

f ers s ho ul d be aut o cl aved o r f i l t er- st eri l i zed , and co nt ai ners shoul d b e l ab el ed and d at ed
. St o ck b uf f ers sh o u ld b e re frige ra te d . S to re d sto c kb u ff e rs sh o u ld b e fre e fro m turb id ity. Q C 4. 4. 3 E ach b at ch (or l o t , i f co m m erci al l yprep ared ) of dilut i o n/ri nse wat er s ho ul d be checked f o r st eri l i tyby add i ng 5 0 m L o f

wat er t o 50 m L o f

a do ub l e strength non-s el ect i ve b roth (e. g. , tryp t i c soy, tryp t i case s o yor tryp t o se broth)
. In c u b a te a t 3 5 °± 0 .5 °C , an d c h e c kfo r g ro w th a fter2 4 a n d 4 8 ho u rs . R e c o rd re su lts. D is c a r d b a tc h i f g ro w th is det ec t ed. 4. 5 G las s war e Was hing 4. 5. 1 D i s t i l l ed or dei oni zed wat ers houl d be us ed f or f i nalr i ns e. 4 .5 .2 L a b o ra to ry g la ssw a re sh o u ld b e w a sh e d w ith a d e te rg e n t d e sig n e d fo r la b o ra to ry u se . Q C 4. 5. 3 A g la ssw a re inh ib ito ry re sid u e te st ( Standard Methods , Sec t i o n 90 2 0 B , unde r Lab oratory Sup pl i es ) s h ou l d be per f ormed bef

or e th e i n i t i alu s e of

a w as h i n g compou n d an d wh en ev er a di f f er en tf ormu l at i on of

w as h i n g c ompou n d, o r w a sh ing p ro c e d u re , i s u se d . R e c o rd re su lts. T h is tes t will en su re tha t gla ssw a re is f re e o f t o x ic re sid u e . Q C 4 .5 .4 E a c h b a tc h o f d ry g la ssw a re us e d fo r m ic r o b ia l an a lysis sh o u ld b e c h e c k e d fo r p H re a c tio n , es p e c ia lly if g l as s wa r e i s s oak ed i n a l k al i ora ci d ( Standard Methods , Se ct i on 9020B , unde r Lab oratory Sup pl i es )
. U s e 0. 04% b ro m thym o l b lue (o r e q u iva le n t p H ind ic a to r) a n d o b se rv e c o lo r re a c tio n . C le a n g la ssw a re w itho u t an a lka li o r a c id res i d ual s ho ul d have a neutral c o l o r react i o n (b l ue- green for b rom thy m o l b l ue). R eco rd res ul t s. T hi s t estwi l l ensur e th a tg l a s s w a r e i s a ta n e u tr a l p H. 

## 5. Analytical Methodol ogy 5. 1 G en eral 5 .1 .1 Fo r com p l i ance sam p l es, l ab o rat o ri es mustus e o nl ythe analyt i cal m etho d o l o gysp eci f i ed i n the T o t al C o l i f

o rm R u le (4 0 C F R 1 4 1 .2 1 (f)) , th e S u rfa c e W a te r T re a tm e n t Ru le (S W T R ) (4 0 C F R 1 4 1 .7 4 (a )) , an d the G ro u n d w a te r R u le ( TBD)
. F o rc o n v e n i e n c e , th e s e r e g u l a t i o n s a r e r e p r o d u c e d i n A p p e n d i x G. 5 .1 .2 A l a bo r a t ory mu s t be c e r t i f i e d f ora l l a n a l y t i c a l me th od s th a ti tu s e s f orc om pl i a n c e purpo s e s . Ata mi n i mu m, the l ab o rat o rym ustbe c ert i f i ed for o ne t o t al c o l i f

o rm m etho d and one f

ecal co l i f

o rm o r E . co lim etho d
. A l ab o rat o ry shoul d a l s o be ce r t i f

i ed f or a s ec on d t ot al col i f

orm meth od i f on e meth od ca n n ot be u s ed f or s ome dr i n k i n g wat ers (e .g., w h e re the w a te r u su a lly p ro d u c e s c o n flue n t g ro w th o n a p la te ). In a d d iti o n , fo r l a b o ra to rie s tha t m a y e n u m e ra te het er otr op hi c bact er i a ( as meas ur ed by the H et er otr op hi c P l at e C ount , H P C ) f or com pl i anc e wi th the Sur f ace W at er T r ea tmen tRul e, th e l ab or at orymu s tbe c ert i f i ed ei th erf

orth e PourPl at e M eth od orth e Si mP l at e meth od f or het er otr op hi c bact er i a.V -9 5 .1 .3 W at er s am p l es s ho ul d be s haken vi go rousl yatl east25 t i m es bef o re anal y zi ng. Q C 5 .1 .4 If d ilutio n b u ff e r is u se d , ch e c kthe a c c u r a c y o f th e b u ffe r v o lum e in o n e d ilutio n b o ttl e in e a c h b a tc h o r lo t. Fora 90- mL or99- mL v ol u me, th e t ol er an ce s h ou l d be ±2 mL. 5 .1 .5 Sa mpl e v ol u me an al y ze d f ort ot alc ol i f

orms i n dr i n k i n g w at ermu s tbe 100 mL. T o as s ur e a ccu r acy an d c on s i s t en cyw i th i nme th ods a n d be twe enme th ods i ti s i m p o rtant tha tthe l ab o rato ry ob t ain p rec i se m e a su re m e n t o f th e vo lum e o f sam p le to b e a n a lyz e d . T o e n su re tha t th e re q u ire d vo lum e o f 1 0 0 m L is an al y ze d, n o ma tt erw h i chof th e appr ov ed meth ods th e l abor at oryw i l l be e mpl oy i n g f ora n al y s i s , g ood la b o ra to ry p ra c tice d ic ta te s tha t a ste rile , ca lib ra te d m e a su rin g ve sse l b e us e d fo r m e a su re m e n t o f th e sa m p le vol ume. I ti s i nappropri at e f or aport i on of

the s amp l e t o be po ur ed t o was t e i n or der t o m eetthe r equi r ed sa m p le vo lum e , as this p ra c tice c o u ld e a sily re su lt in la b o ra to ry e rr o r w h ic h c o u ld the n re q u ire the sa m p le to b e inv a lid a te d
. 5 .1 .6 M edi a ( or def

i ned s ubs tr at e) 5. 1. 6. 1 T h e us e o f d e h yd ra te d o r p re p a re d m e d ia m a n u fac tur e d c o m m e rc ia lly is stro n g ly re c o m m e n d e d d u e to c o n c e rn a b o u t q u a lity c o n tro l. D e h yd ra te d m e d ia sh o u ld b e sto re d in a c o o l, dryl ocat i on, an d di s car ded b ymanu f actur er ' s ex pi r at i on dat e. Caked ordi s col ored dehy drat ed medi a s h ou l d be di s ca r ded. Q C 5 .1 .6 .2 Fo r med i a p repa red i n the l ab o rat o ry , the d at e o f prep arat i o n, typ e o f

m ed i um , l o tnum b er, s t er i l i za t i on t i me an d t empe r atu r e, f i n alpH ( af

t ers t er i l i za t i on ) , a n d th e t ec h n i ci an ' s i n i t i al s s h ou l d be r ec or ded. Q C 5 .1 .6 .3 F o r m e d ia p re p a re d c o m m e rc ia lly, the d a te re c e ive d , typ e o f m e d ium , l o t nu m b e r, a n d (if

id e n tif ie d b y the m a n u fac tur e r o r m e tho d ) p H ve rifica tio n fo r e a c h lo t sho u ld b e re c o rd e d . M e d ia s houl d be di s car ded b ymanu f actur er ' s ex pi r at i on dat e. Q C 5 .1 .6 .4 E a c h ne w lo t o f d e h yd ra te d o r p re p a re d c o m m e rc ia l m e d ium a n d e a c h b a tc h o f l a b o ra to ryp repa red m ed i um s ho ul d be checked be f o re us e f o r s t eri l i tyand wi th po si t i ve and negat i ve cul ture con tr ol s . T h os e l abor at or i es u s i n g c ommer ci al l ypr epa r ed medi a wi th ma n u f ac tu r ers h el f

- l i v es of great er than 9 0 da y s shoul d run p o si t i ve and negat i ve co ntrols each q uart er, i n ad d i t i o n t o runni ng these contr o l s and s t eri l i tychecks on each new l o tof m ed i a. Lab o rat o ri es ar e enco uraged t o pe rf o rm po s i t i ve and n ega t i ve contr ol t es t s on a mo r e f r equen tbas i s . Co ntr ol org ani s ms ( t ot al col i f orms , f eca l co l i f o rms, and / o r E . co li , a s a ppr opr i at e) c anbe s t ockc u l tu r es ( per i odi ca l l yc h ec k ed f orpu r i ty ) or co m m erci al l yavai l ab l e d i sks i m p regnat ed wi th the o rgani sm. R esul t s shoul d be record ed
. T he f o l l o wi ng T ab l e i d ent i f

i es a f ew p o si t i ve and negat i ve cul ture contr o l s thatl ab o rat o ri es mi ght cons i der, al thoug h othercu l tur e c ontr ol s ar e a l s o acce pt abl e.V
- 10 C o ntro lC ult ur es f

o r M i cro biol o g i ca lT est s G r ou p Pos i t i v e Cul tu r e Contr ol N eg at i v e Cul tu r e Contr ol 9 9 T ot al c ol i f orms Esc h e ric h ia c o li

E nt er ob acteraer og enes Staphylococ cus aur eus 1 P ro te u s v u lg a ris 2 P seu d o m o n a s a eru g i n o sa 1 Fecal c ol i f orms Esc h e ric h ia c o li

Kl ebs i ella pne um oni ae (th e rm o to le ra n t) E nt er ob acteraer og enes 3

E . co li Esc h e ric h ia c o li(M U G
- p o si t i ve strai n) E nt er ob acteraer og enes Kl ebs i ella pne um oni ae (th e rm o to le ra n t) 4 E nt er ococci E n te ro c o c c u s fa e c a lis

5 Ent erococ cus f

aec i um Staphylococ cus aur eus 6 E . co li 7 Serr at i a m ar ce s ens 8 S

. a u reu s, P

. a eru g i n o sa - notl act ose f erment er 1 P . vu lg a ris - not l act o se f erm ent er; us es hy d roly zed l act o se, i nd i cat i ng “ o verco o ked ” m ed i um 2 E

. aer og enes - f

erm ent s l act o se, but i s no ttyp i cal l ytherm o t o l erant 3 K. pne um oni ae - f

e rm e n t s l a c t o s e , b u td o e s n o th y d r o l y z e M UG 4D o no tuse c l o sely related strains f ro m ge nu s Str eptococ cus a s a pos i t i v e con tr ol 5 S. aur eus - s ensi t i ve t o nal i d i xi c aci d i n m ed i um 6 E . co li- s ensi t i ve t o s o d i um azi d e i n m ed i um 7 S. m ar ce s ce ns - wi l l not hy d roly ze f l uo rogenic com p o und i n m ed i um 8E xam p l es of

ap p rop ri at e A T C C s trai ns i ncl ud e the f o l l o wi ng: 9 E n tero co cc u s f a ec a l is AT CC 11700 Ent erococ cus f aec i um AT CC 6057 E n tero b a cter a ero g en es AT CC 13048 Esc h e ric h ia c o li AT CC 8739 or 25922 Kl ebs i ella pne um oni ae ( th ermot ol er an t ) AT CC 13883 P ro teu s vu l g a ris AT CC 13315 Ps eudomonas ae r ugi nos a AT CC 27853 Serr at i a m ar ce s ens AT CC 14756 S ta p hylo co cc u s a u reu s AT CC 6538 5 .1 .6 .5 I f

prep ared m ed i um i s st o red aft er s t eri l i zat i o n, i ts ho ul d be m ai nt ai ned i n the d ark, avo i d i ng moi s tu r e l os s , pe rth e f ol l owi n g T abl e. Pr epa r ed pl at es ma ybe s t or ed i n s ea l ed pl as t i c bag s or con t ai n er s . Fore i th erbr oth ora g arme di a, e ac h ba g orc on t ai n ers h ou l d i n clu de th e dat e pr epa r ed or ane x pi r at i on da t e. I f

th e medi u m i s s t or ed i n a r ef r i g er at or , i ts h ou l d be w armed t o r oom te m p e ra tur e b e fo re us e ; tu b e s o r p la te s tha t sho w gr o w th a n d /o r b u b b le s sh o u ld b e d isc a rd e d . L iq u id med i a s houl d be di s car ded i f ev aporat i on ex cee ds 10 % of the ori gi nalvol ume. M a x im u m H o ld in g T im e s a n d T e m p e r a tu r e s fo r P r e p a r e d M e d ia Con t a i n e r M a x s t or a g e t e mp . M a x . s t or a g e t i me P o ured aga r pl at es 1
- 5 °C 2 weeks B roth i n tub es, bo tt l es, or f l asks w i th lo o se-f

i tt i ng clo sure s 1
- 3 0 °C 2 weeks
B r oth i n t i gh t l ycl ose d s cr ew- cap tub es, bo tt l es, or f

l asks 1
- 3 0 °C 3 m o nthsV
- 11

Q C 5. 1. 7 Labo r a t or i e s a r e e n c ou r a g e d t o p e r f orm p a r a l l e l t e s t i n g be tw e e n a ne w l yap pr ov e d t e s tan d a n oth e rEPAa p p ro v e d p ro c e d u re fo r e n u m e ra ting to ta l co li fo r m s f o r a t l e a st se v e ra l m o n ths a n d /o r o v e r se v e ra l se a so n s to a sse ss th e e ff

e c tive n e ss o f th e ne w tes t f o r th e w id e va rie ty o f wa te r typ e s su b m it te d f o r a n a lysis. D u rin g this t es t i n g , s pi k i n g th e s ampl es occ as i on al l y wi th s ew ag e or a pu r e cu l tu r e maybe n ec es s aryt o e n s u r e th ats ome of the t es t s ar e posi t i ve. 5 .1 .8 A l i stof ap p roved anal y t ic a l m e tho d s (or pro p o sed m etho d s, where noted), ap p l i cab l e regul at i o ns, and s ect i on i dent i f

i er s f or e ach method i s provi ded i n the T abl e bel ow.
A p p r o v e d M e th o d s P a r t M e d ia M et hod C i t at i on 1 T CR 2

(D etect) SW TR 2

(C o un t) G WR 2

(D etect) Tot alCol i for ms F e rm e n ta tio n b ro th meth od 5
. 2
. 2 L TB 

### Y B G L B B ro th S M 9 2 2 1 B ,C X X
5 .2 .3 P
- A B roth 

### Y B G LB B ro th SM 9221D X E n z ym e su b stra te meth od 5. 3. 2 Col i l ert ® , Col i l ert - 18® SM 9223 X X 5. 3. 2 Col i s u r e® SM 9223 X 5 .3 .2 R e a d yc u lt® o r F luo ro c u lt L M X® X 5. 3. 2 E * C ol i t e® X 5
. 3
. 2 C o l i t ag® X

M emb r ane f

i l t er meth od 5 .4 .2 M
- Endo orLES- Endo 

### Y L T B , B G L B B ro th S M 9 2 2 2 B ,C X X 5. 4. 2 M I M ediu m SM 9222 X X 5. 4. 2 m- Col i B l u e 24® X 5 .4 .2 C h ro m o c u lt® X 5
. 4
. 2 C o l i scan® X X Fe c alCol i for ms F e rm e n ta tio n b ro th meth od 5 .2 .4 LT B or P /A bro th 

### Y EC b ro th ( S M 9 2 2 1 B, D) SM 9221E X X 5. 2. 4 A- 1 br oth SM 9221E X M emb r ane f

i l t er meth od 5 .2 .4 M
- En do me di u m 

### Y EC b ro th ( S M 9 2 2 2 B) SM 9221E X X 5. 4. 2 mFC SM 9222D XA p p r o v e d M e th o d s P a r t M e d ia M et hod C i t at i on 1 T CR 2

(D etect) SW TR 2

(C o un t) G WR 2

(D etect)

V
- 12 E sc h e ric h ia c o li

E n z ym e su b stra te meth od 5. 3. 2 Col i l ert ® orCol i l ert - 18® SM 9223 X X 5. 3. 2 Col i s u r e® SM 9223 X X 5
. 3
. 2 E* C o l i t e ® X X 5 .3 .2 R e a d yc u lt® o r F luo ro c u lt L M X® X

5 .3 .3 LT B , P/A br oth , M
- Endo


### Y E C
- M UG ( S M 9 2 2 1 B, D; S M 9 2 2 2 B) SM 9221F X X
5
. 3
. 2 C o l i t ag® X

M emb r ane f

i l t er meth od 5. 4. 2 M I M ediu m SM 9222 X X 5. 4. 2 m- Col i B l u e24® X X 5 .4 .2 C h ro m o c u lt® X 5
. 4
. 2 C o l i scan® X 5 .4 .3 M
- Endo orLES Endo 

### Y NA
- M UG (SM 92 2 2 B ) 

### 6 SM 9222G X X E nte rococci 3 E n z ym e su b stra te meth od 5
. 3
. 4 E nt erolert AST M D 6503- 99 X F e rm e n ta tio n b ro th meth od 5 .2 .5 A zi de D extr ose 

### Y BE A /BHI SM 9230B X

M emb r ane f

i l t er meth od 5 .4 .4 m E 

### Y E IA m - E nt eroc o ccus SM 9230C X 5. 4. 4 mEI EP A 1600 X H e te r o tr o p h ic B a c te r ia Pourpl at e meth od 5. 5 Pl at e coun ta g ar SM 9215B X M u l t i pl e e n z y me su b stra te 5. 5 Si mP l at e® X P ourpl at e, s prea d p l at e, or m em b rane f i l t erme th ods 5. 5 R2A X 4A p p r o v e d M e th o d s P a r t M e d ia M et hod C i t at i on 1 T CR 2

(D etect) SW TR 2

(C o un t) G WR 2

(D etect)

V
- 13 M al e- Spe ci f

i c and S omat i c C ol i phage 3 Aga rpl at e meth od 5. 6. 2 T wo- St ep Enr i ch men t EP A 1601 X 5. 6. 3 Si n g l e Aga rLa y er EP A 1602 X SM = Stan da rd Metho ds f or the Exam i na t i on of

W at erandW

astewa ter , 18 , 19 or 2 0 ed i t i o n. 1 th th th T CR =T ot alCol i f

orm Rul e ( 40 CFR 141. 21 (f )) , S W T R = Surf

ac e W at erT r ea tmen tRul e ( 40 CFR 141. 74 ( a) )
. For 2 c o n v e n i e n c e , a n a l y t i c a l m e th o d s a p p r o v e d f o rth e TC R a n d S W TR a r e r e p r o d u c e d i n A p p e n d i x G. G W R = B as ed on pr opos ed G r ou n dwa t erRul e ( 65 FR 30194, da t ed 5/ 10/ 2 0 0 0 )
. U n t i l th e G W R i s pr omu l g at ed, 3 l ab o rato ries wil l no tb e c erti f i ed f o r en t ero co cc i o r co l i p ha ge m etho d s. F orpo s s i bl e u s e i f

s y s t e m o pe r a t e s u n de ra v a r i a n c e t o th e T CR. 4 5. 2 Fer men t at i on br ot h me t hods 5. 2. 1 G ene r al 5 .2 .1 .1 T he wa t er leve l o f the wa t er b ath sho uld b e a b o ve the up p er leve l o f the m ed i um i n the c ulture tub es. 5 .2 .1 .2 A D r i

- bath i n cubat ori s a cc ept abl e i f th e s pec i f

i ed t empe r atu r e r equ i r eme n t ca n be mai n t ai n ed i n a l l tu be l oca t i on s u s ed. 5 .2 .2 M ul t i pl e T ube Ferment at i on T ech ni que (f or det ect i ng t ot al co l i f

orms i n dr i nk i ng wat eran d enu mera t i ng t ot al co l i f o rms i n s o urce wat er) 5 .2 .2 .1 F o r d rin k ing w a te r sam p le s: V a rio u s te sting c o n figu ra tio n s c a n b e us e d (C F R 1 4 1 .2 1 (f)( 3 ), se e A p p e n d ix G ), a s lo n g a s a to ta l sa m p le vo lum e o f 1 0 0 m L is ex a m ine d fo r e a c h tes t. 5. 2. 2. 2 F o r so u rc e w a te r sam p le s: L a b o ra to rie s m u st u se a t le a st 3 se rie s o f 5 tub e s e a c h w ith a p p ro p ria te sa m p le d ilutio n s o f so u rc e w a te r (e .g., 0 .1 m L , 0 .0 1 m L , 0 .0 0 1 m L ). 5 .2 .2 .3 M e d ia 5 .2 .2 .3 .1 Laury l tryp t o se broth ( LT B ) ( al so kno wn as la u ry l s ul f at e b roth) m ustbe us ed i n the p re su m p tive te st a n d 2 % b rilli a n t g re e n la c to se b ile b ro th ( B G L B B ) in the c o n firm e d tes t. L a c to se b ro th ( LB ) maybe us ed i n l i eu of L T B (40 C FR 1 41
. 21 ( f ) ( 3)) i f the l aborat oryconduc t s atl eas t25 p ar al l el t es t s betweenthi s medi um an d LT B us i ng the wat er s normal l yt es t ed an d thi s comp ar i s on demo ns tr at es tha t th e false -p o sitive ra te a n d false -ne g a tive ra te fo r to ta l co lif o rm s, us ing L B , i s le ss th a n 1 0 % . T h is co m p ari son shoul d be do cum ent ed and the reco rds r et ai ned
. T he f i nal pH m ustbe 6.8± 0
. 2 f o r LT B , and 7 .2 ± 0 .2 fo r 2 % B G L B B
. 5 .2 .2 .3 .2 T he t es tmedi um conce ntr at i on mus tbe adju s t ed t o c om pens at e f or the s amp l e v ol ume s o that the re su lti n g m e d ium a ftersa m p le a d d iti o n i s s in g le s tr e n g th. O p tio n a lly, if a sing le 1 0 0 -m L sa m p le vo l um e i s used, the i nvert ed vi al shoul d be repl aced wi th an aci d i nd i cat o r ( b rom cresol purp l e) t o prevent p ro b l em s asso ciated w i th ga s b ub b l es i n large i nv erted tub es. T he m ed i a m ust be au t o clav ed at 1 2 1 °C f o r 1 2
- 1 5 m i nu t es. 5 .2 .2 .3 .3 St er i l e medi um i n tubes mus tb e exami ned t o ens ur e thatthe i nv ert ed vi al s , i f us ed, ar e f

r ee of a i rbubbl es a n d a r e atl ea s ton e- h al f t o two- th i r ds c ov er ed af t erth e wa t ers ampl e i s a dded. 5 .2 .2 .4 A fterthe m e d ium is in o c u la te d , it m u st b e in c u b a te d a t 3 5 °± 0 .5 °C fo r 2 4 ± 2 ho u rs . If n o ga s o r a c id is d e te c te d , i t m u st b e inc u b a te d fo r a n o the r 2 4 ho u rs (to ta l i n c u b a tio n ti m e 4 8 ± 3 ho u rs ).V
- 14 5 .2 .2 .5 E a c h 2 4 - an d 4 8 -ho u r tu b e tha t co n ta ins gr o w th, a c id , o r g a s m u st b e c o n firm e d us ing 2 % B G L B B . A compl et ed t es ti s n otr equ i r ed. 5 .2 .2 .6 For dri nk i ng wat ers amp l es: T est e ac h t o t al col i f orm- po s i t i ve s amp l e f

or the pres enc e of

ei therf eca l co l i f o rms or E . co li
. 5 .2 .2 .7 Inva l i d ati o n o f t o t al co l i f

o rm - ne ga t i ve sam p l es 5 .2 .2 .7 .1 F o r d rin k ing w a te r sa m p le s: A ll sa m p le s tha t p ro d u c e a turb id c u lture (i.e ., he a v y gr o w th) in th e abs en ce of

ga s / ac i d produ ct i on , i n LT B or LB , m u s tbe i n v al i dat ed. T h e l abor at orymu s tc ol l ec t , or r eques tthatthe s y s t em col l ect , an others amp l e wi thi n 24 hour s f r om the s ame l ocat i on as the ori gi nal i nv al i dat ed s amp l e. ( B ef ore i nvali dat i on, the l aborat orymayper f orm a c onf

i rmed t es tan d/ or af

eca l c ol i f orm/E . co lit eston the t o t al co l i f o rm-negat i ve cul ture t o check f o r co l i f o rm supp res si o n. I f

the co nfi rm ed t est i s t o t al co l i f o rm - p o sitiv e o r i f

f ec al co l i f

o rm s/E . co lia r e det ec t ed, th e s ampl e mu s tbe r e po r t e d a s s u c h
. A f

e c a l c ol i f

orm/E . co li

- p osi t i ve r es ul ti s co ns i dere d a t ot al co l i f orm- po s i t i ve, f

eca l c ol i f orm/E . co li

- po s i t i ve s amp l e, ev en i f the pres umpt i ve o r c o nfi rmed t ot al col i f orm t es ti s ne gat i ve. If the f o l l o w- up t est (s ) i s negat i ve, the samp l e m ust b e i nval i d at ed b ecause hi gh l evel s o f non-col i f

o rm ba c t e r i a i n th e pr e s u mp t i v e tu be s ma y h a v e i n j u r e d, k i l l e d, ors u pp r e s s e d th e g r ow th of a n yc ol i f

orms i n the samp l e. ) 5. 2. 2. 7. 2 F o r so u rc e w a te r sa m p le s: A ll sa m p le s tha t p ro d u c e a turb id c u lture (i.e ., he a v y gr o w th) in the a b se n c e o f g a s/a c id p ro d u c tio n , in L T B o r L B , sho u ld b e inv a lid a te d . T h e lab o ra to ry sh o u ld c o llec t, o rr eques tthatthe s y s t em col l ect , an others amp l e f r om the s ame l ocat i on as the ori gi nali nv al i dat ed samp l e. ( B ef

o re i nval i d at i o n, the l ab o rat o rym ay p erf o rm a co nf

i rme d t eston the t o t al co l i f o rm-negat i ve cul tur e. If

the conf i rmed t es ti s t ot al co l i f

orm- po s i t i ve, the M P N sh oul d be r eport ed. I f the t es ti s t ot al co l i f o rm-negat i ve, the samp l e shoul d be i nval i d at ed
. ) 5 .2 .3 P res ence- A b sence (P

- A ) Co l i f o rm T est(f

o r det ect i ng t o t al co l i f

o rms i n dri nki ng wat er) 5 .2 .3 .1 M ed i um 5 .2 .3 .1 .1 Si x - t i mes f

ormu l at i on s tr en g th ma ybe used
. I f th e 6- t i mes f

ormu l at i on i s u s ed, i tmu s tbe f i l t er - s t er i l i ze d r ath erth ana u t ocl av ed. 5 .2 .3 .1 .2 T h e m e d ium m u st b e a u to c la v e d fo r 1 2 m inu te s a t 1 2 1 °C . T o ta l t im e in the a u to c la v e sh o u ld be l es s th an30 mi n u t es . Spa ce s h ou l d be a l l owe d be twe enbott l es . T h e f i n alpH mu s tbe 6. 8±0. 2. 5 .2 .3 .1 .3 I f

prep ared m ed i um i s st o red, i ts ho ul d be m ai nt ai ned i n a cul ture bo tt l e at1°- 3 0 °C i n the d ark f orn o l on g erth anth r ee mon th s . I f e v apor at i on e x ce eds 10% of or i g i n al v ol u me, th e medi u m s h ou l d be d isc a rd e d
. 5 .2 .3 .2 A 10 0- mL sa mp l e mus tbe i nocul at ed i nt o a P
- A cu l tur e bott l e. 5 .2 .3 .3 M ed i um m ust be i nc ub ated at 3 5 °± 0 .5°C an d o b serv ed f o r a yell o w co l o r (ac i d ) aft er 2 4 an d 4 8 ho urs. 5 .2 .3 .4 Ye l l ow c u l tu r e s mu s tbe c on f i rme d i n BG LBB a n d af e c a l c ol i f

orm/E . co lit es tc on du ct ed. 5 .2 .3 .5 Al l s ampl es wh i ch pr odu ce a n on - y el l ow tu rbi d cu l tu r e i n P- A medi u m mu s t be i n v al i dat ed
. T he l aborat orymus tcol l ect , or r eques tthatthe s y s t em col l ect , anothersampl e f r om the s ame l ocat i on as the ori gi nal i nval i d at ed s am p l e. ( B ef o re i nval i d at i o n, the l ab o rat o rym ay p erf o rm a co nf

i rme d t eston the t o t al co l i f o rm n e g a t i v e c u l tu r e a n d/ ora f e c a l c ol i f

orm/E . co lites t. If the c o n firm e d tes t i s to ta l co lif o rm -p o sitive , th e sa m p le mu s tbe r e po r t e d a s s u c h
. I f th e co n f i rme d t e s ti s ne g a t i v e , th e s a mp l e mu s tbe i n v a l i da t e d. A f e c a l c ol i f

orm/E. c o lipo s i t i v e r e s u l ti s c on s i de r e d at ot a l c ol i f orm- po s i t i ve, f

e c a l c ol i f

orm/ E . co lipo si t i ve samp l e, even i f the p res um p t i ve and / o r conf i rme d t o t al co l i f

o rm t esti s negat i ve. )V
- 15 5. 2. 4 F e c a l C o l i f

o rm T e s t( u s i n g E C M e d i u m f o rf e c a l c o l i f

o rm s i n d r i n k i n g w a t e ro rs o u r c e w a t e r , o r A- 1 M edi um fo r fe c a l co lif

o rm s in so u rc e w a te r o n ly) 5 .2 .4 .1 E C M ed i um 5 .2 .4 .1 .1 U s e EC medi um t o t es ta t ot al col i f orm- po s i t i ve c ul tur e f

or f eca l col i f

orms un der the T ot al C o l i f o rm R ul e. T he l ab o rat o rym usttrans f er each t o t al co l i f

o rm-po si ti ve cul ture f

rom a presum p t i ve tub e/b o tt l e, o r ea ch p resu m p t i ve t o t al co l i f o rm - p o si t i ve co l o ny un l ess a c o tt o n sw ab i s used , t o at l ea st o ne tub e co nt ai ni ng E C M ed i um wi th an i nvert ed vi al , as s p eci f i ed by §1 4 1
. 2 1 (f

)( 5 )( Se e A p p end i x G ).

5 .2 .4 .1 .2 E C M ed i um m ay b e used t o e num erat e f ecal co l i f

o rms in source water , in acco rdanc e wi th the S u rfa c e W a te r T re a tm e n t R u le . Initia lly, c o n d u c t a M T F tes t (p re su m p tive p h a se ). T h re e sa m p le v o lum e s o f so u rc e w a te r (e .g., 1 0 , 1 a n d 0 .1 m L ), 5 o r 1 0 tub e s/sa m p le vo lum e , sho u ld b e us e d . A cul ture f rom each t o t al co l i f

o rm-po si t i ve tub e shoul d be trans f erred t o a tub e co nt ai ning E C M ed i um w ith a n inv e rte d via l. 5 .2 .4 .1 .3 Au t ocl av e EC M ediu m f or12- 15 mi n u t es a t121° C. T h e f i n alpH s h ou l d be 6. 9±0. 2. 5. 2. 4. 1. 4 I nv ert ed vi al s s houl d be exa mi ned t o ens ur e thattheyare f r ee of ai rbubbl es . T he i nv ert ed vi al m u st b e a t l e a st o n e -ha lf to two -third s c o v e re d a fterthe sa m p le is ad d e d
. 5. 2. 4. 1. 5 E C M ed i um m ust be i nc ub ated at 4 4 .5°± 0 .2°C f o r 2 4 ± 2 ho urs. 5 .2 .4 .1 .6 An ya mou n tof g as de t ec t ed i n th e i n v ert ed v i alof a tu be th athas turbid g r owth mu s tbe cons i dere d a f

eca l col i f

orm- po s i t i ve t es t , r ega r dl es s of the r es ul tof

an ys ubs equen tt es ton thatcu l tur e. 5 .2 .4 .2 A
- 1 M ed i um 5 .2 .4 .2 .1 A -1 m e d ium m a y b e us e d a s a n a lter n a ti v e t o E C M e d ium to e n u m e ra te fec a l co lif o rm s in s ou r ce wa t er , i n ac cor dan ce wi th th e Su r f ac e W at er T r ea tmen t Rul e. A- 1 M ediu m mu s tn otbe u s ed f or dr i n k i n g w at ers ampl es . T h r ee s ampl e v ol u mes of s ou r ce w at er( e. g . , 10, 1 a n d 0. 1 m L), 5 or 10 tub es/ s amp l e v ol ume, s houl d be us ed. U nl i ke E C M edi um, A- 1 M edi um maybe us ed f or the di r ect iso la tio n o f f

e c a l co lif

o rm s fro m w a te r. 5. 2. 4. 2. 2 A- 1 M ediu m mu s t be s t er i l i ze d bya u t ocl av i n g a t121° C f or10 mi n u t es . T h e f

i n alpH mu s tbe 6 .9 ± 0 .1
. 5. 2. 4. 2. 3 In v e rte d via ls sh o u ld b e e x a m ine d to e n su re tha t th e y a re fre e o f air b u b b le s. 5 .2 .4 .2 .4 A- 1 M e di u m m u s tbe i n c u ba t e d a t35 ° ±0. 5° C f orth r e e h ou r s , th e n at44
. 5° ±0. 2° C f or21 ±2 ho urs. 5. 2. 4. 2. 5 L o o se -ca p tub e s sh o u ld b e s to re d in d a rk a t r o o m te m p e ra tur e n o t m o re th a n tw o w e e k s. A -1 M e di u m m u s tn otbe h e l d m or e th a n th r e e mo n th s i n t i g h t l ycl os e d s c r e w - c a p tu be s i n th e da rka t4° C. 5. 2. 4. 3 A nyamountof

ga s det ect ed i n the i nv ert ed vi al of a tube that has turbi d gr owth mus tbe cons i dere d a f eca l col i f orm- po s i t i ve t es t , r ega r dl es s of the r es ul tof

an ys ubs equen tt es ton thatcu l tur e.
5 .2 .5 A zi d e d extrose m ed i um (f o r det ect i ng f

ecal streptoco cci i n ground wat er) 5 .2 .5 .1 Fo r t est i ng 1 0 0
- m L samp l es, p repa re trip l e strength (3X ) f o rmulat i o n i n a cul ture bo tt l e and aut o cl ave at121° C f

or15 mi n u t es . Fi n alpH s h ou l d be 7. 2±0. 2.V
- 16 5 .2 .5 .2 A d d a 1 0 0 -m L w a te r sa m p le to the ste rilize d m e d ium , an d inc u b a te a t 3 5 °± 0 .5 °C
. 5 .2 .5 .3 C h e c kc u lture fo r tu rb id ity a fter2 4 ± 2 h o u r s . If turb id ity is no t o b se rv e d , re inc u b a te a n d c h e c ka g a in

aft er a t o t al i nc ub ati o n p erio d o f 4 8 ± 3 ho urs. 5 .2 .5 .4 A turb id c u lture m a y b e c o n firm e d a s fec a l stre p to c o c c i b y s tre a k ing a p o rtio n o f th e b ro th o n to b ile

escul i n agar ( B E A ) or bil e escul i n azi d e agar ( B E A A ). (T he co nf i rma t i o n m ed i um i n Standard Methods , P S E M e d ium , i s n o lo n g e r c o m m e rc ia lly a v a ilab le .) 5 .2 .5 .5 B ef

or e s tr ea k i n g , B EA an d B EA A mu s tbe s t er i l i ze d bya u t ocl av i n g a t121° C f or15 mi n u t es . Fi n alpH s h ou l d b e 6. 6±0
. 2 f

orBE A a n d 7
. 1±0
. 2 f

orBE AA. 5 .2 .5 .6 A f

t er strea king, B E A an d B E A A p l ates m ust be i nc ub ated at 3 5 °± 0 .5°C f o r 4 8 ho urs. 5 .2 .5 .7 B ro w n ish -b la c kc o lo n ie s o n B E A o r B E A A w ith b ro w n h a lo s c o n firm th e p re se n c e o f f e c a l stre p to c o c c i. I f r equ i r ed, a n e n t er ococ cit estc an b e p er f ormed on on e ormor e f ec als tr ept ococ cic ol on i es by trans f erri ng them t o b rai n he artinf usi o n b roth supp l em ent ed w i th 6.5% N aC l , and i ncub at i ng the cul ture a t 3 5 °± 0 .5 °C fo r 4 8 ho u rs . G ro w th in d ic a te s the p re se n c e o f en te ro c o c c i. 5 .3 E n zy m e (C h ro m o g en i c/fl u o ro g en i c) su b stra te te sts 5. 3. 1 G ene r al 5 .3 .1 .1 Fordet ec t i n g t ot alc ol i f orms a n d E . co lii n d r i n k i n g w a t e rs a m p l e s , a l a b o r a t o rym a yu s e th e M M O- M UG t e s t( C o l i l e r t ) , C o l i s u r e t e s t , E * C o l i t e t e s t , R e a d y c u l tC o l i f o rm s 1 0 0 P r e s e n c e /A b s e n c e Te s t( o rF l u o r o c u l tL MX B ro t h t est), o rCo l i t ag t es t
. T hes e t es t s maybe av ai l abl e i n v ar i ous conf i gu r at i ons . For enu mera t i ng t ot al c o lif o rm s in so u rc e w a te rs , a la b o ra to ry m a y us e the C o lil e rt te st. If a la b o ra to ry use s a fer m e n ta tio n m e tho d to d et ectt o t al co l i f

o rms i n dri nki ng wat er, and the samp l e i s t o t al co l i f o rm-po si t i ve, the l ab o rat o rym ay trans f er the p o sitive c u lture to the E C + M U G tes t t o d e te c t E . co li , b u t n o t to a n y o the r en z ym e su b stra te te st m e d ium in th is sect i o n. 5 .3 .1 .2 M e d ia 5. 3. 1. 2. 1 M e d ia m u st no t b e p re p a re d fro m b a sic ing re d ie n ts, b u t ra the r p u rc h a se d fro m a c o m m e rc ia lly ava i l abl e s our ce. 5 .3 .1 .2 .2 . T h e m e d ia m u st b e p ro te c te d fro m li g h t. 5. 3. 1. 2. 3 Some l ot s of e n zyme s u bs tr at e medi a h av e bee n k n own t o f l u or es ce . T h er ef

or e, e ac h l otof medi u m s h ou l d be c h ec k ed bef or e u s e wi th a 365- 3 66- n m u l tr av i ol etl i g h tw i th a 6- wa ttbu l b. For ch ec k i n g Col i l ert , Col i l ert - 1 8 , C o l i s u r e, Rea dy cu l t/Fl u or ocu l tLM X , a n d Col i t ag me di a, a pa ck etof m ed i um shoul d b e d i ss o l ved i n st eri l e water i n a no n- f l uo res ci ng vess el
. If the m ed i um exhi b its f ai nt fluo re sc e n c e , th e lab o ra to ry sh o u ld us e a n o the r lo t th a t d o e s n o t f luo re sc e . 5 .3 .1 .2 .4 If the sa m p le s p lus a m e d ium e x h ib it a n ina p p ro p ria te c o lo r c h a n g e b e fo re inc u b a tio n , i t sho u ld b e d i scarded and ano ther l o tof m ed i um us ed
. T he l ab o rat o ry sh o u ld n o tif ythe m ed i um vend o r and reque stano ther wat er s am p l e fro m the wa t er s y st em . B ef o re i ncub at i o n, C o l i l ert , C o l i l ert - 1 8 , and C o l i t ag shoul d ap p ear co l o rl ess t o a s l i ghtt i nge o f co l o r, whi l e C o l isu re and E * C o l i t e are y el l o w and R eady cul t/Fl uor ocul ti s s l i gh t l yy el l ow. 5 .3 .1 .3 G l ass and plas t i c b o tt l es and t esttub es s ho ul d be t est ed b ef o re use wi th a 3 6 5
- 3 6 6
- nm ul travi o l etl i ght s ou r ce w i th a 6- wa ttbu l b t o e n s u r e th eydo n otf l u or es ce . I f

th eyf l u or es ce , u s e an oth er l ot of con t ai n er s th atdo notf l uor es ce.V
- 17 5 .3 .1 .4 I f

a W hi r l

- P ak ® b ag i s us ed t o i ncubat e the C ol i l ertor C ol i t ag med i um or a nyothermedi um whi ch changes to a y el l ow col ort o i n di ca t e a pos i t i v e r es u l t , u s e a type th ath as a ba rr i er( e. g . , B 01417) t o pr ev ent g a se o u s e m issio n s to o the r W h irl-P a k ® b a g s d u rin g inc u b a tio n . Q C 5. 3. 1. 5 I n c u ba t or s , e s pe c i a l l y s ma l l , l ow w a tt a g e a i r - type i n c u ba t or s , ma y n ot br i n g a c ol d 10 0- mL wat er

sa m p le (s ) to th e s p e c ifi e d in c u b a tio n te m p e ra tur e fo r se v e ra l h o u rs . T h e p ro b le m m a y c a u se fa lse -ne g a tive re su lts wi th the enzym e substrat e t est s and p o ss i b l yo ther t est s as wel l
. T heref o re, l ab o rat o ri es wi th ai r- typ e i ncub at o rs sh o u ld o b se rv e the fo llo w ing instru c tio n s fo r c h ro m o g e n ic /flu o ro g e n ic su b stra te tes ts:
T est P re- i ncub at i o n s am p l e i nstruct i o ns 1,2 C o lil e rt (P re se n c e /A b se n c e ) S p e c ifi e d 2 4 -ho u r in c u b a tio n ti m e inc lud e s tim e ittak e s to b rin g sa m p le t em p era ture up t o 3 5 °C 1 C o lil e rt Q u a n ti- T ra y S p e c ifi e d 2 4 -ho u r in c u b a tio n ti m e inc lud e s tim e ittak e s to b rin g sa m p le t e mp e r a tu r e up t o 3 5° C Col i l ert - 18 ( Pre s en ce /Abs en ce ) Pre wa rm s ampl e i n 35° C wa t erba th f or20 mi n u t es or44. 5° C f or 7- 10 mi nu t es C o l i l ert - 1 8 Q uant i

- T ray A l l o w samp l e t o eq ui l i b rat e t o roo m t em p erature ( 2 0
- 3 0 °C ) bef o re beginni ng 18
- h ou ri n c u ba t i ont i me C o l i sur e A l l o w samp l e t o eq ui l i b rat e t o roo m t em p erature ( 2 0
- 3 0 °C ) bef o re beginni ng 24
- h ou ri n c u ba t i ont i me Rea dy cu l tCol i f

orms 100 P res ence/Ab sence T estand F luo ro c u lt L M X B ro th S p e c ifi e d 2 4 -ho u r in c u b a tio n ti m e inc lud e s tim e ittak e s to b rin g sa m p le t e mp e r a tu r e up t o 3 5° ±0. 5° C
C o lit a g S p e c ifi e d 2 4 -ho u r in c u b a tio n ti m e inc lud e s tim e ittak e s to b rin g sa m p le t e mp e r a tu r e up t o 3 5° ±0. 5° C If the l ab o rat o ryplans t o put a l arge load int o a sm al l incubator, sam p l es should b e b rought t o roo m t em p erature before 1 i ncub at i o n. Inform ati o n b ase d o n m an ufac ture r’ s i nstructi o ns. 2 5 .3 .1 .6 I f

a w a t e rba th i s u s e d, th e w a t e rl e v e l s h ou l d b e a bo v e th e upp e rl e v e l of th e me di u m. 5 .3 .1 .7 Fo r E . co lit e s t i n g , th e l a bo r a t orymu s tpl a c e al l t ot a l co l i f orm- po s i t i v e s a mp l e s un de ranu l tr a v i ol e tl a mp (3 6 5 -3 6 6 nm , 6 -w a tt) in a d a rk e n e d a re a . If E . co lii s pre s ent , the med i um wi l l emi ta bl ue f l uor es cen ce. 5. 3. 1. 8 T he enzyme substrat e t est s shoul d no t b e used t o co nf i rm a p res um p t i ve t o t al co l i f o rm-po si t i ve cul ture i n f erm ent at i o n bro th (e. g. , LT B , LB , P
- A co l ifo rm te st) o r o n a m em b rane f i l t er. T he hi gh de nsi t i es ofnonco l i f o rm s o r turb i d i ty i n the ino cu l um m ay eithe r sup p ress co l i f o rm s o r o ve rloa d the e nz ym e sub strate test sup p ressa nt reag en tsy st em an d ca use f alse-p o si t i ve result s. 5 .3 .1 .9 A n y sa m p le tha t p ro d u c e s a n a typ ic a l co lo r c h a n g e (e .g., g re e n ish -b la c ko r b la c k ) i n t h e a b s e n c e o f a ye llo w c o lo r sh o u ld b e in v a lid a te d
. T h e la b o ra to ry m u st c o llec t, o r re q u e st tha t th e syste m c o llec t, a n o the r sa m p le f rom the same l o cat i o n as the o ri gi nal i nval i d at ed samp l e. T he l ab o rat o ry shoul d use ano ther m etho d t o t estthe second s am p l e. A cco rding t o the m anufa c tur e r o f Co l i l ert , wat er wi th hi gh i ron o r mangane se l evel s i n the p res ence o f

hy d rogen sul f

i d e m ay cause a gr eeni sh- b l ack o r bl ack co l o r. T hi s greeni sh- b l ack co l o r d o es no toc cur w he n using R ea d ycult , C o l i sure , o r C o l i t ag , ac co rd i ng t o the i r m an ufac ture rs. 5. 3. 1. 10 A n y re fer e n c e c o m p a ra to r p ro v id e d b y the m a n u fac tur e r sh o u ld b e d isc a rd e d b y the m a n u fac tur e r’s e x p ira tio n d a te
.V
- 18 5 .3 .2 C rite ria fo r sp e c ifi c m e d ia 5 .3 .2 .1 F o r the C o l i l erttes t , sam p l es mustbe i ncub at ed at35 °±0
. 5 °C f o r 24 hours. A y el l o w co l o r i n the m ed i um e q u a l t o o r gre a te r th a n the re fer e n c e c o m p a ra to r i n d ic a te s tha t the sa m p le is to ta l c o lif o rm -p o sitive
. If the sa m p le i s y el l ow, butl i gh t erthanthe com para t or, i tmus tbe i ncubat ed f or a notherf

ourhours (do noti ncubat e more than 28 h ou r s t ot al )
. I f

th e col ori s s t i l l l i g h t erth anth e r ef er en ce c ompar at o r at 2 8 hou r s , th e s ampl e s h ou l d be re p o rte d a s n e g a tive . A c o lif o rm -p o sitive sa m p le tha t f luo re sc e s u n d e r a U V li g h t i n d ic a te s the p re se n c e o f E. c o li
. Labora t ori es thatus e the C ol i l ert - 18 t es tmus ti ncubat e s amp l es f or 18 hour s ( up t o 22 hour s i f s amp l e a f

t er 1 8 hours i s y el l o w, but i s l i ght er than the co m p arat o r)
. 5 . 3 . 2 . 1 . 1 Fo r enum era t i n g t ot alc ol i f orms i n s ou r ce w at erw i th th e Col i l ertt es t , a 5- or10- tu be con f i g u r at i on , Q u an t i

- T r ay , or Q u an t i

- T r ay2000 maybe u s ed f ore ac h s ampl e di l u t i on t es t ed. D i l u t i on water(i f us ed ) may be st eri l e d ei o ni zed or st eri l e d i st i l l ed wat er, but no tbuff ered wat er. QC 5. 3. 2. 1. 2 If

the Q uant i

- T rayor Q uant i

- T ray20 0 0 t esti s used, the seal er s ho ul d be checked m o nthl yby add i ng a dy e ( e. g . , br omcr es ol pu rpl e) t o th e wa t er . I f dy e i s obs erv ed ou t s i de th e we l l s , ei th erpe r f orm mai n t en an ce or u se a n o the r se a le r. 5. 3. 2. 2 Fo r the C o l i sur e t est , s am p l es mustbe i ncub at ed at35 °±0
. 5 °C f o r 24 h o u rs . If

an exam i nat i o n of the res ul t s at24 hours i s no tco nveni en t, the n r e su lts m ay b e exam i ned atanyt i m e up t o 48 hours. I f the m ed i um changes f

rom a y el l o w co l o r t o a red/mage nt a co l o r, the samp l e i s t o t al co l i f o rm-po si t i ve. A co l i f

o rm-po si t i ve sa m p le tha t f

luo re sc e s u n d e r a U V li g h t i n d ic a te s the p re se n c e o f E. c o li
. 5 .3 .2 .3 F o r th e E *C o lit e tes t, sa m p le s m u st b e inc u b a te d a t 3 5 °± 0 .5 °C fo r 2 8 h o u rs . If to ta l c o lif o rm s a re p re se n t, the m ed i um changes f

rom a y el l o w co l o r t o a blue or blue- green co l o r, or a b l ue co l o r i n the co rners o f the b ag. If E . co lii s pre s ent , medi um wi l l f l uor es ce un der a U V l i gh t
. I f no f l uor es cen ce i s observed, r e- i ncubat e f or a n addi t i onal 20 hour s ( f

or at ot al i ncubat i on t i me of 48 h our s ) and aga i n che ck f or f l uor es cen ce. I f

medi um becom es re d in c o lo r, a ssu m e tha t a fa u lty se a l ha s a llo w e d the b a c te ric id e (in the third c o m p a rtm e n t o f th e b a g ) to lea kinto the com partmentcont ai ni ng the med i um. I n thi s ca s e, di s car d the s amp l e, an d re ques tan others amp l e. 5 .3 .2 .4 F o r th e R e a d yc u lt C o lif o rm s 1 0 0 P re se n c e -A b se n c e te st, th e c o n te n ts o f a s n a p p a c ksh o u ld b e a d d e d to a 100- mL w at ers ampl e, f ol l owe d byi n cubat i on a t35° ±05 

### / C f o r 24 ± 1 ho urs . If

c o l i f o rms a re present , the m ed i um c h a n g e s c o lo r fro m a s ligh tly ye llo w c o lo r to b lue -gr e e n . I n a d d iti o n , i f E . co li is p re se n t, the m e d ium w ille m it a b ri ghtl i ght

- b l ue f

l uo res cence w hen subject ed t o a l o ng wave (36 5
- 3 6 6 nm ) ul travio le t ( U V ) l i ght
. I f c o n firm a tio n o f E . co lii s des i r ed, K ov ac ’ s i n dol e r ea g en t s h ou l d be adde d t o th e br oth ; th e i mmedi at e f ormat i on o f a re d rin g c o n firm s the p re se n c e o f E . co li
. 5 .3 .2 .5 F luo ro c u lt L M X b ro th is id e n tica l t o R e a d yc u lt, e x c e p t th a t i t i s a d e h yd r a te d c u lture m e d ium in g r anu l at ed f

orm pa ck ed pr i mar i l yi n a 500 g pl as t i c bott l e. Fort es t i n g a 100- mL w at ers ampl e, s u s pen d 34 g of Fl uor ocul tL M X i n 1L pur i f i e d w a t e ra n d b oi l t o d i s s ol v e c om pl e t e l y . Tr a n s f e r10 0- mL a l i qu ot s t o 2 50
- mL b o t t l es a n d a u t ocl av e f or15 mi n a t121° C. Cool t o r oom t empe r atu r e, a dd th e 100- mL w at ers ampl e, a nd i n cubat e. D o n ota dd E . co li/ Col i f orm S u pp l e me n tt o th e me di u m. 5 . 3 . 2. 6 Forth e Col i t ag t es t , s ampl es mu s tbe i n cubat ed at35 E ± 0 .5 E C f or24±2 h ou r s . D u r i n g i ncub at i o n, trim e thyla m ine -N -o x id e in the C o lit a g m e d ium c a u se s the p H o f th e m e d ium to in c re a se fro m 6 .2 to 6 .8 -7 .2 . A y el l ow col or i n the med i um i ndi cat es the pres enc e of t ot al col i f

orms . A co l i f

orm- po s i t i ve s amp l e thatf l uor es ces u n d e r a U V li g h t i n d ic a te s the p re se n c e o f E. c o li
. 5 .3 .3 E C M e d ium + M U G T e st (fo r d e te c tio n o f E . co li ) 5 .3 .3 .1 I f

E C m ed i um + M U G i s used, a t o t al c o l i f o rm-po si t i ve cul ture m ustb e trans f erred f rom a presum p t i ve tub e/b o tt l e o r col o nyt o E C m ed i um + M U G , as specif i ed by §1 4 1
. 2 1 (f

)( 5 )( Se e A p p end i x G ).V
- 19 5 . 3 . 3 .2 M U G m a y b e a d d e d to E C M e d ium b e fo re a u to c la v ing . E C M e d ium + M U G is also a v a ilab le comme r ci al l y . T h e f

i n alM U G c on ce n tr at i on mu s tbe 50 μg /mL. T h e f i n alpH s h ou l d be 6. 9±0. 2. 5 .3 .3 .3 T he i nv ert ed vi al maybe om i tt ed, becau s e g as product i on i s notr el eva ntt o the E . co lites t. 5. 3. 3. 4 T he med i um mus tbe i ncubat ed at44. 5°±0. 2°C f or 24± 2 hour s , an d t es t ed f or f l uor es cen ce. 5 .3 .4 E nt erolertt est(f o r det ect i o n of ent eroc o cci i n ground wat er) 5 .3 .4 .1 M edi um s houl d be s t ored i n the darkat4- 30 ° C un t i l us e. 5 .3 .4 .2 A d d E ntero l ert rea ge nt t o 1 0 0
- m L wa t er sa m p l e, an d i nc ub ate a t4 1 °± 0 .5°C f o r 2 4
- 2 8 ho urs. F luo re sc e n c e un d e r a U V lam p ind ic a te s the p re se n c e o f en te ro c o c c i. 5 .3 .4 .3 T h e d e v e lo p m e n t o f f luo re sc e n c e a fter2 8 ho u rs is no t a v a lid tes t f o r e n te ro c o c c i. 5. 4 M emb ran e Fi l t er ( M F) met hods 5 .4 .1 G ene r al 5. 4. 1. 1 F o r so u rc e w a te r sa m p le s (S W T R ): T o o p tim iz e c o u n ting , a p p ro p ria te s a m p le d ilutio n s m u st b e u se d to yie ld 2 0 to 8 0 to ta l co lif o rm c o lo n ie s o r 2 0 -6 0 fec a l co lif o rm c o lo n ie s fo r a t l e a st o n e d ilutio n o r v o lum e . Q C 5. 4. 1. 2 A tl eastone m em b rane f i l t er and f

i l trat i o n uni tst eri l i tyc heck shoul d be co nd uct ed atthe b egi nni ng and the end of each f

i l trat i o n s eri es by f

i l t eri ng 20
- 3 0 m L o f

dilut i o n wat er through the m em b rane fi l t er and t est i ng f or g r owth. I f

the contr ol i ndi cat es cont ami nat i on, al l dat a f r om af f ect ed s amp l es mus tbe r ej ect ed an d an i m med i at e r es amp l i ng s houl d be re ques t ed. A f i l tr at i on s er i es en ds whe n 30 mi nu t es or mo r e e l aps e between sam p l e fi l trati o ns. 5 .4 .1 .3 Eac h f

i l tr at i on f

u n n elmu s tbe r i n s ed af t er ea ch s ampl e f

i l tr at i on wi th two orth r ee 20- 30 mL port i on s of s t er i l e r i ns e wa t ert o ens ur e thaten t i r e s amp l e i s r i ns ed of f the f un nelbef

ore the f i l t eri s r emo ved. Af

t erthe f i l t er i s r emo ved, the f

un nelmaybe r i ns ed aga i n wi th two or thr ee 20
- 30 mL po rt i ons of s t er i l e r i ns e wa t eror expos ed to U V li g h t with a 2 5 4 -nm w a v e le n g th fo r a t l e a st tw o m inu te s to p re v e n t c a rr y-o v e r b e tw e e n sa m p le s, e sp e c ia lly fo r su rfa c e w a te r sa m p le s.. 5 .4 .1 .4 Abs orben t pads mu s t be s atu r at ed wi th a l i qu i d medi u m ( at l ea s t 2 mL of br oth ) an d ex ce s s m ed i um re m o v e d b y " d e c a n ting " the p la te
. 5 .4 .2 M F me th od f

orde t ec t i n g t ot alc ol i f

orms a n d E . co lii n dr i nk i ng w ater, e nu mera t i ng t ot al col i f orms or f

eca l co l i f o rms i n s o urce wat er, and de t ect i ng E . co lii n g r ound wat er 5 .4 .2 .1 M edi a f

ort ot alc ol i f

orms , f ec alc ol i f

orms , a n d E . co li

5 .4 .2 .1 .1 M
- Endo M ediu m ag ar or br oth ( al s o k n own as M
- Endo br oth M F a n d M

- Col i f orm B r oth ) or LES Endo a g ar( al s o kn owna s M
- Endo Ag arLES) f orde t ec t i n g t o t al c o l i f

orms i n dr i n k i n g w at eror enum erat i n g t ot alc ol i f orms i n s ou r ce w at er . M ediu m maybe u s ed i n th e s i n g l e s t ep ore n r i ch ment t ec h n i qu es . Ens u r e th at eth an ol u s ed i n th e r eh y dr at i on pr oce du r e i s n otde n atu r ed. M ediu m s h ou l d be p re p a re d in a ste rile fl a sk a n d b ro u g h t ju st to the b o ili n g p o int w ith a b o ili n g w a te r b a th o r, if c o n sta n tly att en ded, a h otpl at e wi th a s t i r bar
. T h e medi u m mu s t n ot be boi l ed. Fi n alpH s h ou l d be 7. 2±0. 2 f or M
- En do Ag a rL ES a n d 7
. 2±0
. 1 f orM
- En do me di u m. 5 .4 .2 .1 .2 m- Col i B l u e24 medi u m f orde t ec t i n g t ot alc ol i f

orms a n d E . co lii n dri nk i ng wat er . Am pul esV
- 20 of broth s houl d be i nv ert ed 2- 3 t i mes t o m i x cont ent s bef ore brea ki ng . T hen cont ent s s houl d be pour ed ev en l yov era bs orben tpa d. U n open ed r ef r i g er at ed a mpu l es m aybe s t or ed i n th e darku n t i l th e ex pi r at i on dat e, bu ts h ou l d be di s ca r ded ea r l i er i f g r owth i s obs erv ed. T h e f i n alpH of

me di u m s h ou l d be 7. 0±0. 2. 5 .4 .2 .1 .3 M I M edi um ( wi th or wi thoutag ar ) f or det ect i ng t ot al col i f

orms an d E
. col i i n dr i nk i ng wat er or e nu mera t i ng t otalcol i f orms i n s our ce wat er . D o notau t ocl ave comm er ci al l ymade, pre s t er i l i zed bo tt l ed M I aga r or broth. M el t bo tt l ed aga r i n aboi l i ng wat erbath ( or byotherproces s es r ecomm ended b y the m a n u fac tur e r) . A s so o n a s c o m p le te m e lti n g ha s o c c u rr e d , co o l sli g h tly a n d p o u r im m e d ia te ly i nt o s t eri l e p l at es. C are s ho ul d be t aken t o prevent o verheat i ng the agar, as excess i ve heatde stroys the ef f ect i ven es s of the ant i bi ot i c, ce f s ul od i n. I f

dehy drat ed cu l tur e medi um i s us ed, i ts houl d be prepar ed and aut o cl aved acco rding t o the m anuf acturer’ s i nstruct i o ns. C o o l the agar, ad d f res hl yprep ared , f

i l t ers t e r i l i z e d c e f

s u l o d i n , a n d p o u r i m m e d i a t e l y i n t o s t e r i l e p l a t e s . Th e f i n a l p H o f M I a g ars h ou l d be 6
. 9 5 + 0
. 2 0 ; th e f

i n a l p H o f

MI b r o th s h o u l d b e 7
. 0 5 + 0
. 2 0
. T h e p r e p a r a t i o n a n d u s e o f MI a g a ra n d MI b roth i s d escrib ed i n the art i cl e, “ N ew m ed i um f o r the si m ul t aneo us d et ect i o n of t o t a l c o l i f o rm and Esc h e ric h ia c o liin w a te r” b y B re n n e r, K .P ., e t a l. , 1 9 9 3 , Appl i ed and Envi r onm entalMi crobi ology 5 9 :3 5 3 4 -3 5 4 4 . E P A M e tho d 1 6 0 4 , w h ic h c a n b e fo u n d o n line a t www. epa. gov/mi cr ob es , i s id e n tica l. 5 .4 .2 .1 .4 Chr omocu l t ® Col i f orm Aga rf

orde t ec t i n g t ot alc ol i f

orms a n d E . co lii n d r i n k i n g w a t e r . Do n o t a u to c la v e o r o v e rh e a t. T h e fina l p H sh o u ld b e 6 .8 + 0 .2
. If a h e a v y b a c k g ro u n d o f h e te r o tro p h ic b a c te ria is ex p e c te d (e sp e c ia lly Ps eudomonas a n d Aer om onas s pp. ) , a dd c ef s u l odi n s ol u t i on t o the cool ed ( 45°
- 50° C) me di u m ( di s s ol v e 10 mg c ef s u l odi n i n 2 mL d ei o n i ze d ordi s t i l l ed wa t er , a n d a dd s ol ut i on t o 1L of

medi um). Checkwi th the manuf actur er , E M D Chemi cal s , I nc. , at w w w . e md c h e mi c a l s . c om , o r c a ll (8 0 0 ) 2 2 2 -0 3 4 2 fo r a d d iti o n a l i n fo rm a tio n o n the p e rfo rm a n c e o f th is te st w ith v a rio u s fil te r typ e s. 5 .4 .2 .1 .5 Col i s ca n ® f orde t ec t i n g t ot alc ol i f

orms a n d E . co li i n dr i n k i n g w a t e r or e n u me r a t i n g t ot al col i f orms i n s our ce wat er . C ol i s can i s av ai l abl e a s a drypowd erag armi x oras a pr es t er i l i zed bot t l ed agar. Fo r r eco nst i tut i o n an d a n t i b i o t i c ad d i t i o n, f o l l o w the p rotoco l of the m anuf acture ( M i crology L a b o ra to rie s, L L C ). D o n o t o v e rh e a t the a n tib io tic, c e fsu lo d in. T h e fi n a l p H o f C o lisca n a g a r sh o u ld be 7. 00+0. 20. 5 .4 .2 .1 .6 m- FC br oth (w i th or wi th ou tag ar ) f oren u mer at i n g f

ec alcol i f

orms i n s ou r ce wat er . D o n ot au t ocl av e. B r i n g me di u m j u s tt o th e boi l i n g poi n t
. T h e f i n alpH s h ou l d be 7. 4±0. 2. 5 .4 .2 .1 .7 W h e n sto re d , p re p a re d m e d ium sh o u ld b e re frige ra te d . P e tri d ish e s c o n ta inin g m e d ium sh o u ld be s t ored i n a pl as t i c bag or t i gh t l ycl ose d cont ai ner , an d us ed wi thi n two week s . B ef ore us e, r ef

r i ger at ed ste rilize d m e d ium sh o u ld b e b ro u g h t t o ro o m tem p e ra tur e . P la te s w ith la b o ra to ry p re p a re d b r o th m ed i um m ustbe dis card e d a fter 9 6 ho urs , po ured M F aga r pl at es di scarded af t er two weeks, and a m p u l e d M
- E n d o b r o th a n d o th e rp r e p a r e d m e d i a d i s c a r d e d i n a c c o r d a n c e w i th th e m a n u f a c tu r e r 's exp i rat i o n da t e. B roth, plat es, or am p ul es shoul d b e d i scarded earl i er i f growth o r ( f

o r M
- E nd o agar) s u r f ac e s h ee n i s obs erv ed. Rec or d da t e an d t i me pr epa r ed. 5. 4. 2. 2 Incuba t i o n co nd i t i o ns and co l o nyco l o r of i no cul at ed m ed i um
M e di um I nc ub at i on Tot alc ol i for ms E. co li 1 M
- Endo me di u m or M
- En d o a g a rL ES 35 ° ±0
. 5°C f or 22- 24 hr s M et al l i c ( gol den) s hee n

col oni es ( pres umpt i ve) N /A

m - C o l i B l ue 2 4 3 5 °± 0 .5°C f o r 2 4 hrs R ed co l o nies B l ue t o p urp l e c o l o nies M I 35 ° ±0
. 5°C f or 24± 2 hr s Fl uor es cen tcol oni es un der

U V l i ght B l ue col oni es un der normal l i ghtM e di um I nc ub at i on Tot alc ol i for ms E. co li 1

V
- 21 C hr om ocul t 36 ° ±1 ° C f or 24± 1 hr s Sal mo n t o re d col oni es D ark- bl ue t o vi ol etcol oni es 2 C ol i s can 32 °
- 37 ° C f or 24- 28 hr s P i nk

- magen t a c ol oni es P urpl e- bl ue col oni es
m- FC 44
. 5°±0. 2°C f or 24± 2 hr s N /A B l ue col oni es ( f eca l co l i f o rm s) W itho u t the p re se n c e o f E . co li
. I f an E . co lico l onyi s pre s ent , as i ndi cat ed by the l as tcol umn, i ts houl d be count ed as 1 a to ta l co lif

o rm -p o sitive c o lo n y. If co n firm a tio n o f E . co lii s de s i r ed, a dd on e dr op of K ov ac ’ s r ea g en t t o ea ch dark

- bl u e t o v i ol etc ol on y ; th e f ormat i on 2 o f a c h e rr y-re d c o lo r w ithin se c o n d s c o n firm s the p re se n c e o f E . co li
. 5 .4 .2 .3 I n v al i dat i on of a t ot alc ol i f

orm- n eg at i v e dr i n k i n g w at ers ampl e: Al l s ampl es r es ul t i n g i n c on f l u en tor T N T C ( t oo nu m ero us t o c ount) gr owth mus tbe i nv al i dat ed un l es s t ot al col i f orms ar e det ect ed. I f

no t ot al c ol i f orms a r e de t e c t e d, r e c or d a s "c on f l u e n tgr ow th " or"T N T C" a n d r e qu e s ta n a dd i t i on a l s a mp l e f r om th e s a me s amp l i ng s i t e. Co nf

l uen tgr owth i s def

i ned a s a cont i nu ous b act er i al growth cover i ng the ent i r e memb r ane f i l t er wi tho utevi d ence o f

t o t al co l i f

o rm-typ e co l o ni es. T N T C i s d ef i ned as great er than 2 0 0 c o l o ni es o n the mem b rane f i l t er i n the abs enc e of det ect abl e col i f orms . Labo r at ori es mus t not i nv al i dat e s amp l es whe n the mem bran e f i l t er c on t a i n s a tl e a s tone c ol i f orm- type c ol on y( i
. e . , s h e e n c ol on yf orM
- En do me di u m, r e d o rblu e c ol on yf ormCol i B l u e 2 4 a gar, flu or es ce n torbl u e col on yf orM I a g ar , s al mon t o r ed orda rk

- bl u e t o v i ol etc ol on i es f or C hrom o cul tC o l i f

o rm agar , pin k/mag ent a orblue/p urple co l o nyf o r Co l i scan). ( B ef

o re i nval i d at i o n, the l ab o rat o ry maype r f orm a v er i f

i ca t i on t es ton th e t ot alc ol i f orm- n eg at i v e cu l tu r e, i
. e. , on c on f l u en tor T N T C gr owth , a n d a f e c a l c ol i f

orm/E . co li t es t
. I f the ver i f i cat i on t es ti s t ot al col i f orm- po s i t i ve, the s amp l e mus tbe r eport ed as t ot al c ol i f orm- po s i t i v e . I f

th e t e s ti s t ot a l c ol i f

orm- n e g a t i v e , th e s a mp l e mu s tbe i n v a l i da t e d. A f e c a l c ol i f

orm/E . co lipo s i t i v e r e s u l ti s c on s i de r e d at ot a l c ol i f orm- po s i t i v e , f

e c a l c ol i f

orm/E . co li

- po s i t i ve sa m p l e, e veni f the i ni t i al and / o r ver i f

i cat i o n t o t al co l i f

o rm t esti s negat i ve. ) 5 .4 .2 .4 In v a lid a tio n o f so u rc e w a te r sa m p le s (S W T R ): L a b o ra to rie s m u st inv a lid a te a n y sa m p le w h ic h re su lts i n co nf l uentgrowth o r T N T C , even when to ta l coli f o rm o r f ecal co l i f

o rm co l o ni es ar e p res ent , be cause co l i f o rm d e n sity m u st b e d e te rm ine d
. 5 .4 .2 .5 For dri nk i ng wat ers amp l es ( t o ver i f ycol oni es on Endo

- type medi um): Atl eas tf i ve typi cals hee n c o lo n ie s a n d f

iv e no n typ ic a l co lo n ie s m u st b e ve rified us ing e ithe r sin g le stre n g th la c to se b ro th ( L B ) o r la u ryl trypt ose broth ( LT B ) and then s i ng l e s tr eng th 2% bri l l i antgr eenl act ose bi l e br oth ( B G LB B )
. Al t ernat i vel y , s hee n col on i es ma ybe v er i f

i ed u s i n g a cyt och r ome ox i das e an d ß- g al ac t os i das e pr oce du r e. I n di v i du alc ol on i es c anbe trans f erred with a s t eri l e need l e o r l o o p , or ap p l i cat o r s t i ck. If no sheen co l o ni es are o b served , veri f yup t o f i ve red q ue st i o na b l e she en co l o nies an d / o r red no n-shee n c o l o nies re p r e se n tin g d i f f ere nt m o rp ho l o gica l typ es. A l t ernat i vel y , wi p e the ent i re s urf ace o f the m em b rane f

i l t er with a s te rile c o tt o n s wab , and i no cul at e the veri f i cat i o n m ed i a (LT B , then B G LB B ).

5 .4 .2 .6 Fo r d ri nki ng w at er sam p l es: T o t al c o l i f o rm-po si t i ve co l o ni es m ustbe t est ed f o r E . co lio r f

ec al co l i f

o rm s. T h e membr an e f

i l t erte s t s a ppr ov ed byEP A t o da t e do n otr equ i r e addi t i on alme di a f ors u cha t es t , e x ce ptf or th os e u s i n g E n do- type medi u m ( M
- Endo medi u m orM
- Endo a g arLES)
. EP A h as a ppr ov ed s ev er alopt i on s f or t est i ng a t o t al co l i f

o rm-po si t i ve co l o ny o n E nd o - typ e m ed i um f o r E . co lior f

ecal co l i f

o rms. W hen E C M ed i um (f o r f ecal co l i f

o rms) or E C M ed i um + M U G (f o r E . co li ) i s used, the co l o ni es mustbe trans f erred b yem p l o y i ng one of the op t i ons s peci f

i ed by the T ot al Co l i f

orm R ul e a t40 C FR 141
. 21 ( f ) ( 5)( See A pp endi x G )
. For the s wab t ech ni que, a s i ng l e s wab canbe us ed t o i nocul at e apre s umpt i ve t ot al col i f orm- po s i t i ve cul tur e i nt o up t o thr ee d iff e re n t m e d ia ( e .g., E C o r E C -M U G M e d ium , L T B , a n d B G L B B , in th a t o rd e r) . If N u trie n t A g a r + M U G is u s ed, r ef ert o pa r ag r aph5. 4. 3. 5 .4 .2 .7 F o r so u rc e w a te r sa m p le s: I n iti a l to ta l c o lif o rm c o u n ts m u st b e a d

ju ste d b a se d up o n ve rified d a ta , as in Standard Methods , Sec t i o n 92 2 2 B (5).V
- 22

Q C 5 .4 .2 .8 For s our ce wat ersampl es (SW T R ) : I f two or more ana l y s t s ar e a vai l abl e, ea ch ana l y s ts houl d countt ot al c o l i f o rm s o rf

e c a lc o l i f

o rm c o l o n i e s o n th e s a m e m e m b r a n e m o n th l y . C o l o n yc o u n t s s h o u l d a g r e e w i th i n 1 0 %. 5. 4. 3 N u trie n t A g a r + M U G T e st (fo r d e te c tio n o f E . co lii n dri nki ng wat er or ground wat er)

5 .4 .3 .1 M ed i um m ustbe aut o cl aved at12 1 °C f o r 15 m inutes
. M U G m ay b e ad d ed t o N utri entA gar before au t ocl av i n g . N u tr i en tAg ar+ M U G i s al s o av ai l abl e comme r ci al l y . T h e f i n al M U G c on ce n tr at i on mu s tbe 100 μg/mL. T h e f

i n alpH s h ou l d be 6. 8±0. 2. QC 5 .4 .3 .2 P o sitive a n d ne g a tive c o n tro ls sh o u ld b e tes te d a s sta te d in p a ra g ra p h 5 .1 .6 .4 . F ilt e r o r s p o t-ino c u la te con tr ol c u l tu r es on t o a me mbr an e f i l t eron M
- Endo ag arLE S orM
- Endo br oth or ag ar , a n d i n cubat e at35° ±0. 5° C f o r 2 4 ho urs. T he n transfer th e fi lte r t o N u tr i en tA ga r + M U G an d i nc ub ate a t3 5 °C f o r an o the r f

o ur ho urs. T h e r es u l t s s h ou l d be r ea d a n d r ec or ded. 5 .4 .3 .3 T he m em b rane f i l t er contai ni ng co l i f

o rm co l o ny (i es) m ust b e trans f erred f rom the t o t al co l i f

o rm m ed i um t o the s ur f

ace o f N utr i entA gar+ M U G m edi um. E ach sh eenco l onysh oul d be mark ed wi th apermanen tmarker o n the l i d
. A l so, the l i d and the b ase s ho ul d be m arked wi th a l i ne t o real i gn the lid s h o u ld it b e remo ved
. ( A p o rtio n o f th e c o lo n y m a y b e tra n sfe rr e d w ith a ne e d le to the to ta l co lifo rm ve rifica tio n tes t b e fo re tra n sfe r to N utr i entAgar+ M U G o raf t erthe 4- houri ncubat i on t i me. Anothermethod i s t o s wab the ent i r e memb r ane f i l t er s ur f ace wi th as t er i l e c ot t on s wab af t erthe 4- houri ncubat i on t i me on N utr i entA gar+ M U G medi um, an d tr ans f er to a to ta l co lif

o rm ve rifica tio n tes t.) 5 .4 .3 .4 Ino cu l ated m ed i um m ust be i nc ub ated at 3 5 °± 0 .5C ° f o r f o ur ho urs. 5 .4 .3 .5 Checkthe f

l uor es cence u si ng an ul tr avi ol etl amp ( 36 5- 36 6 nm) w i th a6- wattbul b i n adarken ed ar ea. A nyam o untof f

l uo res cence in a hal o around a s heen co l o nys ho ul d be co nsi d ered po si t i ve f o r E . co li
. 5 .4 .4 M F method f

or det ect i ng en t er ococci / f eca l s tr ept ococci i n g r ound wat er 5 .4 .4 .1 M e d ia 5 .4 .4 .1 .1 For mE ag ar( SM 923 0C ) f or the det ect i on of

en t ero cocci : Prepar e bas al mE ag ar . T hen a u to c la v e a n d c o o l i n a 4 4 -4 6 °C w a te r b a th. D isso lve 0 .4 8 g n a lid ixic a c id a n d 0 .4 m L 1 0 N N a O H into 10 mL of

r eag ent - gr ade di st i l l ed wat eran d m i x. Fi l t er - s t er i l i ze the s ol ut i on, an d add 5. 2 m L perl i t er o f b a sa l m E a g a r. F o r trip h e n yl t e tra z o lium c h lo rid e (T T C ), a d d 0 .2 5 g o f T T C to 2 5 m L o f re a g e n tg r a de w a t e r , an d war m t o di s s ol v e . F i l t e r - s t e r i l i z e th e s olu t i on , a n d a dd 15 mL pe rl i t e rof ba s a l mE ag ar . Fi n alpH s h ou l d be 7. 1± 0. 2.

5 .4 .4 .1 .2 F o r m - E nt eroc o ccus aga r (SM 9 2 3 0 C ) f o r the d et ect i o n o f

fecal streptoco cci (no tent eroc o cci ): H eatt o dis sol ve i ngredient s, but d o not aut o cl ave. D i spense int o st eri l e p etri plat es ( 9 X 50 m m ) ( ab o ut 4 mL) , a n d a l l ow t o s ol i di f y . Fi n alpH s h ou l d be 7. 2±0. 2. 5 .4 .4 .1 .3 F o r m E I a g a r (E P A M e tho d 1 6 0 0 ) fo r th e d e te c tio n o f en te ro c o c c i: A d d 0 .7 5 g in d o x yl- 

### $

- Dgl uco si d e t o 1L of b asal m E a gar, and pro ceed acco rding t o pa ragraph 5
. 4
. 4
. 1
. 1 , excep tthatthe p repa rat i o n of T T C i s as f o l l o ws: A d d 0
. 1 g o f

T T C t o 10 m L o f

reagent - grade dis t i l l ed wat er, and warm t o di s s ol v e. Fi l t er - s t er i l i ze th e s ol u t i on , an d add 2 mL perl i t erof me di u m. Fi n alpH s h ou l d be 7. 1±0. 2. 5 .4 .4 .2 A f

t erf i l t er i ng a 10 0- mL s amp l e, p l ace m emb r ane i n a p etr i d i s h on one of the aga rmedi a l i s t ed above. Se ri al dilut i o ns shoul d not no rma l l ybe necess aryf o r det ect i ng ent eroc o cci i n ground wat er. 5 .4 .4 .3 I f

m - E nt eroc o ccus a g a r is used , i ncub at e i nvert ed plat e at35 °±0
. 5 °C f o r 48 hours and, usi ng m a g n ifi c a tio n a n d a flu o re sc e n t l a m p , co u n t all li g h t an d d a rk re d c o lo n ie s a s fec a l stre p to c o c c i.V
- 23 5 .4 .4 .4 I f

m E agar i s used, i ncub at e i nvert ed plat e f o r 48 h o u rs a t41 °±0
. 5 °C , and then trans f er f i l t er t o E IA m ed i um . I ncub at e at4 1 °± 0
. 5 °C for 2 0
- 3 0 m i nut es and , usi ng m agni f i cat i o n and a f

l uo res centl am p , exam i ne the col on i es . Pi n kt o r ed col on i es on mE ag arw i th a bl ac k or r eddi s h br ownpr ec i pi t at e on th e u n der s i de of f i l t eron E IA ind ic a te s the p re se n c e o f en te ro c o c c i. 5 .4 .4 .5 If m E I a g a r is u se d , in c u b a te in v e rte d p la te fo r 2 4 ho u rs a t 4 1 °± 0 .5 °C . U sin g m a g n ifi c a tio n a n d sm a ll f l uo res centl am p , exam i ne b o th the t o p and bo tt o m of the p l at e f o r col o ni es wi th a b l ue hal o . A co l o nywi th a blue h a lo , re g a rd le ss o f co lo n y c o lo r, ind ic a te s p re se n c e o f en te ro c o c c i. 5. 5 Heterotr ophic Plate Count ( f o r en u m era t i n gh etero tro p h i c b a cteria i n dri n kin gw a ter) 5 .5 .1 T h e PourPl at e M eth od ( Standard Methods 92 1 5 B ) o r the S i m P l at e M etho d m ustbe us ed f o r det erm i ni ng compl i an ce w i th 40 CFR 141. 74( a) ( 1) ( al s o l i s t ed i n Appen di x G ) a n d s h ou l d a l s o be us ed f ort es t i n g r ea g en tg r ade wat er . For s y s t ems thatha ve been gr ant ed a var i anc e f r om the T ot al Co l i f

orm R ul e' s maxi mum cont ami nan tl eve l ( s ee vari ance cri t eri a i n the p ream b l e o f FR 5 6 : 1 5 5 6
- 1 5 5 7 , January 1 5 , 19 9 1 ), anym etho d i n Standard Methods , Se ct i on 9 2 1 5 , Heterotr ophi c Plate Count , m ay b e used with R 2 A m ed i um , f o r enumerati ng het erotr o p hi c b act eri a i n dri nki ng w a te r. 5 .5 .2 M edi a M et hod M ed i um Fi nalpH P o ur P l at e P l at e co untagar, al so kno wn as tryp t o ne glucos e y eas tag ar 7 .0 ± 0 .2
PourPl at e R2A ag ar 7. 2 ± 0. 2 Spr ea d Pl at e R2A ag ar 7. 2 ± 0. 2 M embr an e Fi l t er R2A ag ar 7. 2 ± 0. 2 Si mP l at e M u l t i pl e en zyme s u bs tr at e 7. 2 ± 0. 2 5 .5 .3 ( For P our P l at e M ethod) M el t ed aga rmus tbe t emp er ed at44°- 46 ° C i n w at erbath bef ore po ur i ng . M el t ed ag ar s houl d be hel d no l ongerthanthr ee hour s . St er i l e a garmedi um s houl d notbe mel t ed mo r e thanonce . 5. 5. 4 (F o r S p rea d P l ate M etho d ) 1 5 m L o f R 2 A ag ar m ed i um (o r o the r m ed i um ) sho uld b e p o ure d i nto a p etri d i sh (1 0 0 x 1 5 m m o r 9 0 x 1 5 m m ) a n d a llo w e d to so lid ify. 5 .5 .5 Ref

r i ger at ed medi um maybe st ored i n bott l es or i n s cr ew- capped tubes f or up t o s i x months , or i n petr i di s hes f o r up t o two weeks. P repa red p etri dis hes wi th R 2 A m ed i um m ay b e st o red f o r up t o one week. 5 .5 .6 Form os tpot abl e wa t ers ampl es , cou n t abl e pl at es ca n be obt ai n ed bypl at i n g 1. 0 m L an d/ or0. 1 m L v ol u mes of the und i l uted sam p l e (d i l uti o ns m ay no tb e n ec essa ry f o r S i m P l ate, wh i ch ha s a co un t i ng ran ge up t o 7 3 8 /m L ). A tl ea st d u p lica te p la te s p e r d ilutio n sh o u ld b e us e d
. 5 .5 .7 ( For P ourP l at e M ethod) T he s amp l e mus tbe as ept i cal l ypi pett ed ont o the bo tt om o f a s t er i l e p etr i di s h. T hen atl eas t10- 12 mL m L of t emp er ed mel t ed ( 44 °
- 46 ° C ) ag armus tbe added t o eachpetr i di s h. T he s amp l e and mel t ed ag ar m ust be m i xe d ca refull y t o av o i d sp i l l ag e. A f t er a ga r p l ates ha ve so l i d i f i ed o n a l ev el surf

ac e, the p l ates m ust be i nv ert ed an d i ncubat ed at35°±0
. 5°C f or 48± 3 hour s . Pl at es s houl d be s t ack ed no m ore thanf ourhi gh an d arr ang ed i n the i ncub at o r t o al l o w p rop er ai r ci rcul at i o n and t o m ai nt ai n uni f o rm i ncub at i o n t em p eratur e . A vo i d excess i ve hum i d i tyi n the i ncub at o r t o reduce the p o ss i b i l i tyo f sp read er f

o rma t i o n on the agar me d i um . A l so avo i d excess i ve dry i n g of

th e ag arme di u m; a g arme di u m i n pl at es s h ou l d n otl os e mor e th an15% b y w ei ghtdu r i n g 48 h ou r s of inc u b a tio n .V
- 24 5 .5 .8 ( Fo r S p read P l at e M etho d ) 0. 1 or 0
. 5 m L o f the samp l e (or dilut i o n) s ho ul d be pipett ed onto the sur f ace o f the p redri ed agar i n the p l at e, and then spread ove r the ent i re s urf ace o f the agar us i ng a s t eri l e b entgl ass rod
. T he i n ocu l u m s h ou l d be a bs orbed compl et el ybyth e ag arbe f or e th e pl at e i s i n v ert ed an d i n cubat ed. T h e pl at e s h ou l d be i nc ub ated at 2 0 °- 2 8 °C f o r 5
- 7 d ays. 5 .5 .9 (For M emb r ane F i l t erT ech ni que) T he vol ume t o be f i l t er ed s houl d y i el d between20- 20 0 col oni es . T he f i l t er i s trans f erred t o a p etri d i sh co nt ai ni ng 5 m L of sol i d i f i ed R 2 A m ed i um , and i ncub at ed at20 °- 2 8 °C f o r 5- 7 da y s. I f p l at es wi th lo o se -f

i tt i ng l i d s are us ed , plat es s ho ul d be placed i n a plas t i c b o x wi th a cl o se f i tt i ng l i d co nt ai ni ng moi s t en ed pa p e r t o w e l s . Pa p e r t o w e l s s h o u l d b e r e w e tt e d a s n e c e s s a ry t o m a i n t a i n m o i s tu r e . Co l o n i e s s h o u l d be c o u n te d us ing a ste re o sc o p ic m ic ro sc o p e a t 1 0 -1 5 X a m p lif ic a tio n . 5. 5. 10 ( ForSi mP l at e M eth od) 5. 5. 10. 1 U ni t D o se (f

o r a si ngl e samp l e). A 1 0
- m L vol um e o f t est samp l e i s ad d ed t o a t est tub e c o n ta ining d ehydrat ed SimP l at e m ed i um . T hen the d i ss o l ved m ed i um sho ul d b e p o ured o nt o the centerof a p l at e co nt ai ni ng 8 4 s m al l wel l s (pro vi d ed b ythe m anuf acturer, ID E X X L ab o rat o ri es, Inc.)
. A l t ernat i vel y , 9 m L o f s t eri l e d i l uent (D
. I. w at er, d i st i l l ed w at er, o r b uf f ered wat er [ Standard Methods , 90 50 C , 1a]) ca n be a dd ed t o the tube, f ol l owed by1- mL s ampl e. T h enf ol l ow th e pr oce du r e as i n di ca t ed abov e f orth e 10- mL s ampl e. T h e mi x tu r e s h ou l d be di s tr i but ed ev enl yt o the 84 wel l s on the pl at e, and the exc es s l i qui d dr ai ned i nt o a n abs orbentpad on the pl at e. T h e pl at e s h ou l d th enbe i n v ert ed ( th e f l u i d i n e ac h w el l i s h el d i n pl ac e bys u r f ac e t en s i on ) , a n d i n cubat ed f or 45
- 72 hour s at35°±0
. 5°C. B act er i al d ens i tyi s det ermi ned bycoun t i ng the nu mb erof wel l s thatf

l uor es ce un der a 3 6 5 -3 6 6 nm U V ligh t, a n d c o n v e rting this va lue to a M o st P ro b a b le N u m b e r u sin g the U n it D o se M P N tab le p ro v id e d b y the m a n u fac tur e r. If 1 0 -m L sa m p le is u se d , r e a d th e U n it D o se M P N /m L d ire c tly. If a 1 -m L sa m p le i s u s ed, th enc orr ec tth e M PN /mL v alu e bymu l t i pl y i n g i tby10. 5 .5 .1 0 .2 M ul t i p l e D o se (fo r 1 0 s a m p le s o f 1 m L eac h): A 10 0
- m L st eri l e d i l uents ho ul d be ad d ed t o the deh y dr at ed Si mP l at e medi u m t o r ec on s t i tu t e, a n d s h ak ent o di s s ol v e. T h ena 1
. 0
- m L t es ts ampl e s h ou l d be pipe tt e d t o th e c ent ero f a pl a t e c on t a i n i n g 84 s ma l l w e l l s , f ol l ow e d b y9. 0 m L of

th e r e c on s t i tu t e d m e di u m. G ent l ys wi r l pl at e t o m i x the s amp l e a nd med i um, and di s tr i but e the mi xtur e eve nl y t o the 84 wel l s on the pl at e. T hencont i nu e wi th th e pr ocedur e i ndi cat ed i n par agr aph 5. 5. 10
. 1 abo ve, ex ceptthatthe M ul t i

- D ose t abl e s uppl i ed b ythe m anuf

acturer shoul d be us ed t o d et erm i ne the M P N /m L. If a d i l ut i o n i s m ad e d uri ng samp l e p repa rat i o n, then m ul t i p l ythe M P N /m L value by the d i l ut i o n f act o r. 5. 5. 11 (Fo r P o ur P l at e and Sp read P l at e T echni q ues) C o l o ni es s ho ul d be co unt ed m anual l yus i ng a da rk- f i el d co l o ny count er . I n det ermi ni ng sampl e count , l aborat ori es m us to nl yco un tp l at es havi ng 3 0 t o 30 0 col oni es , ex ceptf or pl at es ino c u la te d w ith 1 .0 m L o f un d ilute d sa m p le . C o u n ts les s tha n 3 0 fo r su c h p la te s a re a c c e p ta b le . (F u lly a u to m a tic c o lo n y c o u n te rs a re no t suita b le b e c a u se o f th e siz e a n d sm a ll n u m b e r o f co lo n ie s o b se rv e d w h e n p o ta b le w a te r is a n a lyz e d fo r h e te ro tro p h ic b a c te ria .) QC 5. 5. 12 Eac h ba t chorf l as kof a g ars h ou l d be c h ec k ed f ors t er i l i tybypou r i n g a f i n alc on tr ol pl at e. D at a s h ou l d be r ej ec t ed i f

c on tr ol i s c on t ami n at ed. 5 .6 C ol i pha ge (D raf

tM etho d 16 0 1 and 16 0 2 , pro p o sed G round W at er R ul e) N o t e : E P A M etho d 1 6 0 1 and 1 6 0 2 are p erf o rma nce- b ased method s f o r d et ect i ng the p res ence of m al e- specif i c (F ) and + so m a tic c o lip h a g e in g ro u n d w a te r a n d o the r w a te rs . ( Per forman ce - bas ed me t hod : I n r e c o g n i t i o n o f th e v ar i ety of s i tu at i on s t o wh i chs ome meth ods ma ybe a ppl i ed, a n d i n r ec og n i t i on of c on t i n u i n g t ec h n ol og i ca l a dv an ce s , s ome meth ods ar e per f ormanc e- bas ed. A perf ormanc e- bas ed method permi t s l aborat ori es t o mo di f yo ro mi tst eps o rprocedur es , provi ded thatal l per f

ormanc e r equi r ement s s etf orth i n the val i dat ed methods ar e met
. Anys t eps thatmaynotbe mo di f i ed or omi tt ed m u st b e sp e c ifi e d in the m e tho d .) 5. 6. 1 EPA M et hod 1601: M al e- s pe ci f i c ( F ) and S o mat i c C ol i phage i n W at er by Tw o- St ep Enri ch men t + P rocedure M etho d S um m ary: A 1 0 0
- m L (o r 1
- L w ater s am p l e) i s sup p l em en t ed w i th m ag ne si um ch l o ride , lo g -p h a se ho stV
- 25 amp bact er i a (E . co liF f

o r male- specif

i c co l i p hage and E

. co liC N -1 3 f o r s o m a tic c o lip h a g e ), a n d T ryp tic S o y B ro th (T S B ) as an en richm en tst ep f o r co l i p ha ge . A f

t er an o ve rnight i nc ub ati o n, sam p l es are “ sp o tt ed ” o nto a l aw n of ho st b act eri a specif

i c f o r each typ e o f co l i p hage, i ncub at ed , and exam i ned f o r ci rcul ar l y si s zo nes, whi ch i nd i cat e the pres enc e of

col i phage
. 5 .6 .1 .1 M e d ia 5. 6. 1. 1. 1 An t i bi ot i c s t oc k s
— An t i bi ot i c s mu s ta l w a y s be a dd e d t o m e di u m afterthe med i um ha s been a u to c la v e d . S to re fro z e n a t - 2 0 °C fo r u p to o n e ye a r. T h a w a t ro o m tem p e ra tur e o r ra p id ly i n a w a te r b a th up t o 37 ° C an d m i x w ell p r i or t o us e. P lea se no te: A nt i bi ot i cs m ay be toxi c. W ears ui tab le pr ot ecti ve clothi ng, glove s , and e

ye/ f ace pr otec t i on and us e i n a c hemi cal f um e hood. 5. 6. 1. 1. 2 1 0 X T ryp tic S o y B ro th ( T S B )— S to re a t 1 °-5 °C un tilus e . 5. 6. 1. 1. 3 1
. 5 % T ryp t i c So yA gar ( T SA )— If not used i m m ed i at el yaf t er add i ng ant i b i o t i c and l ett i ng the p l ated m ed i um so l i d i f

y, st o re the p l ates inve rted at 1 °- 5 °C f o r up t o 2 we ek s. 5 .6 .1 .1 .4 0. 7% T SA t op aga r tubes wi th appropri at e ant i bi ot i cs
— D i s pens e 5 mL per s t er i l e 10
- mL tube, l abe l , a n d k ee p a t45°
- 48° C un t i l u s e. T u bes mu s tbe u s ed th e dayth eya r e pr epa r ed. 5 .6 .1 .1 .5 Spotpl at es
— C ondens at i on mayac cumul at e a tthe edges of s t ored s po t pl at es an d m aydrip over aga rs ur f ace i f t i l t ed, rui ni ng the s po tp att ern. I f the s t ored spotp l at es ha ve condens at i on, i ncubat e pl at es f or app r oxi mat el y10 mi nu t es t o r educe condens at i on pri or t o i nocul at i on. Spo t pl at es maybe us ed that d ay o r st o red at 1 °- 5 °C f o r up t o f

o ur d ays. 5 .6 .1 .2 Co l i phage s t ock 5 .6 .1 .2 .1 M S2 ( AT CC #15597- B 1, ma l e- s pec i f i c) a n d ph i

- X 174 s t ockc ol i ph ag e ( AT CC #13706- B 1, s omat i c)

— M aybe s t or ed a t2- 8° C f orup t o 5 y ea r s . Ref

ert o http : //www . at cc. o rg f or i n i t i alpr epa r at i on o f p u re c o lip h a g e sto c k
. 5. 6. 1. 2. 2 A nal y si s o f

raw s ewage f

i l trat e shoul d be gi n wi thi n 24 hours of co l l ect i o n. 5. 6. 1. 2. 3 A l l o w the raw sew ag e to sett l e at 1 °- 5 °C f o r 1 t o 3 ho urs. T his w i l l m ak e the f i l trati o n p ro ce ss easi er. 5. 6. 1. 2. 4 H o l d the ass em b l yove r a 15
- m L p o l yp rop y l ene tub e wi th s cre w -ca p o r snap-cap, i nsertthe p l un ge r i nto the syringe b arre l , an d p ush the sew ag e thro ug h the fi l t er into the st eril e tub e. If f i l t er c l o gs, c h a n g e ita s n e c e ssa ry a n d c o n tinu e to fil te r sew a g e u n til a t le a st 1 0 m L o f f ilt e re d se w a g e is o b ta ine d in the 1 5
- m L p o l yp rop y l ene tub e (f i l trat i o n m ay requir e use of num erous f

i l t ers ). 5 .6 .1 .2 .5 I f

f i l tr at e i s s t ored more than24 hour s , i tmus tbe r e- t i t er ed bef ore us e. 5 .6 .1 .3 H ostbact er i a s t ock c ul tur es 5 .6 .1 .3 .1 F ro z e n ho st b a c te ria sto c kc u lture s— A fterp re p a ra tio n , fre e z e h o st b a c te ria sto c kc u lture s a t - 70 ° C , i f poss i bl e. Cul tur es ca n be f r ozen at- 20 ° C i f

the l aborat orydoes notha ve the capabi l i tyt o f r eez e samp l es at-70 °C
. H o stb act eri a st o red at-70 °C m ay b e ret ai ned for up to o ne y ear. I f s t o red at

- 2 0 °C , the ho stb ac t eria m ay b e re t aine d f o r up t o tw o m o nths. 5. 6. 1. 3. 2 O v e rn igh t ho st b a c te ria sto c kc u lture s— A fterp re p a ra tio n , ch illo n w e t ic e o r a t 1 °-5 °C un til r eadyf or u s e. 5. 6. 1. 3. 3 Log- phas e h ostbact er i a s t ock c ul tur es
— A f t erpre para t i on, ch i l l on we ti ce or a t 1 °- 5 °C t o s l o w repli cat i o n unt i l read yfor use. T he sus p ensi o n m ay b e st o red up t o 48 hours. H o wever, the b est result s o cc ur w he n culture s are use d i m m ed i ately (w i thin 6 ho urs). S t o re re m aining b ac t erial ho st cu l ture at 1 °- 5 °C o ve rnight t o i no cu l ate fl ask s f o r the p rep ara t i o n o f

ne w l o g-p ha se b ac t erial ho st s.V
- 26 5. 6. 1. 4 Ge n e r a l QC 5 .6 .1 .4 .1 In itia l d e m o n st r a tio n o f c a p a b ility (ID C ). T h e la b o r a to ry m u st d e m o n stra te th e a b ili ty to g e n e ra te a c c e

p ta b le p e rfo rm a n c e w ith th is m e th o dby p e rfo rm in ga n I D Cte st b e

fo re a n a lyzin ga ny fi e ld sa mp les. T he ID C t estco nsi st s o f ten reagentwat er s am p l es s p i ked wi th enum erat ed s ewage or eq ui val ent at 1
- 2 P FU p er samp l e f o r each colip hage typ e used, acco rding t o the ID C T ab l e b el o w. A m i ni m um nu mb erof

s amp l es mus tbe posi t i ve, dependi ng o n col i phage type us ed ( s ee I D C T abl e)
. Spi ke s amp l es i n “ b ul k” atco ncentrat i o ns i n the T ab l e. T est s m ustbe acco m p ani ed b ya m etho d blank f o r each colip hage type. 5 .6 .1 .4 .2 M e th o d b la n ks . The labor ator y m us tanal

yz e m ethod blanks (

r eage ntw aters ample contai ni ng n o c o lip h a g e

) t o d e m o n s tr a te fre e d o m fro m c o n ta m in a tio n . Fo r each colip hage typ e used, p repa re and a n a lyz e a ste rile re a g e n t wa te r sa m p le c o n ta inin g n o c o lip h a g e u sin g th e s a m e p ro c e d u re us e d fo r a n a lysis of th e f i el d or Q C s ampl es . Ata mi n i mu m, th e l abor at orymu s ta n al y ze on e meth od bl an kf ore ac h s pot pl at e u s ed f

orf i el d s ampl es . I n a n e f

f ortt o de t ermi n e i f cr os s - con t ami n at i on i s an i s s u e, th e s t er i l e meth od b la n ksh o u ld b e sp o tted o n to the law n o f ho st b a c te ria im m e d ia te ly f o llo w ing the p o sitive c o n tro l sp o t.
5 .6 .1 .4 .3 P o sitiv e c o n tr o ls. The labor ator y m us tanal

yz e pos i t i ve c ontr ols to e ns ur e thats tock c ol i phage su sp e n sio n s, h o st b a c te ria l cu ltu re s, a n dg ro w th m e d ia a re p e rfo rm in gp ro p e rly

. Fore ac h c ol i ph ag e type u s ed, a 100- mL r ea g en tw at ers ampl e mu s tbe s pi k ed wi th 20 PFU f r om s ew ag e f

i l tr at e or60 PFU f

r om a p u re c o lip h a g e sto c kc u lture . T h e lab o ra to ry m u st ino c u la te o n e p o sitive c o n tro l sp o t f o r e a c h sp o t p la te u se d fo r field sa m p le s. If m u ltip le sp o t p la te s a re ino c u la te d w ith sa m p le s o n the sa m e d a y , a sing le e n ric h e d p o sitive c o n tro l sa m p le m a y b e us e d to ino c u la te m u ltip le sp o t p la te s o n tha t d a y. 5 .6 .1 .4 .4 M a tr ix sp ike s (M S ). To as s es s m ethod pe r f orm ance i n a gi ve n s our ce waterm atr i x, the lab oratory m ust

ana l yz e one s et of MS s ampl es f or each col i

ph ag e t ype wh en s ampl es ar e f i rs tr ece i ved fro m a g ro u n dw a te r so u rc e fo r w h ic h th e l a b o r a to ry h a s n e v e r b e fo re a n a lyze dsa mp le s. For e ach col i phage type an al y zed, thr ee f i el d s amp l es ar e s pi ked wi th 1- 2 P FU
. A t a mi ni mum, one outof the thr ee M S sa m p le s fo r e a c h c o lip h a g e typ e m u st b e p o sitive fo r m e tho d p e rfo rm a n c e to b e c o n sid e re d a c c e p ta b le f orth atg r ou n d w at ers ou r ce . I f th e M S r es u l t s ar e u n ac ce pt abl e, an d th e O D C s ampl e an d pos i t i v e con tr ol samp l e res ul t s ass o ci at ed wi th thi s b at ch o f s am p l es ar e accep t ab l e, a m atri x i nt erf erence m ay b e causi ng the po or r es ul t s . In addi t i on, the l aborat orymus tan al y ze one s etof M S s amp l es on anongoi ng bas i s af t er ev ery 2 0 f

i eld sam p l e f

o r e ac h gro un d w ater so urc e. (F o r ex am p l e, wh en a lab o rato ry rece i ve s the f i rst th samp l e f rom a s o urce, the l ab o rat o rym usto b t ai n a d d i t i o nal a l i q uo t s o f the f i el d s am p l es t o be us ed f o r the M S tes t. W h e n the lab o ra to ry re c e ive s the 2 0 fi e ld sa m p le fro m this site , ad d iti o n a l aliq u o ts o f th is sa m p le th m ust be c o l l ec t ed a nd sp i ke d .) M S sa m p l es sho uld b e co l l ec t ed a tthe sa m e t i m e as ro uti ne f i eld sam p l es. Spi ke s amp l es i n “ bul k” atthe conce ntr at i ons i ndi cat ed i n the M S and O D C T abl e bel ow. 5 .6 .1 .4 .5 O n g o in g d e m o n st r a ti o n o f c a p a b ili ty (O D C ). The labor ator y m us tde m ons tr ate thatthe an al yt i cal s ys tem is i n c on tr ol on an on go i ng ba s is through ana l ys is of O D Cs ampl es . Fo r eac h co l i p hage type us ed, thr ee r eag entwat ers amp l es ar e s p i ked wi th 1- 2 P FU
. T he O D C t es ts amp l es ar e a nal y zed exa ct l yl i ke f

i el d s amp l es , an d ata mi ni mum, one outof thr ee O D C t es tsampl es m us tbe posi t i ve f or e ach col i ph ag e type u s ed. I f n ot , me th od per f

orman ce i s un ac ce pt abl e, an d an al y s i s of f i el d s ampl es mu s tbe s t opped. I den t i f

ya n d c orr ec tth e pr obl em an d de mon s tr at e ac ce pt abl e per f orman ce th r ou g h a n al y s i s of ano ther O D C t est b ef

o re co nt i nui ng wi th the anal y si s o f f i el d s am p l es. T he l ab o rat o rym ustanal y ze o ne set o f O D C samp l es af t er every 2 0 f

i el d and M S s am p l es o r o ne p er week, whi chever o ccu rs m o re f r equen t l y . Spi ke s amp l es i n “ bul k” atthe conce ntr at i ons i ndi cat ed i n the M S and O D C T abl e bel ow. 5 .6 .1 .4 .6 P e r fo r m a n c e stu d ie s. T he l aborat orys houl d peri od i cal l yan al y ze an ext ernalQ C s amp l e, s uch a s a p e rfo rm a n c e tes ting sa m p le , w h e n a v a ila b le . T h e lab o ra to ry sh o u ld a lso p a rtic ip a te in a v a ilab le i nt er l aborat orype rf

o rm an ce s tudi es conduc t ed by l ocal , St at e, an d f eder al ag enc i es or com merc i al o rganiza tio n s. T he l ab o rat o rys ho ul d revi ew resul t s, co rr ectuns at i sf act o rype rf

o rma nce, and record co rrec t i ve ac t i o ns.V
- 27

I ni t i alde mons t rat i on of l abor at ory cap abi l i t y ( I D C ) f or M et hod 1601
C ol i phage t ype S a m p le

s i ze 1 T a rg et s pike

con ce nt rat i on ( P F U per s am pl e) “ B ulk”

vol ume t o be s pi ke d B ulk spi ke

con ce nt rat i on (P F U p e r b u lk vo l ume) M i ni mum num ber

of pos i t i ve s am pl es out of 10 F 100- mL 1. 3 1000 mL 13 5 + Soma t i c 100- mL 1. 5 1000 mL 15 5 A 10 0
- m L sam p l e i s requir ed und er the G round W at er R ul e. H o wever, f o r other purp o ses , thi s t estm ay b e used with a 1 1
- L s a m p l e v o l u m e . Be c a u s e I DC s a m p l e s s h o u l d b e a n a l y z e d j u s tl i k e f i e l d s a m p l e s , i n c l u d i n g s a m p l e v o l u m e s , th e I DC a n a l y s e s s h o u l d b e p e r f o rm e d a tth e 1
- L s a m p l e v o l u m e w h e n th e l a b o r a t or y i s e v a l u a t i n g 1
- L s a m p l e s . ( Th e I DC p ro c e d u re fo r 1 -L sa m p le s is p ro v id e d in the p ro to c o l t o M e tho d 1 6 0 1 , T a b le 1 .) M S an d O D C s am pl e s pi ki ng requi rement s f or ong oi ng eva l uati on o f M et hod 160 1 performa nce C ol i phage t ype S a m p le

s i ze 1 T a rg et s pike

con ce nt rat i on ( P F U per s am pl e) N umb er of s am pl es

t hat mustbe s pi ke d ( 

### $ 1 mus t be posi t i ve) “ B ulk”

vo l ume t o be s pi ke d B ulk spi ke

con ce nt rat i on (P F U p e r b u lk vo l ume) F 1 0 0 -m L 1 .3 3 3 0 0 -m L 3 .9 + S o m a tic 1 0 0 -m L 1 .5 3 3 0 0 -m L 4 .5 A 10 0
- m L sam p l e i s requir ed unde r the G round W at er R ul e. H o wever, f o r o ther p urpo ses , thi s t estm ay b e used with a 1 1
- L sam p l e v o l um e. B ec au se O D C an d M S sam p l es s ho uld b e ana l yze d just l i ke f i eld sam p l es, i nc l ud i ng sam p l e v o l um es, the O D C an d M S an alyses sho uld b e p erfo rm ed a tthe 1
- L sam p l e v o l um e w he n the lab o rato ry i s eva l ua t i ng 1
- L sam p l es. (T he M S and O D C pro ced ure f o r 1- L sam p l es i s p rovided i n the p rotoco l t o M etho d 16 0 1 , T ab l es 2. ) 5. 6. 2 EPA Me th o d1 6 0 2 : Ma le - s p e c i f ic ( F ) a n dS o m a tic Co li

p h ag e in Waterby Sin g le La y e rAg a r(

S AL ) + P ro ce d u re M etho d Sum m ary : M etho d 16 0 2 i s a p erf o rma nce- b ased m etho d f o r det ect i ng or enum e ra ti ng m al e- specif i c (F ) and + som at i c col i ph ag e i n g r ou n d wa t er an d oth er wa t er s . A 100- mL g r ou n d wa t er s ampl e i s as s ay ed by addi n g m agnesi um
amp chl ori de an d hos tbact er i a (E . co li F f o r F c o l i p h ag e an d E . c o liC N
- 1 3 f o r s o m at i c co l i p hage), and then ad d i ng the + s amp l e/hos tb act er i a m i xtur e t o 10 0 mL of d oubl e- s tr eng th m ol t en T rypt i c SoyAgarcont ai ni ng the appropri at e a nt i bi ot i c. T he s amp l e i s thor ough l ymi xed a nd the t ot al vol ume is p o ure d into 5 to 10 pl at es ( dependenton pl at e s i ze)
. Af t eran overni gh ti ncubat i on, an yci r cul arl y s i s zone s ( pl aques ) i ndi cat e the pres enc e of col i phage
. 5 .6 .2 .1 . M e d ia

-- P le a se re fer to S e c tio n 5 .6 .1 fo r an tib io tic sto c k s, 1 0 X T ryp tic S o y B ro th (T S B ), 1 .5 % T ryp tic S o y A ga r (T S A ), 0 .7% T S A t o p ag ar tub es w i th a p p ro p riat e a nti b i o t i cs, and sp o tp l ates.
5.6.2 .1 .1 D o ub l e Str ength T ryp t i c So yA gar (2 X T SA )— M ed i um m ay b eco m e d arker af t er aut o cl avi ng butthi s s houl d notaf

f ectmedi a per f

ormanc e. 5 .6 .2 .1 .2 2X T SA wi th appropri at e ant i bi ot i cs
— K eep mo l t en at 45 °
- 48 ° C i n wat er bath un t i l us e. Agar m ustbe us ed the d ay o f prep arat i o n. 5.6.2 .2 C o l i p hage st o ck— P l ease ref er t o Sec t i o n 5.6.1. 2 f

o r col i p hage st o ck. 5 .6 .2 .3 H o stb ac t eria stoc k c ulture s — P l ea se re f er to S ec t i o n 5 .6.1 .3 fo r ho stb ac t eria stoc k c ulture s. 5 .6 .2 .4 Ge n e r a l QC 5 .6 .2 .4 .1 I n i t i a l p r e c i s i o n a n d r e c o v e r y ( I PR )
. T h e la b o ra to ry m u std em o n stra te th e a b i l i t y to p erfo rm this m etho d acceptab l

y by p er f orm i ng a n IPR tes tbef

ore ana l yz i ng any f

i eld s ampl es . Fourr eag entwat erV
- 28 sa m p le s fo r e a c h c o lip h a g e typ e a re re q u ire d fo r the I P R te st. I P R s a m p le s m u st b e s p ike d in b u lk to yield a t argets p i ke co ncentrat i o n of 8 0 P FU pe r s am p l e. I P R s am p l es mustbe s p i ked wi th enum erat ed s ewage
r f i l trat e o r equi val ent
. T he rel at i ve st and ard d evi at i o n of the r e c o v e ry (R SD ) and the average p ercent re c o v e ry ( 

### 0 ) b a se d o n a ll fo u r sam p le re su lts fo r ea c h c o lip h a g e typ e sh o u ld m e e t th e a c c e p ta n c e c rite ria i n the Q C ac cept anc e t abl e bel ow. 5 .6 .2 .4 .2 M eth o d b l a n ks. The labor ator y m us tanal

yz e m ethod blanks (

r eage ntwaters ample contai ni ng n o c o lip h a g e

) to d e m o n stra te fre e d o m fro m co n ta m in a tio n . T he l ab o rat o rym ustanal y ze o ne m etho d blank wi th e ach an al y t i calbat ch. For eac h co l i phage type us ed, pre pare an d ana l y ze a s t er i l e r eag entwat er sam p l e co ntaini ng no co l i p ha ge using the sa m e p ro ce d ure as use d f o r an alysi s o f

the f i eld o r Q C sam p l es. A n a nal y t i calbat ch i s def i ned a s al l sampl es anal y zed d ur i ng a s i ng l e day , up t o a maxi mum o f 20 sa mp l es ( f i el d s amp l es an d m atr i x s pi ke s amp l es ) percol i phage type. 5 . 6 . 2 . 4 .3 M a tri x s pikes (M S). T o a ssess m eth o dp erfo rm a n ce i n a g i ve n m a tri x, th e la b o ra to ry m u st anal yz e on e seto

f MSs amples f

ore ach coli

phage t ype when s amples ar e f i rs tr ec ei ve d f

r om a ground w a te r so u rc e fo r w h ic h th e la b o ra to ry h a s n e v e r b e fo re a n a lyze dsa mp le s. T he M S ana l y s i s i s per f ormed o n an ad d i t i o nal (s eco nd ) s am p l e al i q uo tco l l ect ed f rom the gro und w at er source at the same ti m e as the ro uti ne f i eld sa m p l e. If

the lab o rato ry ro uti ne l y a na l yze s sa m p l es f ro m o ne o r m o re gro un d w ater so urc es, o ne M S an alysi s m ust be p erfo rm ed p er 2 0 f i eld sam p l es. Fo r ex am p l e, wh en a lab o rato ry rece i ve s the f i rst s ampl e f r om a s ou r ce , th e l abor at orym u s tobt ai n a s ec on d al i qu otof th i s s ampl e t o be us ed f orth e M S. W h enth e l abor at oryr ec ei v es th e 20 s ampl e f r om th i s s i t e, as epa r at e al i qu otof th i s 20 s ampl e mu s tbe th th c o llec te d a n d sp ike d . C o m p a re the c o lip h a g e re c o v e ry w ith the c o rr e sp o n d ing li m its i n the Q C T a b le bel ow. I f

the r ecove ryf

or col i phage f

al l s out s i de i t s l i mi t , method perf ormanc e i s un acc ept abl e f or that samp l e. I f

the res ul t s f

o r the O P R sam p l e ass o ci at ed wi th thi s b at ch o f s am p l es ar e wi thi n thei r r espec t i ve c o n tro l l im its, a m a trix in te rfe re n c e m a y b e c a u sin g p o o r re c o v e ry. If the re su lts f o r th e O P R a re n o t with in th ei rcon tr ol l i mi t s , m eth od per f orman ce i s un ac ce pt abl e ( s ee Sec t i on 5. 6. 2. 4. 4)
. T h e pr obl em s h ou l d be i d ent i f i ed and co rr ect ed , an d th e m a trix s p i ke and ass o ci at ed f i el d s am p l e(s ) s ho ul d be qua l i f i ed
. T he re c o v e ry sh o u ld b e m a inta ine d o n a c o n tro l ch a rt a n d up d a te d o n a re g u la r b a sis. 5 .6 .2 .4 .4 O ng o i ng pre ci si o n a nd reco v ery (O P R ). T h e la b o ra to ry m u st

, o n a n o n g o i n gb a sis, d e m o n stra te a c c e

p ta b le p e rfo rm a n c e th ro u g h a n a lysis o f a n O PR sa mp le
. Fore ac h c ol i ph ag e type u s ed, a r e a g e n twat er s am p l e i s spi ked wi th ap p roximatel y80 P FU
. T he O P R i s anal y zed exact l yl i ke a f ie ld sa m p le . T h e lab o ra to ry m u st a n a lyz e o n e O P R sa m p le fo r ea c h a n a lyti c a l b a tc h . A n a n a lyti c a l b a tc h is d ef i ned as al l s am p l es anal y zed duri ng a s i ngl e d ay, up t o a m axi m um of 2 0 s am p l es ( f i el d s a m p l es and m a trix s p ike sa m p le s) p e r c o lip h a g e typ e us e d
. P leas e note: the O PR s erve s as the pos i t i ve co n trolf or Method 1602. Compar e th e O PR pe r ce n tr ec ov ery( R) w i th th e corr es pon di n g l i mi t s f oron g oi n g pr ec i s i on a n d re c o v e ry in the Q C T a b le b e lo w . If R m e e ts th e a c c e p ta n c e c rite ria , sys te m p e rfo rm a n c e is ac c e p ta b le a n d a n a lysis o f sam p le s m a y c o n tinu e . If R falls o u tsid e the ra n g e fo r re c o v e r y , m e tho d p e rfo rm a n c e is una c c ep t ab l e, and anal y si s o f f i el d s am p l es mustbe s t o p p ed
. Ident i f yand co rr ectthe p rob l em and d em o nstrat e accep t ab l e p erf o rm a n c e th ro ugh s uccessf ul anal y si s o f

ano ther O P R t estbe f

o re conti nui ng w i th the a na l ysi s o f

f i eld sam p l es. 5 .6 .2 .4 .5 P er fo rm a n ce stu d i es. T he l aborat orys houl d peri od i cal l yan al y ze an ext ernalQ C s amp l e, s uch a s a p e rfo rm a n c e tes ting sa m p le , w h e n a v a ilab le . T h e lab o ra to ry a lso sh o u ld p a rtic ip a te in a v a ilab le i nt er l abora t oryper f

ormanc e s tudi es conduc t ed by l ocal , s t at e, an d f eder al ag enc i es or com merc i al o rgani zat i o ns. T he la b o ratorys ho ul d revi ew resul t s, co rr ectuns at i sf act o rype rf

o rma nce, and record co rrec t i ve ac t i o ns.V
- 29

Q ual i t y c ont rolac ce pt anc e cr i t er i a f or M et hod 1602
P er fo rm a n ce test M a le -s p e c ific accept ance c r ite r ia S o m a tic

accept ance c r ite r ia I n i t i a l pr e c i s i ona n d r e c ov e ry( I PR)

• M e a n p e rc e n t re c o v e ry

• P reci si o n (as m axi m um rel at i ve st and ard d evi at i o n) 9% - 130% 86% - 177% 46% 23% O n g oi n g pr ec i s i on a n d r ec ov ery( O PR ) a s pe r ce n tr ec ov ery 4% - 135% 79% - 183% M atr i x s pi k e ( M S)

• M S pe r ce n tr ec ov ery D et ec t- 120% 48% - 291% M a tr i x s p i k e , m a tr i x s p i k e d u p l i c a t e ( M S /M S D)

• M ea n pe r ce n tr ec ov eryf orM S/M SD D et ec t- 120% 48% - 291%

• Pre ci s i on ( as ma x i mu m r el at i v e per ce n tdi f f er en ce of

M S/M SD ) 57% 28% 

## 6. Sample Collection, Hand ling, an d Preservation P ar agr aphs 6. 1- 6. 5 ar e app l i cabl e t o thos e l aborat ori es thatcol l ects amp l es . H owever , al l l aborat ori es sho uld m ake an e ff o rt to e n su re p ro p e r sa m p le c o llec tio n ; all lab o ra to rie s a re re sp o n sib le fo r p a ra g ra p h 6 .6
. 6. 1 Sample Collec tor T h e sa m p le c o llec to r sh o u ld b e tra in e d i n a s e p tic sa m p ling p ro c e d u re s a n d , i f re q u ire d , ap p ro v e d b y the a p p ro p ria te r egu l at oryau thor i tyor i t s des i gn at ed r epre s ent at i ve. 6. 2 Sampling 6 .2 .1 ( Fo r T C R ) Sa m p l es mustbe represent at i ve o f the wat er di strib ut i o n s y st em . W a te r tap s used f o r s am p l i ng s h ou l d be f

r ee of a er at or s , s tr ai n er s , h os e att ac h men t s , mi x i n g type f au ce t s , a n d pu r i f

i ca t i on dev i ce s . Col d w at ert aps s houl d be us ed. T he s ervi ce l i ne mus t be cl ear ed bef ore s amp l i ng by mai nt ai ni ng a s t eady wat er f l ow f or a tl eas ttwo m i nu t es (un t i l a st ea d y w ater t em p era ture i s ac hiev ed ). A t l ea st 1 0 0 m L o f sam p l e m ust b e co l l ec t ed , all o w i ng at l ea st a 1- i n ch ai r s pac e t o f

ac i l i t at e mi x i n g of

th e s ampl e bys h ak i n g . I mmedi at el ya f t erc ol l ec t i on , a s ampl e i n f ormat i on fo rm sh o u ld b e c o m p le te d (se e p a ra g ra p h 6 .5 ). S e e S e c tio n 3 .1 5 .4 re g a rd ing sa m p le d e c h lo rin a tio n . If a sa m p le b o ttl e i s f i l l ed t o o ful l to al l o w f

o r p rop er m i xi ng, d o no tp o ur of f and dis card a po rt i o n of the samp l e. R ather, po ur the ent i re sam p l e into a l arg er steril e c o ntainer, m i x p ro p erly, an d p ro ce ed with the a na l ysi s. 6 .2 .2 ( ForSW T R) Sou r ce w at ers ampl es mu s tbe r epr es en t at i v e of th e s ou r ce of

s u ppl y , col l ec t ed n ott oo f arf r om the p o int o f i n ta k e , b u t at a r e a so n a b le d istan c e fro m th e b a n ko r sh o re . T h e sa m p le vo lum e sh o u ld b e su ff ic ie n t t o per f orm a l l th e t es t s r equ i r ed. 6 .2 .3 ( For col i phage an al y s i s un der G W R ) A 10 0- mL sa mp l e v ol ume i s r equi r ed f or the as s ay . Co l l ect i on of an ad d i t i o nal 10 0
- m L wa t er s am p l e wo ul d al l o w f o r sa m p l e re- anal y si s, i f necess ary(e. g. , i f

the p o si t i ve o r negat i ve c o n tro ls fail). T o e n su re su ff ic ie n t sa m p le vo lum e , an a d d iti o n a l 5 0 -m L w a te r sa m p le sh o u ld b e c o llec te d
. 6 .2 .4 ( Fo r E . co lia n d e n te ro c o c c i un d e r G W R ) A 1 0 0 -m L sa m p le vo lum e is re q u ire d fo r th e a ssa y.
6. 3 Sample I

cing 6 .3 .1 ( B act eri al s am p l es) Sam p l ers are encouraged , but no trequi red, t o ho l d d ri nki ng wat er s am p l es at< 1 0 °C duri ng tra n sit t o the lab o ra to ry. S o u rc e w a te r sa m p le s re q u ire d b y the S u rfa c e W a te r T re a tm e n t R u le (S W T R ) m u st b e he ldV
- 30 at <1 0°C dur i ng tr ans i t( s ee Standard Methods , Sect i on 906 0B )
. Labora t ori es s houl d re j ects amp l es thatha ve been fro z e n . 6. 3. 2 ( For col i phage anal y s i s underG W R ) S hip s amp l es at<1 0°C us i ng weti ce, B lue I ce ® , o r sim ilarp ro d u c ts to m ai nt ai n t em p erature, and s t o re at1°- 5 °C
. Sam p l es s ho ul d not b e f roze n. Q C 6 .3 .3 For SW T R s amp l es an d col i phage s amp l es , sampl e t emp er atur e upo n re ceiptsh oul d be r ecor ded. A s amp l e that h a s a tem p e ra tur e up o n re c e ip t o f > 1 0 °C , wh e th e r ice d o r n o t, sh o u ld b e fl a g g e d un le ss th e ti m e sinc e sa m p le co l l ec t i o n ha s b ee n less tha n two ho urs. 6.4 S a mp le Ho ldin g

/T ra v e l T im e 6 .4 .1 Forth e an al y s i s of t ot alc ol i f orms i n dr i n k i n g w at er , th e t i me betwe ens ampl e col l ec t i on a n d th e pl ac eme n tof sa m p le in the inc u b a to r m u st no t ex c e e d 3 0 ho u rs (p e r re g u la tio n a t 4 0 C F R 1 4 1 .2 1 (f)( 3 )) . A ll sa m p le s re c e ive d in the l ab o rat o rys ho u ld b e a n a lyzed on the d ay o f recei p t
. I f

the l ab o rat o ryrecei ves the samp l e l at e i n the d ay, the samp l e m ay b e ref

ri gerat ed ove rni ghtas l o ng as anal y si s b egi ns wi thi n 30 hours of s am p l e co l l ect i o n. 6 .4 .2 T he t i me f

r om s amp l e c ol l ect i o n t o p l ac em en tof the s amp l e i n the i ncubat or f or t ot al col i f

orms an d f

eca l col i f o rm s in surf

ace wat ers our ces , an d het er otr op hi c bact er i a i n dr i nk i ng wat er , mus tnotex cee d ei gh thour s ( per re g u la tio n a t 4 0 C F R 1 4 1 .7 4 (a )( 1 ))
. 6 .4 .3 (Fo r co l i p ha ge an alysi s) T he t i m e b etwe en sam p l e c o l l ec t i o n a nd the p l ac em en to f sam p l e i n the i nc ub ato r m ust notex cee d 48 hour s . T he t i me f r om s ewag e s amp l e col l ect i on t o ana l y s i s o f Q C spi ki ng su s pens i ons maynotex cee d 2 4 h o u r s , u n l e s s r e - t i t e r e d a n d t i t e rh a s n o td e c r e a s e d b ym o r e th a n 5 0 %. I f t i t e rh a s n o td e c r e a s e d b ym o r e th a n 5 0 %, the sam p l e c an b e sto red f o r up t o 7 2 ho urs. 6 .4 .4 ( Fo r E . co lia n d e n te ro c o c c i un d e r G W R ) T h e t im e b e tw e e n sa m p le c o llec tio n a n d the p la c e m e n t o f sam p le in the i nc ub ato r m ust no tex ce ed 3 0 ho urs. 6.5 S a mp le In

fo rm a tio n Fo rm A f t er coll ect i o n, the samp l er s ho ul d ent er on a sam p l e i nf o rma t i o n f

o rm, i n i nd el i b l e i nk, the f o l l o wi ng i nf

o rma t i o n:
• N ame of

s y s t em ( publ i c wa t ers y s t em s i t e i dent i f i cat i on nu mb er , i f

av ai l abl e) • S a m p le id e n tif

ic a tio n (if an y)

• Sam p l e si t e l o cat i o n
• Samp l e type ( e. g. , r out i ne di s tr i but i on s y s t em s amp l e, r epeats amp l e, r aw orproces s wat er , others peci al purpo s e s amp l e)

• D at e an d t i me of

c ol l ec t i on
• Anal y s i s r eques t ed

• D i s i nf ect antr es i dual

• N ame of

s amp l er
• A nyremarks 6. 6 Chain- of

- Cus tody S a m p le c o llec to rs a n d lab o ra to rie s m u st fo llo w a p p lica b le S ta te re g u la tio n s p e rta inin g to c h a in-o f- c u sto d y. A n e x a m p le of s u c h a pl a n i s pr ov i de d i n Ap pe n di x A. 

## 7. Quality Assurance 7 .1 A w r i tt enQ A pl an s h ou l d be pr epa r ed an d f ol l owe d ( s ee Chapt erI I I )
. T h e Q A pl ans h ou l d be a v ai l abl e f ori n s pec t i on byth e ce r t i f

i ca t i on of

f i ce r . As s pec i f

i ed byth e Q A pl an , a l abor at oryth atper f orms i t s own ca l i br at i on of e qu i pmen tor su p p lies ( e .g., the rm o m e te rs ) sh o u ld ha v e a S ta n d a rd O p e ra ting P ro c e d u re a v a ilab le fo r re v ie w . If a lab o ra to ry w ish e s to p e rfo rm a d d iti o n a l Q A b e yo n d tho se in th is m a n u a l, the la b o ra to ry m a y re ferto Standard Methods , Se ct i on 9020, Q u a lit y As s ur ance (

Q ual i t y As s ur ance/

Q ual i t y Contr ol , i n 2 0 e d .). th 7. 2 St at es a r e en cour ag ed t o e s t abl i s h pr of i ci en cyt es t i ng (P T ) a s pa rtof th ei rdr i n k i n g w at erc ert i f i ca t i on pr og r am f

or mi cr ob i ol ogy . A l aborat orys houl d s ucc es s f ul l yan al y ze atl eas to ne s etof P T s amp l es once ev ery12 m onths , f or e ach m e tho d fo r w h ic h itis ce rtifi e d
.V
- 31

F o r m e tho d s u se d to te st the p re se n c e o r ab se n c e o f an o rg a n ism in a sa m p le , ea c h P T se t sho u ld c o n ta in te n sa m p le s, a ll ship p ed atthe same ti m e i n ei ther a l y o p hi l i zed , d ehydrat ed , o r aq ueo us st at e. T he setsho ul d i ncl ud e samp l es, i n vari o us c o m b ina tio n s, tha t co n ta in to ta l co lif o rm s, fec a l co lif

o rm s, E . co li , no n -co lif o rm s, a n d a t le a st o n e b la n k
. E a c h se t sho u ld b e used o nl yw i th a si ngl e anal y t i cal m etho d
. T o b e accep t ab l e, a l ab o rat o rys ho ul d co rr ect l yanal y ze a m i ni m um of ni ne o f the t en samp l es, wi th no f al se- negat i ve res ul t(i
. e. , a s i ngl e f al se- p o si t i ve res ul tm ay b e accep t ab l e). B e c a u se e v e n m e tho d s b a se d u p o n th e s a m e p rin c ip le ( e .g., m e m b ra n e filtra tio n ) m a y b e q u ite d issim ilar , a R e g io n o r S ta te s houl d cons i der cert i f

y i ng a l aborat oryonl yf or thos e s peci f

i c methods f

or w hi ch the l aborat oryha s s ucc es s f ul l yan al y zed a s etof P T s amp l es . T he T abl e bel ow r ef l ect s thi s approach , and i dent i f i es the f ew methods that maybe s uf f i ci ent l ys i mi l ar t o all o w a lab o rato ry t o b e c erti f i ed f o r m o re than o ne m etho d up o n suc ce ssf ul co m p l eti o n o f

a si ng l e set o f

P T sam p l es.
M et hod Cate gory Spe ci f i c M et hod 1 F e rm e n t a t i o n b r o th m e th o d L TB o rP
- A b r o th , f o l l o w e d b yBGL B a n d e i th e rE C o rE C
- M UG F e rm e n ta tio n b ro th m e tho d A -1 b ro th ( fec a l co lif o rm , S W T R o n ly) Enz y me s u bs tr at e meth od Col i l ertorCol i l ert18 E nzyme substrat e m etho d C o l i sur e E n z y m e s u b s tr a t e m e th o d R e a d y c u l to rF l u o r o c u l tL MX E n z ym e su b stra te m e tho d E *C o lit e E nzyme s ubs tr at e method C ol i t ag M e m b r a n e f

i l t e rm e th o d M
- En d o o rL ES E n d o , f o l l o w e d b yBGL B a n d e i th e rE C , E C
- M UG, o rNA
- M UG M em b rane f

i l t er method M I M ed i um M emb r ane f

i l t ermethod C ol i s can M embr an e f

i l t erme th od m- Col i B l u e24 M e m b ra n e fi lterm e tho d C h ro m o c u lt M e m b ra n e fi lterm e tho d m F C a g a r (fe c a l co lif o rm , S W T R o n ly) H PC me th od PCA H P C m e tho d S im P la te Separate set of

prof ic iency test sam ple s re com

mende d for

each c ell. A

singleset of PT sam

ple s would cover

ever y met hod within the 1 sa me cell. 

## 8.Re c or ds a nd D at a Reporting 8. 1 L egalD ef

ens ibilit y: C o m p lian c e m o n ito rin g d a ta sh o u ld b e m a d e leg a lly d e fen sib le b y ke e p ing tho ro u g h a n d a c c u ra te r ec or ds
. T h e Q A pl ana n d/ orSO Ps s h ou l d de s cr i be th e pol i ci es a n d pr oce du r es u s ed byth e f ac i l i tyf orr ec or d r et en t i on and s t orag e. I f

s amp l es ar e expec t ed t o bec om e part of a l ega l act i on, cha i n- of

- cus t od y procedur es s houl d be u s ed ( See A p p end i x A ). 8. 2 Mainten anc e of

Rec or ds : Publ i c wa t ers y s t ems a r e r equ i r ed t o ma i n t ai n r ec or ds of mi cr obi ol og i cala n al y s es of co m p l i ance sam p l es f

o r f i ve y ears (40 C FR 1 4 1
. 3 3 ). T he l ab o rat o rysho ul d m ai nt ai n easi l yaccessib l e reco rds f o r f i ve y ears oru n t i l th e n ex t ce r t i f

i ca t i on da t a au di ti s c ompl et e, w h i ch ev eri s l on g er . A c h an g e i n ow n er s h i p, me r g er , orc l os u r e of a lab o rato ry do es no tca nc el this req uirem en t
. T he cli en twa t er syst em sho uld b e n o t i f i ed b efo re d i sp o si ng o f rec o rd s so theymayr eques tcopi es i f ne eded. T hi s i ncludes al l raw dat a, cal cul at i ons , an d qual i tycontr ol dat a. T hes e dat a f i l es may b e ei ther hard co p y , m i crofi che o r el ectronic. E l ectronic d at a shoul d al ways be ba cked up b ypro t ect ed t ap e o r di sk o r har dV
- 32

c o p y. If the lab o ra to ry c h a n g e s its c o m p u te r ha rd w a re o r so ftwa re , it sh o u ld m a k e p ro v isio n s fo r tra n sfe rr ing o ld d a ta to th e n e w s y s t e m s o th a ti tr e ma i n s r e tr i e v a bl e w i th i n th e t i me f r a me s s pe c i f

i e d abo v e . Da t a w h i c h i s e x pe c t e d t o b e c om e partof a l ega l ac t i on wi l l probabl yne ed t o be m ai nt ai ned f or a l onger peri od of t i me. Checkwi th y ourl ega l coun s el
. See G oo d A ut om at ed Lab oratory P r act i ces , E PA 2 1 8 5 , Of f i c e o f I n f o rm a t i o n M a n a g e m e n t , R e s e a r c h Tr i a n g l e P a rk , NC 27711, 8/ 10/ 95.
8. 3 Sampling Rec or ds : D at a s houl d be re corded i n i nkwi th any cha ng es l i ned thr ough s uch that ori gi nal entryi s vi s i bl e. C han ges s h ou l d be i n i t i a l e d a n d da t e d. T h e f ol l ow i n g i n f

orma t i on s h ou l d be r e a di l y a v a i l a bl e i n a s u mma ry or other reco rd(s) : 8 .3 .1 Sam p l e i nf

o rma t i o n f

o rm, f rom 6.5 ab o ve 8 .3 .2 D at e and t i m e o f s am p l e recei p tby the l ab o rat o ry 8 .3 .3 N a m e o f l a b o ra to ry p e rs o n re c e ivin g the sa m p le 8 .3 .4 A ny d efi cien cy i n the c o nd i t i o n o f the sam p l e. A sam p l e sho uld b e inva l i d ated f o r the f o l l o w i ng rea so ns:

- T i me betweens amp l e c ol l ect i on an d re ceiptbyl aborat oryha s been ex cee ded

- P res ence o f

dis i nf ect anti n s am p l e i s no t i ced (e. g. , od o r)

- E vi d ence o f

f reezi ng

- U s e of a c ont ai nernotapproved bythe l aborat oryf or the purpo s e i nt ended

- I n s u f f i ci en ts ampl e v ol u me ( e. g . , <100 mL) - P re se n c e o f i n te rfe rin g c o n ta m ina n t, if n o tice d (e .g., h yd ro c a rb o n s, c le a n se rs , he a v y m e ta ls, etc .) - S a m p le tem p e ra tur e e x c e e d s the m a x im u m a llo w a b le 8. 4 Anal ytical Rec or ds : D at a sh oul d be r ecor ded i n i nkwi th a nych ang es l i ned thr ough s uchthatori gi nalen tryi s vi s i bl e. C han ges s houl d be i ni t i al ed an d dat ed. T he f ol l owi ng i nf

ormat i on s houl d b e rea d i l y a vai l abl e i n as ummaryor other reco rd(s) : 8 .4 .1 Labor at orys ampl e i den t i f i ca t i on 8 .4 .2 D at e and t i m e anal y si s b egi ns 8 .4 .3 L a b o ra to ry a n d a sign a tur e o r in iti a ls o f p e rs o n (s ) p e rfo rm ing a n a lysis 8 .4 .4 Anal y t i calt ech ni que ormethod us ed 8 .4 .5 A l l i t e m s m a rk e d QC 8 .4 .6 Res ul t s of

an al y s es 8. 5 P r eve ntive Maintenance La b o ra to ries s ho ul d m ai nt ai n preventi ve m ai nt enance and repairact i vi t i es r eco rds f o r al l i nstrume nt s and eq ui p m ent ( i n clu di n g pH met er s , an al y t i ca l bal an ce s , i n cubat or s , r ef r i g er at or s , a u t ocl av es , a n d w at erba th s )
. Rec or ds s h ou l d be k ept f o r f i ve y ears i n a m anner thatal l o ws f o r eas yi nspecti o n. 

## 9. Action Respon se to Laboratory Results 9. 1 Tes ting Total

Co li f orm -P ositive C ultur es F o r the T o t al C ol i f

orm R ul e, l aborat ori es mus tt es tal l t ot al col i f orm- po s i t i ve cul tur es f or the pres enc e of

ei therf eca l co l i f o rms or E . co li
. 9 .2 No tifica tio n o

f P o sitive R e su lts 9 .2 .1 Fo r th e T ot a l Col i f orm Ru l e , l a bo r a t or i e s mu s t pr om pt l y n ot i f y th e pr op e r a u th or i ty of a po s i t i v e t ot a l c ol i f orm, f ecal co l i f

o rm, or E . co lir es ul t , so thatappr opr i at e f ol l ow- u p a ct i on s ( e. g . , c ol l ec t i on of r epe ats ampl es ) c anbe c o n d u c te d (se e 4 0 C F R 1 4 1 .2 1 (b ) a n d (e ), a n d 1 4 1 .3 1 , etc .). 9 .2 .2 I f anys am p l e i s f

ecal co l i f

o rm- o r E . co li

- po s i t i ve, “ the s y s t em mus tnot i f ythe St at e bythe end of the daywh en the syste m is no tif

ie d o f th e tes t re su lt, un le ss th e syste m is no tif ie d o f th e re su lt a fterthe S ta te o ff ic e is clo se d , i n w h ic h c a se the syste m m u st no tif y the S ta te b e fo re the e n d o f th e ne x t b u sin e ss d a y.” (4 0 C F R 1 4 1 .2 1 (e )( 1 ))
.V
- 33 9 .2 .3 A t o t al co l i f

o rm-po si t i ve res ul t i s b ased on the co nf i rme d p hase i f

the M ul t i p l e T ub e Fe rme nt at i o n T echni q ue orPr es en ce - Abs en ce ( P- A) Col i f orm T es ti s u s ed, orth e v er i f i ed t es tf orth e M embr an e Fi l t erT ec h n i qu e i f M
- Endo m e d ium o r L E S E n d o a g a r is u se d . N o re q u ire m e n t ex ists to c o n firm a to ta l co lif o rm -p o sitive re su lt u sin g C o lil e rt, C o lisur e , M I a g a r, E *C o lit e , M I a g a r, m -C o liB lu e 2 4 , C h ro m o c u lt, R e a d yc u lt/F luo ro c u lt, C o lisca n , o r C o lit a g tes t. A l so, no requir em ent exi st s t o co nf i rm a p o si t i ve f

ecal co l i f

o rm o r E . co lites t
. In tho se r are cases where a presum p t i ve t ot alcol i f

orm- pos i t i v e c u l tu r e doe s notcon f i rm/v er i f yas s u ch , buti s f

ou n d t o be f

ec alc ol i f

orm orE . co li

- po s i t i ve, th e s a mp l e i s c on s i de r e d t ot a l c ol i f orm- po s i t i v e a n d f

e c a l c ol i f

orm/E . co li

- po s i t i ve. 9. 3 Noti f ication o f Total Co li f orm I nter f erence For the T ot al Co l i f

orm R ul e, the l aborat orymus tpromp t l y no t i f ythe proper author i ty( us ual l ythe wat ers y s t em) whe n r es u l t s i n di ca t e t hatn on - col i f orms ma yh av e i n t er f er ed wi th th e t ot alc ol i f orm a n al y s i s , a s de s cr i bed i n 40 CFR 1 4 1
. 2 1 (c)( 2 ).V
- 34



# Ex amp le C h ecklis tsfo r On -s ite E valu ationof Lab orato riesAn alyzin g D rin kin g W at er

## MICRO CHECKLI STS

V
- 35 G en er alA udi t I nf

ormat i on L a b o ra to ry:
M ai l i ng A d d res s (ma i l i ng ad d res s o f ow ner i f dif f erent ): Str eet
Ci ty , St at e, Zip c ode
A ud i tLo cat i o n (i f

dif f erent ):
T el ephone:
Fa x:
E -m a il:
O ther:
A ud i tO rgani zat i o n:
A ud i t o rs/ S i gn ature s:

A ud i tD at e(s ):

## MICRO CHECKLI STS

V
- 36 L abo ratory P ers onn el
P osi t i on/ T i t l e N am e E ducati on L evel D egr ee / M aj or Speci al i zed Trai ni ng Pre s en t

Sp eci a l ty E xperi ence,

i nc l udi ng # yr s at c ur re nt pos i t i on La b o rat o ry Superv i s or La b o rat o ry C o nsul t ant P rima ry A na l yst
A n a lyst 2
A n a lyst 3
A n a lyst 4
O thers

## MICRO CHECKLI STS

V
- 37 E l em en t N um be r Y es N o N A C o m m en ts 1. PERSO N N EL S u p e r v iso r /C o n su lta n t 1 .1 D o e s the su p e rv iso r o f th e m ic ro b io lo g y lab o ra to ry h a v e a b a c h e lo r’s de g r e e i n mi c r ob i ol og y , bi ol og y , ore qu i v a l e n t ? H as a s upervi s or w i th adegr ee i n as ubj ectotherthanthos e l i s t ed ab o ve ha d at l ea sto ne co l l eg e-l ev el m i cro b i o l o gy l ab o rato ry co urse i n w h i c h e n v i r on me n t a l mi c r ob i ol og yw a s c ov e r e d? In add i t i o n, has the supervi sor had a m i ni m um of two weeks trai ni ng a t a F e d e ra l o r S ta te a g e n c y o r a c a d e m ic institutio n in mi cr obi ol og i ca l a n al y s i s of dr i n k i n g w at eror80 h ou r s of on - th e- j ob tr ai ni ng i n w at ermi cr ob i ol ogyata c ert i f i ed l aborat ory , or other tr a i n i n g a c c e pt a bl e t o th e S t a t e orEPA? I f a s upervi s or i s notav ai l abl e, an d a wai verha s notbeen gr ant ed as p er Secti o n 1.3, i s a co nsul t antwi th the same q ual i f i cat i o ns s u bs t i tu t e d? C an the l ab o rat o rydo cum entthatthe co nsul t anti s accep t ab l e t o the Stat e, and presenton-s i t e f reque nt l yeno ugh t o s at i sf act o ri l ype rf

o rm a sup erv i so r’ s d uti es? C an the l ab o rat o rys up ervi sor dem o nstrat e thatal l l ab o rat o ry p e rs o n n e l ha v e the a b ili ty t o sa tisf a c to rily p e rfo rm the a n a lyse s to wh i chth eya r e as s i g n ed? Can th e l abor at orys u perv i s orde mon s tr at e th ata l l da t a r eport ed by the l ab o rat o rym eet s the requir ed qua l i tyass urance and regul at o ry cr i t er i a? A n a ly st (o r e q u iv a le n t jo b title ) 1 .2 D oes th e an al y s th av e atl ea s ta h i g h s ch ool e du ca t i on , a mi n i mu m of thr e e m o n ths b e n c h e x p e rie n c e in w a te r, m ilk o r fo o d m ic ro b io lo g y, tra inin g in m ic ro b io lo g ic a l an a lysis o f d rin k ing w a te r a c c e p ta b le to the Stat e (or E P A ), and a m i ni m um of 3 0 da y s o n- the- job trai ni ng u n d e r a n e x p e rie n c e d a n a lyst? H as the ana l y s tdemo ns tr at ed acc ept abl e r es ul t s on u nknown s amp l es b efo re a na l yzing co m p l i an ce sam p l es? W ai ver of

A cad emi c Trai ni ng 1. 3 H as the cert i f

i cat i o n autho ri tywai ved the need f o r the ab o ve sp e c ifi e d a c a d e m ic tra inin g fo r h igh ly ex p e rie n c e d a n a lysts in this la b o ra to ry? H as the cert i f

i cat i o n autho ri tywai ved the need f o r the ab o ve sp e c ifi e d tra inin g fo r su p e rv iso rs o f l a b o ra to rie s a sso c ia te d w ith dr i n k i n g w a t e rs y s t e ms th a ton l ya n a l y z e s a mp l e s f r om th a ts y s t e m?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 38 I f y es t o ei therof

the above, does the l aborat oryha ve a c op yof that wri tt en and s i gned wai ver avai l ab l e f o r i nspecti o n? P e r so n n e l R e c o r d s 1 .4 D o es the l ab o rat o rym ai nt ai n pe rs o nnel record s o n l ab o rat o ry anal y st s thati ncl ud e acad em i c b ackgro und , s p eci al i zed trai ni ng cour s es c ompl et ed, a n d types of mi cr obi ol og i ca l a n al y s es c on du ct ed? 2. LAB O R A TO R Y FAC I LI TI ES D o es the l ab o rat o ryhave f aci l i t i es thatare cl ean and t em p erature and hu m i d i ty co ntro l l ed , an d with a d eq ua t e li gh t i ng at the b en ch t o p s? D o e s the lab o ra to ry m a inta in e ff e c tive se p a ra tio n o f i n c o m p a tib le t esti ng are as? D oes the l aborat orycontr ol ac ces s wh er e app r op r i at e, an d m i ni mi ze traff i c fl o w thro ug h the w o rk a rea s? D o e s the lab o ra to ry e n su re tha t co n ta m ina tio n d o e s n o t ad v e rs e ly a ff e c t d a ta q u a lity? D o e s the lab o ra to ry h a v e b e n c h to p s a n d fl o o rs tha t ar e e a sily cl ea n ed an d di s i n f

ec t ed? D o es the lab o rato ry hav e suff i cien tsp ac e fo r p ro ce ssi ng sam p l es; s t or ag e s pac e f

orme di a, g l as s wa r e, a n d port abl e equ i pmen t ; f l oor space f o r s t at i o naryeq ui p m ent ; and areas f o r cl eani ng gl ass ware and st eril i zing m aterials? D oes the l aborat oryha ve provi s i ons f or di s po s al of

mi cr ob i ol ogi cal w astes? 3. LAB O R A TO R Y EQU I PM ENT A N D S U PPLI ES D o e s the lab o ra to ry h a v e the e q u ip m e n t an d su p p lies n e e d e d to perf orm the approved methods f or w hi ch cert i f

i cat i on ha s been r equ es t ed? pH me t er 3. 1 A re a cc ura cy an d sca l e g rad ua t i o ns w i thin ± 0 .1 u nit s? 3 .1 .1 A r e pH buf

f eral i quot s us ed onl yonce ? 3 .1 .2 A re e le c tro d e s m a inta ine d a c c o rd ing to the m a n u fac tur e r’s rec o m m en d ati o ns? 3 .1 .3 QC A re p H m e te rs sta n d a rd iz e d b e fo re e a c h us e p e rio d w ith p H 7 .0 and e i ther4. 0 or 10. 0 s t andar d buf f er s , wh i che vercove r s the des i r ed p H o f th e m e d ia o r re a g e n t? 3 .1 .4

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 39 QC A re bo th the d at e and buff ers us ed record ed i n a l o gb o o k al o ng w i th the a na l yst ’ s i nit i als? QC Is the p H s l o p e reco rded m o nthl y , af t er cal i b rat i o n? 3 .1 .5 QC If the p H m e te r d o e s n o t ha v e a fea tur e to a u to m a tica lly cal cul at e the sl o p e, but can p rovide i n the p H i n m i l l i vo l t s, i s the f ormul a i n Sec t i on 3. 1. 5. 1 us ed t o cal cul at e the s l op e? 3 .1 .5 .1
QC I f the sl o p e i s b el o w 9 5 % or ab o ve 1 0 5 % , are the manu f ac tu r er ’ s i n s tru ct i on s f

ol l owe d f

orme t erore l ec tr ode m ai nt enance and general cl eani ng? 3 .1 .5 .2
QC Are co m m erci al pH buff er s o l ut i o ns d at ed when recei ved and w h e n op e n e d? 3 .1 .6 QC Ar e pH buf

f ers ol ut i ons di s car ded b ythe expi r at i on dat e? B a la n c e (to p lo a d e r o r p a n ) 3 .2 D o es the b al ance have a read ab i l i tyof 0
. 1 g? 3 .2 .1 D oes th e bal an ce h av e a s en s i t i v i tyof a tl ea s t0. 1 g f

ora l oad of

150 g, and 1 m g for a l o ad o f 1 0 g o r l ess? 3 .2 .2 QC Ar e th e bal an ce s c al i br at ed mon th l yu s i n g AST M Cl as s 1, 2, or 3 wei ght s (mini m um 3 traceab l e wei ght s whi ch b racketl ab o rat o ry w e i g h i n g n e e ds , w i th a r e a dabi l i tyof 0. 1 g ) ? 3 .2 .3
QC A re no n -re fer e n c e w e igh ts c a lib ra te d e v e ry six m o n ths w ith refere nc e w eigh t s? QC Are cal i b rat i o ns reco rded i n a l o gb o o k wi th the i ni t i al s o f the i nd i vi d ual pe rf

o rming the cal i b rat i o n? QC Ar e corr ec t i on v alu es on f i l e an d u s ed? QC A re re f

ere nc e w eigh t s re-certi f i ed ev ery f

i ve years? QC Ar e dama g ed orc orr oded we i g h t s r epl ac ed? QC Are servi ce co ntract s o r i nt ernal m ai nt enance p rotoco l s and mai nt ena nce r ecor ds ava i l abl e? 3 .2 .4 QC Is m aintena nc e, ca l i b rati o n, and clea ning c o nd uc t ed at l ea st a n n u a lly b y a q u a lif

ie d ind e p e n d e n t t e c h n ic ia n , un le ss th e ne e d is m o d i f i ed or waived b ythe cert i f i cat i o n off

i cer? T e m p e r a tu r e M o n ito r in g D e v ic e 3 .3 A re gla ss, d ia l, o r e le c tro n ic the rm o m e te rs gr a d u a te d in 0 .5 

### / C inc re m e n ts (0 .2 

### / C i ncr ement s f or t es t s wh i ch ar e i ncubat ed at 4 4 .5 

### / C ) or l ess , excep tas no t ed f o r hotai r ovens ( Se ct i o n 3.6.1) and ref ri gerat o rs (Sect i o n 3.9.1)? 3 .3 .1

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 40 D o e s o b se rv a tio n o f gla ss th e rm o m e te rs ind ic a te no se p a ra tio n in f l uid co l um ns? Ar e on l ydi alth ermomet er s w h i chc anbe a dj u s t ed u s ed? QC A re gl ass and el ectronic therm o m et ers cal i b rat ed annual l yand d i a l th e rm o m e t e r s q u a r t e r l ya tth e t e m p e r a tu r e u s e d , a g a i n s ta N I S Ttra c e a b le re fer e n c e the rm o m e te r o r o n e tha t m e e ts the re q u ire m e n ts of N B S M on og r aphSP 250- 23? 3 .3 .2

QC Ar e both th e ca l i br at i on f ac t ora n d c al i br at i on da t e i n di ca t ed on the therm o m et er? QC I s th e f

o l l o w i n g c a l i b r a t i o n i n f o rm a t i o n r e c o r d e d i n a QC reco rd b o o k?



### S Ser i al nu mb erof

the l aborat orythermo met er



### S Ser i al nu mb erof

the N I ST
- tr ace abl e thermo met er( or other ref erence therm o m et er)



### S T emp er atur e of

the l aborat orythermo met er



### S T emp er atur e of

the N I ST
- tr ace abl e thermo met er( or other ref erence therm o m et er)



### S Cor r ec t i on ( orc al ibr at i on ) f ac t or



### S D at e of

ch eck



### S A na l yst ’ s i nit i als QC Is the therm o m et er di scarded i f i tdif f ers by mo re than 1 

### / C f

r om the ref erence therm o m et er? 3 .3 .3 QC Are ref

erence therm o m et ers recal i b rat ed atl easteveryf i ve yea rs? QC I s r ef

er en ce th ermomet erc al i br at i on doc u men t at i on mai n t ai n ed? QC Ar e con t i n u ou s r ec or di n g de v i ce s u s ed t o mon i t ori n cubat or t emp er atur e r eca l i brat ed atl eas tan nu al l y , us i ng a r ef er enc e th ermomet erth atme et s th e s pec i f i ca t i on s n ot ed i n Se ct i on 3. 3. 2? 3 .3 .4

In c u b a to r U n it 3 .4 D o i ncubat or uni t s ha ve an i nt ernalt emp er atur e moni t ori ng devi ce a n d m a inta in a tem p e ra tur e sp e c ifi e d b y the m e tho d us e d , usu a lly 35 ° ±0. 5° C a n d 4 4. 5° ±0. 2° C? 3 .4 .1

Fo r non- p o rt ab l e i ncub at o rs , are therm o m et ers placed on t o p and bott om s h el v es of

th e u s e ar eaa n d i mmer s ed i n l i qu i d a s di r ec t ed by the m anuf

acturer (excep tf

o r el ectronic therm o m et ers )? W hen al um i num block i ncub at o rs are us ed , do cul ture di shes and tub e s fitsn u g ly?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 41 QC I s the cal i brat i on- corr ect ed t emp er atur e r ecor ded f or e ach therm o m et er being used atl easttwi ce p er day duri ng each d ay the i ncubat or i s i n u s e? 3 .4 .2
QC A re these rea d i ng s sep ara t ed b y atl ea stf o ur ho urs? QC D o es the d o cum ent at i o n i ncl ud e the d at e and t i m e o f reading, t em p era ture , an d t ec hn i cian ’ s i nit i als? I f a c i r cul at i ng wat erbath i s us ed, i s i tequipp ed wi th agabl e c over t o e n s u r e a n i n c u ba t i ont e mp e r a tu r e of 44
. 5° ±0. 2° C? A u to c la v e 3 .5 D o es the aut o cl ave have an i nt ernal heats o urce, a t em p erature gauge wi th as ens or on the exh aus t , a pres s ur e g aug e, an d an oper at i onal s af etyva l ve? 3 .5 .1

C an the aut o cl ave m ai nt ai n a s t eri l i zat i o n t em p erature duri ng the s t er i l i zi ng cy cl e a nd comp l et e a n e nt i r e c y cl e wi thi n 45 mi nu t es wh en a 12
- 15 mi n u t e s t e r i l i z a t i onpe r i od i s u s e d? D o e s the a u to c la v e d e p re ssu riz e slo w ly en o u g h to e n su re tha t m e d ia w i l l no tb o i l o ve r an d b ub b l es w i l l no tf o rm i n inve rted tub es? QC Is the fo llo w ing info rm a tio n re c o rd e d e a c h ti m e the a u to c la v e is u s ed?



### S D a te



### S C o n te n ts



### S Steri l i zat i o n t i m e and t em p erature



### S T o t al t i m e i n the aut o cl ave



### S A n a lyst’ s initia ls 3 .5 .3

QC Ar e c op i es of

the s ervi ce contr act s or i nt ernalmai nt ena nce p ro to c o ls a n d m a inte n a n c e re c o rd s k e p t? QC Is m a inte n a n c e c o n d u c te d a t l e a st a n n u a lly? QC I s a reco rd o f

the m o strecents ervi ce p erf o rme d on f

i l e and avai l ab l e f

o r i nspecti o n? QC Is a m a x im u m -tem p e ra tur e -re g ister ing the rm o m e te r, e le c tro n ic t emp er atur e r eadoutdevi ce, or cont i nu ous r ecor di ng devi ce us ed eac h aut ocl ave cy cl e t o ens ur e thatthe proper t emp er atur e wa s r ea ch ed? 3 .5 .4

QC I s th e t empe r atu r e r ec or ded? QC I s ov er cr owdi n g a v oi ded? QC Ar e s po r e s tr i ps or s po r e a mp ul es us ed mo nthl yas b i o i nd i cat o rs t o co nf

i rm st eri l i zat i o n?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 42 QC Are aut o m at i c t i m i ng m echanis m s checked qua rt erl ywi th a sto p w a tc h o r o the r a c c u ra te ti m e p ie c e o r tim e sign a l, a n d the re su lts r ec or ded an d i n i t i al ed? 3 .5 .5

A r e a ut ocl ave doo rs eal s cl eanan d f r ee of ca r amel i zed medi a? 3 .5 .6 Ar e au t ocl av e dr ai n s cr ee n s c l ea n ed f r equ en t l ya n d de br i s r emov ed? H o t A ir O v e n 3 .6 D oes th e ov enma i n t ai n a s t abl e s t er i l i za t i on t empe r atu r e of 170 

### /

- 180 

### / C f o r at l ea sttw o ho urs? 3 .6 .1
I s ov er cr owdi n g a v oi ded? I s th e ov enth ermomet erg r adu at ed i n 10 

### / C inc re m e n ts o r le ss, w ith the bul b pl ace d i n s and dur i ng us e? QC I s the f

ol l owi ng i nf

ormat i on r ecor ded f

or e ach cy cl e?



### S D a te



### S C o n te n ts



### S Steri l i zat i o n t i m e and t em p erature



### S A n a lyst’ s initia ls 3 .6 .2

QC Are spo re s trip s used m o nthl yt o co nf i rm st eri l i zat i o n? 3 .6 .3 C o lo n y C o u n te r 3 .7 Is a d a rk fi e ld c o lo n y c o u n te r u se d to c o u n t H e te ro tro p h ic P la te C o un tco l o nies? C o n d u c tiv ity M e te r 3 .8 A re meters s ui t ab l e f

o r checki ng l ab o rat o ryreagent - grade wat er and r e a dabl e i n u n i t s of

e i th e rmi c r om h os / c m o rmi c r os i e me n s / c m? 3 .8 .1 QC I s the m et er cal i b rat ed atl eastm o nthl y , f o l l o wi ng the manuf actur er ’ s r ecomm endat i ons an d us i ng anappropri at e c ert i f i ed a n d tr a c e a bl e l ow - l e v e l s t a n da r d? 3 .8 .2
QC If the m e te r c a n n o t b e c a lib ra te d a s n o te d a b o v e , i s the c e ll co nst antde t erm i ned atm o nthl yi nt erval s usi ng a m etho d i n S ta n da rd Methods , Se ct i on 2510? Is a n in-line un it tha t ca n n o t b e c a lib ra te d us e d to c h e c kre a g e n tgrade wat er? 3 .8 .3
R e fr ig e r a to r 3 .9 D o e s the re frige ra to r m a inta in a tem p e ra tur e o f 1 

### / -5 

### / C? 3 .9 .1 Is the re frige ra to r th e rm o m e te r g ra d u a te d in a t l e a st 1 

### / C inc re m e n ts an d th e th ermomet erbu l b i mmer s ed i n l i qu i d?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 43 QC On da y s th e r ef

r i g er at ori s i n u s e, a n d th e l abor at oryi s s t af f ed, i s the cal i brat ed- corr ect ed t emp er atur e r ecor ded atl eas tonce per d a y? 3 .9 .2

I noc ul at i ng Equi pme nt 3. 10 A re steril e m etal o r d i sp o sab l e p l asti c lo o p s, wo o d ap p l i ca t o r st i ck s, s t er i l e s wabs , ors t er i l e pl as t i c di s pos abl e pi pett i ps u s ed? A re w o o d a p p lica to r stic k s, if us e d , ster ili z e d b y d ry h e a t? Ar e met ali n ocu l at i n g l oops a n d/ orn ee dl es ma de of n i ck ela l l oyor pl a t i n u m? M emb ran e Fi l t rat i on ( M F) Equi pme nt 3. 11 Ar e M F u n i t s ma de of

s t ai n l es s s t ee l , g l as s , por ce l ai n , or a u t oc l a v a bl e pl a s t i c ? 3 .1 1 .1
A re they scr at ched , co rr o d ed , or l eaki ng? QC I f graduati o ns o n cl ear or p l ast i c f unnel s are us ed t o m easure s amp l e v ol ume, i s thei rac cur acych eck ed wi th aCl as s B gr aduat ed c ylind e r o r b e tter(o r o the r C la ss B gla ssw a re ) a n d a re c o rd o f th is ca l i br at i on c h ec kr et ai n ed? 3 .1 1 .2

I s a 10x t o 15 x s t er eo mi cr osc op e wi th af l uor es cen tl i gh ts our ce use d t o co un tshee n c o l o nies? 3 .1 1 .3
A r e the mem bran e f

i l t er s approved bythe manuf actur erf or t ot al co l i f o rm wa t er a na l ysi s? 3 .1 1 .4
Ar e membr an e f

i l t er s t o be u s ed ce l l u l os e es t er , w h i t e, g r i dmark ed, 47 mm di ame t er , a n d 0. 45 

### : m p ore s i ze? I f a l t ern at e por e s i ze s a r e u s ed, doe s th e manu f ac tu r erpr ov i de per f orman ce da t a equ alt o orbe tt erth anth e 0. 45 

### : m p ore s i ze? A r e memb r ane f

i l t er s an d pad s pur cha s ed pres t er i l i zed or aut ocl ave d f or10 mi n u t es a t121 

### / C bef ore us e? QC I s the l otnu mb erf or membran e f

i l t er s an d the dat e r ece i ved r ec or ded? 3 .1 1 .5 QC Ar e th e membr an es c h ec k ed t o s ee th atth eya r e n otbr i tt l e or di s t ort ed? QC A re the m a n u fac tur e r’s sp e c ifi c a tio n /c e rtifi c a tio n sh e e ts ava i l abl e? A re the f o rcep s b l untand s m o o th- t i p p ed wi tho utco rrugat i o ns o n the i nn er sid es o f

the t i p s? 3 .1 1 .6
C ul t ur e D i s he s ( l oos e or t i ght l i ds ) 3. 12

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 44 Ar e pr e s t e r i l i z e d p l a s t i c ors t e r i l i z a bl e g l a s s c u l tu r e di s h e s u s e d? 3 .1 2 .1 Is the ste rility o f th e gla ss c u lture d ish e s m a inta ine d b y p la c e m e n t i n st ai nl ess s t eel or al um i num cani st ers or a wrap of heavy al um i num f o i l or char- res i st antpa p er? A re l o o se- l i d pe tri dis hes i ncub at ed i n a t i ght

- f i tt i ng co nt ai ner wi th a m o isten e d p a p e r to w e l? 3 .1 2 .2
A re o p en ed p ac ks o f

d i sp o sab l e c ulture d i she s resea l ed b etwe en use p erio d s? 3 .1 2 .3
F o r m e m b ra n e fi lterm e tho d s, a re c u lture d ish e s o f an a p p ro p ria te s i ze t o al l ow the tr ans f

erof a s i ng l e memb r ane perpl at e? 3 .1 2 .4
Pi pe t s 3. 13 Ar e g l as s pi pet s s t er i l i ze d a n d ma i n t ai n ed i n s t ai n l es s s t ee l or alu mi n u m c an i s t er s orw r appe d i n di v i du al l yi n c h ar - r es i s t an tpa peror a lum inu m fo il? 3 .1 3 .1

D o pi pet s h av e l eg i bl e mark i n g s a n d a r e th eyn otc h i pped ore t ch ed? 3 .1 3 .2 A re o p en ed p ac ks o f

d i sp o sab l e steril e p i p ets resea l ed b etwe en use p erio d s? 3 .1 3 .3
A re pi p et s d el i veri ng volumes of 1 0 m L o r l ess accurat e t o wi thi n a 2. 5% t ol er anc e? 3 .1 3 .4
Ar e c a l i br a t e d m i c r op i pe tt e r s u s e d w i th s t e r i l e t i ps ? 3 .1 3 .5 A re micr o p i p ett ers cal i b rat ed annual l yand ad jus t ed or replaced i f

the p r e c i s i o n o ra c c u r a c yi s g r e a t e rth a n 2
. 5 %? G l as s w are and Pl as t i cw are 3. 14 Is the gl ass ware m ad e o f bo rosi l i cat e gl ass , or o ther corrosi o nresist an tglass, an d f

ree o f ch i p s and cra ck s? 3 .1 4 .1
A r e marki ng s on g r aduat ed cy l i nder s an d pi pet s l egibl e? A re p l asti c it em s clear an d no nto xic to m i cro o rga nism s? QC A re the gr a d u a te d c ylind e rs us e d fo r m e a su re m e n t o f sam p le vol umes , or otherpre cal i brat ed cont ai ner s thatha ve cl ear l ymarked vo l um es us ed i n l i eu o f graduated cyl i nd ers , accurat e t o wi thi n a 2. 5% t ol er anc e? 3 .1 4 .2

Ar e cu l tu r e tu bes a n d c on t ai n er s c on t ai n i n g f ermen t at i on me di u m of suf f i ci ents i ze t o co nt ai n m ed i um plus s am p l e wi tho utbe i ng m o re tha n thr e e q u a rte rs full? 3 .1 4 .3

A r e tube c l osu r es made of s t ai nl es s s t eel , pl as t i c, alumi nu m, or s cr ew ca p s with no nto xic li ne rs? 3 .1 4 .4

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 45 Ar e cott onorf

oam pl u g s u s ed? Sampl e C ont ai ne rs 3. 15 A re sa m p l e c o ntainers w i d e-m o uth p l asti c o r no n-co rro si ve glass b o ttl e s w ith n o n -lea k ing gr o u n d gla ss sto p p e rs o r c a p s w ith n o n to x ic l i ner s , s t er i l e pl as t i c bag s cont ai ni ng s od i um thi osu l f at e, or other ap p ro p riat e sa m p l e c o ntainers? 3 .1 5 .1

Is sa m p le c o n ta ine r c a p a c ity a t l e a st 1 2 0 m L (4 o z ) to a llo w a t l e a st a 1- i nchhe ad s pace? A re gl ass s t o p p ers co vered wi th al um i num f o i l or char- res i st ant p ap er f o r s t eri l i zat i o n? 3 .1 5 .2
A re uns t eri l i zed gl ass and plas t i c b o tt l es s t eri l i zed by aut o cl avi ng or, alt ern ati ve l y, b y dry o ve n for g l ass b o tt l es? 3 .1 5 .3
A re emp tyco nt ai ners m o i st ened wi th s everal dro p s o f wat er before aut ocl avi ng t o prev entan“ ai rl ock” s t er i l i zat i on f ai l ur e? If c h lo rin a te d w a te r is to b e a n a lyz e d , i s su ff ic ie n t so d ium thio su lfate ad d ed t o the samp l e b o tt l es bef o re s t eri l i zat i o n t o neutral i ze any r es i dualch l ori ne i n the wat ers amp l e? 3 .1 5 .4

U l t ravi ol etLamp ( i f

us ed ) 3. 16 I s th e g ermi ci dalu n i tdi s con n ec t ed mon th l ya n d th e l amp cl ea n ed by w ip ing w ith so ft c lo th m o isten e d w ith e tha n o l? 3 .1 6 .1
Is the l o ngwave uni tus ed f o r f l uo rom etri c t est s kep tcl ean? QC I s th e g ermi ci dalu n i tt es t ed qu art er l yw i th a U V l i g h tme t eror a g a rs pr e a d p l a t e ? 3 .1 6 .2 QC I s the l amp r epl ace d i f i temi t s l es s than70% of i t s i ni t i al ou tpu tori f

a n a g ars pr ea d pl at e con t ai n i n g 200 t o 250 mi cr oor g an i s ms , e x pos ed t o th e U V l i g h tf ortwo mi n u t es , doe s n ot s h o w a c o u n tr e d u c t i o n o f 9 9 %? Spect ropho t om et er or col ori mete r ( i f u s ed) 3. 17 A r e wa vel eng ths i n the vi s i bl e r ang e? 3 .1 7 .1 QC I s a c al i brat i on s t andar d and a method- s peci f i c bl ankan al y zed ev ery d ay the i nstrum en ti s used , p rior to sam p l e a na l ysi s? 3 .1 7 .2 QC I s thi s ca l i brat i on s t andar d ob t ai ned f r om anout s i de s our ce? QC D oes the cal i brat i on s t andar d gi ve a r eadi ng i n the des i r ed abs orbanc e r ang e?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 46 4. GEN ERA L LAB O R A TO R Y PRA C TI C ES A re lab o ra to ry p e rs o n n e l aw a re o f ge n e ra l an d c u sto m a ry sa fety pr a c t i c e s f

orl a bo r a t or i e s ? D oes the l aborat oryha ve a s af etypl an ava i l abl e? D oes th e l abor at oryk ee p a c opy , a n d f ol l ow th e per s on alpr ot ec t i on gui d el i nes, of any m at eri al s af etyda t a sheetacco m p anyi ng the re c e ip t o f a to x ic m a te ria l? S te r iliz a tio n P r o c e d u r e s 4 .1 D o es the l ab o rat o ryf

o l l o w the m i ni m um t i m es f o r aut o cl avi ng the mat er i al s l i s t ed bel ow at121 

### / C?



### S M e m b ra n e fi lter s a n d p a d s 1 0 m in



### S C a rb o h yd ra te c o n ta inin g m e d ia 1 2 -1 5 m in 1



### S C o n ta m ina te d tes t m a te ria ls 3 0 m in 2



### S M e m b ra n e fi ltera sse m b lies 1 5 m in



### S S a m p le c o llec tio n c o n ta ine rs 1 5 m in



### S In d ivid u a l gla ssw a re 1 5 m in



### S D ilutio n w a te r b la n k 1 5 m in



### S R ins e w a te r (0 .5 - 1 L ) 1 5 -3 0 m in 2

ex ceptwh er e otherwi s e s peci f i ed by the manuf

actur er 1 t i me depends upon wat ervol ume percont ai neran d aut ocl ave l oad 2 4 .1 .1

A r e a ut ocl ave d m emb r ane f i l t er s an d pad s an d al l medi a r emo ved i mm edi at el yaf

t ercomp l et i on of

the s t er i l i zat i on cy cl e? 4 .1 .2
Is m e m b ra n e fi ltere q u ip m e n t au to c la v e d b e fo re the b e g inn ing o f a f i l trati o n series? 4 .1 .3
I f a U V l i gh t( 25 4 nm) i s us ed t o s ani t i ze equi pm entaf t eri ni t i al au t ocl av i n g f

ors t er i l i za t i on , a r e al l s u ppl i es pr es t er i l i ze d? 4 .1 .4
S a m p le C o n ta in e r s 4 .2 QC I s atl eas tone s amp l e c ont ai ners el ect ed atr andom f r om ea ch bat chof s t er i l e s ampl e bott l es , oroth erc on t ai n er s ( orl otof c o m m e rc ia lly a v a ilab le sa m p le c o n ta ine rs ), a n d the ste rility co nf i rme d by add i ng 25 m L o f a s t eri l e no n- sel ect i ve b roth, i n cubat i n g a t35 

### / ± 0 .5 

### / C, a n d c h ec k i n g f org r owth a f

t er24 a n d 48 ho urs? QC Ar e th es e r es u l t s r ec or ded? QC I f g r ow th i s de t e c t e d, i s th e e n t i r e ba t c h r e s t e r i l i z e d? R e a g e n t- G r a d e W a te r 4 .3

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 47 D oes th e l abor at oryon l yu s e s at i s f ac t or i l yt es t ed r ea g en tw at erf r om st i l l s o r dei o ni zat i o n uni t s t o prep are me d i a, reagent s, and d i l uti o n/ri nse wa t er fo r p erfo rm i ng m i cro b i al ana l yses? 4 .3 .1
QC I s the q ual i tyof reagentwat er t est ed and do es i tm eetthe f ol l owi ng cr i t er i a? C o n d u c tivity > 0 .5 m e g o h m s re sista n c e M o n thly*

or <2 m i cr om hos / cm ( mi cr os i eme n s / cm) a t25 

### / C P b , C d , C r, N o t gr e a te r th a n 0 .0 5 m g /L A n n u a lly C u , N i, Z n p e r c o n ta m ina n t. C o llec tive ly

n o g r eat erth an0. 1 mg / L T o ta l ch lo rin e < 0 .1 m g /L M o n thly re sid u a l* H e te ro tro p h ic < 5 0 0 /m L C F U /m L * M o n thly p la te c o u n t* B a c te rio lo g ic a l R a tio o f gro w th r a te A n n u a lly q u a lity o f 0 .8 to 3 .0 reagentwat er*

* See Sec t i on 4. 3. 2 f

or f oo tnot es 4 .3 .2

D ilu tio n /R in se W a te r 4 .4 Is sto c kb u ff

e r so lutio n o r p e p to n e w a te r p re p a re d a s sp e c ifi e d in Standard Methods , S e c t i on90 50 C? 4 .4 .1
Ar e s t ockbu f

f er s a u t ocl av ed orf

i l t er - s t er i l i ze d? 4 .4 .2 Ar e th es e con t ai n er s l abe l ed, da t ed, a n d r ef r i g er at ed? A re sto re d sto c kb u ff

e rs fre e fro m turb id ity? QC Is ea ch b atch (o r l o t , i f co m m erc i all y pre p are d ) o f d i l uti o n/ri nse w a t e rc h e c k e d f

ors t e r i l i tybya dd i n g 50 mL of w a t e rt o 5 0 m L dou bl e s tr en g th n on - s el ec t i v e br oth , i n cubat i n g a t35 

### / ± 0 .5 

### / C , and ch ec king fo r gro w th a f t er 2 4 ho urs an d 4 8 ho urs? 4 .4 .3

QC Ar e th es e r es u l t s r ec or ded? QC I s th e bat ch/ l otdi s ca r ded i f g r owth i s de t ec t ed? G la ssw a r e W a sh in g 4 .5 I s di s t i l l ed or dei oni zed wat erus ed f or the f i nalr i ns e? 4 .5 .1 I s l abor at oryg l as s war e wa s h ed wi th a de t er g en tde s i g n ed f or l aborat oryus e? 4 .5 .2

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 48 QC I s the gl ass ware i nhi b i t o ryres i d ue t estpe rf o rme d be f

o re the i ni t i al us e o f

a washi ng co m p o und and whenever a d i f f erent f ormu l at i on , orw as h i n g pr oce du r e i s u s ed? 4 .5 .3
QC Ar e th es e r es u l t s r ec or ded? QC Is e a c h b a tc h o f d ry g la ssw a re us e d fo r m ic ro b ia l an a lysis sp o tch ec k ed f

orpH r ea ct i on u s i n g 0. 04% br omth y mol bl u e ( or e qu i v a l e n tpH i n di c a t or ) a n d th e c ol orr e a c t i onr e c or de d? 4 .5 .4

5 . ANA L YT I CAL M E T HO DOLOGY G e n e r a l 5 .1 For comp l i anc e s amp l es , does the l aborat oryus e onl ythe ana l y t i cal m etho d o l o gi es s p eci f

i ed i n the T o t al C o l i f

o rm R ul e (T C R ), the S u rfa c e W a te r T re a tm e n t R u le (S W T R ), a n d the G ro u n d w a te r R u le ( G W R) ? 5 .1 .1

I s th e l abor at oryc ert i f

i ed f ora l l a n al y t i ca l me th ods i tu s es f or co m p l i an ce p urp o ses? 5 .1 .2
A ta m i ni m um , i s the l ab o rat o rycert i f i ed f o r one t o t al co l i f

o rm m etho d and one f

ecal co l i f

o rm o r E . co lime th od? Is the l ab o rat o rycert i f

i ed f o r a s eco nd t o t al co l i f

o rm m etho d i f

one m etho d ca nn o tb e u sed f o r so m e d rinking w aters? Fora l abor at oryth ate n u mer at es h et er otr oph i c bac t er i a f or co m p l i ance with the SW T R , i s the l ab o rat o rycert i f i ed f o r ei ther the P o u r P la te M e tho d o r th e S im P la te m e tho d fo r h e te ro tro p h ic bact er i a? A re wat er s am p l es s haken vi go rousl yatl east25 t i m es bef o re anal y zi ng? 5 .1 .3 QC I f di l ut i on buf

f eri s us ed, does the l aborat orych eckthe buf f er v olu me i n on e di l u t i onbo tt l e of e a c h ba t c h orl ot ? 5 .1 .4 QC F ora 90- mL or99- mL v ol u me, i s th e t ol er an ce ±2 mL? D oes the l aborat oryan al y ze a 100
- mL sa mp l e v ol ume f or t ot al co l i f o rms i n dri nki ng wat er? 5 .1 .5
M edi a ( or def

i ned s ubs tr at e) 5 .1 .6 Ar e deh y dr at ed medi a s t or ed i n a c ool dryl oca t i on a n d di s ca r ded by the manuf

actur er ’ s ex pi r at i on dat e? 5 .1 .6 .1
I s c ak ed ordi s col or ed deh y dr at ed medi a di s ca r ded?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 49 QC Fo r med i a p repa red i n the l ab o rat o ryi s the f o l l o wi ng i n f ormat i on r ec or ded?



### S D at e of

pr epa r at i on



### S T yp e o f

m ed i um



### S Lotnu mb er



### S Steri l i zat i o n t i m e and t em p erature



### S Final pH (af

t er s t eri l i zat i o n)



### S T e c h n ic ia n ’s initials 5 .1 .6 .2

QC F orme di a pr epa r ed comme r ci al l yi s th e f ol l owi n g r ec or ded f

or e a c h lo t?



### S D at e r ece i ved



### S T yp e o f

m ed i um



### S Lotnu mb er



### S pH v er i f

i ca t i on 5 .1 .6 .3

QC A re m e d ia p re p a re d c o m m e rc ia lly d isc a rd e d b y m a n u fac tur e r’s expi r at i on dat e? QC I s each new l o tof d ehydrat ed or p repa red co m m erci al m ed i um an d ea ch b atch o f

l ab o rato ry- p rep are d m ed i um ch ec ke d b efo re u se f o r st eril i ty and with p o si t i ve an d ne ga t i ve cu l ture co ntro l s? 5 .1 .6 .4
QC Ar e th es e r es u l t s r ec or ded? QC F o r la b o ra to rie s u sin g c o m m e rc ia lly p re p a re d m e d iaw ith m anuf acturer shel f

- l i ves of great er than 9 0 da y s, are po si t i ve and neg at i ve contr ol s runea ch quart er , i n a dd i t i on t o thatnot ed above? QC Ar e th es e r es u l t s r ec or ded? QC For c ontr ol org ani s ms, ar e s t ock c ul tur es per i od i cal l ych eck ed fo r p u rity an d the re su lts re c o rd e d , o r a re c o m m e rc ia lly a v a ilab le di s k s i mpr eg n at ed wi th th e or g an i s m u s ed? If prep ared m ed i um i s st o red aft er s t eri l i zat i o n, i s i tm ai nt ai ned i n the d ark as f o l l o w s?



### S po ur ed pl at es 1 

### / -5 

### / C 2 weeks



### S b ro th in c o n ta ine rs w ith 1 

### /

- 30 

### / C 2 weeks l oo s e- f i tt i ng cl osu r es



### S broth i n t i gh t l ycl ose d 1 

### /

- 30 

### / C 3 m o nths co ntainers 5 .1 .6 .5

QC D o e s the lab o ra to ry p e rfo rm p a ra llel tes ting b e tw e e n a ne w ly ap p roved t estand ano ther E P A
- ap p roved pro ced ure f o r enumerati ng to ta l co lif

o rm s fo r a t l e a st se v e ra l m o n ths a n d /o r se v e ra l se a so n s to as s es s the ef

f ect i ven es s of

the new t es tf

or the wi de va r i etyof

wat er types s u bmi tt ed f

ora n al y s i s ? Rec ommen ded. 5 .1 .7

D o e s the lab o ra to ry p e rfo rm the a p p ro v e d m e tho d s listed in this s e c t i onf orth e T CR, S W T R, a n d/ orGW R? 5 .1 .8

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 50 F e r m e n ta tio n b r o th m e th o d s 5 .2 G ene r al 5 .2 .1 Is the wat er l evel of the wat er bath ab o ve the up p er l evel of the m ed i um i n the c ulture tub es? 5 .2 .1 .1
If a D ri

- b ath i ncub at o r i s used, i s the specif i ed t em p erature r equ i r eme n tma i n t ai n ed i n a l l tu be l oca t i on s u s ed? 5 .2 .1 .2
M u l t i pl e T u be F e rme n t a t i onT e c h n i qu e ( f orde t e c t i n g t ot a l c ol i f

orms i n dri nki ng wat er and enum erat i ng t o t al co l i f o rms i n s o urce wat er) 5 .2 .2
Fo rdr i n k i n g w a t e rs a mp l e s , i s th e t ot a l s a mp l e v olu me of 10 0 m L u s e d f ore a c h t e s t ? 5 .2 .2 .1
F o r so u rc e w a te r sa m p le s, a re a t l e a st 3 se rie s o f f ive tub e s e a c h w ith appr opr i at e s ampl e di l u t i on s u s ed? 5 .2 .2 .2
M e d ia 5 .2 .2 .3 I s l aury l trypt os e br oth ( LT B ) u s ed i n th e pr es u mpt i v e t es ta n d 2% b rilli a n t gr e e n lac to se b ile b ro th ( B G L B B ) in the c o n firm e d tes t? 5 .2 .2 .3 .1
If l act o se broth ( LB ) i s used i n l i eu o f LT B , has the l ab o rat o ry c o n d u c t e d a tl e a s t2 5 p a r a l l e l t e s t s b e tw e e n th i s m e d i u m a n d L TB u s i n g th e wa t er s n ormal l yt es t ed? H as thi s co m p ari son d em o nstrat ed thatthe f al se- p o si t i ve rat e and f a l s e - n e g a t i v e r a t e f

o rt o t a l c o l i f

o rm s , u s i n g L B , i s l e s s th a n 1 0 %? I s th i s c ompar i s on i n f

ormat i on doc u men t ed an d th e r ec or ds r et ai n ed? I s th e f i n alpH of

LT B me di u m 6. 8 ± 0. 2? I s th e f i n alpH of

2% B G LB B 7. 2 ± 0. 2? Is the t estm ed i um co ncentrat i o n ad jus t ed t o co m p ensat e f o r the sa m p le vo lum e so tha t th e re su lti n g m e d ium a ftersa m p le a d d iti o n is si ngl e strength? 5 .2 .2 .3 .2

I f a s i ng l e 100
- mL sa mp l e v ol ume i s us ed, i s the i nv ert ed vi al replaced wi th an aci d i nd i cat o r ( b rom cresol purp l e)? I s th e medi u m a u t ocl av ed at121 

### / C f o r 1 2
- 1 5 m i nu t es? I s the s t er i l e medi um i n tubes ex ami ned t o ens ur e thatthe i nv ert ed v i al s , i f u s ed, a r e f

r ee of a i rbubbl es a n d a r e atl ea s ton e- h al f t o twoth i r ds c ov er ed af

t erth e wa t ers ampl e i s a dded? 5 .2 .2 .3 .3

I s th e i n ocu l at ed medi u m i n cubat ed at35 

### / ± 0 .5 

### / C f o r 2 4 ± 2 ho urs? 5 .2 .2 .4 I f n o g as ora ci d de t ec t ed, i s th e i n ocu l at ed medi u m i n cubat ed f or an o the r 2 4 ho urs f

o r a to t al i nc ub ati o n ti m e o f 4 8 ± 3 ho urs?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 51 I s e ac h 24- a n d 48- h ou rtu be th ath as g r owth ori s g as - pos i t i v e or a c i d

- p o s i t i v e c o n f

i rm e d u s i n g 2 % BGL BB? 5 .2 .2 .5
F o r d rin k ing w a te r sa m p le s, is ea c h to ta l co lif o rm -p o sitive sa m p le t est ed f o r the p res ence o f

ei ther f ecal co l i f

o rms or E . co li ? 5 .2 .2 .6
I nv al i dat i on of

t ot al col i f

orm- neg at i ve s amp l es 5 .2 .2 .7 F o r d rin k ing w a te r sa m p le s, a re a ll sa m p le s tha t p ro d u c e a turb id c u lture (i.e ., he a v y gr o w th) in the a b se n c e o f ga s/a c id p ro d u c tio n , i n LT B orLB , i n v al i dat ed? 5 .2 .2 .7 .1

D o e s the lab o ra to ry th e n c o llec t, o r re q u e st tha t th e syste m c o llec t, ano ther s am p l e wi thi n 24 hours f rom the same locati o n as the ori gi nali nv al i dat ed s amp l e? A l tho ugh not requir ed be f o re i nval i d at i o n, do es the l ab o rat o ry pe r f orm ac on f

i rme d t e s ta n d/ ora f

e c a l c ol i f

orm/E . co lit eston the t o t al co l i f

o rm-negat i ve cul ture t o check f o r col i f o rm supp res si o n? A nd i f the conf i rmed t es ti s t ot al col i f

orm- po s i t i ve or f

eca l c ol i f orm/E . co li

- po s i t i ve, does the l aborat oryr eportthe s amp l e a s s u c h ? If the f o l l o w- up t esti s t o t al co l i f o rm-negat i ve, do es the l ab o rat o ry i nv al i dat e the s amp l e? F o r so u rc e w a te r sa m p le s, a re a ll sa m p le s tha t p ro d u c e a turb id c u lture (i.e ., he a v y gr o w th) in the a b se n c e o f ga s/a c id p ro d u c tio n in LTB orL B i n v a l i da t e d? 5 .2 .2 .7 .2

D o e s the lab o ra to ry c o llec t, o r re q u e st tha t th e syste m c o llec t, anothers amp l e f

r om the s ame l ocat i on as the ori gi nali nv al i dat ed s amp l e? A l tho ugh not requir ed be f o re i nval i d at i o n, do es the l ab o rat o ry per f orm a c on f

i rmed t es ton th e t ot alc ol i f orm- n eg at i v e cu l tu r e an d, i f th e con f i rmed t es ti s t ot alc ol i f

orm- pos i t i v e, i s th e M PN r eport ed? If the c o n firm e d tes t i s to ta l co lif o rm -ne g a tive , i s the sa m p le i n v al i dat ed? Pr e s e n c e - Abs e n c e ( P- A) Col i f orm Tes t( f

orde t e c t i n g t ot a l c ol i f

orms i n dri nki ng wat er) 5 .2 .3
M ed i um 5 .2 .3 .1 W hen si x- t i m es f

o rmulat i o n s trength m ed i um i s used, i s i tf i l t ers t er i l i ze d r ath erth ana u t ocl av ed? 5 .2 .3 .1 .1
I s th e medi u m a u t ocl av ed f or12 mi n u t es a t121 

### / C? 5 .2 .3 .1 .2 Is the t o t al t i m e in the au t o clav e less tha n 3 0 m i nu t es?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 52 Ar e th e bo tt l e s pl a c e d i n th e a u t oc l a v e w i th s pa c e be tw e e n th e m? I s th e f i n alpH of

th e medi u m 6. 8±0. 2? If the p re p a re d m e d ium is sto re d , i s it m a inta ine d in a c u lture b o ttl e at1°
- 30 

### / C i n the d ark f o r no m o re than 3 m o nths? 5 .2 .3 .1 .3
I s th e s t or ed medi u m di s car ded i f e v apor at i on e x ce eds 10% of ori gi nalvol ume? I s a 100- mL sa mp l e i nocul at ed i nt o a P
- A cu l tur e bott l e? 5 .2 .3 .2 I s th e s ampl e/medi u m i n cubat ed at35 

### / ± 0 .5 

### / C an d obs erv ed f or yell o w co l o r (ac i d ) aft er 2 4 an d 4 8 ho urs? 5 .2 .3 .3
Ar e y e l l ow c u l tu r e s c on f i rme d i n BG LBB a n d af e c a l c ol i f

orm/E. c o lit es tc on du ct ed? 5 .2 .3 .4
A re a ll sa m p le s w h ic h p ro d u c e a no n -yello w turb id c u lture in P -A medi u m i n v al i dat ed? 5 .2 .3 .5
D o e s the lab o ra to ry c o llec t, o r re q u e st tha t th e syste m c o llec t, anothers amp l e f

r om the s ame l ocat i on as the ori gi nali nv al i dat ed s amp l e? A l tho ugh not requir ed be f o re i nval i d at i o n, do es the l ab o rat o ry p erf o rm a co nf

i rme d t eston the t o t al co l i f o rm-negat i ve cul ture a n d/ ora f

e c a l c ol i f

orm/E . co lit es tan d, i f

the conf i rmed t es ti s t ot al co l i f o rm-po si t i ve, i s the samp l e repo rt ed as such? If the c o n firm e d tes t i s to ta l co lif o rm -ne g a tive , i s the sa m p le i n v al i dat ed? F e c a l C o lif

o rm T e st (u sin g E C M e d ium fo r fe c a l co lif o rm s in d rin k ing o r so u rc e w a te r, o r A -1 M e d ium fo r fe c a l co lif o rm s in so u rc e w a te r o n ly) 5 .2 .4

E C M ed i um 5 .2 .4 .1 I s E C medi um us ed t o t es ta t ot al col i f orm- po s i t i ve cul tur e f

or f eca l col i f orms un der the T ot al Co l i f orm R ul e? 5 .2 .4 .1 .1
Is each t o t al co l i f

o rm-po si t i ve cul ture trans f erred f rom a presum p t i ve tub e/b o tt l e, or each p res um p t i ve t o t al co l i f o rm-po si t i ve co l o ny ( u n l e s s a c o tt o n s w a b i s u s e d ) , t o a tl e a s to n e tu b e c o n t a i n i n g EC M e d ium w ith a n inv e rte d via l? Is E C m e d ium us e d to e n u m e ra te fec a l co lif o rm s in so u rc e w a te r, in a c c or da n c e w i th th e S W T R? 5 .2 .4 .1 .2
W henconduc t i ng a M T F t es t , ar e thr ee s amp l e v ol umes of s our ce wa t erw i th f

i v e ort entu bes / s ampl e v ol u me u s ed?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 53 I s a c u l tu r e f

r om e ac h t ot alc ol i f

orm- pos i t i v e tu be tr an s f err ed t o a tub e c o n ta inin g E C M e d ium w ith a n inv e rte d via l? I s EC M ediu m a u t ocl av ed at121 

### / C f o r 1 2
- 1 5 m i nu t es? 5 .2 .4 .1 .3 I s th e f i n alpH of

EC me di u m 6. 9±0. 2? A re the inv e rte d via ls e x a m ine d to e n su re tha t th e y a re fre e o f air b u b b le s a n d a t l e a st o n e -ha lf to two -third s c o v e re d a fterthe sa m p le is adde d? 5 .2 .4 .1 .4

Is E C M e d ium inc u b a te d a t 4 4 .5 

### / ± 0 .2 

### / C f o r 2 4 ± 2 ho urs? 5 .2 .4 .1 .5 I s an yamountof

ga s det ect ed i n the i nv ert ed vi al of a tube thatha s tu rbi d g r owth c on s i der ed a f ec alc ol i f

orm- pos i t i v e t es t , r eg ar dl es s of the r es ul tof

an ys ubs equen tt es ton thatcu l tur e? 5 .2 .4 .1 .6

A
- 1 M ed i um 5 .2 .4 .2 I f A- 1 M e di u m i s u s e d, i s i tu s e d t o e n u me r a t e on l yf e c a lc ol i f

orms i n s o urce wat er, i n acco rdanc e wi th SW T R , and not f o r dri nki ng w ater sa m p l es? 5 .2 .4 .2 .1

A re three s am p l e vo l um es of s o urce wat er us ed i n a f i ve- or t entub e /sa m p le vo lum e fo rm a t? I s A- 1 M ediu m a u t ocl av ed at121 

### / C f o r 1 0 m i nu t es? 5 .2 .4 .2 .2 ForA- 1 M ediu m, i s th e f i n alpH 6. 9±0. 1? A re inv e rte d tub e s e x a m ine d to e n su re tha t th e y a re fre e o f air b ub b l es? 5 .2 .4 .2 .3
I s A- 1 M ediu m i n cubat ed at35 

### / ± 0 .5 

### / C f or thr ee hour s , an d then inc u b a te d a t 4 4 .5 

### / ±0. 2 

### / C f o r 2 1 ± 2 ho urs? 5 .2 .4 .2 .4
A re l o o se- cap tub es s t o red i n the d ark atroo m t em p erature f o r no l o nger than two weeks, or i n t i ght l ycl o sed screw- cap tub es i n the darka t<30 

### / C f

o r no l o ng er than thre e m o nths? 5 .2 .4 .2 .5

Is a n y a m o u n t o f ga s d e te c te d in the inv e rte d via l o f a tub e w ith turb id gr o w th c o n sid e re d a fec a l co lif o rm -p o sitive tes t? 5 .2 .4 .3
A zi d e d extrose m ed i um (f o r det ect i ng f

ecal streptoco cci i n ground water ) 5 .2 .5
Fo r t est i ng 10 0
- m L sam p l es, i s trip l e strength (3X ) f o rmulat i o n i n a cu l tu r e bott l e pr epa r ed an d th ena u t ocl av ed at121 

### / C f o r 1 5 m i nu t es? 5 .2 .5 .1
I s me di u m f

i n alpH 7. 2±0. 2? Is a 10 0
- m L wa t er s am p l e ad d ed t o the st eri l i zed m ed i um and i n cubat ed at35 

### / ± 0 .5 

### / C? 5 .2 .5 .2

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 54 Is the cu l ture ch ec ke d f o r turb i d i ty af

t er 2 4 ± 2 ho urs? 5 .2 .5 .3 I f turbi di tyi s notobserved, i s the cul tur e r ei ncubat ed an d che cke d ag ain aft er a t o t al i nc ub ati o n p erio d o f 4 8 ± 3 ho urs? A re turbid cul tures co nf i rme d as f

ecal streptoco cci by s treaki ng a p o rtio n o f th e b ro th o n to b ile e sc u lin a g a r (B E A ) o r b ile e sc u lin azi d e agar ( B E A A )? 5 .2 .5 .4

Ar e B EA a n d B EA A au t ocl av ed at121 

### / C f o r 1 5 m i nu t es? 5 .2 .5 .5 I s th e f i n a l pH 6. 6±0
. 2 f

orBE A a n d 7
. 1±0
. 2 f

orBE AA? Af t ers tr ea k i n g , a r e pl at es i n cubat ed at35 

### / ± 0 .5 

### / C f o r 4 8 ho urs? 5 .2 .5 .6 Ar e th e br ow n i s h

- bl a c kc ol on i e s w i th br ow n h a l os onBE A o rBE AA u se d a s c o n firm ing the p re se n c e o f f e c a l stre p to c o c c i? 5 .2 .5 .7
I f r equ i r ed, doe s th e l abor at orype r f orm a n e n t er ococ cit es tby trans f erri ng one or m o re f

ecal streptoco cci co l o ni es t o brai n heart i nf usi o n bro th s up p l em ent ed wi th 6.5% N aC l and i ncub at i ng the cu l tu r e at35 

### / ± 0 .5C f

o r 4 8 hrs? E n z y m e (c h r o m o g e n ic /flu o r o g e n ic ) su b st r a te te st s 5 .3 G ene r al 5 .3 .1 Forde t ec t i n g t ot alc ol i f orms a n d E . co lii n dr i nk i ng wat erbyan e n z ym e su b stra te tes t, d o e s the lab o ra to ry u se o n e o f th e fo llo w ing : M M O -M U G tes t (C o lil e rt), C o lisur e tes t, E *C o lit e tes t, R e a d yc u lt Col i f orms 100 Pr es en ce /Abs en ce T es t , Fl u or ocu l tLM X t es t , or C o lit a g tes t? 5 .3 .1 .1

Fo re n u me r a t i n g t ot a l c ol i f orms i n s ou r c e w a t e r s bya n e n z y me su b stra te tes t, d o e s the lab o ra to ry u se the C o lil e rt tes t? If a lab o ra to ry u se s a fer m e n ta tio n m e tho d to d e te c t t o ta l co lif o rm s in d ri nki ng wat er, and the samp l e i s t o t al co l i f o rm-po si t i ve, do es the l aborat orytr ans f

erthe po s i t i ve cul tur e t o the E C +M U G t es tt o det ect E . co li , bu tn ott o a n yoth ere n zyme s u bs tr at e t es tme di u m i n Se ct i on 5. 3? M e d ia 5 .3 .1 .2 D o e s the lab o ra to ry p u rc h a se m e d ia fro m a c o m m e rc ia lly a v a ilab le so urc e o nly , an d no tp rep are m ed i a fro m b asic ingre d i en t s? 5 .3 .1 .2 .1
A re m e d ia ke p t p ro te c te d fro m li g h t? 5 .3 .1 .2 .2 Is each l o tof m ed i um checked f o r f l uo res cence b ef

o re us e wi th a 365- 366- n m u l tr av i ol etl i g h tw i th a s i x w attbu l b? 5 .3 .1 .2 .3

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 55 I f medi um ex hibi t s f

ai ntf l uor es cen ce, i s an otherl otus ed thatdoes n otf l u or e s c e ? If sa m p le s p lus m e d ium e x h ib it c o lo r c h a n g e s b e fo re inc u b a tio n , i s th e medi u m di s ca r ded an d a n oth erl otof me di u m u s ed? 5 .3 .1 .2 .4
A re gl ass and plas t i c b o tt l es and t esttub es checked be f o re us e wi th a 36 5- 36 6- nm ul tr avi ol etl i gh ts our ce wi th a6- wattbulb t o ens ur e that theydo notf

l uor es ce? 5 .3 .1 .3

If they f l uo res ce, do es the l ab o rat o ryus e ano ther l o tof co nt ai ners thatdoes notf

l uor es ce? I f a W hi r l

- P ak® bag i s us ed t o i ncubat e the C ol i l ertor C ol i t ag m e d ium o r a n y o the r m e d ium w h ic h c h a n g e s to a ye llo w c o lo r to ind ic a te a p o sitive re su lt, is a typ e us e d tha t ha s a b a rr ie r (e .g., B 0 1 4 1 7 ) t o prevent gaseous em i ss i o ns t o otherW hi rl

- P ak® ba gs du r i n g i n c u ba t i on ? 5 .3 .1 .4

QC If a sm a ll a ir-typ e inc u b a to r is u se d , ar e sa m p le s b ro u g h t t o roo m t em p erature before i ncub at i o n? 5 .3 .1 .5
If a wat er bath i s used, i s the wat er l evel ab o ve the up p er l evel of the me di u m? 5 .3 .1 .6
For E. col i t es t i ng , ar e a l l t ot al col i f orm- po s i t i ve s amp l es pl ace d un der a U V l amp ( 36 5- 36 6 nm) i n adarken ed ar ea? 5 .3 .1 .7
D o e s the lab o ra to ry re fra in fro m us ing the e n z ym e su b stra te tes t t o co nf i rm a p res um p t i ve t o t al co l i f o rm-po si t i ve cul ture i n a f erm ent at i o n bro th or o n a m em b rane f i l t er? 5 .3 .1 .8

D oes the l aborat oryi nv al i dat e a nys amp l e thatproduces anatypi cal col orc h an g e ( i n th e abs en ce of a y el l ow col or ) a n d th enc ol l ec t , or r e qu e s tth a tth e s y s t e m c ol l e c t , a n oth e rs a mp l e f r om th e s a me l ocat i on as the ori gi nali nv al i dat ed s amp l e? 5 .3 .1 .9

D oes the l aborat oryus e a nothermethod t o t es tthe s econd s amp l e? I s the r ef er enc e c om para t or pr ovi ded b ythe manuf actur erdi s car ded by the manuf

actur er ’ s ex pi r at i on dat e? 5. 3. 1. 10
C rite ria fo r sp e c ifi c m e d ia 5 .3 .2 Forth e Col i l ertt es t , a r e s ampl es i n cubat ed at35 

### / ± 0 .5 

### / C f or24 ho urs? 5 .3 .2 .1
I s a s amp l e wi th ay el l ow col or i n the med i um equalt o or g r eat er thanr ef er enc e c om para t or r ecor ded as t ot al col i f orm- po s i t i ve? I s a s ampl e wi th a y el l ow col orl i g h t erth anc ompar at ori n cubat ed f or a n o the r fo u r h o u rs b u t no lo n g e r th a n 2 8 ho u rs to ta l?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 56 I s a s ampl e wi th a y el l ow col orl i g h t erth anth e compa r at ora f t er28 h ou r s of i n c u ba t i onr e c or de d a s t ot a l c ol i f orm- n e g a t i v e ? A re coli f o rm-po si t i ve samp l es thatf l uo res ce und er a U V l i ght m ark ed as E . co li- po s i t i ve? Forth e Col i l ert - 18 t es t , a r e s ampl es i n cubat ed f or18 h ou r s ( u p t o 22 ho urs i f the samp l e af

t er 18 hours i s y el l o w, but l i ght er than the co m p arat o r) ? Fo r enumerati ng t o t al co l i f o rms i n s o urce wat ers , do es the l ab o rat o ry u s e th e Col i l ertt es t , a 5- or10- tu be con f i g u r at i on , Q u an t i

- T r ay , or Q u an t i

- T r ay2000 f

ore ac h s ampl e di l u t i on t es t ed? 5 .3 .2 .1 .1

W h e n d ilutio n w a te r is u se d , i s it e ithe r ste rile d e io n iz e d o r ste rile d i st i l l ed wat er, not b uf f ered wat er? QC I f the Q uant i

- T rayor Q uant i

- T ray20 0 0 t esti s used, i s the seal er checked m o nthl yby add i ng a dye t o the wat er? 5 .3 .2 .1 .2
Forth e Col i s u r e t es t , a r e s ampl es i n cubat ed at35 

### / ± 0 .5 

### / C f or24- 48 ho urs? 5 .3 .2 .2
If the m e d ium c h a n g e s fro m a ye llo w c o lo r to a re d /m a g e n ta c o lo r, is the s amp l e n ot ed as t ot al col i f orm- po s i t i ve? Is a co l i f o rm-po si t i ve samp l e thatf l uo res ces under a U V l i ght m ark ed as E . co li

- po s i t i ve? Forth e E* Col i t e t es t , i s th e s ampl e i n cubat ed at35 

### / ± 0 .5 

### / C f or28 ho urs? 5 .3 .2 .3
I f the med i um ch ang es f

r om a y el l ow col or t o a bl ue or bl ue- gr een col or, or a bl ue col or i n the corn er s of the bag, i s the s amp l e marked as t ot al col i f

orm- po s i t i ve? I f the med i um f

l uor es ces un der a U V l i gh t , i s the s amp l e c ons i dere d as E . co li

- po s i t i ve? I f f l uor es cen ce i s notobserved, i s the s amp l e r ei ncubat ed f or a n ad d i t i o nal 20 hours ( f

o r a t o t al i ncub at i o n t i m e o f 48 hours) and che cke d aga i n f

or f l uor es cen ce? If the m ed i um be co m es r ed i n co l o r, i s the samp l e d i scarded and an oth ers ampl e r equ es t ed? Fo r the R ead y cul tC o l i f o rms 10 0 P res ence- A b sence t est , are the cont ent s of

a s nap packadded t o a 100
- mL sa mp l e a nd then i n cubat ed at35 

### / ± 0 .5 

### / C f o r 2 4 ± 1 ho urs? 5 .3 .2 .4

I f the med i um ch ang es col or f r om a s l i gh t l yy el l ow col or t o bl uegr een , i s the s amp l e marked a s col i f orm- po s i t i ve?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 57 If the m e d ium flu o re sc e s a b rig h t l igh t-b lue c o lo r w h e nsu bje c te d to l o ng wa ve U V (3 6 5
- 3 6 6 nm ) l i gh t , i s the sam p l e m ark ed as E . co li

po s i t i ve? Fo r the Fluoro cul tLM X t est , i s the m ed i um ad d ed t o puri f i ed wat er, m i xed , and the m i xture then b o i l ed t o dis sol ve the m ed i um c om pl e t e l yi n th e w a t e r ? 5 .3 .2 .5

A r e 100
- mL al i quot s tr ans f err ed t o 25 0- mL b ott l es an d then au t o clav ed f

o r 1 5 m i nu t es? A r e the aut ocl ave d bo tt l es cool ed bef ore addi ng the 10 0- mL wat er s amp l e? I s the E . co li/ Col i f

orm S u pp l e me n tn ota dd e d t o th e me di u m? I s th e s ampl e th eni n cubat ed at35 

### / ± 0 .5 

### / C f o r 2 4 ± 1 ho urs? I f the med i um ch ang es col or f r om a s l i gh t l yy el l ow col or t o bl uegr een , i s the s amp l e marked a s col i f orm- po s i t i ve? If the m e d ium flu o re sc e s a b rig h t l igh t-b lue c o lo r w h e nsu bje c te d to l o ng wa ve U V (3 6 5
- 3 6 6 nm ) l i gh t , i s the sam p l e m ark ed as E . co li

po s i t i ve?

Forth e Col i t ag t es t , a r e s ampl es i n cubat ed at35 E ± 0 .5 E C f or24 ±2 ho urs? 5 .3 .2 .6
I f the med i um ch ang es t o a y el l ow col or, i s the s amp l e marked a s col i f orm- po s i t i ve? I f the med i um f

l uor es ces un der a U V l i gh t , i s the s amp l e marked a s E. c o li

- po s i t i ve? E C M e d ium + M U G (fo r d e te c tio n o f E . co li ) 5 .3 .3 If E C m ed i um + M U G i s used, i s a t o t al co l i f o rm-po si t i ve cul ture tr a n s f e rr e d f r om a pr e s u mp t i v e tu be /bo tt l e orc ol on yt o th i s me di u m? 5 .3 .3 .1

I s th e f i n alpH of

EC me di u m + M U G 6. 9±0. 2? 5 .3 .3 .2 Is the m e d ium p lus sa m p le inc u b a te d a t 4 4 .5 

### / ± 0 .2 

### / C f o r 24 ± 2 hours and thent es t ed f

or f l uor es cen ce? 5 .3 .3 .4
E nt erolertt est(f

o r det ect i o n of ent eroc o cci i n ground wat er) 5 .3 .4 Is the m e d ium sto re d in the d a rk a t 4 

### /

- 30 

### / C un t i l u s ed? 5 .3 .4 .1 Is E nt erolertreagentad d ed t o a 10 0
- m L sam p l e and the s ampl e/medi u m i n cubat ed at41 

### / ± 0 .5 

### / C f o r 2 4
- 2 8 ho urs? 5 .3 .4 .2

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 58 I s f l u or es ce n ce u n dera U V l amp u s ed t o i n di ca t e th e pr es en ce of e n te ro c o c c i? M e m b r a n e F ilte r (M F ) m e th o d s 5 .4 G ene r al 5 .4 .1 For s our ce wat ers amp l es ( SW T R ) , do d i l ut i ons y i el d 20 t o 80 t ot al col i f orm c ol on i es or20 t o 60 f ec alc ol i f

orms f ora tl ea s ton e di l u t i on or v ol ume? 5 .4 .1 .1
QC I s atl eas tone membran e f i l t eran d f

i l tr at i on un i ts t er i l i tych eck con du ct ed atth e beg i n n i n g a n d th e en d of e ac h f i l tr at i on s er i es by f i l t eri ng 20
- 3 0 m L o f

dilut i o n wat er through the m em b rane f i l t er and t est i ng f o r gr o wth? 5 .4 .1 .2

QC If the co ntrol i nd i cat es cont am i nat i o n, do es the l ab o rat o ry reje c t all d a ta fro m a ff

e c te d sa m p le s a n d re q u e st a n im m e d ia te res am p l i ng? QC D oes the l aborat orycons i der a f i l tr at i on s er i es as en ded when 3 0 m i nu t es o r m o re h as e l ap sed b etwe en sam p l e fi l trati o ns? Ar e f i l tr at i on f

u n n el s r i n s ed af

t ere ac h s ampl e f

i l tr at i on w i th two or three 20

- 3 0 m L p o rt i o ns o f s t eri l e ri nse wat er t o ensur e thatthe ent i re samp l e i s ri nsed o f

f the f unnel onto the f

i l t er? 5 .4 .1 .3

A re abso rbent p ad s saturat ed wi th atl east2 m L o f bro th and the exc es s medi um r emo ved by“ decan t i ng ” the pl at e? 5 .4 .1 .4
M F me th od f

orde t ec t i n g t ot alc ol i f

orms a n d E . co lii n dri nki ng water , enum erat i ng t o t al co l i f o rms or f

ecal co l i f

o rms i n s o urce wat er, and de t ect i ng E . co lii n g r ound wat er 5 .4 .2

M edi a f ort ot alc ol i f

orms , f ec alc ol i f

orms , a n d E . co li 5 .4 .2 .1 I f ei therM
- E ndo aga ror broth orM
- E ndo aga rLE S i s us ed t o det ect to ta l co lif

o rm s in d rin k ing w a te r o r e n u m e ra ting to ta l co lif o rm s in source water , i s ei ther the si ngl e st ep or the enri chm entt echni q ue u s ed? 5 .4 .2 .1 .1

I s dena tur ed ethan ol us ed i n the r ehy drat i on procedur e? Is the m ed i um prep ared i n a s t eri l e f l ask? Is a b o ili n g w a te r b a th o r a c o n sta n tly a tten d e d ho t p la te w ith a stir baru s ed t o br i n g th e medi u m j u s tt o th e boi l i n g poi n tbu tn otboi l ed? I s th e f i n a l f o rM- E n d o m e d i u m p H 7
. 2 ± 0
. 1 a n d th e f i n a l p H f o rMEndo a g arLES 7. 2±0. 2? I s M
- Endo me di u m orM
- Endo a g arLES i n cubat ed at35° ±0. 5° C f or 22
- 24 h r s ? ( 5. 4. 2. 2)

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 59 A re coloni es wi th a m et al l i c (gol d en) s heen reco rded as p res um p t i ve t o t al co l i f

o rm s? ( 5. 4. 2. 2)
I f m- Col i B l u e24 medi u m i s u s ed t o de t ec tt ot alc ol i f orms a n d E . co li in d rin k ing w a te r, a re the a m p u le s o f b ro th in v e rte d 2 -3 ti m e s to m ix cont ent s bef

ore brea ki ng an d the cont ent s thenpoure d eve nl yoveran abs orben tpa d? 5 .4 .2 .1 .2

A re unop ened ref

ri gerat ed am p ul es s t o red i n the d ark? Ar e u n open ed ampu l es di s ca r ded bef or e th e ex pi r at i on da t e, or ea r l i eri f c on t ami n at i on i s obs erv ed? I s th e medi u m f

i n alpH 7. 0±0. 2? I s m- Col i B l u e 24 i n c uba t e d a t35 ° ±0. 5° C f or24 h r s ? ( 5. 4. 2. 2) A re re d c o lo n ie s re c o rd e d a s to ta l co lif o rm s, a n d b lue to p u rp le co l o nies rec o rd ed as E . co li ? ( 5. 4. 2. 2)
I f M I me di u m ( w i th orw i th ou ta g a r ) i s u s e d t o d e t e c tt ot a l c ol i f orms an d E . co lii n dr i nk i ng wat eror enu mera t e t ot al col i f orms i n s our ce wa t er , i s c ommer ci al l ypr epa r ed pr es t er i l i ze d bott l ed M I a g aror br oth n ota u t ocl av ed? 5 .4 .2 .1 .3

I s thi s pre s t er i l ai zed bot t l ed ag armedi um mel t ed i n aboi l i ng wat er b ath (or by other pro cess es r eco m m end ed by the m anuf acturer) , and c a r e t a k e n n ott o o v e rh e a tth e a g a r ? Is the m e d ium the n c o o le d sligh tly a n d p o u re d im m e d ia te ly i n to st eril e p l ates? I f dehy drat ed cu l tur e medi um i s us ed, i s i tpre pare d and au t ocl ave d ac co rd i ng t o m an ufac ture r’ s i nstructi o ns? Is thi s agar me d i um co o l ed be f o re add i ng f

res hl yprep ared , f

i l t erst eril i ze d ce f

sulo d i n, and the n p o ure d i m m ed i ately i nto st eril e p l ates? Is the fi n a l p H o f M I a g a r 6 .9 5 ± 0 .2 0 a n d the fi n a l p H o f M I b ro th 7. 05±0. 20? I s M I me di u m i n c u ba t e d a t35 ° ±0. 5° C f or24 ±2 h r s ? ( 5. 4. 2. 2) A re fluo resc en tco l o nies un d er U V l i gh trec o rd ed as total co l i f o rm s, an d b l ue co l o nies un d er n o rm al l i gh trec o rd ed as E . co li ? ( 5. 4. 2. 2)
If C hrom o cul t ® C o l i f

o rm A gar i s used t o de t ectt o t al co l i f o rms and E . co lii n dr i n k i n g w at er , i s th e ag arme di u m a u t ocl av ed or ov erh ea t ed? 5 .4 .2 .1 .4

I s th e f i n alpH of

th i s me di u m 6. 8±0. 2?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 60 If a he a v y b a c k g ro u n d o f he te ro tro p h ic b a c te ria is ex p e c te d , i s ce f s u l odi n s ol u t i on a dded t o 1L of c ool ed ( 45 

### /

- 50 

### / C ) m ed i um as a s ol ut i on of

10 m g c ef

s ul od i n di s s ol ved i n 2- mL d ei oni zed or di s t i l l ed water ? Is C hro m o cu l ti nc ub ated at 3 6 °± 1 °C f o r 2 4 ± 1 hrs? ( 5. 4. 2. 2) A re s al m o n t o red co l o ni es r eco rded as t o t al co l i f o rms, and da rk- b l ue t o vio l et co l o nies rec o rd ed as E . co li ? ( 5. 4. 2. 2)
I f Col i s ca n ® i s u s ed t o de t ec tt ot alc ol i f orms a n d E . co lii n dri nki ng wateror enum erat e t o t al co l i f o rms i n s o urce wat er, i s the manu f ac tu r er ’ s pr ot ocolf

orr ec on s t i tu t i on a n d a n t i bi ot i c addi t i on f ol l owe d? 5 .4 .2 .1 .5

I s th e an t i bi ot i c, c ef

s u l odi n , ov erh ea t ed? I s th e f i n alpH of

Col i s ca n a g ar7. 00±0. 20? Is C o l i sca n incu b ated at 3 2 °- 3 7 °C f o r 2 4
- 2 8 hrs? ( 5. 4. 2. 2) A r e pi nk

- magen t a c ol oni es r ecor ded as t ot al col i f orms , an d purpl eb l ue co l o nies rec o rd ed as E . co li ? ( 5. 4. 2. 2)
I f m- FC broth, wi th orwi thoutag ar , i s us ed t o enu mera t e f eca l c ol i f orms i n s ou r c e w a t e r , i s th e me di u m a u t oc l a v e d? 5 .4 .2 .1 .6
Is m -F C b ro th j u st b ro u g h t t o the b o ili n g p o int? I s th e f i n alpH of

m- FC medi u m 7. 4±0. 2? Is m
- F C b ro th incu b ated at 4 4 .5°± 0 .2°C f o r 2 4 ± 2 hrs? ( 5. 4. 2. 2) A re b l ue co l o nies rec o rd ed as f ec al co l i f

o rm s? ( 5. 4. 2. 2) Is the p re p a re d m e d ium re frige ra te d w h e n sto re d a n d b ro u g h t t o r oo m t emp er atur e bef

ore us e? 5 .4 .2 .1 .7
A re p e tri d ish e s c o n ta inin g m e d ium sto re d in a p la stic b a g o r tigh tly clo sed co ntainer, an d used within 2 we ek s? Ar e pl at es w i th l abor at ory - pr epar ed br oth me di u m di s car ded af t er96 hour s , poure d aga rpl at es af t er2 week s , an d amp ul ed broth di s car ded bef ore the manuf

actur er ’ s ex pi r at i on dat e? Ar e th e dat e an d t i me of me di u m pr epa r at i on r ec or ded? For i nv al i dat i on of

a t ot al col i f

orm- neg at i ve dri nk i ng wat ers amp l e, a re a ll sa m p le s re su lti n g in c o n flue n t gr o w th o r T N T C gr o w th i n v al i dat ed u n l es s a tl ea s ton e t ot alc ol i f orm c ol on yi s de t ec t ed? 5 .4 .2 .3

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 61 If no co l i f o rms are detect ed , i s the samp l e reco rded as “conf l uent grow th” o r “ T N T C ” and an ad d i t i o nal s am p l e reque st ed f rom the s ame s amp l i ng s i t e? D oe s th e l a bo r a t orype r f orm av e r i f

i c a t i ont e s tonth e t ot a l c ol i f ormnegat i ve cul ture before i nval i d at i o n? If the veri f

i cat i o n t esti s t o t al co l i f o rm-po si t i ve, do es the l ab o rat o ry r eportthe s amp l e a s t ot al col i f orm- po s i t i ve? If the ve rifica tio n tes t i s to ta l co lif o rm -ne g a tive , i s the sa m p le i n v al i dat ed? Fo r i nval i d at i o n of source waters am p l es ( SW T R ), where co l i f o rm d ensi tym ustbe de t erm i ned , do es the l ab o rat o ryi nval i d at e any s amp l e thatr es ul t s i n c onf l uen tgr owth orT N T C , ev en whe n t ot al c o lif o rm o r fe c a l co lif

o rm c o lo n ie s a re p re se n t? 5 .4 .2 .4

For dr i nk i ng wat ers amp l es on M
- E ndo type medi a, ar e a l l s hee n col on i es , up t o a ma x i mu m of f i v e, v er i f i ed byu s i n g e i th erLB or LTB a n d th e n 2% B G LBB or , a l t e rn a t i v e l y , byu s i n g a c y t oc h r om e ox i das e an d 

### $

- gal act osi das e pr ocedur e? 5 .4 .2 .5

If no sh e e n c o lo n ie s a re o b se rv e d , ar e up to fi v e re d q u e stio n a b le sheen coloni es and/ o r r ed non-s heen co l o ni es r ep res ent i ng dif f erent morph ol og i ca l types v er i f i ed? For dr i nk i ng wat ers amp l es , ar e t ot al col i f orm- po s i t i ve col oni es t est ed f o r E . co lio r f

ec al co l i f

o rm s? 5 .4 .2 .6
W h enEC M ediu m orEC + M U G i s u s ed, a r e col on i es tr an s f err ed by emp l oy i ng one of

the op t i ons s peci f

i ed by the T ot al Co l i f

orm R ul e? W henthe s wab t ech ni que i s us ed, i s a s i ng l e s wab us ed t o i nocul at e a p r e s u m p t i v e t o t a l c o l i f o rm - p o s i t i v e s a m p l e i n t o E C o rE C + M UG f i r s t , LT B s ec on d, a n d B G LB B th i r d? F o r so u rc e w a te r sa m p le s, a re the initial to ta l co lif o rm c o u n ts adju s t ed bas ed upon ve r i f i ed dat a? 5 .4 .2 .7 QC Fo r s o urce wat er s am p l es when two or m o re anal y st s are avai l ab l e, do es each anal y stco untthe t o t al co l i f o rm o r f ecal co l i f

o rm col oni es on the s ame memb r ane mo nthl yan d do the count s ag r ee w i th i n 1 0 %? 5 .4 .2 .8

N u trie n t A g a r + M U G T e st (fo r d e te c tio n o f E . co lii n dri nki ng wateror ground wat er) 5 .4 .3
I s th e medi u m a u t ocl av ed at121 

### / C f o r 1 5 m i nu t es? 5 .4 .3 .1 I s th e f i n alM U G c on ce n tr at i on 100 

### : g / L? I s th e f i n alpH of

N A + M U G 6. 8±0. 2?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 62 QC A re p o sitive a n d ne g a tive c u lture c o n tro ls tes te d a s sta te d in 5. 1. 6. 4? 5 .4 .3 .2 QC Ar e cu l tu r e con tr ol s f i l t er ed ors pot - i n ocu l at ed on t o a m em b rane f

i l t er on M
- E nd o bro th or agar, or M
- E nd o agar LE S, and i n cubat ed at35 

### / ± 0 .5 

### / C f o r 2 4 ho urs? QC Is the fi lterthe n tra n sfe rr e d to N A + M U G a n d inc u b a te d a t 35 

### / ± 0 .5 

### / C f

o r an o the r f

o ur ho urs? QC Ar e th es e r es u l t s r ea d a n d r ec or ded? I s the mem bran e f

i l t ercont ai ni ng t ot al col i f orm col oni es tr ans f

err ed t o th e s u r f

a c e of th e Nu tr i e n tAg a r+ M U G me di u m? 5 .4 .3 .3
Is the p re se n c e o f ea c h sh e e n c o lo n y m a rk e d o n the p e tri d ish li d w ith p e rm a n e n t m a rk e r, a n d the li d a n d b a se m a rk e d to re a lign the li d wh enr emov ed? Fo r the t o t al co l i f

o rm veri f

i cat i o n t est , i s a p o rt i o n of each co l o ny trans f erred with need l e b ef

o re the M F tr ansf

er or af t er the f o ur- ho ur N A + M U G i ncubat i on t i me? A lter n a tive ly, is the m e m b ra n e fi ltersu rfa c e sw a b b e d w ith a ste rile co tt o n s wab af

t er the f o ur- ho ur i ncub at i o n t i m e o n N A + M U G and the n tra n sfe rr e d to ato ta l co lif o rm ve rifica tio n tes t? I s th e i n ocu l at ed N A + M U G me di u m i n cubat ed at35 

### / ± 0 .5 

### / C f or f o ur ho urs? 5 .4 .3 .4
Is f l uo res cence che cked by us i ng a U V l am p (36 5
- 3 6 6 nm ) wi th a six -w a tt b u lb in a d a rk e n e d ro o m a n d a n y flu o re sc e n c e in the ha lo around a s heen co l o nyco nsi d ered po si t i ve f o r E . co li ? 5 .4 .3 .5

M F m etho d f

o r det ect i ng ent eroc o cci / f ecal streptoco cci i n ground wat er 5 .4 .4
M e d ia 5 .4 .4 .1 W h e n mE a g a ri s u s e d f orth e de t e c t i onof

e n t e r oc oc c i , i s ba s a l mE ag arpr epa r ed, a u t ocl av ed, a n d c ool ed bef or e th e addi t i on of nal i di xi c a ci d (ori t s s od i um s al t ) an d tr i pheny l t etr azol i um ch l ori de, both of w h i cha r e adde d s epa r at el yt o th e medi u m a n d mi x ed? 5 .4 .4 .1 .1

I s th e f i n alpH of

mE ag ar7. 1±0. 2? W henm- E nt er ococcus ag ari s us ed f or the det ect i on of

f eca l s tr ept ococ ci( n ote n t er ococ ci ) , i s th e medi u m h ea t ed, n ota u t ocl av ed, t o d i sso l ve the i ng red i en t s? 5 .4 .4 .1 .2

I s th e f i n alpH of

m- Ent er ococ cu s a g ar7. 2±0. 2?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 63 W h enmEI a g ari s u s ed f orth e det ec t i on of

e n t er ococ ci , i s 0. 75g ind o x yl- 

### $

- D
- glucos i de added t o 1L bas al mE ag aran d thenpr epar ed acco rding t o 5.4.4. 1
. 1 excep tthatonly0.02 g/L trip henylt etrazo l i um ch l or i de i s a dded? 5 .4 .4 .1 .3

I s th e f i n alpH of

mEI a g ar7. 1±0. 2? Is a 10 0
- m L sam p l e f

i l t ered and the m em b rane placed on o ne o f the ag arme di a pr ev i ou s l yl i s t ed? 5 .4 .4 .2
I f m- E nt er ococcus ag ari s us ed, ar e the pl at es i ncubat ed i n a n i n v ert ed pos i t i on a t35 

### / ± 0 .5 

### / C f or48 h ou r s ? 5 .4 .4 .3
U s i ng magn i f

i cat i on an d a f

l uor es cen tl amp , ar e a l l l i gh tan d darkr ed c o lo n ie s c o u n te d a s fec a l stre p to c o c c i? I f mE ag ari s u s ed, a r e th e pl at es i n cubat ed i n a n i n v ert ed pos i t i on f or 48 h ou r s a t41 

### / ± 0 .5 

### / C? 5 .4 .4 .4
I s the mem bran e f

i l t erthentr ans f

err ed t o E I A medi um an d i ncubat ed at41 

### / ± 0 .5 

### / C f

o r 2 0
- 3 0 m i nu t es? U s i ng magn i f

i cat i on an d a f

l uor es cen tl amp , ar e a l l pi nkt o re d co l o ni es on m E agar wi th a black or redd i sh b row n precipi t at e o n the u n d e rs id e o f th e fi ltero n E IA a g a r c o u n te d a s e n te ro c o c c i? I f mEI a g ari s u s ed, a r e pl at es i n cubat ed i n a n i n v ert ed pos i t i on f or 24 h ou r s a t41 

### / ± 0 .5 

### / C? 5 .4 .4 .5
U s i n g ma g n i f

i ca t i on a n d a f

l u or es ce n tl amp, i s th e pl at e ex ami n ed, t o p and bo tt o m , f

o r col o ni es wi th a blue hal o , and anyco l o nywi th a bl u e h al o ( r eg ar dl es s of c ol on yc ol or ) c on s i der ed as pos i t i v e f or e n te ro c o c c i? H et er ot rop hi c Pl at e C ount (fo r e n u m e ra ting he te ro tro p h ic b a c te ria i n dri nki ng wat er) 5 .5
D o e s the lab o ra to ry u se the P o u r P la te M e tho d o r th e S im P la te M etho d f

o r enumerati ng het erotr o p hi c b act eri a i n dri nki ng wat er and f o r t est i ng reagentgrade wa t er? 5 .5 .1

Fo r s y st em s grant ed a vari ance f rom the T C R ’ s m axi m um cont ami nan tl eve l , does the l aborat oryus e R2 A medi um wi th a m etho d i n Standard Methods , Sec t i o n 92 1 5 f o r enumerati ng het erotr o p hi c b act eri a i n dri nki ng wat er? M e d ia 5 .5 .2 I s the f i nalpH r ecor ded f

or pl at e c ountag arpH 7. 0± 0. 2, R2 A ag ar 7. 2±0. 2, a n d Si mP l at e 7. 2±0. 2? Forth e PourPl at e M eth od, i s me l t ed ag art empe r ed at44 

### /

- 46 

### / C i n a waterba th and m ai nt ai ned no m o re than 3 hours bef o re po uri ng? 5 .5 .3

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 64 I s thi s s t er i l e medi um mel t ed onl yonce ? For the Spread Pl at e M ethod, i s 15 m L of R2 A medi um ( or other m e d ium ) p o u re d into a ste rile p e tri d ish a n d a llo w e d to so lid ify? 5 .5 .4
Is ref ri gerat ed m ed i um i n bo tt l es or s crew- cap p ed tub es s t o red f o r no l o nger than si x m o nths, or i n pe tri dis hes f o r no l o nger than 2 weeks (one we ek f

o r prep ared pe tri dis hes wi th R 2 A m ed i um )? 5 .5 .5

Fo rc ou n t a bl e pl a t e s of mo s tpo t a bl e w a t e rs a mp l e s , a r e 1. 0 m L a n d/ or0. 1 m L v olu me s of th e u n di l u t e d s a mp l e pl a t e d? 5 .5 .6
Ar e atl ea s tdupl i ca t e pl at es pr epa r ed perdi l u t i on t es t ed? F o r th e P o u r P la te M e tho d , i s the sa m p le p ip e tted a se p tica lly o n to the bo tt om of

a s t er i l e petr i di s h a nd thenatl eas t10- 12 mL t emp er ed mel t ed ag ara dded? 5 .5 .7

I s the s amp l e a nd mel t ed ag armi xed, av oi di ng s pi l l age ? A f t erthe aga rpl at es ha ve s ol i di f i ed on a l eve l s ur f

ace , ar e they i n v ert ed an d i n cubat ed at35 

### / ± 0 .5 

### / C f o r 4 8 ± 3 ho urs? A re pl at es s t acked no m o re than f o ur hi gh and arr anged i n the i ncub at o r t o al l o w p rop er ai r ci rcul at i o n and t o m ai nt ai n a uni f o rm i ncubat i on t emp er atur e? D o es the lab o rato ry ensure tha ti nc ub ato r d o es no tha ve ex ce ss hum i d i tyand thatthe p l at es do not l o se mo re than 1 5 % by wei ght d uri ng the 4 8 hours of

i ncub at i o n? Forth e Spr ea d Pl at e M eth od, i s 0. 1 or0. 5 mL of th e s ampl e ( or d ilutio n ) p ip e tted o n to the su rfa c e o f th e p re d rie d a g a r in the p la te an d th ens pr ea d ov erth e en t i r e s u r f ac e u s i n g a s t er i l e ben tg l as s r od? 5 .5 .8

Is the i no cul um ab sorb ed co m p l et el ybe f o re i ncub at i ng? Ar e th e pl at es i n cubat ed i n a n i n v ert ed pos i t i on a t20 

### /

- 28 

### / C fo r 5 -7 d ays? F o r th e M e m b ra n e F ilt e r T e c h n iq u e , d o e s the fi lter e d vo lum e yield b etwe en 2 0
- 2 0 0 co l o nies? 5 .5 .9
I s th e f i l t ertr an s f

err ed t o a pe tr i di s h c on t ai n i n g 5 mL s ol i di f i ed R2A medi u m a n d th eni n cubat ed at20 

### /

- 28 

### / C f o r 5
- 7 d ays? A r e pl at es wi th l oo s e- f i tt i ng l i ds pl ace d i n apl as t i c box wi th acl ose fitting li d a n d m o isten e d p a p e r to w e ls, an d re w e tted a s n e c e ssa ry? Ar e col on i es c ou n t ed u s i n g a s t er eos copi c mi cr os cope a t10- 15X m agni f i cat i o n? Si mP l at e M eth od 5. 5. 10

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 65 F o r a single sam p l e U nitD o se, i s a 1 0
- m L t est sam p l e a d d ed t o a test tub e c o n ta inin g d e h yd ra te d S im P la te m e d ium a n d the n p o u re d o n to the ce nter o f

a p l ate c o ntaini ng 8 4 sm all we l l s? 5 .5 .1 0 .1

Al t ern at i v el y , i s 9- mL of s t er i l e di l u en ta dded t o th e t es ttu be co nt ai ni ng the d ehydrat ed m ed i um , f o l l o wed by a 1- m L sam p l e, and the m e d ium p lus sa m p le the n p o u re d o n to the c e n te r o f a p la te co ntaini ng 8 4 sm all we l l s? Is this m i xture d i stribu t ed ev en l y t o the 8 4 we l l s and i s the ex ce ss l i qui d drai ned i nt o the abs orbentpad on the pl at e? I s th e pl at e i n v ert ed an d i n cubat ed at35 

### / ± 0 .5 

### / C f o r 4 5
- 7 2 ho urs? I s bact er i al dens i tydet ermi ned bycoun t i ng the nu mb erof wel l s that fluo re sc e un d e r a 3 6 5 -3 6 6 -nm U V li g h t, a n d c o n v e rting this v a lue to a M o stP ro b ab l e N um b er/m L using the m an ufac ture r’ s U nitD o se M P N t abl e? If a 1 0 -m L sa m p le is use d , i s the U n it D o se M P N /m L re a d d ire c tly or , i f a 1- mL s ampl e i s u s ed, i s th e M PN /mL v alu e corr ec t ed by mu l t i pl y i n g i tby10? Fo rth e M u l t i pl e Dos e f or10 s a mp l e s of

1 mL e a c h , i s a 10 0- mL s t er i l e di l uen tadded t o the dehy drat ed Si mP l at e medi um an d s hak en t o di s s ol ve? 5 .5 .1 0 .2

I s a 1. 0- mL t es ts amp l e thenpi pett ed t o the cen t erof a pl at e, f ol l ow e d b y9 mL of

th e r e c on s t i tu t e d m e di u m? Is the p l at e then gent l ys wi rl ed t o m i x and dis trib ut e the samp l e and med i um mi xtur e e ven l yt o the 84 wel l s , wi th the exc es s l i qui d then bei ng dra i ned i nt o the abs orbentpad on the pl at e? I s th e pl at e i n v ert ed an d i n cubat ed at35 

### / ± 0 .5 

### / C f o r 4 5
- 7 2 ho urs? I s bact er i al dens i tydet ermi ned bycoun t i ng the nu mb erof wel l s that fluo re sc e un d e r a 3 6 5 -3 6 6 -nm U V li g h t, a n d c o n v e rting this v a lue to a M o stP ro b ab l e N um b er/m L using the m an ufac ture r’ s M ult i

- D o se M P N t abl e? If s am p l e d i l ut i o ns were m ad e d uri ng s am p l e p repa rat i o n, i s the M P N /m L value mul t i p l i ed by the d i l ut i o n f act o r? For the P ourP l at e a nd Sprea d P l at e T ech ni ques , ar e c ol oni es co unt ed m anual l yus i ng a da rk f i el d co l o nyco unt er? 5. 5. 11
A r e onl ypl at es ha vi ng 30 t o 30 0 col oni es coun t ed, ex ceptf or pl at es i nocul at ed wi th 1. 0 m L of un di l ut ed s amp l e wh er e c ount s of l es s than 30 ar e a cce pt abl e?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 66 QC I s each b at ch o r f l ask of agar checked f

o r s t eri l i tyby po uri ng a f i nalcontr ol pl at e? 5. 5. 12 QC Does th e l abor at oryr ej ec tda t a i f th e con tr ol i s c on t ami n at ed? C ol i phage ( D r a f

tM e th od 16 01 a n d 1 60 2 f orth e GW R) 5 .6 E P A M e tho d 1 6 0 1 : M a le -sp e c ifi c (F + ) a n d S o m a tic C o lip h a g e in W at er by T wo - Step E nri chm ent 5 .6 .1
Is the 1 0 0
- m L (o r 1- L) wat er s am p l e supp l em ent ed wi th m agnesi um chl ori de, l og- phas e h ostbact er i a, an d trypt i c s oy broth ( T SB ) as an enr i chmen ts t ep f

or c ol i phage ? A f t er incu b ati o n o ve rnight, are sam p l es sp o tt ed o nto a law n o f ho st b ac t eria, i nc ub ated , an d the n e xa m i ne d f o r circu l ar ly si s zo ne s? M e d ia 5 .6 .1 .1 Fo r ant i b i o t i c st o cks, are ant i b i o t i cs al ways ad d ed t o the m ed i um af terth e medi u m h as be ena u t ocl av ed? 5 .6 .1 .1 .1
Ar e an t i bi ot i c s t ock s s t or ed f r oze n a t- 20 

### / C f

o r no l o nger than o ne y ear? A re ant i b i o t i c st o cks thawed atroo m t em p erature or rapi d l yi n a wa t erba th up t o 37 

### / C and m i xed wel l be f o re us i ng? Is 1 0 X tryp tic so y b ro th ( T S B ) sto re d a t 1 

### / -5 

### / C un t i l u s ed? 5 .6 .1 .1 .2 A re 1 .5 % tryp tic so y a g a r (T S A ) p la te s a ftera n tib io tic su p p le m e n ta tio n a n d so lid ifi c a tio n sto re d inv e rte d a t 1 

### / -5 

### / C f o r no lo n g e r th a n two w e e k s, if no t use d im m e d ia te ly? 5 .6 .1 .1 .3

A r e 0. 7% T SA t op ag artubes wi th app r op r i at e a nt i bi ot i cs , di s pens ed as 5- mL pe rs t er i l e 10- mL tu be, l abe l ed, a n d k epta t45 

### /

- 48 

### / C un til u s ed? 5 .6 .1 .1 .4

A re these tub es us ed the d ay o f prep arat i o n? For s po tpl at es thatdeve l op conden s at i on dur i ng s t orag e, ar e pl at es i n cubat ed f

ora ppr ox i mat el y10 mi n u t es t o r edu ce c on den s at i on pr i or t o i no cul at i o n? 5 .6 .1 .1 .5

A re sp o t p la te s u se d tha t d a y o r sto re d a t 1 

### / -5 

### / C f o r up t o f

o ur d ays? C ol iphage s t ock 5 .6 .1 .2 A r e M S2 (AT C C #1 55 97
- B 1 m al e- s peci f i c) an d phi

- X 174 s t ock c o lip h a g e (A T C C # 1 3 7 0 6 -B 1 , so m a tic) sto re d a t 2 

### / -8 

### / C f o r up t o f

i ve yea rs? 5 .6 .1 .2 .1

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 67 D oes a n aly s i s of

r aw s ew ag e f

i l tr at e beg i n w i th i n 24 h ou r s of co l l ect i o n? 5 .6 .1 .2 .2
I s a tl ea s t10 mL of

f i l t er ed s ew ag e obt ai n ed? 5 .6 .1 .2 .4 I f the f i l tr at e i s s t ored more than24 hour s , i s i tr e- t i t er ed bef ore us e? 5 .6 .1 .2 .5 H ostbact er i a s t ock c ul tur es 5 .6 .1 .3 A fterp re p a ra tio n , ar e ho st b a c te ria sto c kc u lture s h e ld a t a t empe r atu r e betwe en- 20 

### / C an d - 70 

### / C? 5 .6 .1 .3 .1
Ar e bac t er i a s t or ed n o l on g erth antwo mon th s a t- 20 

### / C or no l onger th anon e y ea ra t- 70 

### / C? A r e pr epar ed overni gh thos tbact er i a s t ock c ul tur es ch i l l ed on weti ce o r a t 1 

### / -5 

### / C un t i l r eadyt o us e? 5 .6 .1 .3 .2
A f t erpre para t i on, ar e l og- phas e h ostbact er i a s t ock c ul tur es ch i l l ed o n w e t i c e o r a t 1 

### / -5 

### / C t o s l ow r epl i cat i on un t i l r eadyf or u s e? 5 .6 .1 .3 .3
W he n store d , are the se susp en si o ns st o red no m o re than 4 8 ho urs? A re re m a inin g b a c te ria l ho st c u lture s sto re d a t 1 

### / -5 

### / C o v e rn igh t t o i no cu l ate fl ask s f

o r the p rep ara t i o n o f

ne w l o g-p ha se b ac t erial ho st s? Ge n e r a l QC 5 .6 .1 .4 I n i t i a l de mo n s tr a t i onof c a pabi l i ty( I D C) 5 .6 .1 .4 .1 QC D id the lab o ra to ry d e m o n stra te d a b ili ty t o ge n e ra te a c c e p ta b le p erf o rma nce wi th thi s m etho d by perf o rming an ID C t estbe f o re an alyzing an y f

i eld sam p l es? QC D i d the I D C t es tcons i s tof t en r eag entwat ers amp l es s pi ked wi th e nu mera t ed s ewag e f i l tr at e orequi val entat1- 2 P FU mL per sa m p le fo r e v e ry sa m p le fo r e a c h c o lip h a g e typ e us e d , ac c o rd ing to the I D C T abl e? QC W er e thes e I D C t es t s ac com pani ed by a method bl ankf or e ach col i ph ag e type u s ed? M etho d B l anks 5 .6 .1 .4 .2 QC Is a m etho d blank ( reagentwat er s am p l e co nt ai ni ng no co l i p hage) anal y zed t o de m o nstrat e f reed o m f rom co nt am i nat i o n? F o r e a c h c o lip h a g e typ e us e d , i s a ste rile re a g e n t wa te r sa m p le pr epa r ed an d a n al y ze d u s i n g th e s ame pr oce du r e u s ed f ora n al y s i s of f i eld an d Q C sam p l es? QC I s atl eas tone method bl ankan al y zed f or e verys po tpl at e u s ed f o r f i eld sam p l es?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 68 P o sitive C o n tro ls 5 .6 .1 .4 .3 QC A re po si t i ve co ntrols anal y zed t o ensur e thats t o ck co l i p hage sus p ensi o ns, hostba ct eri al cul tures , and growth m ed i a are p e rfo rm ing p ro p e rly? QC For ea ch col i phage type us ed, i s a 100 m L r eag entwat er s a mp l e s pi k e d w i th 20 PFU f r om s e w a g e f

i l tr a t e or60 PFU f

r om a pur e c ol i phage s t ock c ul tur e? QC I s on e pos i t i v e con tr ol a n al y ze d f ore v erys potpl at e u s ed f or f i eld sam p l es? QC I f mu l t i pl e s po tpl a t e s a r e i n oc u l a t e d w i th s a mp l e s onth e s a me d a y, i s a sing le e n ric h e d p o sitive c o n tro l sa m p le us e d to ino c u la te m u ltip le sp o t p la te s tha t d a y? M atr i x s pi k es ( M S) 5 .6 .1 .4 .4 QC I s m e th o d p e r f

o rm a n c e a s s e s s e d b ya n a l y z i n g o n e s e to f MS s amp l es f

or e ach col i phage type us ed whe n s amp l es ar e f i r s tr ece i ved f r om a n ew g r ound wat ers our ce? QC For ea ch col i phage type an al y zed, ar e thr ee f i el d s amp l es s pi ked wi th 1- 2 P FU , wi th a tl eas tone outof the thr ee M S s amp l es bei ng posi t i ve f

or e ach col i phage type? QC I s one s etof

M S s amp l es an al y zed on a n ong oi ng bas i s af t er eve ry20 f

i el d s amp l e f

or e ach gr ound wat ers our ce? th QC A re these M S sam p l es col l ect ed atthe same ti m e as r o ut i ne f i eld sam p l es? QC A re these s am p l es s p i ked i n “ b ul k” atthe co ncentrat i o ns i ndi cat ed i n the M S and O D C Samp l e Spi ki ng T abl e? O n g oi n g de mo n s tr a t i onof c a pabi l i ty( O D C) 5 .6 .1 .4 .5 QC D o e s the lab o ra to ry d e m o n stra te tha t th e a n a lyti c a l syste m is i n co ntro l o n a n o ng o i ng b asis thro ug h a na l ysi s o f O D C sam p l es? QC A r e thr ee r eag entwat ers amp l es s pi ked wi th 1- 2 P FU f or e ach col i ph ag e type u s ed? QC A re O D C t est sam p l es a na l yze d ex ac t l y l i ke f i eld sam p l es? QC I s a mi n i mu m of

on e ou tof

th r ee O D C s ampl es pos i t i v e f or ea chc ol i ph ag e type u s ed? QC Is one s etof O D C s am p l es anal y zed af t er every20 f

i el d and M S sa m p le s o r o n e p e r w e e k , w h ic h e v e r o c c u rs m o re fre q u e n tly?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 69 QC A re sa m p le s sp ike d in “b u lk” a t th e c o n c e n tra tio n s ind ic a te d in the M S and O D C Samp l e Spi ki ng T abl e? P er f ormanc e s tudi es 5 .6 .1 .4 .6 QC D oes the l aborat oryper i od i cal l yan al y ze ext ernalQ C s amp l es whe n a vai l abl e? QC D o es the l ab o rat o rypa rt i ci p at e i n avai l ab l e i nt erl ab o rat o ry p erfo rm an ce stud i es? QC D o es the l ab o rat o ryrevi ew resul t s, co rr ectuns at i sf act o ry p erfo rm an ce , an d rec o rd co rrec t i ve ac t i o ns? E P A M e tho d 1 6 0 2 : M a le -sp e c ifi c (F + ) a n d S o m a tic C o lip h a g e in W at er by Si ngl e A gar Layer(SA L) P roce d ure 5 .6 .2
Is the 1 0 0
- m L gro und wat er s am p l e supp l em ent ed wi th m agnesi um chl ori de an d hos tbact er i a, an d thenadded t o 10 0- mL o f doubl estre n g th m o lten tryp tic so y a g a r c o n ta inin g the a p p ro p ria te ant i bi ot i c? I s the s amp l e thor ough l ymi xed a nd the t ot al vol ume thenpoure d i nto f i ve t o t en p l ates? A f t eroverni gh ti ncubat i on, ar e c i r cul arl y s i s zone s r ecor ded as col i phage ? M e d ia 5 .6 .2 .1 Fo r ant i b i o t i c st o cks, are ant i b i o t i cs al ways ad d ed t o the m ed i um a f ter th e medi u m h as be ena u t ocl av ed? Ar e an t i bi ot i c s t ock s s t or ed f r oze n a t- 20 

### / C f

o r up t o one y ear? A re ant i b i o t i c st o cks thawed atroo m t em p erature or rapi d l yi n a wa t erba th up t o 37 

### / C and then m i xed wel l be f o re us i ng? Is 1 0 X T S B sto re d a t 1 

### / -5 

### / C un t i l u s ed? A re 1.5% T SA plat es, af t er ant i b i o t i c supp l em ent at i o n and so lid ifi c a tio n , sto re d inv e rte d a t 1 

### / -5 

### / C f o r no l o ng er than tw o we ek s, if n o t use d im m e d ia te ly? A r e 0. 7% T SA t op ag artubes wi th app r op r i at e a nt i bi ot i cs , di s pens ed as 5- mL pe rs t er i l e 10- mL tu be, l abe l ed, a n d k epta t45 

### /

- 48 

### / C un til u s ed? Ar e th es e tu bes u s ed th e dayth eya r e pr epa r ed? A r e s po tpl at es thatdeve l op conden s at i on dur i ng s t orag e, i ncubat ed fo r a p p ro x im a te ly 1 0 m inu te s to re d u c e c o n d e n sa tio n p rio r to i no cul at i o n?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 70 A re the se p la te s u se d tha t d a y o r sto re d a t 1 

### / -5 

### / C f o r up t o f

o ur d ays? I s 2X T SA, w i th a ppr opr i at e an t i bi ot i cs , k eptmol t ena t45 

### /

- 48 

### / C i n a waterba th? 5 .6 .2 .1 .2
Is thi s agar us ed onlyon the d ay o f prep arat i o n? C ol iphage St ock 5 .6 .2 .2 A r e M S2 (AT C C #1 55 97
- B 1 m al e- s peci f i c) an d phi

- X 174 s t ock c o lip h a g e (A T C C # 1 3 7 0 6 -B 1 , so m a tic) sto re d a t 2 

### / -8 

### / C f o r up t o f

i ve yea rs? D oes a n aly s i s of

r aw s ew ag e f

i l tr at e beg i n w i th i n 24 h ou r s of co l l ect i o n? I s a tl ea s t10 mL of

f i l t er ed s ew ag e obt ai n ed? I f the f i l tr at e i s s t ored more than24 hour s , i s i tr e- t i t er ed bef ore us e? H ostbact er i a s t ock c ul tur es 5 .6 .2 .3 A fterp re p a ra tio n , ar e ho st b a c te ria sto c kc u lture s h e ld a t a t empe r atu r e betwe en- 20 

### / C an d - 70 

### / C? Ar e bac t er i a s t or ed n o l on g erth antwo mon th s a t- 20 

### / C , or no l onger th anon e y ea ra t- 70 

### / C? A f t erpre para t i on, ar e ove rni gh thos tbact er i a s t ock c ul tur es ch i l l ed o n w e t i c e o r a t 1 

### / -5 

### / C un t i l r eadyf or u s e? A f t erpre para t i on, ar e l og- phas e h ostbact er i a s t ock c ul tur es ch i l l ed o n w e t i c e o r a t 1 

### / -5 

### / C t o s l ow r epl i cat i on un t i l r eadyt o us e? W he n store d , are the se susp en si o ns st o red no l o ng er than 4 8 ho urs? A re re m a inin g b a c te ria l ho st c u lture s sto re d a t 1 

### / -5 

### / C o v e rn igh t t o i no cu l ate fl ask s f

o r the p rep ara t i o n o f

ne w l o g-p ha se b ac t erial ho st s? Ge n e r a l QC 5 .6 .2 .4 I n i t i a l pr e c i s i ona n d r e c ov e ry( I PR) 5 .6 .2 .4 .1 QC H a s the lab o ra to ry d e m o n stra te d the a b ili ty t o p e rfo rm this m etho d accep t ab l yby perf o rming an IP R t estbe f o re anal y zi ng any f i eld sam p l es? QC D oes the I P R T es tcons i s tof f ourr eag entwat ers amp l es t es t ed fo r e a c h c o lip h a g e typ e us e d , sp ike d w ith e n u m e ra te d se w a g e fi ltra te or e qui val entt o y i el d a t ar gets pi ke conce ntr at i on of 80 P FU per s amp l e?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 71 QC D o es the rel at i ve st and ard d evi at i o n of the reco very(RS D ), and the a v e ra g e p e rc e n t re c o v e ry ( 

### 0 ) ba s ed on a l l f ou rs ampl e r es u l t s f

or e a c h c o l i p h a g e typ e u s e d , m e e tth e a c c e p t a n c e c r i t e r i a i n th e QC A cce pt anc e Cri t er i a t abl e? M etho d blanks 5 .6 .2 .4 .2 QC Is one m etho d blank ( reagentwat er s am p l e co nt ai ni ng no c o lip h a g e ) a n a lyz e d w ith e a c h a n a lyti c a l b a tc h to d e m o n stra te f reed o m f rom co nt am i nat i o n? QC For ea ch col i phage type us ed, ar e s t er i l e r eag entwat ers amp l es p re p a re d a n d a n a lyz e d us ing the sa m e p ro c e d u re s u se d fo r a n a lysis o f the f i eld an d Q C sam p l es? QC Is an anal y t i cal ba t ch d ef i ned as al l s am p l es anal y zed duri ng a sin g le d a y, up to a m a x im u m o f 2 0 sa m p le s (fie ld sa m p le s a n d m a trix s pi k e s ampl es ) pe rc ol i ph ag e type u s ed? M atr i x s pi k es ( M S) 5 .6 .2 .4 .3 QC I s m e th o d p e r f

o rm a n c e a s s e s s e d b ya n a l y z i n g o n e s e to f MS s amp l es f

or e ach col i phage type us ed whe n s amp l es ar e f i r s tr ece i ved f r om a n ew g r ound wat ers our ce? QC I s on e s etof

M S s ampl es r ou t i n el ya n al y ze d a f t ere v ery20 th f i el d s amp l e f

or e ach gr ound wat ers our ce? QC I f th e r e c o v e ryf

o rc o l i p h a g e f

a l l s o u t s i d e i t s l i m i t( s e e th e QC A cce pt anc e Cri t er i a t abl e) , i s method perf ormanc e c ons i dere d un acc ept abl e? QC I f O P R r es ul t s ar e n otwi thi n c ontr ol l i mi t s , i s the probl em i den t i f i ed an d c orr ec t ed an d th e dat a qu al i f i ed? QC D o es the l ab o rat o rym ai nt ai n a co ntrol charton reco veryand up d ate ito n a reg ular b asis? O n g oi n g pr e c i s i ona n d r e c ov e ry( O PR) 5 .6 .2 .4 .4 QC D oes the l aborat orydemo ns tr at e a cce pt abl e per f ormanc e thro ug h a na l ysi s o f

an O P R sam p l e o n a n o ng o i ng b asis? QC For ea ch col i phage type us ed, i s a r eag entwat ers amp l e s pi ked w i th a p p r o x i m a t e l y8 0 P F U? QC I s the O P R s amp l e a nal y zed ex act l yl i ke a f i el d s amp l e? QC Is one O P R s am p l e anal y zed f o r each anal y t i cal ba t ch? QC D oe s th e l a bo r a t oryc om pa r e th e OP R pe r c e n tr e c ov e ry( R) wi th the corr es po ndi ng l i mi t s i n the Q C Accept anc e Cri t er i a t abl e?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 72 QC I f R f al l s out s i de the r ang e f or r ecove ry , i s an al y s i s s t op ped u n tilthe p ro b le m is i d e n tif ie d , co rr e c te d , an d a n o the r O P R tes t i s s u cc es s f u l l ype r f ormed? P er f ormanc e s tudi es 5 .6 .2 .4 .5 QC D o e s the lab o ra to ry p e rio d ic a lly a n a lyz e a n e x te rn a l Q C sa m p le whe n a vai l abl e? QC D o es the l ab o rat o rypa rt i ci p at e i n avai l ab l e i nt erl ab o rat o ry p erfo rm an ce stud i es? QC D o es the l ab o rat o ryrevi ew these r esul t s, co rr ectuns at i sf act o ry p erfo rm an ce , an d rec o rd co rrec t i ve ac t i o ns? 6 . S A M P L E C OL L E C T I ON , HA N D L I N G , A N D P R E S E R V A T I ON S a m p le C o llec to r 6 .1 Is the sa m p le c o llec to r tra ine d in a se p tic sa m p ling p ro c e d u re s a n d , i f re q u ire d , ap p ro v e d b y the a p p ro p ria te re g u la to ry a u tho rity o r its des i gn at ed r epre s ent at i ve? S a m p lin g 6 .2 A re the d ri nki ng wat er s am p l es col l ect ed unde r the T o t al C o l i f o rm Ru l e r e pr e s e n t a t i v e of

th e w a t e rdi s tr i bu t i ons y s t e m? 6 .2 .1
A re the w ater tap s used f o r sam p l i ng f

ree o f ae rato rs, straine rs, ho se att ac hm en t s, m i xing typ e fauc ets, an d p urif i ca t i o n d ev i ce s? Ar e on l yc ol d w at ert aps u s ed? Ar e s erv i ce l i n es c l ea r ed bef or e s ampl i n g byma i n t ai n i n g a s t ea dy waterf l o w f o r atl east2 m i nut es or unt i l a s t ead ywat er t em p erature i s r ea ch ed? I s a tl ea s ta 100- mL s ampl e v ol u me col l ec t ed, a l l owi n g a tl ea s ta 1- i n cha i rs pac e i n th e con t ai n ert o f ac i l i t at e mi x i n g of

th e s ampl e by shaki ng? Is a sa m p le info rm a tio n fo rm c o m p le te d im m e d ia te ly afte r sa m p le co l l ect i o n? If a s am p l e b o tt l e i s f

i l l ed t o o f

ul l t o al l o w f

o r prop er mixi ng, i s the ent i re s am p l e p o ured i nt o a l arger s t eri l e co nt ai ner and m i xed be f o re p ro ce ed i ng with the a na l ysi s? Fo r the SW T R , are the source wat er s am p l es r ep res ent at i ve o f the source o f

s up p l yand co l l ect ed not t o o f ar f rom the i nt ake p o i nt , but ata r eas onabl e di s t anc e f r om the bank or s hor e? 6 .2 .2

I s th e s ampl e v ol u me s u f f i ci en tt o pe r f

orm a l l th e t es t s r equ i r ed?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 73 F o r th e a n a lysis o f co lip h a g e , E . co li , ore n t e r oc oc c i u n de rth e GW R, i s a tl ea s ta 100- mL s ampl e v ol u me col l ec t ed? 6 .2 .3 6 .2 .4 S a m p le Ic in g 6 .3 F o r d rin k ing w a te r b a c te ria l sa m p le s, is the sa m p le r e n c o u ra g e d to h ol d s ampl es a t<10 

### / C d u rin g tra n sit t o the lab o ra to ry? 6 .3 .1
Fors ou r ce w at erba ct er i als ampl es , a r e s ampl es h el d a t<10 

### / C duri ng tra n sit t o the lab o ra to ry? D o es the l ab o rat o ryrej ects am p l es thathave b een f roze n? For col i phage an al y s i s un der the G W R , ar e s amp l es s hipp ed at <10 

### / C , sto re d a t 1 

### / -5 

### / C , and not f roze n? 6 .3 .2 QC Fo r SW T R s am p l es and co l i p hage samp l es, do es the l ab o rat o ry re c o rd sa m p le tem p e ra tur e up o n re c e ip t? 6 .3 .3 QC Does th e l abor at oryf l ag s ampl es th ath av e a t empe r atu r e u pon r ec eiptof

>10 

### / C , whether i ced or no t , unl ess the t i m e si nce the sam p l e c o l l ec t i o n is l ess than tw o ho urs? S a m p le H o ld in g /T r a v e l T im e 6 .4 Fo rth e a n a l y s i s of

t ot a l c ol i f

orms i n dr i n k i n g w a t e r , do e s th e t i me b etween sam p l e co l l ect i o n and placem entof the samp l e i n the i nc ub ato r no tex ce ed 3 0 ho urs? 6 .4 .1

A re a ll sa m p le s a n a lyz e d o n the d a y o f re c e ip t? A re sa m p le s re c e ive d late in the d a y re frige ra te d o v e rn igh t o n ly i f anal y si s can b egi n wi thi n 30 hours of co l l ect i o n? Fo r t o t al co l i f

o rms and f

ecal co l i f

o rms i n s urf

ace wa t er s o urces, and fo r h e te ro tro p h ic b a c te ria in d rin k ing w a te r, is the ti m e fro m sa m p le co l l ec t i o n to p l ac em en ti n the incu b ato r l ess than eight ho urs? 6 .4 .2

F o r c o lip h a g e a n a lysis, i s the ti m e fro m sa m p le c o llec tio n to p l ac em en to f

sam p l e in the i nc ub ato r l ess than 4 8 ho urs? 6 .4 .3
F o r c o lip h a g e a n a lysis, i s the ti m e fro m se w a g e sa m p le c o llec tio n to ana l y s i s of

Q C s pi ki ng s us pens i on l es s than24 hour s , un l es s r et i t e r e d a n d th e t i t e rh a s n o td e c r e a s e d b ym o r e th a n 5 0 %? I f the t i t erha s notdecr eas ed by mo r e than50% , i s the s amp l e s t ored no l o ng er than 7 2 ho urs? Fo r E . co lia n d e n t e r oc oc c i a n a l y s i s u n de rth e GW R, i s th e t i me b etween sam p l e co l l ect i o n and the p l acem entof samp l e i n the i ncub at o r l ess than 3 0 hours 

## ? 6 .4 .4

S a m p le In fo r m a tio n F o r m 6 .5

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 74 A fterc o llec tio n , d o e s the sa m p le r e n te r th e fo llo w ing info rm a tio n , i n i n de l i bl e i n k , ons a mp l e i n f orma t i onf

orm?



### S N ame of

s y s t em ( P W SS i dent i f i cat i on nu mb eri f

av ai l abl e)



### S S a m p le id e n tif

ic a tio n (if an y)



### S Sa mpl e s i t e l oca t i on



### S Sa mpl e type ( e. g . , a r ou t i n e di s tr i bu t i on , r epe at , r aw orpr oce s s , or others peci al purpo s e)



### S D at e an d t i me of

c ol l ec t i on



### S A nal y s i s r eques t ed



### S D i s i nf ect antr es i dual



### S N ame of

s amp l er



### S A ny rem ark s C h a in -o f- C u st o d y 6 .6 Ar e appl i cabl e St at e r eg u l at i on s pe r t ai n i n g t o c h ai n - of

- cu s t ody fo llo w e d b y sa m p le c o llec to rs a n d the lab o ra to ry? 7 . Q UAL I T Y AS S URANCE D o e s the lab o ra to ry h a v e a w ritten Q A P la n p re p a re d a n d a v a ilab le f o r i nspecti o n? 7 .1
D o es the l ab o rat o ryf

o l l o w the wri tt en Q A P l an? D o e s the lab o ra to ry h a v e a S ta n d a rd O p e ra ting P ro c e d u re a v a ilab le f o r rev i ew p ertaining to it s o w n c ali b rati o n o f eq uip m en to r sup p l i es? Doe s th e l a b o r a t o rys u c c e s s f u l l ya n a l y z e a tl e a s to n e s e to f PT sa m p le s o n c e e v e ry 1 2 m o n ths fo r e a c h m e tho d fo r w h ic h itis ce r t i f i ed? 7 .2

Fo r method s used t o t estthe p res ence o r absence o f an organi sm i n a samp l e, do es the l ab o rat o ryanal y ze each P T s am p l e setus i ng a s i n g l e a n a l y t i c a l me th od on l y ? 8 . RE CORDS AND DAT A RE PORT I NG L e g a l D e fe n sib ili ty 8 .1 A re com p l i ance m o ni t o ri ng da t a b ei ng m ai nt ai ned by the l ab o rat o ry bo th thor ough an d accu r at e, an d thu s l ega l l ydef ens i bl e? D o es the l ab o rat o ry ’ s Q A plan and/or SO P s d escrib e the p o l i ci es and procedur es us ed by the f aci l i tyf or r ecor d re t ent i on an d s t orag e? If s am p l es ar e exp ect ed t o be co m e p artof l egal act i o n, do es the l ab o rato ry f

o l l o w ch ain-o f

- cu st o d y pro ce d ure s? M a in te n a n c e o f R e c o r d s 8 .2 D oes the publ i c wa t ers y s t em mai nt ai n r ecor ds of mi cr ob i ol ogi cal a n a l y s e s f

orf i v e y e a r s ?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 75 D o es the l ab o rat o rym ai nt ai n easi l yaccessib l e reco rds f o r f i ve y ears o r u n tilthe ne x t ce rtifi c a tio n d a ta a u d it is c o m p le te d , w h ic h e v e r is l o nger? D o es the l ab o rat o rynoti f ythe cl i entwat er s y st em be f o re di spo si ng of r ec or ds s o th eyma yr equ es tc opi es i f n ee ded? D oes the l aborat orybackup al l el ectr oni c dat a byprot ect ed t ape, d isk , o r h a rd c o p y? W hen the l ab o rat o rychanges i t s co m p ut er hardware o r s o f tware, are provi s i ons i n pl ace f

or tr ans f err i ng ol d dat a t o the new s y s t em s o that d ata re m ain retri ev ab l e w i thin the sp ec i f i ed t i m e fram es? S a m p lin g R e c o r d s 8 .3 A r e a l l dat a r ecor ded i n i nk , wi th a nych ang es l i ned thr ough s uchthat the ori gi nalen tryi s vi s i bl e? Ar e ch an g es i n i t i al ed an d da t ed? D o e s the lab o ra to ry h a v e the fo llo w ing sa m p le info rm a tio n re a d ily ava i l abl e?



### S D at e and t i m e o f

s am p l e recei p tby the l ab o rat o ry



### S N a m e o f th e lab o ra to ry p e rs o n re c e ivin g the sa m p le



### S In fo rm a tio n o n a n y d e ficie n c y in the c o n d iti o n o f th e sa m p le 8 . 3 . 1 -4

A re sa m p l es inva l i d ated f o r the f o l l o w i ng rea so ns?



### S T i m e b etween sam p l e co l l ect i o n and recei p tby l ab o rat o ry exc eeded



### S Pre s en ce of

di s i n f ec t an ti n s ampl e n ot i ce d, e . g . , odor



### S E vi d ence o f

f reezi ng



### S U se o f a c o ntainer n o tap p ro ve d b y the l ab o rato ry f o r the p urp o se i nt ended



### S I n s u f f i c i e n ts a mp l e v olu me , e . g . , <1 00 mL



### S P rese nc e o f

i nterfering co ntam i na nts no t i ce d , e.g., hyd ro ca rb o ns, cl ean s er s , he avymet al s , et c.



### S S a m p le tem p e ra tur e e x c e e d ing the m a x im u m a llo w a b le 8 .3 .4

A n a ly tic a l R e c o r d s 8 .4 A r e a l l r ecor ded d at a i n i nkwi th a nych ang es l i ned thr ough s uchthat ori gi nalen tryi s vi s i bl e? Ar e th es e ch an g es i n i t i al ed an d da t ed?

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 76 A r e the f ol l owi ng r eadi l yav ai l abl e?



### S La bor at orys ampl e i dent i f i ca t i on i n f

ormat i on



### S Inf o rma t i o n co ncerni ng da t e and t i m e anal y si s b egi ns



### S N am e o f

the l ab o rato ry and a signa ture o r i nit i als o f the p erso n(s) p e rfo rm ing a n a lysis



### S I n f ormat i on c on ce rn i n g th e an al y t i ca l t ec h n i qu e orme th od u s ed



### S I n f orma t i onc on c e rn i n g a l l i t e ms ma rk e d “ Q C”



### S R esu l t s o f

the an alyses 8 . 4 . 1 -6

P r e v e n tiv e M a in te n a n c e 8 .5 D o e s the lab o ra to ry m a inta in p re v e n tive m a inte n a n c e a n d re p a ir re c o rd s fo r a ll ins tru m e n ts a n d e q u ip m e n t? A re these rec o rd s kep tf o r f i ve years in a m an ne r tha tall o w s f o r ea sy i nspecti o n? 9. AC TI O N RESPO N SE TO LAB O R A TO R Y RESULTS T e st in g T o ta l C o lifo r m -P o sitiv e C u ltu r e s 9 .1 For the T ot al Co l i f

orm R ul e, does the l aborat oryt es tal l t ot al c ol i f orm- po s i t i v e c u l tu r e s f orth e pr e s e n c e of

e i th e rf e c a l c ol i f

orms o r E . co li ? N o tific a tio n o f P o sitiv e R e su lts 9 .2 Fo r T o t al C o l i f

o rm R ul e, do es the l ab o rat o rypro m p t l ynoti f ythe p rop er autho ri tyof a p o si t i ve t o t al co l i f o rm, f ecal co l i f

o rm, or E . co li r es u l t , s o th ata ppr opr i at e f ol l ow- u p a ct i on s c anbe c on du ct ed? 9 .2 .1

Fo r the T o t al C o l i f

o rm R ul e, i f

a s am p l e i s f

ecal co l i f

o rm- o r E . co li

pos i t i v e, doe s th e s y s t em n ot i f yth e St at e as s oon a s i ti s n ot i f i ed of the tes t re su lt, i. e ., at the e n d o f th a t d a y o r, if the S ta te o ff ic e is c lo se d , b y the e n d o f th e ne x t b u sin e ss d a y? 9 .2 .2

D o es the l ab o rat o ryba se a t o t al co l i f o rm-po si t i ve res ul ton the con f i rmed ph as e i f

th e M u l t i pl e T u be Fe rmen t at i on T ec h n i qu e or P res ence- A b sence C o l i f o rm T esti s used, o r the veri f i ed t estf o r the M e m b r a n e F i l tr a t i o n Te c h n i q u e i f M
- En d o m e d i u m o rM
- En d o L ES ag ari s u s ed? 9 .2 .3

I f a pr es u mpt i v e t ot alc ol i f orm- pos i t i v e cu l tu r e does n ot con f i rm/v er i f ya s s u ch , bu ti s f

ou n d t o be f

ec alc ol i f

orm orE . co li

po s i t i ve, i s the s amp l e c ons i dere d t ot al col i f orm- po s i t i ve and f

eca l c ol i f orm/E . co li

- po s i t i ve? N o tific a tio n o f T o ta l C o lifo r m In te r fe r e n c e 9 .3

## MICRO CHECKLI STS E l em en t N um be r Y es N o N A C o m m en ts

V
- 77 Fo r the T o t al C o l i f

o rm R ul e, do es the l ab o rat o rypro m p t l ynoti f ythe p rop er autho ri tywhen res ul t s i nd i cat e no n- co l i f o rms m ay have i nterfered with to t al co l i f o rm an alysi s?V I-1 

## Chapter VI Critic al Elem ents for Radiochemistry 1. P ersonnel 1. 1 Labor ator

y Supe r vis or A ta m i ni m um , the l ab o rat o rysup ervi sor sho ul d ha ve a b achelor’ s d egree i n chem i stryor an eq ui val entde gree, and one ye a r o f ex p e rie n c e in the m e a su re m e n t o f ra d io a c tive a n a lytes in d rin k ing w a te r. T h e la b o ra to ry su p e rv iso r is r e q u ire d to hav e a work i ng kn owl edge of Q ual i ty A s s ur anc e ( Q A ) and Q ual i ty C ontr ol ( Q C ) pri ncipl es an d app l yi tt o al l r adi ochemi cal p ra c tice s a n d p ro c e d u re s c o n d u c te d in h is o r h e r la b o ra to ry. T h e lab o ra to ry su p e rv iso r is re sp o n sib le fo r e n su rin g tha t all l ab o rat o rype rs o nnel have d em o nstrat ed thei r ab i l i tyto sat i sf act o ri l yp erf

o rm the anal y ses t o whi ch they are ass i gned and thatal l dat a r eport ed by the l aborat orymeetthe r equi r ed qual i tyas s ur anc e c r i t er i a. 1 .2 L a b o ra to ry A n a l

yst A ta mi ni mum, the l aborat oryan al y s tsh oul d hav e a bache l or’ s d egr ee i n che mi s tryo ran eq ui val entd egr ee, and one y ear o f exp eri ence i n the m ea su re m e n t o f dri nki ng wat er f o r r ad i o chem i cal pa ram et ers . If the anal y sti s res p o nsib l e f o r the o p erat i o n of anal y t i cal i nstrume nt at i o n, he or she i s re q u ire d t o have co m p l et ed s p eci al i zed trai ni ng off ered by the m anuf acturer, ano ther quali f

i ed trai ni ng f

aci l i ty , or served a pe ri o d o f ap p rent i ceshi p unde r an experi enced anal y st
. T he d u ra tio n o f th is a p p re n tice sh ip is p ro p o rtio n a l to th e s o p h istica tio n o f th e instru m e n t. C o m p le tio n o f th is a p p re n tice sh ip peri od f or i ns trument at i on s houl d be do cumen t ed an d m ai nt ai ned i n atr ai ni ng f i l e. 1. 3 Tec hnician A ta m i ni m um , th e la b o ra to ry t echni ci an shoul d have a hi gh s cho o l dipl o m a o r i t s eq ui val ent
. P ri o r t o wo rki ng i ndependen t l yo n dri nk i ng wat ersampl es , t ech ni ci ans sh oul d hav e a tl eas t6 mo nths benc h e xper i enc e i n dr i nk i ng wat er ana l y s es , an d hav e c om p l eted m etho d trai ni ng progr ams i n the methods theywi l l us e on adai l ybas i s . T hei rcomp l et ed m e tho d tra inin g sh o u ld b e re c o rd e d in a tra inin g fi le
. 1. 4 Sam pling P ers onn el If the l ab o rat o ryal so co nd uct s f i el d sam p l i ng a ct i vi t i es, the l ab o rat o rype rs o nnel who co l l ects am p l es s ho ul d have trai ni ng i n the proper col l ect i on t ech ni que f or al l the types of s amp l es they col l ect
. T hei r t ech ni que wi l l be r evi ewed byex peri enc ed samp l i ng pe rs o nnel pri o r t o i n d ep end ent l yco l l ect i ng dri nki ng wat er com p l i ance m o ni t o ri ng s am p l es. B o th trai ni ng and t ec hn i q ue rev i ew s sho uld b e d o cu m en t ed i n sam p l i ng p erso nn el’ s training fi l es. 1. 5 I nitial and Ongoing D em ons tr ations of Pr of icien cyf orAnal ys t s and Tec hnicians B ef ore begi nn i ng the ana l y s i s of com pl i anc e s amp l es , al l ana l y s t s , and t ech ni ci ans mus tdemo ns tr at e the abi l i tyt o conduct thei rm eas ur ement s wi th a cce pt abl e a ccu r acy , pre ci s i on, an d f r eedom f r om i nt er f

er enc es an d dem ons tr at e thi s prof i ci enc y annu al l y . T h es e demon s tr at i on s of pr of i ci en cy s h ou l d be r ec or ded i n ea ch an al y s t ’ s an d t ec h n i ci an ’ s tr ai n i n g f i l e. T he b e n c h sh e e ts a n d instru m e n t p rin to u ts m a d e d u rin g the se d e m o n stra tio n s o f p ro ficie n c y sh o u ld b e re ta ine d a n d b e a v a ilab le fo r in sp e c tio n . D e m o n stra tio n s o f P ro ficie n c y m u st b e d o n e , b y c o n d u c ting a n M D L s tud y a s d e sc rib e d in 4 0 C F R p a rt 1 3 6 , A p p e n d ix B , or b ythe f

o l l o wi ng a l t ernat e p roce d ure. T he anal y stsho ul d p repa re and m easure a samp l e setof atl eastf o ur r eagent bl an k s a n d f

ou rl abor at oryf

ort i f i ed bl an k s th ath av e th e r adi oan al y t e of i n t er es tadde d t o th em ata kn ownc on ce n tr at i on f o r the rel evantm etho d
. T o de m o nstrat e p rof i ci ency a t q u a n tit at i o n l evel s ap p rop ri at e f o r dri nki ng wat er s am p l es, the ac t i v i ty l ev el adde d t o th e l abor at ory f ort i f i ed bl an k s s h ou l d be be twe enth e r adi oan al y t e’ s M CL a n d i t s r equ i r ed det ec t i on l i m i t
. T o be de em ed an accep t ab l e d em o nstrat i o n of p rof i ci e n c y , the m ean reco veri es and the st and ard d evi at i o n of the r ecove r i es of

the r epl i cat e meas ur ement s s houl d be cons i s t entwi th the r equi r ement s f or accu r acyan d preci s i on des cr i bed in S e c tion 7
. 7 , and reagentblank mea sur em ent s m usthave a m ean resul tbe l o w the d et ect i o n l i m i tf o r each ana lyte mea s u r ed wi th th e meth od. C o nti nu i ng D em o nstrati o ns o f P ro f i cien cies m ay b e d o ne b y rep ea t i ng the stud i es d esc ribe d ab o ve an nu all y, o r ca n c o nsist o f d o c u m e n ting b a tc h Q C sa m p le s the a n a lyst h a s p ro c e sse d d u rin g the ye a r sinc e the la st D e m o n stra tio n o f P ro ficie n c y. T he d ata for at l ea stf

o ur re ag en tb l an ks in d i f f ere nt b atch es a nd m ea sure d o n no n-co nse cu t i ve d ays i s used t o assess sens i t i vi ty , and da t a f

rom f o ur l ab o rat o ryf

o rt i f i ed b l anks no tin the same b at ch and in samp l e b at ches m easured o n noncon s ec u t i v e day s , i s u s ed t o a s s es s a cc u r ac ya n d pr ec i s i on . T h e amo unto f the anal y t e adde d t o th e s ampl e pr epa r at i on b atch l ab o rato ry f

o rti f i ed b l an ks sho uld f

o l l o w the guid an ce f

o r the stud i es d esc ribe d i n the p rec ed i ng p ara gra p h, and use th e ac ce pt an ce c r i t er i a f ou n d f orth em i n Se ct i on 7. 7.V I-2

A l l Ini t i al and O ngo i ng D em o nstrat i o ns o f P rof i ci ency shoul d b e reco rded i n a trai ni ng f i l e specif

i c f o r each anal y stand t ec h n i ci an . Rec or ds r el at ed t o me th ods tr ai n i n g a n d e x t ern altr ai n i ng rel evant t o l abor at oryope r at i on s s h ou l d a l s o be do cumen t ed i n the tr ai ni ng f i l e. 1. 6 D ata Pr oduc ed by Anal ys t s and Tec hnicians in Tr aining D at a pr odu ce d by an al y s t s an d t ec h n i ci an s wh o h av e n ot compl et ed th ei r tr ai n i n g , or do n ot h av e a curr en tde mon s tr at i on of pr of i c i e n c y o n r e c o r d , a s w e l l a s i n s tru m e n t o p e r a t o r s s t i l l i n th e p r o c e s s o f o b t a i n i n g th e r e q u i r e d tr a i n i n g or exp eri ence, are accep t ab l e o nl ywhen revi ewed and val i d at ed b ya f ul l yqua l i f

i ed anal y st , or the l ab o rat o rys up ervi sor. T h e y sh o u ld p ro v id e a p e rm a n e n t re c o rd o f th e ir re v ie w a n d the d a ta ’s a c c e p ta b ili ty b y p la c ing the ir sig n a tur e a n d d a te in i nk on any benc h s heet s, cal cul at i o n s heet s, or repo rt s generat ed by s ta ff w h o l ack suf

f i ci enttrai ni ng or a current d e m o n stra tio n o f p ro ficie n c y. 1. 7 W aive rof Acade m ic Tr aining T he cert i f

i cat i on of

f i cermaywai ve the nee d f or s peci f i ed aca dem i c tr ai ni ng , on a cas e- by

- cas e bas i s , f or h i gh l yex peri enc ed an alyst s. 

## 2. Laboratory F acilities 2. 1 G en eral T he anal y si s o f

co m p l i ance sam p l es mustbe co nd uct ed i n a l ab o rat o rywhere the securi tyand i nt egri tyof the samp l es and the d at a can b e m ai nt ai ned
. T he l ab o rat o ryfaci l i t i es should b e cl ean, ha ve ad eq uat e t em p erature and hum i d i tyco ntrol and adequat e l i gh t i ng atthe bench t op . T he l aborat orym us thave p r ovi s i ons f or the proper di s po s al o f chemi calan d ra di ol ogi cal w astes, i nc l ud i ng l i q uid scint i l l ati o n c o ck t ail m i xture s. S i nc e m an y rad i o ch em i stry pro ce d ure s ca n i nv o l ve ro uti ne use of s tr on g mi n er ala ci ds , s u l f i des a n d/ oror g an i c l i qu i ds , th e a ppr opr i at e type of ex h au s th oods a r e r equ i r ed. T h es e h oods m ustbe ab l e t o pro vi d e the anal y stor t e c h n ic ia n w ith a s uf f i ci enthoo d vel o ci tyt o remo ve hazard o us f um es f rom the wo rki ng area, and ad eq uat e b ench space i nsi d e them t o p erf o rm s am p l e p repa rat i o n act i vi t i es saf el y and wi tho utco ncern o f cr o ss-c o n ta m ina ting sa m p le s. H o o d ve lo c iti e s sh o u ld b e c h e c k e d a n n u a lly. A nal y t i calan d s amp l e s t orag e a r eas mus tbe i s ol at ed f r om al l p ot ent i al sour ces o f co nt ami nat i on. A nysampl e h avi ng an em i ss i o n rat e i n e xcess of 0
. 5 m rem /hr mustbe s t o red i n a s ecured l o cat i o n away f rom dri nki ng wat er s am p l es. T here s h ou l d be s u f

f i ci en ts t or ag e s pac e f

orc h emi ca l s , g l as s wa r e, a n d port abl e e qu i pmen t ; s u f f i ci en tf l oora n d be n chs pac e f

or st ati o na ry eq uip m en t ; an d are as for c l ea ning m aterials. 2. 2 I ns tr um en tation I ns trument s s houl d be properl ygr ounded. Co un t i ng i ns trument s mus t be l ocat ed i n a r oo m o therthanone i n w hi ch s amp l es and s t andar ds ar e bei ng prepar ed, or whe r e other types of che mi cal ana l y s es ar e bei ng perf ormed. A n un i nt errupt ed po wer su p p ly (U P S ) sh o u ld b e a v a ilab le fo r all ra d ia tio n m e a su re m e n t ins tru m e n ts a n d the ir a sso c ia te d c o m p u te rs us e d fo r d a ta acq ui si t i o n and reducti o n. 2. 3 Pr eparation of

Standards I n a r eas wh er e radi oact i ve s t andar ds ar e b ei ng p r epar ed, care sh oul d be t ake n t o mi ni mi ze cont ami nat i on o f su r f ace s , other sam p l es, and p erso nn el. B en ch su rf ac es o f a n i m p erv i o us m aterial c o ve red w i th ad so rb en tp ap er, o r p l asti c o r f i b erg l ass tr ay s l i ned wi th a dsorbentpaper ar e a cce pt abl e. 

## 3. Laboratory Equipm ent and In strum entation T h e l abor at oryi s r equ i r ed t o h av e th e equ i pmen t , s u ppl i es a n d i n s tru men t at i on n ec es s aryt o pe r f orm th e appr ov ed meth ods f orw h i chi ti s c ert i f

i ed. 3 . 1 R a diati o n C o un ti ng I nstrum en ts S p ec i f i c typ es o f

ra d i ati o n co un t i ng syst em s a re ne ce ssary to c o nd uc tthe m ea sure m en t s a s d esc ribe d in the re gu l ati o ns. A l l meas ur ementi ns trument s us ed t o meas ur e r adi oact i vi ty i n com pl i anc e mo ni t ori ng s amp l es mus tha ve a document ed h isto ry o f stab ili ty with re g a rd to the ir a b ili ty to m a k e m e a su re m e n ts a c c u ra te ly, w ith a c c e p ta b le p re c isio n , an d su ff ic ie n tly fre e fro m inte rfe rin g b a c k g ro u n d ra d ia tio n so the d e te c tio n lim its c a n b e m e t. T h e fo llo w ing su b s e c ti o n s s p e c ify ins tru m e n ta tio n tha t m a n y o f th e a p p ro v e d m e tho d s u se fo r th e ir m e a su re m e n ts, wh e n c a lib ra tio n o r re c a lib ra tio n is re q u ire d , an d sta b ili ty ch e c k s tha t ar e d o c u m e n te d a n d re c o rd e d to m o n ito r th e lab o ra to rie s a b ili ty t o m a k e m e a su re m e n ts re liab ly.V I-3 3. 1. 1 Li qu i d Sc i nt i l l at i on C ou nt i ng ( LSC) s ys t e m: A n LSC s y s t em i s es s ent i al i f the l aborat oryi s t o be cert i f i ed f or the meas ur ementof

tr i t i um i n dr i nk i ng wat ers amp l es . I ti s r ecomm ended thatthe l i qui d s ci nt i l l at i on s y s t em have spec tral anal y si s cap ab i l i t i es t o est ab l i sh p rop er regi o ns o f e nergyd i scr i m i nat i o n. T he sy st em s ho ul d have ad eq ua t e se nsit i vity t o m ee tthe d etec t i o n li m i to f the m etho d s. P ri o r t o thei r us e t o m easure com p l i ance m o ni t o ri ng s am p l es, LS C s are i ni t i al l ycal i b rat ed f o r ef f i ci ency usi ng N IST
- traceab l e sources, and the b ackgro und m easured. A f t erwards, an ef

f i ci ency check source and an i nstrume nt b l ank t o check b ackgro und s are me asur ed on ea ch d ay o f us e p rio r to a n y sam p l e m easurements t o veri f ythe i nstrume nt is rea d yt o m easure s am p l es. T hese pri o r- t o - use ef f i ci ency check source and the i nstrume ntblank mea s u r eme n t s a r e r ec or ded an d u s ed t o es t abl i s h con tr ol l i mi t s f orth e i n s tru men t ’ s pe r f orman ce . D u r i n g pe r i ods w h e n L S C s a re id le , th e e ff ic ie n c y c a lib ra tio n a n d b a c k g ro u n d sh o u ld b e c h e c k e d w e e k ly to c o n firm th e L S C ’s re a d y sta tus fo r sa m p le m e a su re m e n ts. W arni ng l i m i t s and co ntrol l i m i t s shoul d be cal cul at ed f rom the ef

f i ci ency and ba ckground checks. I f any e ff ic ie n c y c h e c kso u rc e m e a su re m e n t o r in stru m e n t b la n km e a su re m e n t ex c e e d s its co n tro l l im it s , th e L S C is pl ac ed ou tof

s erv i ce u n t i l th e r ea s on f orth e ou tof

c on tr ol me as u r eme n tc anbe i den t i f i ed an d c orr ec t ed. 3. 1. 2 Gas - f

l ow p r op or t i on alc ou nt i ng s ys t e m: A gas- f l o w p rop o rt i o nal co unt i ng s y st em m ay b e used f o r the mea s u r eme n t of g r os s alph a an d g r os s bet a ac t i v i t i es , r adiu m- 226, r adiu m- 228, s tr on t i u m- 89 , s t r on t i u m- 90, r adi oac t i v e ce s i u m, i odi n e- 131, an d u r an i u m as des cr i bed i n th e r ef er en ce s i n 40 CFR part141. 25( a)
. T h e det ec t or m ay b e ei ther a "windo wl ess " (i nt ernal pro p o rt i o nal co unt er) o r a "thi n wi nd o w" typ e. A co m b i nat i o n of shi el d i ng an d a cos mi c ( g u ar d) det ec t or oper at ed i n an t i coi n ci den ce wi th th e mai n det ec t or s h ou l d be u s ed t o a ch i ev e l ow b ackgro und be t a- co unt i ng cap ab i l i ty . T he al p ha and b et a b ackgro und co unt o f the sy st em shoul d b e l o w eno ugh s o that the s ens i t i vi ty of the r adi oanal y s i s of wat er s amp l es wi l l meet the r equi r ement of 40 C FR part141
. 25 ( c) wi th reas o nab l e co unt i ng t i m e (notm o re than 1 , 0 0 0 m i nut es)
. S o l i d s ab so rp t i o n c urv es that relate the att en ua t i o n of alp ha o r b eta p arti cle e m i ssi o n w i th incre asing sa m p l e m ass i n the p l an ch et are essen t i all y i nit i al ca l i b rati o ns f o r sp ec i f i c an alyt es m ea sure d w i th ga s p ro p o rti o na l co un t ers. T heyar e e i therr emeas ur ed an d a new curve gen er at ed, or the curr entac cur acyof the ori gi nalcu rve r eve r i f i ed o n a re g u la r b a sis. E P A re q u ire s this b e d o n e a n n u a lly. EP A r ec ommen ds t he f ol l owi n g a s a n a cc ept abl e appr oac h t o s ol i ds a bs orpt i on c u rv e r ev er i f i ca t i on . Sol i ds ab sorp t i o n s t and ards are s t o red i n a de si ccator, i sol at ed f rom o ther s am p l es af

t er thei r i ni t i al us e i f

the l ab o rat o ry pl an s t o u s e th em l at er t o r ev er i f yth ei rs ol i ds a bs orpt i on c u rv es . W h ens ol i ds a bs orpt i on c u rv es n ee d t o be reveri f i ed , at l east three o f the o ri gi nal sol i d s ab sorp t i o n st and ards m ad e t o p rod uce the sol i d s ab sorp t i o n curve curr ent l yi n u s e f

or dat a r educt i on ar e s el ect ed f or r emeas ur ement
. T heys houl d s pan the r ang e of wei gh t s us ed i n the ori gi nals ol i ds abs orpt i on cu rve. For the r eve r i f i cat i on meas ur ementt o b e ac ce p t ab l e, the ori gi nal mea s u r eme n tof

th e s ol i ds s t an dar d s h ou l d oc cur wi th i n th e r an g e def i n ed byth e u n ce rt ai n tyof th e r ev er i f

i ca t i on m e a su re m e n t ca lc u la te d a t th e 9 5 p e rc e n t c o n fid e n c e lev e l. If an y o f th e ve rifica tio n c h e c k s a re no t with in th is ac ce pt abi l i tyr an g e wh enc ompar ed t o th ei ror i g i nal m easure men t s , a l l s ol i ds a bs orpt i on s t an dar ds mu s tbe r eme as u r ed f

orboth s ampl e we i g h t an d ac t i v i ty , th en u s ed t o g en er at e a n ew s ol i ds a bs orpt i on c u rv e t o u s e f or d a ta re d u c tio n . If a so lid s a b so rp tio n sta n d a rd w e igh t i s n o t with in five p e rc e n t o f i ts p re v io u sly re c o rd e d w e igh t, then i ti s p o ss i b l e the samp l e has been co nt am i nat ed , o r suf f ered loss of samp l e, and theref o re i s d i scarded and a new soli d s ab sorp t i o n s t and ard m anuf actured t o repl ace i t
. I f m o re than o ne sol i d s ab sorp t i o n s t and ard we i ght i s notwi thi n thi s ac cept anc e r ang e, thenthe cu rr ent l yus ed s ol i ds abs orpt i on cu rve maybe ve r i f i ed as des cr i bed bel ow, or an ent i r el yne w s eto f sol i ds abs orpt i on st andar ds canb e m anu f actur ed t o meas ur e a nd ge ner at e ane w sol i d s ab sorp t i o n curve t o us e f o r dat a reduc t i o n. Fors h ortl i v ed r adi oan al y t es , ori f th e or i g i n als ol i ds a bs orpt i on s t an dar ds ar e s u s pec t ed of bei n g c ompr omi s ed, atl eas tthr ee new s ol i ds abs orpt i on s t andar ds ( one eac h r epre s ent at i ve of the l ow, med i um and hi gh wei gh tr ang es o f the o rigi na l so l i d s ab so rp t i o n c urv e) c an b e m an ufac ture d an d used as so l i d s ab so rp t i o n ve rif i ca t i o n stand ard s. Fo r the veri f

i cat i o n m easurement t o b e accep t ab l e, the ef f i ci ency f o r the veri f

i cat i o n s t and ard ca l cul at ed f rom the s ol i ds abs orpt i on curve pres ent l yi n us e sh oul d occurwi thi n the r ang e d ef i ned b ythe un cert ai ntyof the ef f i ci enc y ver i f i cat i on meas ur ement cal cul at ed at the 95 perc entconf i dence l eve l
. I f

an yve r i f

i cat i on s t andar d’ s meas ur ed ef f i ci enc yi s notwi thi n thi s ac cept anc e r ang e, thenad di t i onal sol i ds abs orpt i on st andar ds ar e manu f actur ed that s pan the r ang e of

s amp l e wei gh t s us ed atthe l aborat ory . T he comp l et e se to f s ol i ds s t andar ds i s thenmeas ur ed t o pro d uce a new soli d s ab sorp t i o n curve t o us e f o r dat a reduc t i o n.V I-4 Fo r s t a b ili ty c h e cks, an ef f i ci ency cal i b rat i o n check source and the i nstrume ntba ckground i s m easured and re c o rd e d p rio r to m e a su rin g s a m p le s o n e a c h d a y o f use to ve rify th e instru m e n t i s re a d y to m a k e sa m p le mea s u r eme n t s . D u r i n g pe r i ods w h eng as pr opor t i on alc ou n t er s a r e i dl e, th e ef f i ci ency cali br at i on s h ou l d be c h e c k e d d a ily a n d the b a c k g ro u n d c h e c k e d w e e k ly t o c o n firm the re a d y sta tus o f th e instru m e n t f o r sa m p le m e a su re m e n ts. If a ga s p ro p o rtio n a l co u n te r m e a su re s sa m p le s se q u e n tially w ith a n a u to m a tic sam p le c h a n g e r, it is p o ssib le b o th the s i ze of the bat ch and l ong c ountt i mes cr eat es a s amp l e c ount i ng s etthatr equi r es s eve r al day s t o com pl et e. In these cases , m easurements of samp l e p repa rat i o n ba t ch Q ual i ty C o ntrol (Q C ) s am p l es may s ub st i tut e f o r the i ns trumentch eck s . Af

t er24 hour s of

cont i nu ous l ymeas ur i ng s amp l es , a meas ur ement of an ypre para t i on bat ch Q C s ampl e wi th a kn owna mou n tof th e an al y t e adde d t o i t(S ee s ec t i on 7. 7) m aybe s u bs t i tu t ed f orth e ca l i br at i on ch ec k s o urc e m ea sure m en t , an d an y Q C sam p l e m ea sure m en t ex p ec t ed t o ha ve no ac t i vity i n i t(rea ge nt b l an ks) m a y su b stitute fo r a n instru m e n t b la n km e a su re m e n t. W arn i n g l i mi t s a n d c on tr ol l i mi t s f ori n s tru men ts t abi l i tyme as u r eme n t s s h ou l d be c al cu l at ed. I f i n s tru men tc on tr ol m easurements exceed thei r control l i m i t s, the p rop o rt i o nal co unt er i s p l aced o ut o f servi ce unt i l the reason f o r the c h a n g e in e ff

ic ie n c y o r b a c k g ro u n d c a n b e d e te rm ine d a n d c o rr e c te d . If th e o u t o f c o n tro l m e a su re m e n t i s a sa m p le b at ch Q C s am p l e subst i tut i ng f o r an i nstrume ntco ntrol m easurement, the ent i re batch i s rem easured af t er the r ea s on f orth e ou tof

c on tr ol e v en ti s i den t i f i ed an d a ddr es s ed. I f

a g as proport i on alcou n t eri s mov ed, s erv i ce d, or h ad an i n t errupt i on i n e i th erg as f l ow ore l ec tr i ca l pow er , th e pl at eau v ol t ag e f orboth a l ph a an d be t a i s v er i f i ed, i t s c r os s t al kf

ac t or s r eme as u r ed, an d th e s ol i ds abs orpt i on curv es f or ea ch an al y t e r ev er i f

i ed or r eg en er at ed pr i or t o m ea suring a ny co m p l i an ce m o nit o ring sam p l es. 3. 1. 3 A l ph a s c i nt i l l at i on c ou nt i ng s ys t e m: Fo r m easurement o f gross al p ha act i vi t i es and radium- 2 2 6 , a s ci nt i l l at i on s y s t em de s i g n e d f ora lph ac ount i ng maybe s ubs t i tut ed f or the gas - f

l ow propo rt i onal coun t erdes cr i bed above. In s ucha s y s t e m, a M y l a rdi s c c oa t e d w i th a ph os ph or( s i l v e r - a c t i v a t e d z i n c s u l f i de ) i s e i th e rpl ace d d ire c tly o n th e s a m p le o r o n the fac e o f a p h o to -m u ltip liertub e , en c lo se d w ithin a li g h t-t igh t co n ta ine r, a lo n g w ith th e appr opr i at e el ec tr on i cs ( h i g h v ol t ag e s u p p l y , p re- ampl i f i er , a mpl i f

i er , t i mer , a n d s ca l ar )
. Sol i ds a bs orpt i on cu rve s f o r ea ch an alyt e m ea sure d b y al p ha scint i l l ati o n c o un t i ng sy st em s a r e ge ne rated o r rev erif i ed at l ea st a n n u a lly a s d e sc rib e d a b o v e fo r g a s p ro p o rtio n a l co u n te rs . P ho t o M ul t i p l i er T ub e (P M T ) ef f i ci ency and ba ckground checks a re m e a su red and record ed pri o r t o the mea s u r ementof

a s etof c ompl i an ce mon i t or i n g s ampl es , ora tl ea s tw ee k l yt o v er i f yth e alph a s ci n t i l l at i on syste m ’s re a d y sta tus to m e a su re sa m p le s. W a rn ing lim its a n d c o n tro l l im its f o r b o th typ e s o f i n stru m e n t stab ili ty m easurements s ho ul d be cal cul at ed
. I f a nyP M T e f

f i ci ency o r b ackgro und c heck exceed s i t s co ntrol l i m i t s, the i ns trumenti s pl ace d outo f servi ce un t i l su ch t i me the r eas on f or the outof

contr ol meas ur ementca n be i dent i f i ed an d c orr ec t ed. 3. 1. 4 Sc i nt i l l at i on c e l ls ys t e m: An al pha sc i nt i l l at i on sy s t em des i gn ed t o acce ptalpha s ci nt i l l at i on f l as ks ( "Lucas ce l l s ") s h ou l d be u s ed f orth e s pec i f

i c mea s u r eme n tof

r adiu m- 226 by th e r adon ema n at i on me th od. T h e det ec t i on s y s t e m c o n s i s t s o f a l i g h t - t i g h t e n c l o s u r e c a p a b l e o f a c c e p t i n g L u c a s c e l l s , a p h o t o m u l t i p l i e r tu b e de t ec t or (p h o to tub e ), a n d the a p p ro p ria te e le c tro n ic s (h igh v o ltag e s u p p ly, a m p lif ie r, tim e rs , a n d s c a la rs ). T h e L u c a s c e lls n e e d e d fo r this m e a su re m e n t m a y e ithe r b e p u rc h a se d fro m c o m m e rc ia l su p p lier s o r c o n stru c te d b y the lab o ra to ry. Luca s ce l l ef

f i ci enc yca l i brat i on cons t ant s ar e c hec ked a nd r ecor ded a t l eas t quart er l y or eve ry 10 us es ( whi che ver o c c u rs m o re f

reque nt l y ) us i ng a s t and ardized so l ut i o n of R a- 2 2 6 and a bub b l er excl usi vel yde d i cat ed t o th is act i vi ty . Al l ce l l cons t ant s ar e meas ur ed r el at i ve t o a s peci f i c s ci nt i l l at i on ce l l coun t i ng s y s t em. Al s o, ea ch Luca s ce l l s h ou l d h av e ac ce pt abi l i tyl i mi t s f orre s u l t s th atar e c al cu l at ed f r om th e pr ev i ou s c el l c on s t an t s de t ermi n ed f or eac h s peci f

i c c el l
. I f

a l aborat oryha s mo r e thanone s ci nt i l l at i on c ell c o un t i ng sys t em, the ef f i ci enc yof

ea ch count i ng s y s t em s houl d be det ermi ned u s i ng a c al i brat ed alpha s ci nt i l l at i on s our ce. T hei ref f i ci enc i es ar e c hec ked atl eastweekl yt o veri f ythe read ys t atus o f the sy st em t o m easure s am p l es, or p ri o r t o us e (whi chever i s m o re f reque nt ). C el l b ackgro und s al so are m easured and record ed pri o r t o the m easurement of co m p l i ance m o ni t o ri ng sa m p le s. C o n tro l l im its f o r b o th e ff

ic ie n c y a n d b a c k g ro u n d c h e c k s a re c a lc u la te d
. If a cel l co nst antor ef

f i ci ency check exceed s the d et erm i ned co ntrol l i m i t s, the af f ect ed Lucas cel l or co unt i ng s y s t em mus tnotbe us ed un t i l the outo f co ntr ol p ar amet eri s i nv es t i gat ed. I f , af t err emeas ur i ng the che ck s our ce, the cha ng e i n c el l cons t antor count i ng s y s t em ef f i ci enc yi s conf

i rmed , the l aborat orys upervi s or mayal l ow themV I-5 t o be u s ed, u s i n g th e n ew c el l c on s t an torc ou n t i n g s y s t em ef f i ci en cyw h enr edu ci n g c ou n t i n g da t a f r om com pl i anc e moni t ori ng s amp l e meas ur ement s . I f the out of contr ol che ck i s the backg r ound of the Luca s ce l l , then the cel l mus tbe pl ace d outof s ervi ce un t i l the exc es s back gr o un d decay s t o l eve l s thatal l ow the r equi r ed d etec t i o n li m i t s b e a ch i ev ed within rea so na b l e c o un t i ng t i m es. 3 .1.5 G a m m a sp ec tro m ete r sy stem s: E ithe r a so lid -state , l ithiu m -d rifted ge rm a n ium d e te c to r o r a hig h -p u rity i ntr i ns i c g ermani um det ect or c onnec t ed t o a mul t i cha nn el an al y zeri s ne eded i f the l aborat oryi s t o be cert i f i ed f o r an alyses o f

p ho t o n e m i tt i ng rad i o nu cli d es. A s y s t em wi th a l i th i u m- dr i f t ed g erman i u m, ora h i g h

- pu r i tyi n tr i n s i c g erman i u m det ec t orma ybe u s ed f or mea s u r eme n tof

ph ot on e mi tt i n g r adi on u cl i des i f th e e f f i ci en cyof

th e de t ec t ori s ade qu at e t o m ee tth e det ec t i on l i mi t s l i s t ed at40 CFR part141. 25( c)
. T h es e det ec t or s s h ou l d be s h i el ded wi th a mi n i mu m of 10 cm of i r on or eq uiva l en t
. In a d d i t i o n to ap p ro p riat e e l ec tro nics, the m ult i ch an ne l an alyzer sh o uld co ntain a m em o ry of no tl ess than409 6 cha nn el s an d atl eas tone r eadoutdevi ce. U p o n recei p t , af

t er s ervi c ing o r if

the i nstrume nt ’ s sett i ngs are changed f o r anyreas o n, gam m a spectr o m etry sy st em s m ustbe ef

f i ci ency a n d e nergycal i b rat ed , and the b ackgro und of the d et ect o r cave me asur ed f o r a suf f i ci entt i m e so thatanypo t ent i al l yi nt erf eri ng pho t o - p eaks f

rom natural l yoc curri ng gam m a em i tt ers i n the c o u n t ro o m e n v iro n m e n t ca n b e id e n tif ie d , as w e ll a s the gr o ss g a m m a a c tivity. C a lib ra tio n so u rc e s m u st b e in the s ame ge om etrythe l aborat orych oo s es t o m eas ur e dr i nk i ng wat ersampl es , and cont ai n N I ST
- tr ace abl e s our ces fo r se v e ra l ga m m a e m itt e rs w h o se p h o to -p e a k s sp a n the w id th o f th e ga m m a sp e c tra l win d o w the instru m e n t i s s e tt o d e t e c t
. The c a v e ba c k g r ou nd m us tb e r e me a s u r e d a tl e a s tmo n th l y , a n d th e ba c k g r ou n d g r os s g a mma ac t i v i tyr ec or ded. A cal i b rat i o n check source i s m easured pri o r t o s am p l e m easurements on ea ch d ay o f us e. If gam m a d et ect o rs are i d l e, the cal i b rat i o n check source i s m easured atl eastweekl yt o ve ri f ythe d et ect o r’ s read ys t atus f o r measuri ng co m p l i an ce m o nit o ring sam p l es. Cal i br at i on ch ec ks ou r ce s f ori n tr i n s i c g erman i u m g amma s pec tr omet er s m u s tcon t ai n a m i n i mu m of two ph ot opeaks
. O ne phot o- peak i s l ocat ed aten er gi es bel ow the maxi mum ef f i ci enc yen er gyof

the det ect or ( kn own as the “ knee” o f

the d et ect o r energyvs ef f i ci ency curve) where the curve has a po si t i ve sl o p e, a nd ano ther abo ve the d et ect o r ef

f i ci ency knee, where the ef f i ci ency curve has a negat i ve sl o p e. E ach p eak m ustb e eval uat ed f o r the d et ect o r ef

f i ci ency atthatenergy , the channel num b er of the p eak centr o i d , and the FullW i d th atH al f M axi m um (FW H M ). W a rn i n g l i m i t s a n d c o n tr o l l i m i t s f o rth e e f f i c i e n c y , th e c h a n n e l n u m b e ro f th e p e a kc e n t r oi d , a n d th e F W HM o f each p ho t o - p eak i n the check source, and the gross t o t al act i vi tyof the d et ect o r cave bac kground m easurement i s ca l cu l ated
. If

an y m o nit o red p ara m eter i n the eff i cien cy ch ec k s o urc e m ea sure m en t ex ce ed s the i r co ntro l l i m i t s, the ga m m a d e te c to r m u st b e re c a lib ra te d . If the gr o ss g a m m a a c tivity o f th e c a v e b a c k g ro u n d c h e c ke x c e e d s its co ntrol l i m i t , the source o f the i ncreased b ackgro un d m u stbe i d ent i f i ed , remo ved i f

po ss i b l e, and the cave b a c k g ro u n d re m e a su re d p rio r to a n a lyz ing co m p lian c e m o n ito rin g sa m p le s. 3 .1 .6 A l ph a Sp ectro m eter Sy st em s: A lp h a sp e c tro m e try syste m s sh o u ld b e u se d fo r ura n ium m e a su re m e n ts if specif i c urani um i sot o p e act i vi t i es ar e o f i nt erest
. A l p ha spectr o m etrys y st em s co nsi stof a sol i d s t at e d evi ce (a s i l i con s ur f

ace barr i erdet ect or, a pas s i vel yi mp l ant ed pl ana rs i l i con det ect or, et c. ) m ount ed i n ava cuu m chamber a n d c o n n e c te d to a m u lti c h a n n e l an a lyz e r sys te m . T h e d e te c to rs in a lp h a sp e c tro m e try syste m s a re e n e rg y a n d e ff ic ie n c y c a lib ra te d , an d the b a c k g ro u n d o f i ts s amp l e chamber meas ur ed whe n i ti s p l ace d i nt o s ervi ce. E ner gycal i brat i ons are che cke d week l y , an d ef f i ci enc y cal ib rati o ns ch e c k e d m o n thly to ve rify th e a lp h a sp e c tro m e try syste m ’s stab ili ty. T h e sa m p le c h a m b e r b a c k g ro u nd i s checked at l east m o nthl y , and shoul d b e checked af t er each day of

s am p l e m easurements t o m o ni t o r f o r alp ha rec o i l co ntam i na t i o n from sam p l es. T he alpha peak s i n the che ck s our ce ar e e val ua t ed fo r the det ect or e f f i ci enc y , the cha nn el nu mb erof the peak centroid, and the FW H M
. T hese alp ha p eak p rop ert i es ar e reco rded and co ntro l lim its c a l cul at ed
. T he b ackgro und act i vi tycheck i s f o r t o t al al p ha act i vi tyf o r each detect o r s am p l e cham b er and i s al so reco rded and u s ed t o s etc on tr ol l i mi t s . I f a n ymon i t or ed alph a pe akprope rtyi s ou tof c on tr ol , th e det ec t ori s t o be r ec al i br at ed.V I-6 If a ba ckground check i s o ut o f co ntrol, the d et ect o r and cham b er i s p l aced out o f s ervi ce unt i l the source o f the b a c k g ro u n d inc re a se is i d e n tif ie d a n d re m o v e d . If ex c e ss a lp h a b a c k g ro u n d c a n n o t b e re m o v e d , th e d e te c to r is pl ac ed ou tof

s e rv i c e u n t i l th e c o n t a m i n a t i o n d e c a y s t o a l e v e l th a ta l l o w s th e r e q u i r e d d e t e c t i o n l i m i t s t o be achi eved wi thi n reas o nab l e co untt i m es f o r the anal y t e b ei ng i nt erf ered wi th by the co nt am i nat i o n. 3 . 1 . 7 O ther Instrum en tati o n: O the r ra d ia tio n m e a su re m e n t d e v ic e s a s m e n tio n e d in l e g islatio n o r re fer re d to i n th e a ppr ov ed meth ods ca n al s o be us ed f orme as u r i n g r adi oan al y t es i n dr i n k i n g w at er . T h ei ri n i t i alc al i br at i on p e rio d s a n d c h e c k s sh o u ld b e c o n sis te n t w it h the m e tho d s the y a re us e d fo r a n d the m a n u fac tur e r’s re c o m m e n d a tio n s. 

## 4. Ge neral Laboratory P ractices 4 .1 Ch e m ic a ls/re ag e n ts C hemi cal s an d re age nt s s houl d m eetthe s peci f i cat i ons i n the methods. I f not s peci f i ed, then "A nal y t i cal R eag entG r ade" ( AR) orAmer i ca n Che mi ca l Soc i ety( ACS) g r ade c h emi ca l s orbe tt ersho ul d b e u s ed. Radi oac t i v e s t an dar ds a r e t o be c e rtifi e d b y the N a tio n a l In stitute o f S ta n d a rd s a n d T e c h n o lo g y (N IS T ) o r tra c e a b le to a N IS T -ce rtifi e d so u rc e . 4. 2 R eagentWater T he l ab o rat o rysho ul d ha ve a source o f reagentwat er mee t i ng the requir em ent s o f be i ng an A ST M T yp e 1 , 2 , or 3 reagent water , havi ng a m i ni m um res i st i vi tyof 1 0 m ego hm s/ cm at25 C
. I ts ho ul d be m o ni t o red d ai l yby me asur i ng the reagent o wa t er ’ s c on du ct i v i tyorr es i s t i v i tya n d doc u men t ed. R a d io a c tive c o m p o n e n ts ha v e b e e n kn o w n to b re a kthr o u g h re a g e n t wa te r m a n u fac tur ing un its b e fo re a n inc re a se in r es i s t i vi tyi s not ed. T o m oni t or the backg r ound r adi oact i vi tyof the r eag entwat er , i t i s t o be s cr een ed f or r adi oact i vi tyaf

t er each t i m e th e tr e a tm e n t uni ti s servi ced , and pe ri o d i cal l ythereaf t er dep end i ng on vo l um e o f reagentwat er us ed atthe l ab o rat o rybe tween uni ts ervi ci ng. 4 .3 G la ssw a re/P la sticw a re S p e c ifi c re q u ire m e n ts in the m e tho d s fo r th e c le a n ing o f gla ssw a re sh o u ld b e fo llo w e d . T h e p u rp o se o f th e se re q u ire m e n ts ar e t o mi n i mi ze th e pos s i bi l i tyth atg l as s wa r e ca n con t ami n at e s ampl es , an d s h ou l d i n clu de ac i d r i n s i n g . Aci d r i n s i n g n ot onl ymo bi l i zes an ymet al s remai ni ng ad her i ng t o thei rsu r f ace s , b utal s o hy drat es the out ersi l i ca l ay ero n g l as s war e wh i ch i nh i bi t s co nt ami nat i on wi th ra di oact i ve mat er i al s . I f ther e a r e n o s peci f

i cat i ons f

or c l ean i ng gl as s war e i n the method, then g la ssw a re sh o u ld fi rs t b e w a sh e d in d e te rg e n t so lutio n , the n tho ro u g h ly rin se d in t a p w a te r f o llo w e d b y a se c o n d rin se in a d ilute a c id so lutio n , an d fi n a lly rin se d w ith re a g e n t wa te r a n d d rie d
. La bor at oryg l as s wa r e s h ou l d a l so b e ch ec k ed per i odi ca l l yf orc r ac k s , s cr at ch es , a n d a br as i on s . I f th eya r e f

ou n d t o be d am aged , they are to b e d i scarded
. Sc rat ched glas sware i ncreases the l i kel i ho o d of samp l e co nt am i nat i o n or l o ss es due to the inc re a se o f sur fac e a re a tha t i s e x p o se d to the sa m p le
. 4 .4 Sa fe ty G uid eli ne s i n the L ab o rato ry S afetyM an ua l , the C he m i ca l H ygi en e P l an , o r the S t an d ard O p era t i ng P ro ce d ure s (S O P s) s h ou l d i n clu de s af

etytr ai n i n g a n d pr ot ec t i on i n f ormat i on s pec i f

i c t o a r adi och emi s tryl abor at ory . T h i s s h ou l d i n clu de i dent i f y i ng wh ato pera t i ons o f the l aborat oryus e haza r do us m at er i al s , i dent i f ysame, an d det ermi ne the hea l th r i s ks that ar e pos s i bl e i f

s omeon e i s e x pos ed t o th em. T h i s s h ou l d a l s o i n clu de wh atw ork er s s h ou l d do t o pr ot ec tth ems el v es f r om expos ur e a nd po s s i bl e i nj ury . For r adi ochemi s try l aborat ori es , thi s s houl d i nclude whe n and how r adi at i on s hi el di ng i s us ed t o pro t ectanal y st s and t echni ci ans f rom harmf

ul l evel s o f

radi o act i vi ty . I ts ho ul d al so d ef i ne when ci rcum st ances warrant the u se o f

pro t ect i ve eq uip m ent , and s ho ul d i ncl ud e the use of gl o ves, l ab o rat o ryco at s, ey e p rotect i o n and ap p ro p ria te p i p ett i ng t ec hn i q ue s t o av o i d ex p o sure an d p o ssi b l e injury f ro m ch em i ca l s and rad i o ac t i ve sub st an ce s. 

## 5.Analytic al M eth ods T he approved methods ci t ed at4 0 C FR p art1 41
. 25 ( a) and ( b) m us tb e us ed f or the an al y s i s o f

dri nk i ng wat ercomp l i anc e sa m p le s. T h e se a re li ste d in T a b le V I-1
. 5. 1 Stand ard O

per ating P r ocedures A l abor at ory - s pec i f

i c St an dar d O per at i n g Pr oce du r e ( SO P) s h ou l d be w r i tt enf orea chm eth od us ed byth e l abor at oryf or meas ur i ng r egu l at ed r adi o- ana l y t es i n c om pl i anc e mo ni t ori ng s amp l es . T hes e SO P s s houl d be cons i s t entwi th ar ef er enc ed appr ov ed meth od. An yEP A- appr ov ed modi f i ca t i on s s h ou l d be n ot ed.V I-7



## 6. Sample Collection, Hand ling, an d Preservation Samp l e c ont ai ner s , pre s ervat i ves , an d hol di ng t i mes s peci f i ed i n the methods s houl d be f ol l owed. T abl e V I
- 2 l i s t s cr i t i cal el ement s f

or s amp l e h andl i ng , i ncludi ng pre s ervat i on. Samp l e pr es ervat i ves provi ded by the l aborat orys houl d be s cr een ed fo r ra d io a c tive c o n te n t b y lo t nu m b e r p rio r to th e ir u se in th e la b o ra to ry a n d d o c u m e n te d . S a m p le s p re se rv e d w ith re a g e n ts n otpr ov i de d b yth e l a bo r a t orya r e t o be accomp a n i e d b ya bl a n k , r a di oa c t i v e f r e e s a mp l e th a ti s pr e s e rv e d i n th e s a me manne ras the s ubmi tt ed s amp l e. 6. 1 Compo s ited Sam ples If de em ed accep t ab l e b ythe st at e, s am p l es may be co m p o si t ed by the ut i l i tyo r the l ab o rat o ryas an al t ernat e samp l i ng prot ocol , provi ded that al l s amp l e al i quot s ar e properl y pres erved at the t i me of col l ect i on. Si nce the r equi r ed comp l i anc e m o ni t o ri ng m easurement i s “t o t al a ct i vi ty ” samp l es ar e no tf i l t ered be f

o re preservat i o n, as i tm ustrepresentthe m axi m um p o t ent i al exp o sur e f

rom dri nki ng wat er. Sam p l es mustbe drawn o n a qua rt erl yba si s and where co m p o si t i ng i s no tdo ne byth e l abor at ory , th er e mu s tbe doc u men t at i on s u bmi tt ed wi th th e compos i t e s ampl e det ai l i n g on w h atda y s e ac h a l i qu ot was obt ai ned, i t s vol ume, an d wheni twas pre s erved. A s amp l e o f the p rese rva t i ve s houl d accomp anythe com po s i t ed samp l e t o the l ab o rat o ryt o de t erm i ne the co ntrib ut i o n of radioacti vi ty , i f any, f rom the ad d i t i o n of the p res ervat i ve t o the sa m p le . A n a lysis o f co m p o site s a m p le s m u st b e c o m p le te d w it h in o n e ye a r a fterthe fi rs t sa m p le is co llec te d o r w ithin n o rm a l ho ld ing ti m e s if the c o m p o siting p e rio d is l e ss th a n 9 0 d a ys. W h e re p o ssib le , th e la b o ra to ry sho u ld b e re sp o n sib le f o r m an ag i ng the co m p o si t i ng o f sam p l es. 

## 7. Quality Assurance La b o rat o ryQ ual i tyA ss urance sy st em s are al l the p rotoco l s, po l i ci es and p roce d ures i m p l em ent ed a ta l ab o rat o ryt o ensur e the generat i o n o f

d at a o f kno wn q ual i ty . C o m p rehensi ve p rogram s shoul d enco m p ass everyaspect i n the samp l e p roce ss i ng p a th th ro u g h a lab o ra to ry, fro m sa m p le re c e ip t, sto ra g e , m e a s u r e m e n t, to p ro d u c ing the fi n a l re p o rt fo r th e sa m p le m ea sure m en t s as we l l as rec o rd k e e p in g fo r a ll sam p l i ng i nfo rm ati o n, raw d ata a nd rep o rts f o r sam p l es. Itsho uld also addres s l aborat orys uppo r tf un ct i ons , s uchas s peci f y i ng procedur es t o tr a i nan d qu a l i f yt e c h n i c i an s , i ns trumentmai nt ena nce and c al i brat i on procedur es , s uppo r tequipm entmai nt ena nce , pur cha s i ng s uppl i es , and probl em s ol vi ng prot ocol s . Each a re a d isc u sse d sh o u ld a d d re ss p ro v id ing d o c u m e n te d e v id e n c e o f co m p lian c e w ith the sp e c ifi c a re a s o f th e Q u a lity A ssurance sy st em . Sp eci f i c areas f

o r Q ual i ty A ss urance sy st em s are ad d res sed i n C hap t er II I, and area s sp e c ifi c a lly r el eva nti n r adi ochemi s tryl aborat ori es ar e di s cus s ed i n the s ect i ons bel ow. 7 .1 G e n e ra l R e q u ire m e n ts 7 .1.1 A v a i l a b i l i ty o f R ec o rd s a n d D o cu m en ts: T h e a n a lyti c a l m e tho d s re fer e n c e s, Q u a lity A ssu ra n c e M a n u a l, an d SO Ps a r e t o be r ea di l ya v ai l abl e t o th e an al y s t s . Al l Q C da t a an d r ec or ds a r e t o be a v ai l abl e f ori n s pec t i on b ythe cert i f

i cat i o n off

i cer. 7 .2 B a la n c e a n dWe ig h ts R adi ochemi s tryl aborat ori es s houl d hav e bal anc es wi th the appropri at e r ang es f or thei ro pera t i ons . Si nce wei gh t s ar e u s ed t o cal cul at e gravi m etri c reco veri es us ed i n the calcul at i o n o f a nal y t i cal resul t s f

o r som e m etho d s, ba l ances us ed t o wei gh sam p l es sho ul d hav e adocument ed hi s t oryof ac cur acyan d preci s i on. Ata mi ni mum, the l aborat orys houl d hav e a n an al y t i ca l ba l an ce w i th a tl ea s ta . 01 mg s en s i t i v i tyt o me as u r e samp l e wei g h t s . Al s o, a t op l oadi n g ba l an ce s h ou l d be av ai l abl e t o wei g h outch emi ca l s f orre ag en tpre par at i on . B al an ce s m u s tbe re

- ca l i br at ed a tl ea s tan n u al l ywi th AST M T ype 1 wei ght s. A tl eastonc e every31 cal end ar days , the anal yt i cal ba l ance’ s range ap p rop ri at e f o r s am p l e m easurements sho uld b e c he ck ed using thre e trac ea b l e T yp e 1 , C l ass 1 , o r C l ass 2 we i gh t s tha tsp an the ran ge o f sam p l e w eigh i ng ne ed s. P rior to use, the ca l i b rati o n o f the b alan ce sho uld b e ch ec ke d w i th the A S T M T yp e 1 , C l ass 1 , o r C l ass 2 we i gh tclo sest i n wei gh t t o most s amp l e meas ur ement s . M ost com mo nl y thi s i s the 10 gr am wei gh t becau s e t ar ed s t eel pl anc het s o f t en we i g h be twe en8 and 1 0 gram s. T h es e ca l i br at i on c h ec k s s h ou l d be r ec or ded i n a pe rman en tl og . W ei g h t s u s ed f or ca l i b rati o n c he ck s sho uld b e re ce rti f i ed ev ery f

i ve years.
7. 3 MethodSens itivit

y Studies T he l ab o rat o rys ho ul d de t erm i ne the st and ard ana l y t i cal co nd i t i o ns f o r eac h m etho d for m easuri ng co m p l i ance m o ni t o ri ng sa m p le s tha t ca n p ro d u c e d e te c tio n li m its th a t ar e e q u a l t o o r le ss th a n tho se sp e c ifi e d in 4 0 C F R p a rt 1 4 1 .2 5 (c )( 1 ) T a b le B , an d 40 CF R part141
. 25 ( c) ( 2) T abl e C. T hes e pr ocedur es s houl d be cons i s t entwi th A p p en d i x C , of “ P r es cr i bed P roce d ures f

o r the M easurement of R ad i o act i vi tyi n D ri nki ng W at er ( E P A
- 6 0 0 /4
- 8 0
- 0 3 2 ). O nce the m etho d s t and ard a n a lyti c a l co n d iti o n s a re d e te rm ine d , th e lab o ra to rie s sh o u ld the n ins titu te a m o n ito rin g p ro g ra m to e n su re the se n sitivityV I-8

of ea ch meth od u s ed t o an al y ze compl i an ce mon i t or i n g s ampl es does n ote x ce ed th e det ec t i on l i mi t s de f i n ed i n th e CFR r ef er enc es ci t ed above. M ethod backg r ound l eve l s , as meas ur ed f r om a r eag entbl ank , mo ni t or f or pot ent i al cont ami nat i on f r om s eve r al pot ent i al so u rc e s o f i nt erf

erences i n the samp l e p repa rat i o n l ab o rat o ry i n ad d i t i o n t o the b ackgro und o f the d et ect o r used fo r its m ea sure m en t
. C o nse q ue ntl y, sensit i vity m o nit o ring f o r rad i o ch em i ca l m etho d s used t o an alyze d rinking w ater sho uld use th e r es u l t s of

th e r ea g en tbl an k
. T h e ac t i v i tya n d u n ce rt ai n tyof e ac h r ea g en tbl an kpr epa r ed wi t h s ampl e pr epa r at i on b a tc h e s sh o u ld b e c a lc u la te d in the sa m e c o n c e n tra tio n un its as the d e te c tio n li m its l isted in the re g u la tio n s re fer re d to ab o ve, record ed , then the activity p lott ed on a co ntrol chartt o pro vi d e a record thatwi l l s erve as a co nt i nuo us c o n ta m ina tio n a n d se n sitivity m o n ito r fo r th e m e tho d a s it is p e rfo rm e d a t th e lab o ra to ry. 7. 4 P r oficiency Tes t

( P T ) Studies T o b e c erti f

i ed fo r the a na l ysi s o f

a ra d i o - an alyt e in d rinking w ater, a lab o rato ry m ust s uc ce ssf ull y pa rti cip ate in at l ea st o ne P T s tud yf

o r the anal y t e f

o r which they are seeki ng c ert i f i cat i o n. T o m ai nt ai n cert i f i cat i o n f o r an anal y t e, the l ab o rat o ry m ustpass o ne stud ype r y ear thereaf t er. Sco ri ng cri te ria fo r P T sam p l es can be f o und i n the C FR as ci t ed ab o ve. T he l aborat orymaych oo s e t o part i ci pat e i n ei thertype of studyf r om a provi der acc ept abl e t o the s t at e o rE P A thati s des cr i bed bel ow. 7 . 4 . 1 M i xed A l ph a an d M i xed B eta/G a m m a P T Stu dies : T h es e PT s tu dys ampl es c on t ai n e i th era mi x tu r e of

kn own alpha e mi tt i ng r adi onucl i des wh ose conc entr at i ons ar e un kn own t o the l aborat ory , or a mi xtur e of kn own b e ta /ga m m a e m itt ing ra d io a n a lytes w h o se c o n c e n tra tio n s a re a lso un k n o w n to the lab o ra to ry. 7 .4.2 O th er P T S tu d i es: T hes e a r e PT s tud i es w ho se sam pl es cont ai n as i ng l e a nal y t e a tanun kn own conce ntr at i on, o rvar i ed nu mb ero f anal y t es atunkn own concentr at i ons thatar e of t en as s oci at ed wi th e ach other i n dr i n k i n g w at er . T h es e s tu di es a r e di s tr i bu t ed s ev er alt i mes pe ry ea r . I f th e an al y t e th e l abor at ory wi s h es t o be ce r t i f i ed f ori s n otc on t ai n ed i n a M i x ed PT s ampl e, th eymu s ts u cc es sf ul l y a nal y ze i ta tl ea s ton ce a y ea rby part i ci pat i ng i n one of the se o t her P T s tudi es thatcont ai n the r adi oanal y t e f or w hi ch the l aborat orywi s hes cert i f i cat i o n.
7 . 4 . 3 A dd i ti o na lP ro fi ci en cy T est i ng Stu dies : T he l aborat ori es may al s o part i ci pat e i n as econd P T s tudyea ch y earf or each anal y t e f

or whi ch theysee k cert i f i cat i on. V ol un t aryi nt er l aborat oryr ound r ob i n s tudi es coul d m eet th i s r ec ommen dat i on . T h es e addi t i on alP T s tu di es s h ou l d be pe r f ormed i n th e h al f

of th e y ea rth atth e bl i n d or an al y t e- s pec i f

i c PT s tu dyth e l abor at orypa rt i cipat es i n f ora n n u alc ert i f

i ca t i on i s n otpe r f

ormed. T h i s w i l l pr ov i de a bi annualch eckof

the l aborat ory ’ s per f

ormanc e. 7 .5 O p e ra tin gMa n u a ls O p era t i ng m an ua l s and ca l i b rati o n p ro t o co l s f o r co un t i ng i nstrum en t s sho uld b e a va i l ab l e to an alyst s and t ec hn i ci ans . T he se do cumen t s s houl d be s t ored n earthe i ns trument s f or e as yr ef

er enc e. 7. 6 Mainten anc e of

Rec or ds C al i b rat i o n d at a and m ai nt enance reco rds on allradi at i o n i nstrume nt s and anal y t i cal ba l ances s ho ul d be m ai nt ai ned i n a perman entr ecor d. I f

thi s i s a har d copy r ecor d, i tsh oul d be mai nt ai ned i n a p ermanen t l yb ound not eboo k a nd al l en tr i es made i n i n k
. I f

i ti s a n e l ec tr on i c r ec or d, th e i n f ormat i on s h ou l d be s t or ed on a c ompu t ers y s t em th ati s pa s s wor d pr ot ec t ed, a n d b a c k e d up w e e k ly with a c o p y o f th e b a c kup d a ta sto re d o ff site to b e c o n sid e re d a p e rm a n e n t re c o rd
. 7 .7S a mp le Me a su re m e n t Q u a lit y Co n tro l R e q u ire m e n ts Justas the st ab i l i tyof the m easurement i nstrume nt s shoul d be m o ni t o red t o veri f ythe m easurements they p rod uce wi l l have re a so n a b le a c c u ra c y, p re c isio n a n d fre e d o m fro m i n te r fer e n c e s, sim ilarc o n tro ls sh o u ld b e us e d to m o n ito r sa m p le pr epa r at i on a ct i v i t i es . I n or dert o a s s es s th e ef f ec tof s ampl e pr epa r at i on a ct i v i t i es on th e ac cur ac y , pr ec i s i on , a n d f r ee dom f r om i nt er f

er enc es of

dri nk i ng wat er com pl i anc e mo ni t ori ng meas ur ement s , the l aborat oryi ncludes the Q C s amp l es l i s t ed bel ow atthe s peci f

i ed f r equen ci es f

or e ach pre para t i on bat ch of s amp l es thatar e meas ur ed f or the r adi oanal y t es f

or w hi ch the l aborat oryi s ce r t i f

i ed, un l es s otherwi s e n ot ed f or s om e method s. A prepar at i on bat ch of s amp l es ar e thos e s amp l es pr epa r ed s equ en t i al l yt og eth erw i th i n a 24 h ou rper i od. T h e n u mberof c ompl i an ce mon i t or i n g s ampl es i n a pr epa r at i on b at ch shoul d not exceed 20 (excl ud i ng the Q C s am p l es describ ed be l o w).V I-9 7 . 7 . 1 A ss ess m en t o f p rep a ra ti o n b a tch p recis i o n: T h e lab o ra to ry sh o u ld a sse ss th e p re c isio n o f ea c h sa m p le p repa rat i o n ba t ch. T hi s can b e d o ne i n o ne o f two al t ernat i ve ways . Fi rs t , the l ab o rat o rym ay m easure a second al i q uo tof a samp l e f

o r a dup l i cat e m easurement (D U P ). A l t ernat el y , i f nond et ect s f

o r a method are f

reque nt , the l ab o rato ry m ay elec tt o m ea sure a m atri x sp i ke (M S )/ma trix spike d up l i ca te ( M S D ) p air i nstea d
. A n M S i s a se c o n d a liq u o t o f a sa m p le tha t ha s a k n o w n a m o u n t o f th e ra d io a n a lyte b e ing m e a su re d a d d e d to it . A n M S D is a third a l i q uo to f

the sam e sa m p l e that ha s the sam e am o un to f rad i o an alyt e a d d ed t o i tas the M S
. R eg ard l ess o f sam p l e ch o i ce , the D U P o r M S /M S D p air m ust b e p rep are d w i th the o the r sam p l es w i th a f req ue nc y no l ess tha n 1 ev ery 1 0 sam p l es o r l ess. W i th the excep t i o n o f gross al p ha p art i cl e act i vi ty , gross b et a p art i cl e act i vi ty , and tri t i um m easurements , i f there are i nsuff

i cien tsam p l e v o l um es f o r a d up l i ca t ed sa m p l e o r a d up l i ca t e m atri x sp i ke sam p l e in a set o f sam p l es, two al i quot s f

r om the l aborat ory ’ s t ap wat erm aysubst i tut e f or the s amp l e a l i quot s an d be us ed f or the dupl i cat ed s amp l e. P re c isio n sh a ll b e a sse sse d b y c a lc u la ting the re la tive p e rc e n t d iff e re n c e (R P D ) fo r e ithe r th e sa m p le a n d it s d u p lica te , o r fo r th e M S /M S D p a ir. A n R P D re su lt fo r eithe r p re c isio n a sse ssm e n t m e a su re m e n t th a t ex c e e d s its c a lc u la te d c o n tro l l im it, w h ic h , i d e a lly sh o u ld b e 2 0 p e rc e n t o r le ss, ind ic a te s the p re c isio n o f the sa m p le p re p a ra tio n b a tc h is q u e stio n a b le , a n d d a ta re p o rte d fro m the se re su lts sho u ld b e fl a g g e d a s h a v ing q u e stio n a b le p reci si o n. If a dup l i cat ed s am p l e m easurement has an act i vi tythati s l ess than 5 t i m es the radioanalyt e’ s d et ect i o n li m i t , and exceed s 2 0 p ercent R P D when com p ared t o the f i rs t m easurement f

o r the samp l e, i tm ay b e reeval uat ed us i ng the tw o m e a su re m e n t’s re p lica te e rr o r ra tio (R E R ). T h e R E R o f tw o m e a su re m e n ts m a d e fro m th e s a m e s a m p le is ana s s es s men tof

wh eth erth eyar e wi th i n two s t an dar d de v i at i on s of th ei ra g g r eg at e mea s u r eme n tu n ce rt ai n tyof eac h other
. T he R E R i s ca l cul at ed us i ng the f ol l owi ng f

ormul a;

a b RER = | A - B| /S Q RT ( s + s ) < 2 2 2 whe r e; A = N etA ct i vi tyof the f i rs tm easurement B = NetAc t i v i tyof

th e s e c on d m e a s u r e me n tma de f r om a di f

f e r e n ta l i qu otf

r om th e s a me s amp l e. A s = T he uncert ai ntyof the f i rs tm easurement B s = T he uncert ai ntyof the second m easurement If the R E R i s l ess than o r equal t o two , then the two m easurements are wi thi n 2 st and ard d evi at i o ns o f each o ther, and s o are accep t ab l e. I f the R E R exceed s 2 , i ti s unaccep t ab l e si n ce i tm eans there i s great er than 2 s t and ard d ev i ati o ns o f

d i f f ere nc e b etwe en the tw o m ea sure m en t s. I f precisi o n as s es s ment s ex cee d thei rl i mi t s , ca l cul at i ons an d procedur es s houl d be exa mi ned a nd s amp l es re c o u n te d . If the p re c isio n a sse ssm e n ts a re stil l un sa tisf a c to ry afte r th e sa m p le s a re re c o u n te d , th e n a ll sa m p le re su lts i n the p re p a ra tio n b a tc h a re to b e re p o rte d w ith a q u a lif ie r t o ind ic a te the m e a su re m e n t ha s q u e stio n a b le prec i s i on. I f the cl i ent r equi r es un qual i f i ed r es ul t s , then al l the s amp l es i n the di s car ded s amp l e prepar at i on bat ch ar e th enr e- mea s u r ed u s i n g n ew a l i qu ot s of th e s ampl e i f

h ol d t i mes a n d s u f

f i ci en tv ol u me i s a v ai l abl e or r es ampl ed an d r ea n al y ze d. 7 . 7 . 2 A ss ess m en t o f P rep a ra ti o n B a tch A ccu ra cy : M atr i x Spi k e ( M S) s ampl es ar e pr epa r ed t o mon i t orh ow samp l e p repa rat i o n pro ced ures i m p actthe acc uracyo f m easurements of the samp l es in a s am p l e p repa rat i o n ba t ch. A M S s ampl e i s pr epa r ed bya ddi n g a kn owna mou n tof th e r adi oan al y t e be i n g m ea s u r ed t o a s ec on d a l i qu otof a samp l e. T he M S sam p l e has r ad i o anal y t e act i vi t i es add ed to th em thatare app roximatel y10 t i m es the a n t i c i p a t e d l e v e l o f th e s a m p l e a c t i v i ty , o r th e d e t e c t i o n l i m i t , a s a p p r o p r i a t e f o rth e m e a s u r e d r a d i o a n a l y t e . MS s ampl es s h ou l d be pr epa r ed an d pr oce s s ed wi th th e s ampl es a ta f r equ ency o f

on e M S pe rs ampl e pr epa r at i on b at ch. M S perf ormanc e i s as s es s ed us i ng the perc ent r ecove ry of the kn own act i vi ty of r adi oanal y t e a dd ed t o the s amp l e.V I
- 10 T he perc entr ecove ry shoul d be wi thi n the contr ol l i mi t s ca l cul at ed f r om pre vi ous M S meas ur ement s , wh i ch sh o u ld id e a lly b e + 2 0 p e rc e n t o f th e a m o u n t o f a c tivity a d d e d to th e M S s a m p le
. F o r g ro ss a lp h a p a rtic le a c tivity, gr oss bet a part i cl e a ct i vi tymeas ur ement s , an d R a- 22 8 m ethods of an al y s es , wh er e e xper i enc e h as s hown l ower ac cur ac i es c anbe ex pec t ed, c on tr ol l i mi t s c al cu l at ed f r om pr ev i ou s M S r es u l t s s h ou l d be w i th i n + 30 pe r ce n tof th e a mo u n tof

a c t i v i tya dd e d t o th e M S f orth e a c c u r a c yr e l a t i v e t o th e s a mp l e ma tr i c e s t o b e c on s i dere d acc ept abl e. M S sa m p le s n e e d no t b e inc lud e d in p re p a ra tio n b a tc h e s fo r c e rta in m e tho d s. S inc e the re is no w a y c u rr e n tly to q ua nti t ati ve l y ad d an i ne rt ga s t o a li q uid , M S sam p l es w i l l no tb e re q uire d fo r r ad o n in wa t er m ea sure m en t s. A l so , ga m m a sc ree ns o f d r i nk i ng wa t er n ee d no ti nc l ud e M S sam p l es in the i r p rep ara t i o n b atch es o f sam p l es. D ensi ty , as exp res sed b ythe average at o m i c wei ght , o r Z , o f the m at eri al s urr o und i ng a gam m a d et ect o r i s the pri ncipl e f

act or i n s catt er i ng or ab so rb i ng g am ma r ay s . W i th r es pectt o thi s property , ther e i s l i tt l e di f f er enc e betweendei oni zed wat eran d dri nk i ng wat ersampl e m atr i ces . C ons equen t l y , the s ame i nf ormat i on wi th r es pect t o accuracy can b e ass ess ed wi th the l ab o rat o ryf o rt i f i ed blank. (LF B ). W i th the excep t i o n of gross al p ha p art i cl e act i vi ty , gro ss be t a p art i cl e act i vi ty , and tri t i um m easurements , i f there ar e i ns uf f i ci ents amp l e v ol umes f or amatr i x s pi ke i n as etof s amp l es , two al i quot s f r om the l aborat ory ’ s t ap wat er m ay subst i tut e f

o r the samp l e al i q uo t s and us ed f o r the unspi ked / spi ked M S sam p l e p ai r. I f M S a s s es s men t s of

a cc u r ac ye x ce ed th ei rc al cu l at ed con tr ol l i mi t s , c al cu l at i on s a n d pr oce du r es s h ou l d be exa mi ned a nd s amp l es recoun t ed. I f the as s es s menti s st i l l un s at i s f act oryaf t erthe s amp l es ar e r ecoun t ed, then a ll sa m p le re su lts i n th e b a tc h sh o u ld b e fl a g g e d a s p o ssib ly b ia se d lo w o r h igh (a s the re su lt ind ic a te s) d u e to ma tr i x e f f e c t s . I f th e L FB a s s e s s me n tof a c c u r a c y i n de pe n de n t of ma tr i x e f

f e c t s i s a l s o un s a t i s f

a c t oryi n th e s a me p repa rat i o n ba t ch (s ee b el o w), then allthe sam p l es i n the d i scarded s am p l e p repa rat i o n ba t ch are r em easured usi ng n e w a liq u o ts o f th e sa m p le if ho ld ti m e s a n d su ff

ic ie n t vo lum e is av a ilab le
. 7 . 7 . 3 A ss ess i ng the pr ep a ra ti o n b a tch a ccu ra cy i nd ep en de nt o f ma tri x eff ects: In o rd e r to a sse ss the m e tho d ’s prepar at i on bat ch acc ur acyi ndependen to f m atr i x ef

f ect s , an (LFB ) sh al l b e pr epar ed wi th e ach pre para t i on bat ch of s ampl es . T h e LFB i s pr epa r ed bya ddi n g a kn owna mou n tof th e r adi oan al y t e bei n g m ea s u r ed t o a n a l i qu ot of de i on i ze d w at erth ati s pr epa r ed u s i n g th e s ame pr oce du r es u s ed t o me as u r e th e s ampl es i n th e pr epar at i on b a tc h . L F B s a re p re p a re d a t a fre q u e n c y o f o n e p e r p re p a ra tio n b a tc h
. LFB ac cur acyi s as s es s ed us i ng the perc ent r ecove ryof the kn own ac t i vi tyof

r adi oanal y t e a dd ed t o the s amp l e. T h e p e rc e n t re c o v e ry sh o u ld b e w ithin th e c o n tro l lim its c a lc u la te d fro m p re v io u s re su lts, w h ic h sh o u ld id e a lly b e w i thin ± 1 0 p erc en to f the am o un t o f ac t i vity ad d ed t o the L F B sam p l e. F o r gro ss alph a p arti cle a cti vity, gro ss bet a part i cl e ac t i v i ty , a n d Ra- 228 me as u r eme n t s , w h er e l owe ra cc u r ac i es ca n be ex pec t ed, th e ca l cu l at ed con tr ol lim its sho u ld no t ex c e e d + 2 0 p e rc e n t re c o v e ry o f th e a m o u n t o f ac tivity ad d e d to it
. I f LFB as s es s men t s e x ce ed th ei rl i mi t s , c al cu l at i on s a n d pr oce du r es s h ou l d be ex ami n ed an d s ampl es r ec ou n t ed. I f the LFB as s es s menti s s t i l l un s at i s f act oryaf

t erthe sa m p l es ar e r ecoun t ed, the bat ch mus tbe cons i dere d con t ami n at ed. Al l s ampl es i n th e s ampl e pr epa rat i o n b at ch mu s tth enbe di s ca r ded. Af t erth e s ou r ce of cont ami nat i on i s i dent i f i ed an d add r es s ed, al l the s amp l es i n the di s car ded sa mp l e pr epar at i on bat ch mus tthen be r emeas ur ed us i ng ne w al i quot s of the s amp l e i f

hol d t i mes and s uf

f i ci entvol ume i s av ai l abl e. 7 .7.4 A ssessi n g i n stru m en t d rift d u rin g sa m p l e m ea su re m en ts: W h i l e mo s tr a di a t i onme a s u r e me n t s s y s t e ms ar e n ot e w orth y f

or th e i r s t a bi l i ty , s u dd e n c h a n g e s c a n oc c u r du e t o i n s tru me n t c om po n e n t f a i l u r e , l os s of

ga s p re ssu re , v a c u u m , o r co n ta m ina tio n o f a d e te c to r o r sam p le c h a m b e r f ro m a h igh a c tivity sa m p le c a n p o s sib ly occurwh i l e s amp l es ar e bei ng coun t ed. I ns trumentdri f ti n det ect or e f

f i ci enc yan d backg r ound mus tbe ch eck ed b o th be f o re and af

t er mea sur i ng co m p l i ance m o ni t o ri ng s am p l es t o veri f ybo th the cal i b rat i o n and ba ckground di d n otc h an g e s i g n i f

i ca n t l yw h i l e th e s ampl es w er e bei n g me as u r ed. A s amp l e meas ur ement , or “ count i ng ” bat ch i s a s econd type of sampl e b at ch thatcan be def i ned i n r adi ochemi cal meth ods of

a n al y s i s . A s ampl e mea s u r eme n tba t chi n clu des th os e s ampl es th ata r e mea s u r ed wi th i n 24 h ou r s of each o ther, and thei r m easurements o ccur in b etween the d et ect o r ef f i ci ency and ins trume ntba ckground checks m ad e p ri o r t o and atthe end of m easuri ng a s etof co m p l i ance m o ni t o ri ng s am p l es. O f t en a samp l e m easurementV I
- 11 b at ch and a samp l e p repa rat i o n b at ch co nsi stof the same num b er of s am p l es. In som e cases, how ever, i tm ay t ake severalda y s t o m easure al l the samp l es i n a s am p l e p repa rat i o n b at ch. In these i nst ances, ef f i ci ency and i nstrume nt b a c k g ro u n d c h e c k s sh o u ld no t wa it u n tilthe las t sa m p le in a p re p a ra tio n b a tc h h a s b e e n m e a su re d . T o d o so c o u ld p o ssi b l y resulti n sev era l d ays o f l o st co un t i ng t i m e i f the y are o ut o f co ntro l
. Instea d , a sa m p l e c o un to rd er m ust b e arranged so an ef

f i ci ency check and an i nstru m e n t b a c k g ro u n d check i s t o be m ad e atl eastevery24 hours w h ile sa m p le s a re b e ing c o u n te d
. T o f ac i l i t at e th es e dai l yc h ec k s , th e s ampl e or derm aybe arr an g ed s o a s ampl e s pi k ed wi th a kn owna ct i v i tyof the ra d io a n a lyte b e ing m e a su re d (i.e a n L F B , M S , o r M S D sa m p le ) an d a sa m p le n o t e x p e c te d to ha v e m e a su ra b le act i vi tyi n i t(RB ) i s co unt ed atthe end of a 2 4 hour p eri o d , or i f the t o t al s am p l e m easurement bat ch co unting t i me does n ot ex ce ed 24 h ou r s , af t er al l th e oth er s ampl es i n th e s ampl e pr epa r at i on ba t chh av e bee n me as u r ed. I f thes e Q C s amp l es meetthei rac cept anc e c r i t er i a, then i t may be as s umed l i tt l e or no ch ang e i n the ef f i ci enc y o r backgro und oc curred whil e samp l es were b ei ng m easured. If ho wever, sam p l e co unt i ng requi rem ent s p revent thes e s amp l es f

r om bei ng count ed at the end of eac h 24 hour s s amp l e meas ur ementper i od or as the l as ts amp l es meas ur em en t s o f

the s amp l e pr epar at i on bat ch, the ef f i ci enc yca l i brat i on ch ecks our ce us ed f or the ef f i ci enc y ca l i b rati o n c he ck s, an d an i nstrum en tb l an k m ust be m ea sure d i n their p l ac e. B o th o f the se m ea sure m en t s m ust b e w ithin the c o n tro l ra n g e s p ro d u c e d fro m the ir p re v io u s m e a su re m e n ts. If eithe r the Q C sam p l es or the i nstrum en t eff i cien cy o r b ac kg ro un d ch ec k exc ee d the i r ca l cu l ated co ntro l l i m i t s, th e i n s tru men t i s pl ac ed ou t of s erv i ce u n t i l th e s ou r ce of th e ou tof c on tr ol c on di t i on i s i den t i f i ed an d c orr ec t ed. If the f ai l ed i nstrume ntQ C check o ccurs wi th a m easurement mad e at the end of m easuri ng a s am p l e m easurement bat ch , th ena l l th e s ampl es i n th e mea s u r eme n t bat cha r e r ec ou n t ed af t erth e s ou r ce of

th e ou tof

c on tr ol c on di t i on is id e n tif ie d a n d c o rr e c te d
.. 7. 8 I ns tr um en t

and Met

hod Per f orm anc e Char t

s / Rec or ds Q ual i tyco ntrol p erf

o rma nce reco rds or co ntrol chart s shoul d b e m ai nt ai ned f o r each i nstrume ntand m etho d us ed by the l aborat oryf

or c om p l i anc e moni t ori ng s amp l e meas ur ement s . For i ns trument s , the i ni t i al ca l i brat i ons an d al l ef f i ci enc y cal i b rat i o n a nd ins trume ntb ackgro und checks s ho ul d be m ai nt ai ned i n a pe rma nentrecord
. T he ef f i ci ency cal i b rat i o n and i nstrume ntba ckground checks s ho ul d have co ntrol l i m i t s cal cul at ed , and be co n tro l c h a rte d sep arat el yt o m o ni t o r the i nstrume nt s read ys t atus t o m easure s am p l es. Fo r e a c h m etho d us ed f o r com p l i ance m o ni t o ri ng m easurements , the a sse ssm e n ts o f sam p le p re p a ra tio n b a tc h p reci sio n (D U P o r M S D ), a c c u ra c y (M S a n d M S D ), a n d inte rfe re n c e s (R B ) sh o u ld be r ec or ded s epa r at el yf r om s ampl e r es u l t s i n a perman en tre cor d. Labor at ory - s pec i f i c per f orman ce w arn i n g a n d c on tr ol l i mi t s f orea chpar ame t erm on i t or ed f orboth i n s tru men t s an d m eth ods s h ou l d be re ca l cu l at ed e v ery20 me as u r eme n t s f or ea chQ C par ame t er . I f

a Q C r es u l te x ce eds i t s c al cu l at ed con tr ol l i mi t , al l m ea s u r eme n t s us i n g th e as s oci at ed meth od or ins tru m e n t m u st c e a se un tilthe so u rc e o f th e e x c e e d a n c e is i d e n tif ie d a n d c o rr e c te d
. 7. 9 Q ualit

y As s uranc e (

QA) Plan T he l ab o rat o rys ho ul d prep are and f o l l o w a wri tt en Q A plan ( see C hap t er I II ). 

## 8.Re c or ds a nd D at a Reporting 8 .1 L eg a l D e

fe n sib ility La b o rat o ri es are t o ret ai n suf f i ci ent d at a and d o cum ent at i o n f o r co m p l i ance m o ni t o ri ng s am p l es s o thatthei r r eceiptand a n ym e a s u r e m e n tm a d e f o rth i s p u rp o s e c a n b e r e p r o d u c e d i f v a l i d a t i o n o f

th e d a t a i s r e q u i r e d
. Th e QA p l a n a n d / o rS OPs s houl d desc r i be the po l i ci es an d pro cedur es us ed by the f aci l i tyf or r ecor d re t ent i on an d s t orag e. Si nce al l comp l i anc e mo ni t ori ng m eas ur ement s are us ed t o i nt erpretand enf orce l ega l r equi r ement s , ch ai n of cu s t od yprocedur es s houl d be us ed (See A p p end i x A ). 8. 2 Mainten anc e of

Rec or ds Publ i c wa t ers y s t ems ar e r equ i r ed t o ma i n t ai n r ec or ds of r adi on u cl i de an aly s es of c ompl i an ce s ampl es f

or10 y ea r s ( 40 CFR part141. 33)
. T h e l abor at o ry sho ul d m ai n t ai n e as i l ya cc es s i bl e r ec or ds f or10 y ea r s . T h e cl i en tw at ers y s t em s h ou l d be n o t i f i e d b e f o r e d i s p o s i n g o f

r e c o r d s s o th e ym a yr e q u e s tc o p i e s i f n e e d e d
. T h i s i n c l u d e s a l l r a w da t a , c a l c u l a t i o n s , a n d QC dat a. T h es e da t a f

i l es m aybe ei th erha r d copy , m i cr of i ch e, or el ec tr on i c. El ec tr on i c dat a s h ou l d a l wa y s be ba ck ed u p by p rotect ed t ap e, dis k, or hard co p y . If the l ab o rat o ry changes i t s co m p ut er hardware o r s o f tware, i ts ho ul d m ake p rovis i o ns f or tr ans f err i ng ol d dat a t o the new s y s t em s o thati tr emai ns r etr i evabl e wi thi n the t i me f r ames speci f

i ed above. D at a wh i ch is e x p e c te d to b e c o m e p a rt o f a le g a l ac tio n sh o u ld b e m a inta ine d fo r a lo n g e r p e rio d o f t im e .V I
- 12

8. 3 Sampling Rec or ds D at a s h ou l d be r ec or ded i n i n kw i th a n yc h an g es l i n ed th r ou g h s u chth ator i g i n ale n tryi s v i s i bl e. C hanges s h ou l d be i ni t i al ed and da t ed
. T he f o l l o wi ng i nf

o rma t i o n s ho ul d be readil yavai l ab l e i n a s um m aryor o ther r eco rd(s) : 8 .3 .1 D at e, l ocat i on ( i ncludi ng na me of ut i l i tyand P W SID ) , si t e wi thi n the s y s t em, t i me of s amp l i ng , na me, or g an i za t i on a n d ph on e n u mberof th e s ampl er , a n d a n al y s es r equ i r ed. 8 .3 .2 I dent i f

i cat i on of

the s amp l e a s t o whetheri ti s a r out i ne di s tr i but i on s y s t em s amp l e, che ck s amp l e, r aw or f i ni s hed wa t ers amp l e, r epeator conf i rmat i on s amp l e orothers peci al purpo s e s amp l e. 8 .3 .3 D at e of

r ece i ptof

the s amp l e. 8 .3 .4 S a m p le vo lum e /w e igh t, c o n ta ine r typ e , p re se rv a tio n a n d ho ld ing ti m e a n d c o n d iti o n o n re c e ip t. 8 .3 .5 p H and dis i nf ect antres i d ual att i m e o f s am p l i ng (f

rom plantrecord s)
. 8 .3 .6 T rans p o rt at i o n and de l i veryof the samp l e (person/carri er, co nd i t i o ns)
. 8. 4 Anal

ytical Rec or ds D at a mus tbe r ecor ded i n i nkwi th any cha ng es l i ned thr ough s uch that ori gi nalen tryi s vi s i bl e. Chang es mus tbe i ni t i al ed and dat ed. T he f

ol l owi ng i nf

ormat i on mus tbe r eadi l yav ai l abl e: 8 .4 .1 L ab o rato ry and p erso ns resp o nsib l e fo r p erfo rm i ng an alysi s. 8 .4 .2 Ana l y t i ca l t ec h n i qu es /meth ods u s ed. 8 .4 .3 D ate a nd t i m e o f an alysi s. 8 .4 .4 R esu l t s o f

sam p l e a nd q ua l i ty co ntro l an alyses. 8 .4 .5 C al ib rat i o n and s t and ards i nf o rma t i o n. 8 .4 .6 C ount i ng dat a; 8 .4 .7 Res u l t s of

a n al y s es , i n clu di n g th e ac t i v i ty , u n ce rt ai n tyof th e mea s u r eme n t , a n d th e meth od de t ec t i on lim it. 8. 5 D ata Reporting R ep o rt s t o dri nki ng wat er s up p l i ers and pro gram s shoul d c o n ta in su ff i ci enti nf

o rma t i o n t o est ab l i sh the val i d i tyof the repo rt ed res ul t s f

o r the requir ed anal y ses . I ts ho ul d be de si gned as a summ aryf o rm tha t c o n ta ins the samp l i ng and anal y t i cal i nf

o rma t i o n de scr i b ed ab o ve. A ta m i ni m um , these r ep o rt s shoul d co nt ai n the f o l l o wi ng i nf

o rma t i o n: 8 .5 .1 Na m e a n d l o c a t i o n o f th e w a t e rs u p p l y , i n c l u d i n g i t s P W S I D. 8 .5 .2 Lo cat i o n i n the wat er s up p l ywhere the samp l e was t aken. 8 .5 .3 D at e and t i m e o f s am p l i ng. 8 .5 .4 T he name of

the pers on r es po ns i bl e f or t aki ng the s amp l e. 8 .5 .5 S a m p le re c e ip t d a te a t th e lab o ra to ry. 8 .5 .6 D ate a nd t i m e o f an alysi s. 8 .5 .7 T he m etho d used f o r the an alysi s. 8 .5 .8 T he l ab o rato ry and the i nit i als o f the p erso n re sp o nsib l e fo r p erfo rm i ng the an alysi s.V I
- 13 8 .5 .9 T h e a n a lyti c a l re su lt, inc lud ing the c a lc u la te d un c e rta inty o f th e m e a su re m e n t. 8. 5. 10 If the resu l t i s b elo w the ca l cu l ated d etec t i o n l i m i t f o r tha t sam p l e, the resu l tsho uld b e re p o rted as less tha n the c a lc u la te d d e te c tio n li m it. 8 . 6 Co mp u te rp r og r a ms Comput er pr og r ams s h ou l d be v er i f i ed i n i t i al l y an d per i odi ca l l y by manu al ca l cu l at i on s an d th e ca l cu l at i on s s h ou l d be a v a ilab le fo r in sp e c tio n . A c c e ss to c o m p u te r p ro g ra m s a n d e le c tro n ic d a ta sh o u ld b e li m ited to a p p ro p ria te p e rs o n n e l. 

## 9. Action Respon se to Laboratory Results W hena l aborat ory i s r es po ns i bl e, ei ther by contr act or s t at e po l i cy , t o r eport s amp l e r es ul t s whi ch woul d cau s e as y s t em t o b e ou tof

c om pl i a n c e , th e pr op e ra u th or i tymu s tbe pr om pt l yno t i f i e d a n d a r e qu e s tma de f orr e s a mp l i n g f

r om th e s a me sa m p ling p o int.V I
- 14



## T a b le V I-1 M e th o d s fo r R a d io n u c lid e A n a ly sis C FR 141. 25

C on t am i n an t M eth od ol ogy R ef eren ce (m eth od or page n u m ber) E PA E PA E PA E PA E PA SM AS T M U SG S D O E O ther 1 2 3 4 5 5 6 7 8 N a tu r a lly occ ur ri ng Gr o s s alp ha and

11 b e ta Eva p o r at i o n 900. 0 p 1 00- 01 p 1 302, 7110 B
. . . . . . . . . . . . . R- 1120- 76
.......
G ros s alph a C o- preci pi t at i on . . . . . .

11

....... 0 0
- 0 2
........ 7 1 1 0 C
............. .......... ....... Radi um 226 Rado n em ana t i o n, Rad i ochemi cal 9 0 3 .1 9 0 3 .0 p 16

p 13 Ra- 04

Ra- 03 p 19

........ 75 00
- Ra C 304, 305, 75 00
- Ra B D 3454- 97

D 2460- 97 R- 1141- 76

R- 1140- 76 Ra- 04

....... N. Y. 9

Ra di um 22 8 Ra di o c he mi c a l 90 4. 0 p 24 Ra
- 05 p 19 30 4, 75 00
- Ra D
........... ............. R- 1142- 76
. . . . . . . N. Y. 9 N
. J. 10 U ra ni um Rad i ochemi cal

12 Flu o ro m e tric

A lph a spectrometry L as er Ph osphori m etry I C P
- MS 9 0 8 .0 9 0 8 .1

......

......
2 0 0 .8 13 .......

.......

.......

....... ......

...... 00- 07

...... ........

........ p 33

........ 7 5 0 0
- U B 7 5 0 0
- U C (1 7 th E d .) 7500- U C ( 18 , th 19 , o r20 th th E d .)

............. 3125 .............

D 2907- 97 D 3972- 97 D 5174- 97 D 5673- 03 ..........

R- 1180- 76 R- 1181- 76 R- 1182- 76

.......... .......

U
- 04 U
- 02

.......

M an- m ade R ad i oacti ve ce s i um Rad i ochemi cal Gamm a r ay spectrometry 9 0 1 .0 9 0 1 .1 p 4

....... ......

...... ........

p 92 7 5 0 0
- C s B 7120 D
- 2459- 72

D 3649- 91 R- 1111- 76

R- 1110- 76 .......

4 .5 .2 .3
Rad i oac t i ve i odi ne Rad i ochemi cal

Gamm a r ay spectrometry 9 0 2 .0

9 0 1 .1 p 6

p 9

....... ......

...... ........

p 92 7 5 0 0 -I B 75 00
- IC, 7 5 0 0 -I D 7 1 2 0 ( 1 9 th E d .) D 3649- 91

D 4785- 93 ..........

.......... .......

4 .5 .2 .3

R ad i oacti ve Str o nt i um 89, 90 R ad i och em i cal 9 0 5
. 0 p 2 9 Sr- 0 4 p. 6 5 3 0 3 , 7 5 0 0
- Sr B
............. R- 1160- 76 Sr
- 01 Sr
- 02 Tri t i um L i q ui d s ci nt i l l at i o n 906. 0 p 34 H
- 02 p . 87 306, 7500- 3H B
........... D 4107- 91 R- 1171- 76
. . . . . . .
Gamm a emi tt er s Gamm a r ay Spe ctrometry 9 0 1 .1 9 0 2 .0 9 0 1 .0 ....... ...... p 92 7120 7 5 0 0
- C s B 7 5 0 0 -I B D 3649- 91

D 4785- 93 R- 1110- 76 GA- 01- RV I
- 15

T he pr oce du r es s h al l be per f ormed i n ac cor dan ce wi th th e docu men t s l i s t ed bel ow. T h e i n corpor at i on by r ef er en ce of th e f

ol l owi n g docu men t s wa s appr ov ed by the D i r ect or of

the Federa l Regi s t eri n a ccor dance wi th 5 U
. S. C
. 552 ( a) an d 1 C FR part51. Co pi es o f the do cumen t s maybe obt ai ned f r om the s our ces l i s t ed bel ow. In fo rm a tio n re g a rd ing o b ta inin g the se d o c u m e n ts c a n b e o b ta ine d fro m the S a fe D rin k in g W a te r H o tl in e a t 8 0 0 -4 2 6 -4 7 9 1 . D o c u m e n ts m a y b e insp e c te d a t E P A ’s D rin k ing W a te r D o c k e t, 4 0 1 M S tre e t, S W ., W a sh ing to n , D C 2 0 4 6 0 (T e le p h o n e : 2 0 2 -2 6 0 -3 0 2 7 ); o r a t th e O ff ic e o f F e d e ra l R e g ister , 8 0 0 N o rth C a p ito l S tre e t, N W ., Su i t e 70 0, W a s h i n g t on , DC. 1. “ P r es cr i bed P r ocedur es f or M eas ur emento f

R adi oact i vi tyi n D r i nk i ng W at er ", E P A 6 00 /4- 80
- 03 2 , A ug us t1 98 0. A vai l abl e atU
. S. D epartmentof Co mm er ce, N at i on alT ec h n i ca l I n f ormat i on Se rv i ce ( N T I S) , 5285 Por tRoy alRoad, Spr i n g f i el d, V A 22161 ( T el eph on e 800- 553- 6847) , PB 80- 224744. 2. “ I n t er i m Radi och emi ca l M eth odol og yf orD r i n k i n g W at er ”, EP A 600/4- 75- 008( r ev i s ed) , M ar ch1976. Av ai l abl e atN T I S, i bi d. PB 253258. 3. "Radi och emi s tryPr oce du r es M anu al ", EP A 520/ 5- 84- 006, D ec embe r1987. Av ai l abl e atN T I S, i bi d. PB 84- 215581. 4. "Radi och emi ca l An al y t i ca l Pr oce du r es f orAn al y s i s of

Env i r on men t alSa mpl es ", M ar ch1979. Av ai l abl e atN T I S, i bi d. EM SL L V 053917. 5
. " S ta n d a rd M e tho d s fo r the E x a m ina tio n o f W a te r a n d W a ste w a te r" , 1 3 th, 1 7 th, 1 8 th, 1 9 th E d iti o n s, o r 2 0 th e d iti o n . 1 9 7 1 , 1 9 8 9 , 1 9 9 2 , 1 9 9 5 , 1 9 9 8 . A v a ilab le a t A m e ric a n P u b lic H e a lth A sso c ia tio n , 1 0 1 5 F ift e e n th S tre e t N .W ., W a sh ing to n , D .C
. 2 0 0 0 5 . M e tho d s 3 0 2 , 3 0 3 , 3 0 4 , 3 0 5 , an d 3 0 6 a re o n ly i n the 1 3 th ed i t i o n. M etho d s 7 1 1 0 B , 71 1 0 C , 75 0 0
- R a B , 75 0 0 R a C , 75 0 0
- R a D , 75 0 0
- U B , 75 0 0
- C s B , 75 0 0
- I B , 75 0 0
- I C, 75 0 0
- I D , 75 0 0
- Sr B , 7 5 0 0
- 3 H B are i n the 1 7 th, 1 8 th, 1 9 th a n d 2 0 th e d iti o n s. M e tho d 7 5 0 0 -U C F luo ro m e tric U ra n ium is o n ly in th e 1 7 th e d iti o n , an d 7 5 0 0 -U C A lp h a sp e c tro m e try is o n ly i n the 1 8 th 1 9 th a n d 2 0 th e d iti o n s. M e tho d 7 1 2 0 is o n ly i n the 1 9 th a n d 2 0 th e d iti o n s. 6
. A nnual B o o k of A ST M Standard s, V o l
. 11
. 0 2 , 19 9 9
. A vai l ab l e atA m eri can S o ci etyf o r T est i ng and M at eri al s, 10 0 B arr H arbo r D ri ve, W estC o nshoho cken, PA 19428. 7. "M etho d s forD et ermi nat i on of Rad i oact i ve Subs t anc es i n W at eran d Fl uv i al Sedi ment s ", Chapt erA5 i n B oo k 5 of T ech ni ques of

W at er - R es our ces Inves t i gat i o ns o f the U ni t ed Stat es G eo l o gi cal Survey, 1 9 7 7
. A vai l ab l e atU
. S. G eo l o gi cal Survey (U SG S) Inf o rma t i o n Servi ces, B o x 25 2 8 6 , Fed eral C ent er, D e n v e r, C O 8 0 2 2 5 -0 4 2 5
. 8
. " E M L P roce d ures M anual ”, 2 8 (1 9 9 7 ) o r 2 7 (1 9 9 0 ) E d i t i o n, V o l um e I and II. A vai l ab l e atthe E nvi ronm ent al M easurements L ab o rat o ry , U
. S. D ep artm ent th th o f E n e rg y (D O E ), 3 7 6 H u d so n S tre e t, N e w Y o rk , N Y 1 0 0 1 4 -3 6 2 1
. 9
. " D et erm i nat i o n of R a- 2 2 6 and R a- 2 2 8 (Ra

- 0 2 )”, January19 8 0 , R evi sed June 1 9 8 2
. A vai l ab l e atR ad i o l o gi cal S ci ences Inst i tut e C ent er f o r L ab o rat o ri es and Res ea r ch , N ew Y orkSt at e D epa r tmen tof H ea l th , Empi r e St at e Pl az a, Al ban y , N Y 12201. 10. "D et ermi n at i on of Radi u m 228 i n D r i n k i n g W at er ", Au g u s t1980. A vai l abl e atSt at e of N ew J er s ey , D epa rtmen tof Env i r on men t alPr ot ec t i on , D i v i s i on of Env i r on men t alQ u al i ty , B u r eau of Radi at i on a n d I n or g an i c Ana l y t i ca l Se rv i ce s , 9 Ewi n g Str ee t , T r en t on , N J 08625. 11
. N a tu r a l ur a n i u m a n d th or i u m- 23 0 a r e ap pr ov e d a s gr os s a l ph a c a l i br a t i ons t a n da r ds f org r os s a l ph a w i th c o- pr e c i pi t a t i ona n d e v a po r a t i onme th od s ; a me r i c i u m2 4 1 is ap p ro v e d w ith c o -p re c ip itatio n m e tho d s. 1 2
. If ur a n ium (U ) i s d e te rm ine d b y m a ss, a 0 .6 7 p C i/ 

### : g of u r an i u m c on v er s i on f

ac t ormu s tbe u s ed. T h i s c on s erv at i v e f ac t ori s ba s ed on th e 1: 1 a ct i v i tyr at i o of U -2 3 4 to U -2 3 8 tha t i s c h a ra c te ristic o f na tur a lly o c c u rr ing ur a n ium
. 13. "M eth ods f

orth e D et ermi n at i on of M et al s i n Env i r on men t alSa mpl es - Suppl eme n tI , " EP A- 600/R- 94- 111, M ay1994. Av ai l abl e atN T I S, PB 94- 184942.V I
- 16

Tabl e V I - 2: Samp l e H andl i ng, Pre s er vat i on, and I ns t ru men t at i on
P aram et er P res ervat i ve C o nt ai ner M axi m um 1 2 H o l d i ng T i me 3 I n s tru men t at i on 4

3 G ross A l p ha C o nc. H C l or H N O t o pH < 2 P or G 6 m o A , B , or G 5

3 G ross b et a C o nc. H C l or H N O t o pH < 2 P or G 6 m o A or G 5

3 Str o nt i um - 8 9 C o nc. H C l or H N O t o pH < 2 P or G 6 m o A or G 5

3 Str o nt i um - 9 0 C o nc. H C l or H N O t o pH < 2 P or G 6 m o A or G 5

3 R ad i um - 2 2 6 C o nc. H C l or H N O t o pH < 2 P or G 6 m o A , B , D or G 5

3 R ad i um - 2 2 8 C o nc. H C l or H N O t o pH < 2 P or G 6 m o A or G 5 Ces i u m- 134 Conc
. H Cl t o pH <2 5 P or G 6 m o A , C or G Iod i ne- 1 3 1 N o ne P or G 8 da A , C or G T r i t i u m N on e G 6 m o E
3 U ra n ium C o n c . H C l o r H N O to p H < 2 P o r G 6 m o A , B , F , H 5

3 Ph ot one mi tt e r s Con c . HCl orHN O t o p H <2 P o rG 6 m o C 5 It is re c o m m e n d e d tha t th e p re se rv a tive b e a d d e d to the sa m p le a t the tim e o f c o llec tio n u n le ss sus p e n d e d so lid s a c tivity 1 is to b e m e a su re d . If the s a m p le h a s to b e sh ip p e d to a lab o ra to ry o r sto ra g e a re a un p re se rv e d , ac id ifi c a tio n o f th e sa m p le ( i n i t s ori gi nalcont ai ner ) maybe del ay ed f or aper i od nott o exc eed 5 day s . A mi ni mum of 16 hour s mus tel aps e between a c id ifi c a tio n a n d a n a lysis. P = P la stic, h a rd o r so ft; G = G la ss, h a rd o r so ft. 2 H ol di ng t i me i s def

i ned a s the peri od f

r om t i me of s amp l i ng t o t i me of ana l y s i s . I n a l l ca s es , s amp l es s houl d be ana l y zed 3 as so o n a f

t er c o l l ec t i o n a s p o ssi b l e. If a c o m p o si t e sa m p l e is pre p are d , a h o l d i ng t i m e c an no tex ce ed 1 2 m o nths. A = Low ba ck g r ou n d pr oport i on als y s t em; B = Al ph a a n d bet a s ci n t i l l at i on s y s t em; C = G amma s pec tr omet er[ G e( H p) 4 o r G e(Li )] ; D = Sci nt i l l at i o n cel l s y st em ; E = Li q ui d s ci nt i l l at i o n s y st em (s ect i o n C
. 2
. a); F = Fluorom et er ( s ect i o n C
. 1
. 1 ); G = L ow ba c k g r ou n d a l ph a a n d b e t a c ou n t i n g s y s t e m o th e rth a n g a s - f l ow pr op ort i on a l ; H=Al ph a s pe c tr om e trys y s t e m. I f H Cl i s u s ed t o ac i di f

y s ampl es wh i ch ar e t o be an al y ze d f org r os s a l ph a org r os s be t a ac t i v i t i es , th e ac i d s al t s mu s tbe 5 c o n v e rte d to nitra te sa lts b e fo re tra n sfe r o f th e sa m p le s to p la n c h e ts.V I
- 17



# Ex amp le C h ecklis tsfo r On -S ite E valu ationof Lab orato riesAn alyzin g D rin kin g W at er fo r Radioch emis tryV I
- 18



## G eneralA udi tI nf

orm at i on L abor at ory

____________________________________________________ Str eet

________________________________________________________ C i ty______________________________________St at e_______________ SurveyB y

_____________________________________________________ A ffi l i at i on___________________________________________________ D at e________________________________Tel ephone N o.

_____________V I
- 19

La bor at ory_______________________________D at e_________________ Loc at i on_________________________________Eva l u at or

____________
PER SO N N EL
P o sitio n /T itl e N a m e A c a d e m ic T rai ni ng P re se n t

S p e c ia lty Y ears

E xper i enc e (c h e m istry) Y ears

E xper i enc e (r a d io c h e m istry) La bor at oryD i r ec t or Q ual i tyAss ur anc e O f f i cer Se ct i on /D i v i s i on C hi ef /D i rect o r ( i f appl i cabl e) S up erv i so ry A na l yst C he m i ca l A na l yst (s)

C hemi cal T ec hn i cian (s)

C o m p ut er Sup p o rt T ech ni ci an E l ectr oni cs Suppo r tT ech ni ci anV I
- 20

La bor at ory_______________________________D at e_________________ Loc at i on_________________________________Eva l u at or

____________ LA B O R A TO R Y FA C I LI TI ES

Ite m A v a ilab le C o m m e n ts Ye s No L a b o ra to ry E le c tric a l o u tlets 120 V ac . gr ounded D i s t i l l ed or dei oni zed wat er o r AS T M typ e 1 , 2, or 3 Exh au s tH ood V acuum s our ce C o u n ting R o o m - se p a ra te fro m w e t ch emi s try , s ampl e an d s t an dar ds prepar at i on ar ea R e g u la te d p o w e r su p p ly R e a g e n tsV I
- 21

La bor at ory_______________________________D at e_________________ Loc at i on_________________________________Eva l u at or

____________ GEN ERA L LAB O R A TO R Y EQU I PM ENT A N D I N STRU M ENTS I t em N o. of U n its M anuf

acturer M o d el Sa t i sf act o ry Ye s No A nal y t i calB al anc e 0 .1 m g se n sitivity st ab l e b ase AST M type 1 or 2 wei gh t s or bett er pH met er ± 0 .5 un its re a d a b ili ty ± 0 .1 un its l i ne o r batt ery C onduct i vi tymet er Rea dabl e i n oh ms ormh os Ran g e of

2 oh ms ormh os Li ne o r batt ery D ry i ng oven g r av i tyorc on v ec t i on c on tr ol l ed f

r om r oom t emp t o 18 0°C or h i gh er ( ± 2 °C ) I n f r a r e d l a mp maybe s ubs t i tut ed f

or dry i ng oven D es i cc at or G la ss o r p la stic H o t p la te t empe r atu r e con tr ol Ref r i g er at or M agn et i c St i rr er v ar i abl e s peed T ef l on coat ed s t i rbar B al ance, t o p l o ad i ng G l ass ware T herm o m et ers M uff l e furna ce t o 4 5 0 °C C entri f uge t o 3000 rpm t o h ol d 4 x 50 mLV I
- 22

La bor at ory_______________________________D at e_________________ Loc at i on_________________________________Eva l u at or

____________

A L L I N S T R U M E N T A T I ON

Ite m Y e s N o C o m m e n ts A re o p e ra ting m a n u a ls re a d ily av ai l abl e t o th e oper at or A re c a lib ra tio n p ro to c o ls a v a ilab le t o th e oper at or A re cal i b rat i o ns kep ti n a p erm anentco ntrol chart A r e permanen ts ervi ce mai nt ena nce r ec or ds k eptV I
- 23

La bor at ory_______________________________D at e_________________ Loc at i on_________________________________Eva l u at or

____________
TH I N W I N D O W GA S- FLO W PRO PO R TI O N A L CO U N TER

Instrum ent nu m ber M anufacturer M odel Y ear Sam pl e Chang i ng M a n u a l A u to m a tic C a p a c ity

Co u n t i n g Ga s W i n d o w D e n s ity (g / cm )

2 Instrum entB ac k g r o und

O p erat i ng V o l t age A l pha

cpm O p e ra tin

g V o l t age B e ta

cpm

C al i br at i o n St anda r d T ype: A lpha B e ta S u p p l i er: A lpha B e ta C al i b rat i on Freq u en cy S ervi ce M ai n t en an ce F req u en cy C on d i t i on 1 2 3 D W M O ther Q S A O ther G R N

W I N D O W LESS GA S- FLO W PRO PO R TI O N A L CO U N TER

Instrum ent nu m ber M anuf ac tur er M o del Y ea r Sam pl e Chang i ng M a n u a l A u to m a tic C a p a c ity

Co u n t i n g Ga s W i n d o w D e n s ity (g / cm )

2 Instrum entB ac k g r o und

O p erat i ng V o l t age A l pha

cpm O p erat i ng

V o l t age B e ta

cpm C al i b rat i on S t an d ard T ype: A lpha B e ta S u p p l i er: A lpha B e ta C al i b rat i on Freq u en cy S ervi ce M ai n t en an ce F req u en cy C on d i t i on 1 2 3 D W M O ther Q S A O ther G R N

1 . D aily, W ee kl y, M o n th ly. 2
. Q u a rte rly, S em ian n u al ly, A n n u a l ly. 3 . G o o d, o per at i ng butnee ds R ep air, N o to per at i ngV I
- 24

La bor at ory______________________________D at e_____________________ Loc at i on________________________________Eva l u at or

________________ LI QU I D S C I N TI LLATI O N CO U N TER Instrum ent nu m ber M anuf ac tur er M o del Y ea r Sam pl e Chang i ng M a n u a l A u to C a p a c ity

C al i b rat i on S t an d ard T ype: S u p p l i er: C al i b rat i on Freq u en cy S ervi ce M ai n t en an ce F req u en cy C on d i t i on 1 2 3 D W M O ther Q S A O ther G R N

A LPH A S C I N TI LLATI O N CO U N TER Ins trume nt nu mb er M anufacturer M odel Y ear Sam pl e Chang i ng M a n u a l A u to C a p a c ity

C al i b rat i on S t an d ard T ype: S u p p l i er: C al i b rat i on Freq u en cy S ervi ce M ai n t en an ce F req u en cy C on d i t i on 1 2 3
D W M O ther Q S A O ther G R N

R A D O N - GA S C O U N TI N G SYSTEM Sy s t em nu mb er G as coun t i ng ce l l s / s y s t em M anu f actur erof

ga s c o u n ting c e ll C o unt i ng Ins trume nt M ake M od el Y ear

C al i b rat i on Freq u en cy S ervi ce M ai n t en an ce F req u en cy C on d i t i on 1 2 3 D W M O ther Q S A O ther G R N

1 . D aily, W ee kl y, M o n th ly. 2
. Q u a rte rly, S em ian n u al ly, A n n u a l ly. 3 . G o o d, o per at i ng butnee ds R ep air, N o to per at i ngV I
- 25

La bor at ory______________________________D at e_____________________ Loc at i on________________________________Eva l u at or

________________ GA M M A S PEC TRO M ETER SYSTEM D et ect or Sy s t em Sy s t em N umber T ype M ake M od el Y ear Si ze
A nal y zerSy s t em M a k e M o d e l Y e a r C h a n n e ls Cal i br at i on Standa rd T ype Suppl i er Cal i br at i on Fr equ en cy Se rv i ce M ai n t en an ce Fr equ en cy Condi t i on D W M O ther Q S A O ther G R N

OT HE R A P P R OV E D D E T E C T OR D et ect or Sy s t em Sy s t em N umber T ype M ake M od el Y ear Si ze
A nal y zerSy s t em M a k e M o d e l Y e a r C h a n n e ls Cal i br at i on Standa rd T ype Suppl i er Cal i br at i on Fr equ en cy Se rv i ce M ai n t en an ce Fr equ en cy Condi t i on 1 2 3 D W M O ther Q S A O ther G R N

1 . D aily, W ee kl y, M o n th ly. 2
. Q u a rte rly, S em ian n u al ly, A n n u a l ly. 3 . G oo d, o perat i ng bu tneeds R ep air, N o to per at i ngV I
- 26

La bor at ory______________________________D at e_____________________ Loc at i on________________________________Eva l u at or

________________ S A M P L E HA N D L I N G A N D P R E S E R V A T I ON

P aram et er C on tai n er U sed P reser va tive U sed C om m en ts S ati sfactory Y e s No G ro s s A lp h a A c tivity G ro s s B e ta A c tivity St r o nt i um - 89 St r o nt i um - 90 R adi um - 226 R adi um - 228 C es i um - 134 I o d i n e - 1 3 1 NON E T r i t i u m NON E U r ani um P h ot on E m i tt ers
a. b.

c. d.

e.V I
- 27

La bor at ory______________________________D at e_____________________ Loc at i on________________________________Eva l u at or

________________ M E T HO DOLOGY

P aram et er S a m p le L o a d / M o M et hod U se d - C i t e Ed i t i on, Year , and P age 1 S ati sfactory E PA S M I AS T M US GS DOE Ot h e r Ye s No G ross A l p ha A c tivity G ro ss B e ta A c tivity Str on t i u m- 89 Str on t i u m- 90 Radi u m- 226 Radi u m- 228 Ces i u m- 134 I odi n e- 131 T ri t i um U rani um Phot on E m i tt ers Id e n tif y:
a. b.
c. d.
e. 1 - M eth ods u s ed mu s tbe r ef er en ce d i n th e N at i on alPr i maryD r i n k i n g W at erReg u l at i on s ( 40 CFR 141. 25)V I
- 28

La bor at ory______________________________D at e_______________ Loc at i on________________________________Eva l u at or

___________ Q U A L I T Y C ON T R OL It em P erf orm an ce T est i n g S tud i es A B 1 2 O ther P E S tud i es A B 1 2 P a rtic ip a tion in perf orm ance ev aluat i o n and Bl i n d PE stud i es

R epo r t i ng Per i o d:

_______________ to

_______________ G r o ss A lpha G r o ss A lpha G ro s s B e ta G ro s s B e ta Sr- 89 Sr- 89 Sr- 90 Sr- 90 R a- 226 R a- 226 R a- 228 R a- 228 U r ani um U r ani um C s- 134 C s- 134 C s- 137 C s- 137 C o - 60 C o - 60 B a- 133 W r i tt en Q A Pl an i m pl em ent ed a nd ava i l abl e f

orre vi ew Zn- 65 Tri t i um I
- 131 Freq u en cy Y es N o C om m en t S at i sfactory Ye s No D up l i cat e analy ses S pi kes R eagen tB l an ks LaboratoryForti fi ed B l an ks Fai l ed P E stud i es C o n tro lc h a rts C al i br at i o n and M ai nt ena nce r ec o r ds
1 - S ch ed u l ed frequ en cy of p art i ci p at i on b yth e l ab orat ory , t i m es p er y ear. 2
- N u m b er of a ccep t ab l e p erform an ce resu l t s i n th e p ast year, w h ere an acce p t ab l e resu l t i s a n orm al i zed d evi at i on from th e kn ow n valu e of # 3
. 0 s i g maV I
- 29

La bor at ory______________________________D at e_____________________ Loc at i on________________________________Eva l u at or

________________ DAT A RE PORT I NG

I t em C om ments : s yst ems us ed, frequency, et c. R ec o rd s kep tf

o r 1 0 years A c tua l l a b o ra to ry re p o rts T ab ul ar Sum m ary I nf ormat i on i ncluded D a te P l ace o f s am p l i ng T i m e o f s am p l i ng Samp l er D at e of s ampl e r ec eipt D a te o f an a lysis T yp e o f an a lysis La b o rat o ry& pe rs o n re sp o n sib le O the r re p o rte d d a ta O the r re p o rte d d a ta M ethod(s ) us ed R e su lts

# Appendic esA -1 A pp en dix A C hai n- of

- C us t ody Eval uat i ons A . I nt rod uc t i on W r i tt en p r ocedur es f

or s amp l e h andl i ng s houl d be ava i l abl e a nd f ol l owed when eve rs amp l es ar e col l ec t ed, tr an s f err ed, st o red, anal y zed or d estroyed. Fo r the p urpo ses o f l i t i gat i o n, i t i s necessary t o have an accurat e wri tt en reco rd t o trace the p o sse ssio n a n d ha n d ling o f sam p le s fro m c o llec tio n thr o u g h re p o rting . T h e p ro c e d u re s d e fine d he re re p re se n t a m e a n s to sa tisf y this re q u ire m e n t. A sa m p le is i n so m e o n e ’s “c u sto d y” if: 1
. Iti s i n one ’ s actual phys i cal po ss ess i o n; 2
. Iti s i n one ’ s vi ew, af t er being i n one ’ s p hy si cal po ss ess i o n; 3
. It is o n e ’s p h ysic a l p o sse ssio n a n d the n lo c k e d up so tha t no o n e c a n tam p e r w ith it; 4
. It is ke p t i n a se c u re d a re a , re stric te d to a u tho riz e d p e rs o n n e l o n ly. B . Samp l e C ol l ec t i on, H andl i ng and I de nt i f i cat i on 1. I ti s i mp ort antthata mi ni mum nu mb ero f p er s ons b e i nv ol ved i n s amp l e col l ect i on and han dl i ng . G ui del i nes es t abl i s hed i n s t andar d manua l s f

or s amp l e col l ect i on p r es ervat i on and han dl i ng s houl d be us ed ( e. g. , E P A N P D E S C om pl i anc e S a m p ling In sp e c tio n M a n u a l, M C D 5 1 , Stan da rd Metho ds f or Exam i na t i on of

W at erandW

astewa ter)
. Fi el d r ec or ds s houl d be com pl et ed atthe t i me the s amp l e i s col l ect ed an d s houl d be s i gn ed or i ni t i al ed, i ncludi ng the dat e a nd t i me, b ythe samp l e co l l ect o r( s)
. Fi el d record s shoul d co nt ai n the f o l l o wi ng i nf

o rma t i o n:
a. U ni que s amp l e orl og nu mb er 

### ; b. D at e a nd t i me;
c. So urce of samp l e (i ncl ud i ng nam e, l o cat i o n and s am p l e typ e); d. Pre s erv at i v e u s ed;

e. Ana l y s es r equ i r ed; f
. N am e o f

co l l ect o r( s) ; g . P e rtine n t f

ie ld d a ta (p H , D O , C l re sid u a l, e tc .); h. S erial num b er o n sea l s and tran sp o rtat i o n c ase s; i
. C o m m en t s. 2
. E a c h sa m p le is id e n tif ie d b y a ff

ixin g a p re ssu re se n sitive gu m m e d lab e l o r sta n d a rd iz e d tag o n the c o n ta ine r( s) . T h is la b e l sho u ld c o n ta in th e sa m p le nu m b e r, so u rc e o f sam p le , p re se rv a tive us e d , an d the c o llec to r( s’) initia ls. T h e a n a lysis re q u ire d sh o u ld b e id e n tif ie d . W h e re a lab e l i s n o t a v a ilab le , th e s a m p le in fo rm a tio n s h o u ld b e w ritten o n the sa m p le con t ai n erw i th a n i n del i bl e mark i n g pe n . An e x ampl e of a s ampl e i den t i f

i ca t i on t ag i s i l l u s tr at ed i n Fi g u r e A- 1. 3. T h e cl os e d s a mp l e co n t a i n e rs h ou l d th e n be pl a c e d i n a tr a n s po rt a t i onc a s e a l on g w i th th e c h a i n - of

- c u s t od yr e c or d f

orm, p e rtine n t f

ie ld re c o rd s, a n d a n a lysis re q u e st fo rm . T h e tra n sp o rta tio n c a se sh o u ld the n b e se a le d a n d lab e le d . A ll r ec or ds s h ou l d be f

i l l ed ou tl eg i bl yi n w at erpr oof pe n . T h e u s e of

l ock ed ors ea l ed ch es t s w i l l e l i mi n at e th e n ee d f or clo se c o ntro l o f

i nd i vid ua l sam p l e c o ntainers. H o w ev er, the re w i l l u nd o ub t ed l y be o cc asio ns w he n the u se o f a c he st w illb e inc o n v e n ie n t. O n the se o c c a sio n s, the sa m p le r sh o u ld p la c e a se a l a r o u n d the c a p o f th e ind ivid u a l sa m p le con t ai n erw h i chw ou l d i n di ca t e t ampe r i n g i f r emov ed.A -2

C . Tran s fer of

C us t ody and Shi pme nt 1
. W h e n tra n sfe rr ing the p o sse ssio n o f th e sa m p le s, the tra n sfe re e m u st sign a n d re c o rd the d a te a n d tim e o n the c h a in-o fcus t od yr ecor d. Cus t od ytr ans f er s , i f made t o a sampl e cus t od i an i n the f i el d, s houl d accoun tf

or e ach i ndi vi dual s amp l e, al th ou g h s ampl es ma ybe tr an s f err ed as a g r ou p. Eve ryper s on wh o t ak es cu s t odym u s tf i l l i n th e appr opr i at e s ec t i on of th e ch ai n - of

- cu s t odyr ec or d. 2
. T he f

i el d cust o d i an (or f

i el d s am p l er i f

a cust o d i an has notbe en ass i gned ) i s res p o nsib l e f o r prop erl ypa ckagi ng and dis p at chi ng s am p l es t o the ap p rop ri at e l ab o rat o ryf o r anal y si s. T hi s res p o nsib i l i tyincl ud es f i l l i ng out, da t i ng, a n d sign ing the a p p ro p ria te p o rtio n o f th e c h a in-o f- c u sto d y r e c o rd
. A r e c o m m e n d e d c h a in-o f- c u sto d y fo rm a t i s i l l u s tr at ed i n Fi g u r e A- 2. 3
. A l l pa ckages s entt o the l ab o rat o rys ho u ld b e a cco m p ani ed by the chai n- o f

- cust o d yrecord and otherpe rt i nent f o rms. A co p yof these f o rms s ho ul d be ret ai ned by the f i el d cust o d i an (ei ther carbo n or p ho t o co p y ). 4
. M a iled p a c k a g e s c a n b e re g ister e d w ith re tur n re c e ip t re q u e ste d
. If p a c k a g e s a re se n t b y c o m m o n c a rr ie r, re c e ip ts shoul d be ret ai ned as p artof the p erm anentchai n- o f

- cust o d ydo cum ent at i o n. 5. Sa mpl es t o be tr an s por t ed mu s t be pac k ed t o pr ev en t br ea k ag e. I f s ampl es ar e s h i pped by mai l or by oth erc ommon carr i er , the s hipp ermus tcomp l ywi th a nya pp l i cabl e D epartmentof T r ans po rt at i on r egu l at i ons . ( M ostwat er s ampl es ar e ex empt u n l es s qu an t i t i es of pr es erv at i v es u s ed ar e g r ea t erth anc ert ai n l ev el s . ) T h e pac k ag e mu s tbe seal ed or l o cked t o p reventtamp eri ng. A nye vi d ence o f tamp eri ng sho ul d b e read i l yde t ect ed i f ad eq uat e seal i ng dev i ce s a r e u s ed. 6
. If the f i el d s am p l er deli vers s am p l es t o the l ab o rat o ry , cust o d ym ay b e rel i nq ui shed t o lab o rat o rype rs o nnel
. I f ap p rop ri at e p ers o nnel are notpresentt o recei ve the samp l es, they sh o u ld be l o cked i n a de si gnat ed area of the l abor at oryt o pr ev en tt ampe r i n g . T h e per s on de l i v er i n g th e s ampl es s h ou l d ma k e a l og e n trys t at i n g w h er e an d h ow the samp l es were del i vered and s ecured
. La b o rat o rype rs o nnel m ay then recei ve cust o d yby not i ng i n a l o gb o o k, the a b se n c e o f ev id e n c e o f t a m p e rin g , un lo c k ing the se c u re d a re a , an d sign ing the c u sto d y sh e e t. D . La bora t ory Sa mpl e C ontrolP rocedures S a m p le c o n tro l p ro c e d u re s a re ne c e ssa ry in the lab o ra to ry fro m the ti m e o f sam p le re c e ip t t o the ti m e the sa m p le is d isc a rd e d . T h e fo llo w ing p ro c e d u re s a re re c o m m e n d e d fo r th e lab o ra to ry: 1
. A sp e c ifi c p e rs o nm u st b e d e sig n a te d a s c u sto d ia n a n d a n a lter n a te d e sig n a te d to a c t as c u sto d ia n in the c u sto d ia n ’s ab sence. A l l i nco m i ng s am p l es mustbe rec e ive d b y th e cust o d i an, who m usti nd i cat e recei p tby s i gni ng the ac co m p an yi ng cu st o d y/ co ntro l f o rm s and wh o m ust ret ain the signe d f o rm s as p erm an en trec o rd s. 2. T he cus t od i an mus tmai nt ai n ap ermanen tl ogbo ok t o r ecor d, f or e ach s amp l e, the pers on del i ver i ng the s amp l e, the p e rs o n re c e ivin g the sa m p le , d a te a n d ti m e re c e ive d , so u rc e o f sam p le , d a te the sa m p le w a s ta k e n , sa m p le i dent i f i cat i on l o g nu m b er, how tr ans mi tt ed t o the l aborat ory , an d condi t i on r ece i ved ( s eal ed, un s eal ed, broke n co nt ai ner, or o ther pert i nentremarks). T hi s lo g s h o u l d al so show the m o vem entof each sam p l e wi thi n the l ab o rat o ry ; i
. e. , who remo ved the samp l e f rom the cust o d y area, when i t was r em o ved , when i twas r eturned, and w he n itwa s d estro yed
. A st an d ard i ze d f o rm at sho uld b e e st ab l i she d f o r l o gb o o k e ntri es. 3
. A clea n, d ry, i so l ated ro o m , b uil d i ng , an d / o r refri ge rated sp ac e tha tca n b e se cu rely l o ck ed f ro m the o utsi d e m ust b e d e sig n a te d a s a “c u sto d y ro o m .” 4
. T h e c u sto d i an m ustensur e tha t he a t-sen sitive sa m p le s, li g h t-sen sitive sa m p le s, ra d io a c tive sa m p le s, o r o the r sa m p le mat er i al s ha vi ng unus ualphy s i calch ar act er i s t i cs , or r equi r i ng s peci al ha ndl i ng , ar e pr op er l ys t ored an d m ai nt ai ned p rior to an alysi s. 5
. D i stri b ut i o n of samp l es t o the anal y stpe rf o rming the anal y si s m ustbe m ad e b ythe cust o d i an. 6
. T h e lab o ra to ry a re a m u st b e m a inta ine d a s a se c u re d a re a , re stric te d to a u tho riz e d p e rs o n n e l o n ly.A -3

7
. L ab o rato ry pe rso nn el are resp o nsib l e fo r the ca re and cu st o d y o f the sam p l e o nc e iti s rec eive d b y the m an d m ust be pr epa r ed t o t es t i f

yth atth e s ampl e wa s i n th ei rpos s es s i on an d vi ew or s ec u r ed i n th e l abor at oryata l l t i mes f r om th e momen ti tw as r ec ei v ed f r om th e cu s t odi anu n t i l th e t i me th atth e an al y s es a r e compl et ed. 8
. O nc e the sam p l e a na l yses are co m p l eted , the unu sed p o rti o n o f the sam p l e, t o ge the r with a l l i d en t i f yi ng l ab els, m ust be r etu rn ed t o th e cu s t odi an . T h e r etu rn ed t ag g ed s ampl e mu s t be r et ai n ed i n th e cu s t odyr oom u n t i l pe rmi s s i on t o de stroy the samp l e i s recei ved by the cust o d i an. 9
. Sa m p l es will b e d estroyed o nl yupo n the o rder o f the res p o nsib l e l ab o rat o ryoff i ci al when i ti s cert ai n thatthe info rm a tio n is no lo n g e r re q u ire d o r th e sa m p le s h a v e d e te rio ra te d . (F o r e x a m p le , stan d a rd p r o c e d u r e s sh o u ld inc lud e d isc a rd ing sa m p le s a fterthe m a x im u m ho ld ing ti m e h a s e lap se d .) T h e sa m e p ro c e d u re is tru e fo r sa m p le t ag s . T h e l og books h ou l d s h ow wh ene ac h s ampl e wa s di s ca r ded ori f a n ys ampl e t ag w as de s tr oy ed. 1 0
. P ro c e d u r e s s h o u ld b e e sta b lishe d fo r in te rn a l au d its o f sam p le c o n tro l i n fo rm a tio n . R e c o rd s sh o u ld b e e x a m ine d to d e te rm ine tra c e a b ili ty, c o m p le te n e ss, a n d a c c u ra c y.A -4A -5

F i gure A - 2 C hai n-of - C us t ody R ecord
S u rv ey S a m p l er s: Sign ature Stati on Number Stati on Locat ion Dat e Time Samp leTy pe Seq. No. No. Of Conta in ers Analys is

Required Water Air CompGrab.

Relin quis h edby: Sign ature Re ceivedby: Sign ature D at e/T i me
Relin quis h edby: Sign ature Re ceivedby: Sign ature D at e/T i me
Relin quis h edby: Sign ature Re ceivedby: Sign ature D at e/T i me
Relin quis h edby: Sign ature Receivedby M ob ile Lab oratory fo r Fieldan alysis: Sign ature D a t e/T i me
Di sp atc h edby: D a t e/T i me Re ceivedfo r Lab o rato ry by: Si gnat ure D at e/T i me
Meth o do f S hip ment:
Distribution : Orig. --Accom pany Shipment, 1 Cop y--Survey Coordinato r Field Fil esB -1 A pp en dix B R ec ommen de d Prot ocolf or R egi ons C onduc t i ng O n- Si t e Laborat ory Eval uat i ons B ef or e con du ct i n g th e on - s i t e ev alu at i on , th e Reg i on s h ou l d:



### C P la n a ll the re q u ire d a c tivities to b e c o m p le te d d u rin g the a sse ssm e n t.



### C H ol d a pre

- eva l uat i on conf er enc e wi th app r op r i at e l aborat ory and f i el d act i vi tyr epre s ent at i ves t o es t abl i s h as che dul e that w o uld ha ve a m i nim um i m p ac to n the lab o rato ry acti vit i es.



### C R eq ue stan d rev i ew ap p ro p riat e re co rd s.



### C R eq uestthata vari etyof t est s b e scheduled d uri ng the o n- si t e eval uat i o n.



### C A rr a n g e fo r th e lab o ra to ry sta ff to b e a v a ilab le d u rin g the o n -sit e visit. D u rin g the o n -sit e visit, the tea m sh a ll:



### C Co nductanopeni ng conf er enc e oren tr anc e i nt ervi ew.



### C E val uat e the p roce d ures and e q ui p m entused for those spe ci f i c anal y ses f

o r whi ch the l ab o rat o ryhas reque st ed cert i f i cat i o n, u sin g the c rite ria in this m a n u a l.



### C R e v ie w the re c o rd s a n d w ritten sta n d a rd o p e ra ting p ro c e d u re s fo r c o m p lian c e w ith the re q u ire d sa m p ling fre q u e n c y, sa m p le co l l ect i o n, s am p l e ho l d i ng t i m es, and i f ap p rop ri at e, res am p l e no t i f i cat i o n.



### C P erf o rm a d at a aud i ton at l eastone s am p l e and one P E s am p l e f o r atl eastone m etho d but p refe ra b ly fo r eac h m etho d the l ab o rato ry pe rf

o rm s.



### C In su re tha t th e lab o ra to ry h a s a Q A p la n in e ff e c t b y:

- D e te rm inin g if

the lab o ra to ry h a s w ritten p ro c e d u re s ( Q A p la n o r e q u iva le n t) fo r c o n d u c ting it s q u a lity a s s u r a n c e pr og r a m.

- Exa mi n i n g th e qu al i tya s s u r an ce da t a t o de t ermi n e i f th e qu al i tya s s u r an ce pr og r am i s be i n g i mpl eme n t ed.



### C C o m p l et e the o n- si t e checkli st s and othereval uat i o n f o rms duri ng the vi si t(s ee C hap t ers IV , V , and V I)
.



### C Con du cta c l os i n g c on f er en ce ore x i ti n t erv i ew i n w h i chth e au di t or s r ev i ew th e r es u l t s of th e ev alu at i on wi th th e di r ec t orof th e l abor at ory , th e di r ec t orof St at e wa t ers u ppl ya ct i v i t i es , a n d a ppr opr i at e s t af f me mber s . T h e r ev i ew s h ou l d:

- D i scu ss any d ev i ati o ns in the o b serv ed p ro ce d ure s and rec o rd s.

- R e c o m m e n d c h a n g e s in e q u ip m e n t an d su p p ly ne e d s, sta ff ing re q u ire m e n ts, an d fac il it y i m p ro v e m e n ts, i f n e c e ssa ry.

- D isc u ss p o ssib le a ssista n c e the R e g io n c a n p ro v id e the lab o ra to ry.

- D i s cus s a t i me f

r ame f or c orr ect i ve act i ons an d re s po ns e. E va l uati on R eport for P ri nci pal St ate L abo ratori es and La bora t ori es i n N on-P ri ma cy Stat es A ftera n o n -sit e insp e c tio n , th e e v a lua tio n tea m sh o u ld p re p a re a na rr a tive re p o rt a n d a c tio n m e m o ra n d u m . T h is re p o rt sh o u ld co nt ai n al l i nf

o rma t i o n p ert i nentto the eval uat i o n a nd a l so reco m m end the cert i f i cat i o n st atus f

o r al l a nal y ses e val uat ed
. T he re p o rt sh o u ld the n b e fo rw a rd e d fo r e v a lua tio n to the C e rtif ic a ti o n P ro g ra m M a n a g e r fo r re v ie w . A fterre v ie w ing a n d , i f nec es s ary , r evi s i ng the r eport , i ts houl d be f orwa r ded t o the C ert i f

i cat i on Author i tyf

or s i gn atur e.B -2

T h e C e rtifi c a tio n A u tho rity sh o u ld d e c id e the c e rtifi c a tio n s ta tus o f th e la b o ra to ry w ithin tim e c o n stra ints o n p a g e I II -7 a n d no tif y the St at e. T he St at e s houl d be s entthe com pl et e r eport
. I f the r eporti ndi cat es thatthe l aborat orys houl d notbe ce rt i f i ed f or a n an alysi s, the C erti f

i ca t i o n A utho rity s ho uld give the sp ec i f i c re aso ns. T he narrat i ve repo rts ho ul d be att ached t o each co p yof the co m p l et ed e val uat i o n f o rm. Its ho ul d i ncl ud e the general head i ngs and i nf ormat i on l i s t ed bel ow. Ti t l e Page T he t i t l e p age shoul d co nt ai n the f o l l o wi ng: T i t l e: R ep o rtof an o n- si t e eval uat i o n of the (n a m e o f l a b o ra to ry) A t : ( ci ty , s t at e, an d zi p cod e) O n: ( dat e) B y : (name, t i t l e, organi zat i o n, and a dd r e s s of

th e c e r t i f

i c a t i ont e a m) 

### Certification Statu s List e ither "C ertified", "Provisionally Certified", "Administratively /Interim C ertified", or "Not C ertified" for each contaminant evaluated or if applicab le (for VOCs,for instance ) for each class o f com pounds evaluated. Li s t of D evi at i ons L i stea ch d ev i ati o n b y i t em num b er u sed o n the e va l ua t i o n c he ck l i st s. D esc ribe the e xa ct d ev i ati o n a nd rec o m m en d ed ch an ge s. R em a rks R eco m m end imp rovem ent s whi ch, w hi l e no ta f f ect i ng c ert i f

i cat i o n s t atus, wo ul d i m p rove l ab o rat o ry o p erat i o n. O ther r em arks mi g h ti n clu de r ea s on s f orf ai l i n g th e on - s i t e ev alu at i o n, sp ec i alr ec og n i t i on f orou t s t an di n g pe r f

orman ce , a n d de s cr i pt i on of un usua l t ests. L i s t of P ers onn el Lis tnam e and t i t l e o f

pe rs o nnel al o ng wi th the i nd i vi d ual t est s thateach no rma l l ype rf o rms. A l so, i d ent i f

ythe cri t i cal l ab o rat o ry p e rs o n n e l. Si gna t ure T e a m m e m b e rs sh o u ld sign the re p o rt.
D i s t ri but i on C o p i es o f

this re p o rt sho uld b e d i stribu t ed t o the S t ate re q ue st i ng the ev alua t i o n. Fo r l o ca l l ab o rato ries i n no n-p rima cy S t ates, re p o rts sh o u ld b e d istrib u te d to a p p ro p ria te R e g io n a l p e rs o n n e l. A nnual l y , each R egi o n s ho ul d s ub m i tt o O G W D W a l i st i ng of l ab o rat o ri es i n the R egi o n havi ng U
. S. E P A cert i f i catio n . T he l i st i ng s ho ul d i ncl ud e the nam es and l o cat i o n of each l ab o rat o ry , and i t s cert i f i cat i o n s t atus f

o r al l regul at ed co nt am i nant s. I n addi t i on, Regi ons s houl d not i f yO G W D W of

al l cha ng es i n s t atus s oo n af t er they occur s o thatO G W D W ca n mai nt ai n a n upd at ed l i sto f ce rti f i ca t i o n status.C -1 A pp en dix C D ef i ni t i ons and A bbr evi at i ons A STM : A m e ric a n S o c ie ty o f T e sting a n d M a te ria ls A WWA : Amer i ca n W at erW ork s As s oci at i on N E R L -C i: U
. S. E P A N at i o nal E xp o sur e R esearch Lab o rat o ryi n C i nci nnat i , O hi o (O R D ).

N P D WR : N ati o na l P rima ry D rinking W ater R eg ulati o ns. O G WD W: U
. S. E P A O f f i ce o f G round W at er and D ri nki ng W at er. ORD : U .S . E P A O ff

ic e o f R e se a rc h a n d D e v e lo p m e n t. SD W A : T h e S a fe D rin k ing W a te r A c t as a m e n d e d (4 2 U .S .C . 3 0 0 f et se q .). A cc ur acy: A m easure of the cl o senes s o f

an i nd i vi d ual m easurement or the average o f a n u m b e r o f

m easurements t o the true v a lue . A c c u ra c y inc lud e s a c o m b ina tio n o f ra n d o m e rr o r (p re c isio n ) a n d s yste m a tic e rr o r (b ia s) c o m p o n e n ts w h ic h a re d u e to samp l i ng and anal y t i cal op erat i o ns. R ef er t o Standard M etho d s, D at a Q ual i tySec t i o n f o r a mo re detai l ed exp l anat i o n. A dmi ni s t rator: T h e a d m inistra to r o f th e U .S . E P A o r h e r/h is a u tho riz e d re p re se n ta tive . S e e 4 0 C F R 1 4 2 .2
. A g en cy : T h e U .S . E P A . S e e 4 0 C F R 1 4 2 .2
. A udi t or - A p er s on wh o eva l uat es l aborat ori es t o det ermi ne i f they meet the cr i t er i a t o be cert i f i ed. T hi s pers on s houl d be an exp eri enced pro f

ess i o nal , who has ef

f ect i ve co m m uni cat i o ns k i l l s , e xperi en ce inqu al i tyass urance, the anal y t i cal t echni q ues bei ng e v a lua te d , an d fam ili a rity w ith the d rin k ing w a te r re g u la tio n s a n d this m a n u a l. B ach el or D egr ee or Equi val en t : A co l l ege d egree wi th an eq ui val ent 3 0 s em est er hours i n a s p eci f i c d i scip l i ne. E q ui val ent i s atl eas tf

oury ear s of

experi enc e i n as peci f

i c s ci ent i f

i c di s ci pl i ne. B i a s: T he sy st em at i c o r persi st entdis t o rt i o n of a m easurement proc ess whi ch causes errors i n one dir ect i o n. C er t i f i cat i on A ut hor i t y: ( C A ) T he pers on or des i gn ee who h as the au thor i tyt o cert i f yl aborat ori es conduc t i ng dri nk i ng wat er an al y s es a n d t o c ert i f

yth e of f i ci al s of th e St at e r es pon s i bl e f

orth e St at e' s ce rt i f

i ca t i on pr og r am i n ac cor dan ce w i th Se ct i on 1412 of th e Sa f e D r i n k i n g W at erAct
. T h i s a u th or i tyi s de l eg at ed t o th e Reg i on alAdmi n i s tr at orbu tma ybe r ede l eg at ed. C e r t i f i c at i on Pr og r am M ana ger : ( C P M ) T he pers on r es po ns i bl e f or man agi ng the cert i f i cat i on progr am whi ch i ncludes tr ack i ng the cert i f

i cat i on st atus o f

the St at e l aborat ori es , ens ur i ng thatthe r egi onal an d St at e c ert i f i cat i on of

f i cer s ar e qua l i f

i ed a n d re v ie w ing the c e rtifi c a tio n e v a lua tio n re p o rts. C erti fi ca ti o n O ff

i cer: (C O ) A S t ate o r F ed era l l ab o rato ry au d i t o r w ho ha s p asse d the N E R L ce rti f i ca t i o n o f

f i ce rs training c o urse ( l i mi t ed atth i s t i me t o c h emi s trya n d mi cr obi ol og y )
. T h i s pe r s on pr ov i des i n f ormat i on t o th e CA orCP M f orth e pu rpos e of m a k ing d e c isio n s o n the c e rtifi c a tio n sta tus o f a la b o ra to ry. CF R : C o d e o f F e d e r a l R e g u la tio n s - A c o m p ilatio n o f re g u la tio n s is re v ise d e a c h ti m e a re g u la tio n is p ro m u lga te d . It i s p u b lishe d e v e ry ye a r in J u ly.C -2

C onf i rmat i on: V eri f

i c a t i o n of the p res ence o f a co m p o nentthrough the use of an anal y t i cal t echni q ue b ased o n a dif f erent sc ie n tif ic p rin c ip le fro m the o rig ina l m e tho d (e .g., se c o n d c o lum n , alte rn a te w a v e le n g th o r d e te c to r, e tc .) C o nfl i ct of I ntere st : a personal i nteres tor rel at i o nship , as d ef i ned by l aw o r r egul at i o n, thatco nf l i ct s wi th the f

ai thf ul p e rfo rm a n c e o f yo u r o ff ic ia l d u ty. D a ta A ud i t: A q u a li ta ti v e a n d q u a n tit a tive e v a lua tio n o f th e d o c u m e n ta tio n a n d p ro c e d u re s a sso c ia te d w ith m e a su re m e n ts to ver i f ythatthe r es ul t i ng dat a a r e a cce pt abl e. D a ta Q u a li ty O bjecti v es: qual i t at i ve and qua nt i t at i ve s peci f i cat i ons us ed t o desi gna s tudythatwi l l l i mi tun cert ai ntyt o an a c c e p ta b le lev e l. D a ta R ed uc ti o n: T he p ro ce ss o f tran sf o rm i ng the nu m b er o f d ata i t em s b y arithm eti c o r st ati st i ca l ca l cu l ati o ns, s t an d ard cu rve s, co nc en trati o n facto rs, etc. and co l l ati o n int o a m o re u seful f o rm . D ata red uc t i o n i s irrev ersib l e an d g en era l l y re sult s in the l o ss o f d e ta il. D et ec t i on: A n y c o n c e n tra tio n o f a n a n a lyte wh ic h e q u a ls o r e x c e e d s the lab o ra to ry' s d e te c tio n li m it. F o r V O C s, d e te c tio n li m it i s de f i n ed as 0. 0005 mg / L. D ri nking W a ter L a bo ra tory : A la b o ra to ry tha t a n a lyse s sa m p le s a s p a rt o f co m p lian c e m o n ito rin g fo r a p u b lic w a te r su p p ly. H ol di ng ti me: T h e al l owe d t i me f r om wh ena s ampl e wa s t ak en( ore x tr ac t ed) u n t i l i tmu s tbe a n al y ze d. I DC : In iti a l D e m o n stra tio n o f C a p a b ili ty - b e fo re a n a lyz ing c o m p lian c e s a m p le s a n a n a lyti c a l te a m m u st d e m o n stra te a c c e p ta b le pr ec i s i on , a cc u r ac y , s en s i t i v i ty , a n d s pec i f i ci tyf orth e meth od t o be u s ed. L R B - L a bo ra tory R ea g en t B l a nk: (M e tho d b la n k ) A n a liq u o t o f re a g e n t w a te r o r o the r b la n k m a trix th a t i s tre a te d e x a c tly a s a sa m p le to d e te rm ine if m e tho d a n a lytes o r o the r in te rfe re n c e s a re p re se n t. L F B - L a bo ra tory F o rti fi ed B l a nk: ( Spi k e) An al i qu ot of r ea g en t wa t eroroth erbl an kma tr i x t o w h i chkn ownqu an t i t i es of th e meth od a n al y t es a r e adde d i n th e l abor at ory . T h e LFB i s a n al y ze d e x ac t l yl i k e a s ampl e t o de t ermi n e wh eth erth e meth od is in c o n tro l. M C L: M axi m um co nt am i nantl evel m eans the m axi m um pe rmis sib l e l evel of a co nt am i nantin waterw hi ch i s d el i vered to any u s erof a publ i c wa t ers y s t em. Se e 40 CFR P art141. 2. M C L G : M ax i mu m con t ami n an tl ev elgoa l m ea n s th e max i mu m l ev elof a con t ami n an ti n dri n k i n g wat eratwh i chno kn ownor ant i ci p at ed ad vers e ef

f ecton the heal th of p ers o ns wo ul d oc cur, and whi ch allo w s an ad eq uat e m argi n of saf ety . M axi m um con t ami n an tl ev elg oal s a r e n on en f or ce abl e h ea l th g oal s . Se e 40 CFR 141. 2. M etho d D etecti o n L i m i t: ( M D L) th e mi n i mu m c on ce n tr at i on of a s u bs t an ce th atc anbe me as ured and r eport ed wi th 99% c o n fid e n c e tha t th e a n a lyte c o n c e n tra tio n is gr e a te r th a n z e ro . T h e M D L is d e te rm ine d fro m a n a lysis o f a sa m p le in a giv e n m a trix c o n ta inin g this a n a lyte. S e e 4 0 C F R 1 3 6 A p p . B
. M e th o d R e p o r tin g L im it : ( M RL) th e l owe s tc on ce n tr at i on of s t an dar d u s ed f

orc al i br at i on M oni t ori ng T ri gg er : T h e c o n c e n tra tio n o f a r e g u la te d c o n ta m ina n t wh ic h trigg e rs a d d iti o n a l m o n ito rin g . (S e e D e te c tio n L im it 1 4 1
. 2 4 (h) (i i i )( 1 8 )) NE L AC : N at i onal E nv i r onment al L aborat oryA ccr edi t at i on C onf er enc e - a v ol un t aryorg ani zat i on of St at e, Feder al an d other g ro u p s to e sta b lish m u tua lly a c c e p ta b le sta n d a rd s fo r a c c re d iti n g e n v iro n m e n ta l l a b o ra to rie s.C -3

P recis i o n: T he m ea sure o f m utua l ag ree m en tam o ng i nd i vid ua l m ea sure m en t s. P ri m a cy : P r i maryr es po ns i bi l i tyf or a dm i ni s tr at i o n an d e nf orce mentof

pri marydri nk i ng wat err egu l at i ons an d re l at ed re q u ire m e n ts a p p lica b le to p u b lic w a te r sys te m s w ithin a S ta te . P ri nc i pa lSta te L a bo ra tory Sy st em : A l l f aci l i t i es, whether p art o f the Stat e l ab o rat o ry o r co ntract ed b ythe Stat e, pro d uci ng d a ta fo r th e S ta te a n d c e rtifi e d b y the E P A , f u lfi lli n g the re q u ire m e n ts fo r P rim a c y a s listed in the 4 0 C F R 1 4 2 .1 0 (b )( 4 ). P ro feci en cy T est i ng Sa m ples (P T s) : A s am p l e p rovided t o a l ab o rat o ryf o r the p urpo se of

de m o n strat i ng thatthe l ab o rat o ry ca n s u cc es s f

u l l yana l y ze th e s ampl e wi th i n s pec i f i ed ac ce pt an ce l i mi t s s pec i f i ed i n th e r eg u l at i on s . T h e qu al i t at i v e an d/ or q uant i t at i ve co m p o si t i o n of the ref erence m at eri al i s unkno wn t o the l ab o rat o ryatthe t i m e o f the a n a lysis. S e e 4 0 C FR P art 141. 2. Pub l i c W at e r S ys t e m: A syst em fo r the p ro visi o n t o the p ub l i c o f p i p ed wa t er fo r hum an co nsum p t i o n, i f such sy st em ha s at l ea st f i f t een servi ce co nnect i o ns o r regul arl yserves an average o f a tleas ttwenty - f

i ve i nd i vi d ual s d ai l yatl east60 da y s o utof the y ear. Se e 40 CFR P art141. 2. Q ua l i ty A ss ur a nc e: A n inte g ra te d syste m o f m a n a g e m e n t ac tivities in v o lvin g p la n n ing , q u a lity c o n tro l, q u a lity a sse ssm e n t, r eport i n g a n d qu al i tyi mpr ov eme n tt o e n s u r e th ata pr odu ctor s erv i ce m ee t s de f i n ed s t an dar ds of

qu al i tyw i th a s t at ed l ev elof conf i dence
. Q u a lity C o n tr o l: T he over al l sy s t em of t ech ni calact i vi t i es whose p urpo s e i s t o meas ur e and contr ol the qual i tyof a pr od uct or s er v i ce s o th a t i t m e e t s th e n e e d s o f th e u s e r s ; o p e r a t i o n a l t e c h n i q u e s a n d a c t i v i t i e s th a t a r e u s e d t o f u l f i l l r e q u i r e m e n t s f

or q u a lity. Q A P l a n: A c o m p re h e n siv e p la n d e ta il in g t h e a sp e c ts o f q u a lity a ssu ra n c e ne e d e d to a d e q u a te ly f u lfi ll the d a ta ne e d s o f a pr og r am. T h i s doc u men ti s r equ i r ed bef or e th e l abor at oryi s c ert i f i ed. R egul atory L evel : A c o n c e n tra tio n o f a c o n ta m ina n t wh ic h is cite d in the F e d e ra l R e g u la tio n s (e .g., M C L , d e te c t, e tc .) Sh a l l : D e n o te s a m a n d a to ry re q u ire m e n t. Shoul d: D eno t es a gui d el i ne o r r eco m m end at i o n. Sta nd a rd O pe ra ti ng P ro ced ur e: A writt en d o cu m en twh i ch d e ta ils the m etho d o f an o p era t i o n, ana l ysi s o r ac t i o n w ho se te c h n iq u e s a n d p ro c e d u re s a re tho ro u g h ly p re sc rib e d a n d w h ic h is o ff ic ia lly a p p ro v e d a s t h e m e tho d fo r p e rfo rm ing c e rta in ro uti ne o r rep eti t i ve t ask s. T hird P a rty A ud i tor: P e rs o n o r p e rs o n s, n o t a ff ili a te d w ith a R e g io n o r S ta te , w h o is d e sig n a te d b y the R e g io n o r S ta te to a u d it a l abor at ory . T h i s pe r s on mu s tpa s s th e ce r t i f i ca t i on tr ai n i n g c ou r s e pr i ort o a u di t i n g a n yl abor at oryu n l es s h e ors h e i s a pa rtof an aud i tt eam whi ch in c lud e s a R e g i o nal / Stat e cert i f i cat i o n off

i cer. T he thi rd p artyaud i t o r mustal so m eetthe ed ucat i o nal / exp eri ence req ui rem ent s specif i ed i n thi s m anual
. T he cert i f i cat i o n de ci si o n remains wi th the R egi o n. T hird P a rty E xpe rt: A ny p erso n no td esigna t ed as a c erti f i ca t i o n o f

f i ce r o r au d i t o r, w ho i s req ue st ed b y the R eg i o n to assis t i n the aud i tof a l ab o rat o rybe cause of hi s o r her e xp ert i se i n a pa rt i cul ar area ( e. g. , asbest o s)
. T hi s p ers o n i s no trequi red t o t ake the cert i f i cat i o n off

i cers'co urs e i f

he or she i s p artof an aud i tt eam whi ch i ncl ud es a cert i f i cat i o n off

i cer. Tri balN at i on: A rea s wh i ch f o r reg ulato ry pu rp o ses are trea t ed as ind ep en d en tS t ates. O n these l an d s, the In d ia n t r ib e h a s a F ed era l l y reco gn i ze d go ve rning b o d y carrying o ut go ve rnm en td uti es a nd p o w ers.

" U n re g u l a ted " C o n ta m i n a n ts: Con t ami n an t s f orw h i chmon i t or i n g i s r equ i r ed bu tw h i chh av e n o M CL.C -4D -1 Appendix D

UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460

January 16,1997

MEMORANDUM SUBJECT: The Use of "Third-Parties" in the Drinking Water Laboratory Certification

Program FROM: Cynthia Dougherty, Director

Office of Ground Water and Drinking Water TO: Water Supply Representatives, Regions I-X

Certification Authorities, Regions I-X Quality Assurance Officers, Regions I-X Regional Laboratories, Regions I-X
Purpose This memorandum updates and clarifies the guidance memorandum from Michael Cook dated December 5, 1989 on "Third-Party Certification for Laboratories in Primacy States." Action U n d e r 40 CFR 142.10(b) (3), if a State does not perform all analytical measurements in its own laboratory, it must establish and maintain a program for the certification of laboratories as a c o n d i t i o n for receiving and maintaining authority to administer the Saf e D rinking Water Act in lieu of EPA (primacy). This memorandum notifies States with primacy that they may contract w i t h other organizations (third parties) to assist the State in fulfilling this requirement. The authority for m a k i n g certification decisions however, must remain with the State.
Discussion Several States have asked USEPA its position on the use of third-parties, i.e., private sect o r o r g anizations which assist the States with their certification program. OGWDW realizes that dwindling State resources may necessitate assistance from third-parties in the St a t e c e r tification programs. Consistent with the regulatory requirement at 40 CFR 142.10(b), providing for the "establishment and maintenance of a State program for the certification of laboratories," the State must retain ultimate authority to decide whether individual laboratories will be certified: this decision may not be abdicated to the third party.

This Off i c e w i l l n o t pass judgment on any specific third-party program. It is the responsibility of each p r i m acy State to assess the qualifications of the third-party. In assessing whether to choose a particular third-party, the State should consider, as a minimum, the following items wh i c h are described in theD -2

Manual for the Certification of Laboratories Analyzing Drinking Water :

o Ability to provide technical assistance and training
o Availability of records for review by the State
o Quality assurance program
o Freedom from conflicts of interest
o EPA po l i c y , which provides that the auditor should pass an

appropriate course on how to audit in the discipline for which he or she will be auditing.
o Experience of the auditor.

The auditor should be an experienced professional with at least a bachelor's degree or equivalent education/experience in the discipline for which he or she audits. The auditor should have recent laboratory experience Any State certification program using third part y a s s i s tance should meet the re q u i rements in the Manual for the Certification of Laboratories Analyzing Drinking Waterjust as it would if it were using State employees to perform these functions. T h e R egions should assist the State and third-party agent to assure that the certification program meets EPA guidelines. Regions and States should be sensitive to potential conflict-of-interest problems between a third-parties and evaluated l a b o r a t ories. For instance, inspectors employed by firms that provide analytical services in the drinking water area should not be put in the position of passing judgement on their competitors. Further Information If you have questions or need additi o n a l i n formation or assistance, please contact the OGWDW Technical Support Center at 513-569-7904.E -1 A pp en dix E R e qu i r e d A na l yt i c alCapa bi l i t y f or Pr i nc i pa lS t at e Lab or at or y S ys t e ms I NORGANI CS ( 40 CFR 141. 23) As bes t os Cy an i de Fl u or i de N itra te N itrit e A nt i m o ny A rs e n ic B ari um B ery l l i um C ad m i um C hrom i um M ercury Se l eni um T hal l i um ( 40 CFR 141. 89) C op per Lead C o n d u c tivity C al ci um A lka linity O rth o p h o sp h a te Si l i ca

VOLAT I L E ORGANI CS ( 40 CFR 141. 24) T H Ms B enzene Carbon t etr ac h l or i de C hl o rob enzene o - D i chl o rob enzene p - D i chl o rob enzene 1 , 2
- D i chl o roethane 1 , 1
- D i chl o roethy l ene ci s- 1 , 2
- D i chl o roethy l ene trans - 1 , 2
- D i chl o roethy l ene D i chl o rom ethane 1 , 2
- D i chl o rop rop ane E thy l b enzene Styrene T etrachl o roethene T o l uene 1 , 2 , 4
- T ri chl o rob enzene 1 , 1 , 1
- T ri chl o roethane 1 , 1 , 2
- T ri chl o roethane T ri chl o roethene V i n y l c h l or i de X y l ene s DBPs 5 H AA B ro m a te C h lo rite

S OCs ( 40 CFR 141. 24) Al ac h l or A trazi ne B enzo (a)pyr ene C arbo f ur an C hl o rdane 2 ,4 -D D i(2 -eth ylhe x yl)a d ip a te D i(2 -eth ylhe x yl)p h tha la te D i b rom o chl o rop rop ane D al apon D i nos eb Di o x i n ( 2 , 3 , 7 , 8
- TC DD) D i quat E n d o tha ll E n d rin Eth y l en edibr omi de G lyp h o sa te H ept ac h l or H ept ac h l ore pox i de H exachlorob enzene H exachloroc y cl o p ent ad i ene Lindane M eth ox y ch l or O x a m yl P C B s (a s d e c a c h lo ro b ip h e n yl) Pen t ac h l or oph en ol P i cl oram Simaz i ne T o xap hene 2 , 4 , 5
- TP R A D I O N U C LI D ES ( 40 CFR 141. 25) G ross A l p ha U rani um G r oss B et a Ce s i u m- 134 S tr on t i u m- 89 I odi n e- 131 S tr on t i u m- 90 T ri t i um O ther beta/p ho t o n em i tt ers Radi u m- 226/ 228

M I C R O O R G A N I S M S ( 40 CFR 141. 21) T ot a l c ol i f

orms Esc h e ric h ia c o lior f

eca l c ol i f orms H e te ro tro p h ic b a c te riaF -1 A pp en dix FF -2G -1 A pp en dix G A nalyt i calM et hods f

or M i cr obi ol ogy N o t e: Inf

o rm ati o n in b rac ke t s i s no tyeti nc l ud ed i n the C o d e o f F ed era l R eg ulati o ns 1 . T o tal C o l i form R ule ( 4 0 C F R 14 1 . 2 1 (f ) ) (f ) Anal yti cal m ethodology . (1) T h e s t an dar d s ampl e v ol u me r equ i r ed f ort ot alcol i f

orm an al y s i s , re g ar dl es s of a n al y t i ca l me th od u s ed, i s 100 mL. (2 ) P ubl i c wa t ers y s t ems nee d onl ydet ermi ne the pres enc e orabs enc e of t ot al col i f

orms ; a det ermi nat i on of t ot al col i f orm de n s i tyi s n otr equ i r ed. (3) P ub l i c water sy st em s m ustco nd uctt o t al co l i f o rm analy ses i n acco rdanc e wi th one of the anal y t i cal m etho d s i n the fo llo w ing tab le
. O r g an i s ms M eth odol og y Ci t at i on 12 1 T ot al Col i f orms 2 T o ta l C o lif o rm F e rm e n ta tio n T e c h n iq u e ... .... .... .... .... .... .... .... .. 3,4 ,5 T o ta l C o lif

o rm M e m b ra n e F ilt e r T e c h n iq u e ... .... .... .... .... .... .... .... 6 P re se n c e -A b se n c e (P -A ) C o lif o rm T e st
.... .... .... .... .... .... .... .... .... .

5,7 O N P G -M U G T e st
... .... .... .... .... .... .... .... .... .... .... .... .... .... .... .... .... .... . 8 C o l i sure T est 9 E *C o l i t e T est

® 1 0 m - C o l i B l ue 2 4 T est

® 11 Rea dy cu l t Col i f

orms 100 ® 1 3 M emb r ane Fi l t erT ech ni que u s i ng Chromo cul t Co l i f orm Agar ® 1 4 C o l i t ag T est 15 9 2 2 1 A , B 92 22 A, B, C 9 2 2 1 D 9223

T he p roce d ures s hal l be do ne i n acco rdanc e wi th the d o c u m e n ts l i st ed be l o w. T he i nco rpo rat i o n by r ef erence o f

the f ol l owi ng do cumen t s l i s t ed i n f oo tnot es 1, 6, 8, 9, 10 , 11 , 13 and 14 was approved by the D i r ect or of the Federa l Regi s t er i n acc ordanc e wi th 5 U
. S. C
. 55 2(a) and 1 CFR P art 51
. C op i es of the do cumen t s may be ob t ai ned f r om the s our ces l i s t ed bel ow. I n f

ormat i on r eg ar di n g obt ai n i n g th es e docu men t s ca n be obt ai n ed f r om th e Sa f

e D r i n k i n g W at erH ot l i n e at800- 4 2 6 -4 7 9 1 . D o c u m e n ts m a y b e insp e c te d a t E P A ’s D rin k ing W a te r D o c k e t, 1 3 0 1 C o n stitutio n A v e n u e , N W ., E P A W e st, Room B 102, W as h i n g t on , D C 20460 ( T el eph on e 202- 566- 2426) ora t th e O f f i ce of Fe der al Reg i s t er , 800 N orth Capi t ol Str ee t , N W , Su i t e 700, W as h i n g t on , D C 20408. Stan da rd Metho dsf

or the Exam i na t i on of

W at erandW

astewa ter , 18 edi t i on (1992) , 19 edi t i on ( 1995) , or20 e di t i on 1 th th th ( 1998)
. Amer i ca n Publ i c H ea l th As s oci at i on , 1015 Fi f t ee n th Str ee tN W , W as h i n g t on , D
. C. 20005. T h e ci t ed meth ods pu bl i s h ed i n a n yof

th es e th r ee e di t i on s ma ybe u s ed. T h e t i m e f

r o m s a m p l e c o l l e c t i o n t o i n i t i a t i o n o f a n a l y s i s m a y n o t e x c e e d 3 0 h o u r s . S y s t e m s a r e e n c o u r a g ed bu tn ot 2 re q u ire d to ho ld sa m p le s b e lo w 1 0 d e g . C d u rin g tra n sit. La ct os e br oth , a s commer ci al l yav ai l abl e, ma ybe u s ed i n l i euof l aury l trypt os e br oth , i f th e s y s t em con du ct s a tl ea s t25 3 par al l e l t e s t s b e tw e e n th i s m e d i u m a n d l a u ry l tryp t o s e b r o th u s i n g th e w a t e rn o rm a l l yt e s t e d , a n d th i s c o mpar i s on demon s tr at es th atth e f

al s e- pos i t i v e r at e a n d f al s e- n eg at i v e r at e f

ort ot alcol i f

orm[ s ] , us i n g l ac t os e br oth , i s l es s th an10 p e rc e n t. I f i nv ert ed tubes ar e u s ed t o det ectga s product i on, the med i a s houl d coverthes e tubes atl eas tone- hal f t o two- thi r ds af

t er 4 th e s ampl e i s a dded. N o req uirem en tex i st s t o run the co m p l eted p ha se o n 1 0 p erc en to f all t o t al co l i f

o rm - p o si t i ve co nfi rm ed tub es. 5 M I agar al so m ay b e used. P repa rat i o n and us e o f M I agar i s setforth in the art i cl e, `` N ew m ed i um f o r the si m ul t aneo us 6 det ec t i on of

t ot alc ol i f

orm[ s ] a n d Esc h e ric h ia c o liin w a te r' ' b y B re n n e r, K .P ., et a l., 1 9 9 3 , A p p l. E n v iro n . M ic ro b io l. 5 9 : 3 5 3 4
- 3 5 4 4
. A l s o a v a i l a b l e f r o m th e Of

f i c e o f W a t e r R e s o u r c e C e n t e r( R C
- 4 1 0 0 T) , 1 2 0 0 P e n n s y l v a n i a A v e n u e , NW , W as h i n g t on , D C 20460, EP A/ 600/ J- 99/ 225. V er i f i ca t i on of

c ol on i es i s n otr equ i r ed.G -2

Si x - t i mes f

ormu l at i on s tr en g th ma ybe u s ed i f th e medi u m i s f

i l t er - s t er i l i ze d r ath erth ana u t ocl av ed. 7 T h e ON PG
- M U G T e s ti s a l s o kn ow n a s th e Au t oa n a l y s i s Col i l e rtS y s t e m. 8A des cr i pt i on of

the C ol i s ur e T es t , Feb 28 , 199 4, maybe obt ai ned f r om ID E X X L aborat ori es , I nc. , O ne I D E X X D r i ve, 9 W estb ro o k, M aine 0 4 0 9 2
. T he C o l i sure T est m ay b e re ad aft er a n incu b ati o n ti m e o f 2 4 ho urs. A des cr i pt i on of th e E* Col i t e T es t , ` ` Pre s en ce /Abs en ce f orCol i f orms a n d E . C o liin W a te r,' ' D e c 2 1 , 1 9 9 7 , i s a v a ilab le 10 ® f r om Cha rm Sc i en ce s , I n c. , 36 Fr an k l i n Str ee t , M al den , M A 02148- 4120. A des cr i pt i on of

the m- C ol i B l ue24 T es t , Aug 17, 19 9 9 , i s avai l abl e f r om the H ach Co mp any , 100 D ay t on Avenu e, 11 ® Ames , I A 50010. EP A s tr on g l yr ec ommen ds th atl abor at or i es e v alu at e th e f al s e- pos i t i v e a n d n eg at i v e r at es f orth e meth od( s ) th eyu s e f or 12 m o n ito rin g to ta l co lif

o rm s. E P A a lso e n c o u ra g e s la b o ra to rie s to e sta b lish f a lse -p o sitive a n d false -ne g a tive ra te s w ithin the i r o w n lab o rato ry and sam p l e m atri x (d rinking w ater o r so urc e w ater) w i th the int en ttha t i f the m etho d the y cho o se has anunac cept abl e f

al s e- po s i t i ve or neg at i ve r at e, an othermethod can be us ed. T he A gen cys ug ges t s thatl aborat ori es p erf o rm these s tud i es on a m i ni m um of 5 % of al l t o t al co l i f o rm-po si t i ve samp l es, excep t fo r tho se m etho d s where ver i f i cat i on/ conf

i rmat i on i s al r eadyr equi r ed, e. g. , the M
- E ndo and LE S E ndo M emb r ane Fi l t erT es t s , St andar d T ot al C o l i f o rm F erm ent at i o n T echni q ue, and P res ence- A b sence C o l i f o rm T est
. M etho d s f

o r es t ab l i shi ng f

al se- p o si t i ve and negat i ve- rat es may be b ased o n l act o se f erm ent at i o n, the rapid t estf o r 

### $

- ga l ac t o si d ase an d cyto ch ro m e o xid ase , m ult i

- t est id e n tif ic a tio n syste m s, o r e q u iva le n t c o n firm a tio n te sts. F a lse -p o sitive a n d false - ne g a tive info rm a tio n is o ften a v a ilab le i n pub l i shed stud i es and/ o r f rom the m anuf

acturer( s)
. T h e Rea dy cu l t ® Col i f orms 100 Pr es en ce /Abs en ce T es ti s de s cr i b ed i n th e docu men t , “ Rea dy cu l t ® Col i f orms 100 13 Pre s en ce /Abs en ce T es tf orD et ec t i on a n d I den t i f i ca t i on of

Col i f orm B ac t er i a an d Esc h e ric h ia c o liin F inish e d W a te rs ,” ( N ovemb er2 0 0 0 , V ersio n 1. 0) a nd i s av ai l abl e f r om E M Sci enc e [ now EM D Chemi cal s , I nc. ] , anaf f i l i at e of M er ck K G g A, D arms t adtG erman y ) , 480 S. D emoc r atRoad, G i bbs t own , N J 08027- 1297. T el eph on e n u mberi s ( 800) 222- 0342, E -M a il a d d re ss is: a de l l e n bu s c h @ e ms c i e n c e . c om . [ E
- M ai l addres s i s now a de l l e n bu s c h @ e md c h e mi c a l s . c om . W e b site i s w w w . e md c h e mi c a l s . c om ] M em b rane Fi l t er T echni q ue usi ng C hrom o cul t ® C o l i f o rm A gar i s d escrib ed i n the d o cum ent , “ C hrom o cul t ® C o l i f o rm 14 Aga rPr es en ce /Abs en ce M embr an e Fi l t erT es tM eth od f orI den t i f

i ca t i on of

Col i f o rm B ac t er i a an d Esc h e ric h ia c o liin Fi n i s h ed W at er s , ” N ov embe r2000, V er s i on 1. 0, a v ai l abl e f r om EM Sci en ce [n ow EM D C h emi ca l s , I n c. ] ( ana f f i l i at e of M er ckK G g A, D arms t adtG erman y ) , 480 S. D emoc r atRoad, G i bbs t own , N J 08027- 1297. T el eph on e n u mberi s ( 800) 2 2 2 -0 3 4 2 , E -M a il a d d re ss is: a de l l e n bu s c h @ e ms c i e n c e . c om . [E
- M ai l ad dres s i s now a de l l e n bu s c h @ e md c h e mi c a l s . c om . W eb si t e is ww w. emdc h emi ca l s . com ] Col i t ag ® pr odu ctf

orth e det ermi n at i on of

th e pr es en ce / abs en ce of t ot alc ol i f

orms a n d E . co liis d escrib ed i n ` ` C o l i t ag® 15 Produ cta s a T es t f

or D et ec t i on a n d I den t i f i ca t i on of

Col i f orms a n d E . co liB act er i a i n D r i nk i ng W at eran d So ur ce W at er as Requ i r ed i n N at i on alPr i maryD r i n k i n g W at erReg u l at i on s , ' ' Au g u s t2001, a v ai l abl e f r om CP I I n t ern at i on al , I n c. , 5580 S kyl an e B l vd ., S an t a R o sa, C A , 9 5 4 0 3 , t elep ho ne (8 0 0 ) 8 7 8
- 7 6 5 4 , F ax (7 0 7 ) 5 4 5
- 7 9 0 1 , Intern et ad d ress h ttp ://w w w .c p iinte rn a tio n a l.co m . ( 4) [ Res erv ed] (5) P ub l i c wat er s y st em s m ust co nd uct f ecal co l i f

o rm anal y si s i n acco rdanc e wi th the f o l l o wi ng pro ced ure. W hen the M T F T echni q ue o r P res ence- A b sence (P A ) C o l i f o rm T esti s used t o t estf o r t o t al co l i f

o rms, s hake the l act o se- p o si t i ve p re su m p tive tub e o r P -A vig o ro u sly a n d tra n sfe r th e gr o w th w it h a s te r il e 3 -m m lo o p o r ste rile a p p lica to r stic kinto br i l l i an tg r ee n l ac t os e bi l e br oth a n d EC me di u m t o de t ermi n e th e pr es en ce of t ot ala n d f

ec alc ol i f

orms , r es pec t i v el y . For E P A
- ap p roved anal y t i cal m etho d s whi ch use a mem b rane f i l t er, trans f

er the t o t al co l i f

o rm-po si t i ve cul ture b y o ne o f the f ol l owi n g me th ods : r emov e th e membr an e con t ai n i n g th e t ot alc ol i f orm c ol on i es f

r om th e s u bs tr at e wi th a s t er i l e f or ce ps an d c ar ef

u l l yc u r l a n d i n s ertth e membr an e i n t o a tu be of EC medi u m ( th e l abor at oryma yf i r s tr emov e a s mal l port i on of sel ect ed co lo n ie s fo r v e ri f i cat i o n), s wab the ent i re mem b rane f i l t er s urf ace with a s t eri l e co tt o n s wab and trans f er the i no cul um to E C m ed i um (d o no tleave the co tt o n s wab i n the E C m ed i um ), or i no cul at e i nd i vi d ual t o t al co l i f o rm-po si t i ve c o lo n ie s into E C M e d ium . G e n tly sh a k e th e in o c u la te d tu b e s o f E C m e d ium to insu re a d e q u a te m ixin g a n d inc u b a te in a w a t e rb a th a t4 4
. 5 ± 0
. 2 ° C f or 2 4 ± 2 h o u r s . Ga s p r o d u c t i o n o f a n ya m o u n ti n th e i n n e rf e rm e n t a t i o n tu b e o f

th e EC medi u m i n di ca t es a pos i t i v e f ec alcol i f

orm t es t
. T h e pr epa r at i on of E C medi u m i s de s cr i bed i n M eth od 9221E ( par ag r aph 1 a ) in Stan da rd Metho ds f or the Exam i na t i on of

W at erandW

astewa ter , 18 th ed i t i o n (19 9 2 ), 19 th ed i t i o n (19 9 5 ), and 2 0 th e d iti o n (1 9 9 8 ); the c ited m e tho d i n a n y o n e o f th e s e thr e e e d iti o n s m a y b e us e d . P u b lic w a te r sys te m s n e e d o n ly det ermi n e th e pr es en ce ora bs en ce of f ec alc ol i f

orms ; a de t ermi n at i on of f ec alc ol i f

orm de n s i tyi s n otr equ i r ed. ( 6) Publ i c wa t er s y s t ems mu s t con du cta n al y s i s of Esc h e ric h ia c o lii n a ccor dance wi th one of the f ol l owi ng an al y t i calG -3

m etho d s: ( i ) E C m edi um s uppl ement ed wi th 50 μg/mL o f 4- methy l umbel l i f

ery l

- bet a- D
- glucur oni de ( M U G ) ( f i nal c o n c e n tra tio n ), a s d e sc rib e d in M e tho d 9 2 2 2 G in S ta n d a rd M e tho d s fo r th e E x a m ina tio n o f W a te r a n d W a ste w a te r,1 9 th edi t i on ( 19 95 ) an d 20 th e di t i on ( 19 98 )
. Ei theredi t i on maybe us ed. A l t ernat i vel y , the 18 th e di t i on ( 19 92 ) maybe us ed i f a tl e a s t1 0 m L o f

E C m e d i u m , a s d e s c r i b e d i n p a r a g r a p h ( f ) ( 5 ) o f th i s s e c t i o n , i s s u p p l e m e n t e d w i th 5 0 μ g /m L o f M UG b ef o re aut o cl avi ng. T he i nner i nvert ed f erm ent at i o n tub e m ay b e o m i tt ed
. If the 1 8 ed i t i o n i s used, ap p l ythe p roce d ure th i n p a r a g r a p h ( f

) ( 5 ) o f th i s s e c t i o n f

o rtr a n s f e rr i n g a t o t a l c o l i f

o rm - p o s i t i v e c u l tu r e t o E C m e d i u m s u p p l e m e n t e d w i th M UG, i ncub at e the tub e at 4 4
. 5 ± 0.2°C f o r 24 ± 2 ho urs , and then o b serve f l uo res cence w i th an ul travi o l etl i ght(36 6 nm ) i n the d a rk . If fluo re sc e n c e is visib le , E . co lia re p re se n t. ( i i ) N utri entag ars uppl ement ed wi th 100 μ g/mL 4
- methy l umbel l i f ery l

- bet a- D
- glucur oni de ( M U G ) ( f i nal c o n c e n tra tio n ), a s d e sc rib e d in M e tho d 9 2 2 2 G in S ta n d a rd M e tho d s fo r th e E x a m ina tio n o f W a te r a n d W a ste w a te r,1 9 th edi t i on ( 19 95 ) an d 20 th e di t i on ( 19 98 )
. Ei theredi t i on maybe us ed f or det ermi ni ng i f a t ot al col i f orm- po s i t i ve s amp l e, as d eterm i ne d b y a m em b ran e fi l t er tec hn i q ue , co ntains E . co li
. A l t ernat i vel y , the 1 8 ed i t i o n (19 9 2 ) m ay b e used i f the th membr an e f

i l t erc on t ai n i n g a t ot alc ol i f orm- pos i t i v e col on y ( i es ) i s tr ansf err ed t o n u tr i en ta g ar , a s de s cr i bed i n M eth od 9 2 2 1 B (p a ra g ra p h 3 ) o f S ta n d a rd M e tho d s (1 8 th e d iti o n ), su p p le m e n te d w ith1 0 0 μ g /m L o f M U G . If the 1 8 th e d iti o n is used, i ncub at e the agar plat e at35 °C f o r 4 ho urs and then o b serve the co l o ny (i es) unde r ul travi o l etl i ght(36 6 nm ) i n the d a rk fo r fluo re sc e n c e . If fluo re sc e n c e is visib le , E . co lia re p re se n t. ( i i i ) M i n i malM ediu m O N PG
- M U G ( M M O
- M U G ) T es t , a s s etf orth i n th e art i cl e ` ` N at i on alFi el d Eva l u at i on of a D ef i n ed Subs tr at e M eth od f orth e Si mu l t an eou s D et ec t i on of T ot alC ol i f

orms a n d Esc h e ric h ia c o lif rom D ri nki ng W a te r: C o m p a riso n w ith P re se n c e -A b se n c e T e c h n iq u e s' ' (E d b e rg e t a l.), A p p lied a n d E n v iro n m e n ta l M ic ro b io lo g y, V o lum e 5 5 , p p . 1 0 0 3 -1 0 0 8 , A p ril 1 9 8 9 . (N o te : T h e A u to a n a lysis C o lil e rt S yste m is an M M O -M U G tes t). If the M M O
- M U G t esti s t o t al co l i f o rm-po si t i ve af

t er a 24
- h o u r inc u b a ti o n, t estthe m ed i um f o r f l uo res cence w i th a 3 6 6
- nm ultrav i o l et l i gh t (p refera b l y w i th a 6
- w att l am p ) i n the d ark
. If f l uo resc en ce i s o b ser v e d , th e sa m p le is E. c o li

- p o si t i ve. I f

f l uo res cence is que st i o nab l e (canno tbe de f i ni t i vel yread) af

t er 24 hours i ncub at i o n, i ncub at e the cul ture f o r an add i t i o nal f

o ur hours (butnot t o exce e d 2 8 h o u rs to tal ), and agai n t estthe m ed i um f o r f l uo res cence. T he M M O- M UG T e st w ith h e p e s b u ff

e r in li e u o f p h o sp h a te b u ff e r is the o n ly ap p ro v e d fo rm u la tio n fo r th e d e te c tio n o f E . co li
. ( i v) T he C o l i sur e T est
. A de scr i p t i o n of the C o l i sur e T e st m a y b e ob t ai ned f rom the M i l l i p o re C o rpo rat i o n, T ec hn i ca l S erv i ce s D ep artm en t , 8 0 A shb y R o ad , B ed f o rd , M A 0 1 7 3 0
. [N o t e: M an ufac ture r i s no w ID E X X L ab o rato ries. S e e fo o tno te 9 to the tab le in p a ra g ra p h (f)( 3 ) o f th is se c tio n .] (v ) T h e membr an e f

i l t erm eth od wi th M I ag ar , a des cr i pt i on of wh i chi s ci t ed i n f

ootn ot e 6 t o th e t abl e i n pa r ag r aph (f )( 3 ) of thi s sect i o n. (vi ) E * C o l i t e T est , a de scr i p t i o n of whi ch i s ci t ed in f o o tno t e 1 0 to the t ab l e atp aragraph (f )( 3 ) of thi s sect i o n. ® (v ii) m -C o liB lue 2 4 T e st, a d e sc rip tio n o f w h ic h is c ited in f o o tno te 1 1 to the ta b le in p a ra g ra p h (f)( 3 ) o f th is ® sect i o n. ( v iii ) R e a d yc u lt® C o lif o rm s 1 0 0 P re se n c e /A b se n c e T e st, a d e sc rip tio n o f w h ic h is c ited in fo o tno te 1 3 to the tab le atpa ragraph (f

)( 3 ) of thi s sect i o n. (ix ) M e m b ra n e F ilt e r T e c h n iq u e u sin g C h ro m o c u lt C o lif o rm A g a r, a d e sc rip tio n o f wh ic h is cite d in fo o tno te ® 1 4 t o the t ab l e atpa ragraph (f )( 3 ) of thi s sect i o n. ( 7) As anopt i on t o parag r aph ( f ) ( 6)( i i i ) of

thi s s ect i on, a s y s t em wi th a t o t al c o l i f o rm - po s i t i ve, M U G
- neg at i ve, M M O -M U G te st m a y fu rth e r a n a lyz e the c u lture fo r th e p re se n c e o f E . co li b y tr a n s f e rr i n g a 0
. 1 m L , 2 8
- h o u rM M O- M UG cul tur e t o E C M edi um + M U G wi th api pet
. T he f ormu l at i on a n d i n cubat i on c on di t i on s of EC M ediu m + M U G , a n d obs erv at i on of th e r es u l t s ar e des cr i bed i n pa r ag r aph (f )( 6 )( i ) of thi s sect i o n.G -4

(8) T he f

o l l o wi ng m at eri al s are i nco rpo rat ed by r ef erence i n thi s sect i o n wi th the ap p roval o f the D i rect o r of

the F e d e ra l R egi st er i n acco rdanc e wi th 5 U
. S.C
. 5 5 2 (a) and 1 C FR p art 5 1
. C o p i es o f the anal y t i cal m etho d s ci te d in Stan da rd Metho ds f

or the Exam i na t i on of

W at erandW

astewa ter( 18th , 19th , a n d 20th e di t i on s ) ma ybe obt ai n ed f r om the A m eri can P ub l i c H eal th A ss o ci at i o n et al
. ; 1 0 1 5 Fif t eenth Str eet N W
. , W ashi ngt o n, D C 2 0 0 0 5 2 6 0 5
. C o p i es of the M M O
- M U G T estas setf o rth i n the art i cl e ` `N a tio n a l F ie ld E val uat i o n of a D ef i ned Sub strat e M etho d f o r the Si mu l t an eou s Enu mer at i on of T ot alCol i f

orms a n d Esc h e ric h ia c o lif rom D ri nki ng W at er: C o m p ari son wi th the Standard M ul t i p l e T ub e Fe rme nt at i o n M etho d ' ' (E d b erg e t a l. ) ma ybe obt ai n ed f r om th e Amer i ca n W at er W o rk s As s oci at i on R es ear ch Foundat i on, 66 66 W es t Q ui ncy A venue, D env er , C O 80 23 5. A des cr i pt i on of the C ol i s ur e T es t maybe obt ai ned f r om th e M i l l i por e Corp. , T ec h n i ca l Se rv i ce s D epa rtmen t , 80 As h by Road, B edf or d, M A 01730 [ N ot e: N ow a des cr i pt i on o f the C o l i sur e T estm a ynow be ob t ai ned f rom ID E X X Lab o rat o ri es, Inc. , O ne ID E X X D ri ve, W estb roo k, M ai ne 04 09 2. ]
. Co pi es maybe i ns pect ed atE P A ' s D r i nk i ng W at erD ocke t ; 401 M St
. , SW
. ; W as hi ng t on, D C 204 60 [ N ot e: curr en tl oca t i on of

EP A' s D r i n k i n g W at erD ock eti s 1301 Con s t i tu t i on Av enu e, N W
. , EP A W es t , Room B 102, W ashi ngt o n, D C 20 4 6 0 ], or at the O f f i ce o f the Fe d eral R egi st er; 80 0 N o rth C ap i t o l Str eet , N W
. , Suit e 7 0 0 , W ashi ngt o n, D C. 2 . Su rfac e W a ter T rea tmen t R ule ( 4 0 C F R 14 1 . 7 4 (a)) (a) A n a ly tica l re q u ire m e n ts . On l yth e a n a l y t i c a l me th od ( s ) s pe c i f i e d i n th i s pa r a g r a ph , oroth e rw i s e a pp r ov e d b yEPA, m ay b e used t o de m o nstrat e co m p l i ance with §§ 1 4 1
. 7 1 , 14 1
. 7 2 a nd 1 4 1
. 7 3
. M easurements f o r pH , turbidi ty , t em p erature a n d re sid u a l d isinfe c ta n t c on c e n tr a t i on s mu s tbe co n du c t e d b ya pe r s ona pp r ov e d b yth e S t a t e . M e a s u r e me n tf ort ot a l c ol i f

orms , f e c a l c ol i f

orms and H PC mu s tbe c on du ct ed bya l abor at oryc ert i f i ed byth e St at e orEP A t o do s u cha n al y s i s . U n t i l l abor at o ry cert i f i cat i on cr i t er i a a r e dev el op ed f or the ana l y s i s of

f eca l col i f

orms an d H P C , an yl aborat orycert i f i ed f or t ot al co l i f

orms anal y s i s b ythe St at e o rE P A i s deemed cert i f i ed f o r f ecal co l i f

o rm s a n d H P C a n a l y si s. T he f o l l o wi ng pro ced ures s hal l be co nd uct ed i n acco rdanc e wi th the p u b lica tio n s listed in the fo llo w ing se c tio n . T hi s i ncorpo r at i on byr ef er enc e wa s approved bythe D i r ect or of the Federa l Regi s t eri n a ccor dance wi th 5 U
. S. C
. 552 ( a) and 1 C FR p art5 1
. C o p i es of the m etho d s p ub l i shed i n Stan da rd Metho dsf or the Exam i na t i on of

W at erandW

astewa ter maybe obt ai n ed f

r om th e Amer i ca n Publ i c H ea l th As s oci at i on et al
. , 1015 Fi f t ee n th Str ee t , N W
. , W as h i n g t on , D C 20005; copi es of the M i ni mal M edi um O N P G
- M U G M ethod as s etf orth i n the art i cl e ` ` N at i onal Fi el d E valuat i on of a D ef i ned Sub strat e M eth od f

orth e Si mu l t an eou s Enu mer at i on of T ot alCol i f

orms a n d Esc h e ric h ia c o lif rom D ri nki ng W at er: C om pari s on w i th th e S t a n da r d M u l t i pl e T u be F e rme n t a t i onM e th od ' ' ( Edbe r g e ta l
. ) , Ap pl i e d a n d En v i r on me n t al M i cr ob i ol ogy , V ol ume 54, pp . 1 5 9 5
- 1 6 01, J u n e 1988 ( as a men ded u n derEr r atu m, Appl i ed an d Env i r on men t alM i cr obi ol og y , V ol u me 54, p. 3 1 9 7 , D ec em b er, 1 9 8 8 ), m ay b e o b t aine d fro m the A m erica n W ater W o rks A sso ciati o n R ese arc h F o un d ati o n, 6 6 6 6 W est Q u i n cyAv enu e, D en v er , Col or ado, 80235; a n d c opi es of th e I n di g o M eth od as s et f orth i n th e art i cl e ` ` D et ermi n at i on of O zo ne i n W at er by the Indi go M etho d ' ' (B ad er and H o i gne), m a y be ob t ai ned f rom O zo ne Sc i ence & E ngi neeri ng, P er gamon P r es s Lt d. , Fai rvi ew Park , E l msf ord, N ew Y ork 10 52 3. C op i es may be i ns pect ed atthe U
. S. E nv i r onment al P rotect i on Ag en cy , Room EB 15, 401 M St
. , SW
. , W as h i n g t on , D C 20460 [ N ot e: c u rr en tl oca t i on of EP A' s D r i nki ng W at erD ock eti s 1301 Con s t i tu t i on Av enu e, N W
. , EP A W es t , Room B 102, W as h i n g t on , D C 20460; ] ora tth e O f f i ce of th e F e de r a l Re g i s t e r , 80 0 North Ca pi t olS tr e e t , NW
. , s u i t e 70 0, W a s h i n g t on , DC. ( 1) P ubl i c wat er s y s t ems mus t conductan al y s i s of pH an d t emp er atur e i n a ccor dance wi th one of the methods l i s t ed at§14 1. 23 ( k) ( 1). Publ i c wa t ers y s t ems mus tconduc tan al y s i s of t ot al col i f orms , f eca l col i f

orms , het er otr op hi c bact er i a, and turbi di tyi n a ccor dance wi th one of the f ol l owi ng an al y t i cal methods and by us i ng ana l y t i calt es tprocedur es cont ai ned i n T e c h n i c a l No te s o n Dr i n k i n g Wa t erMethods , E P A
- 6 0 0 /R
- 9 4
- 1 7 3 , O ct o b er 19 9 4 , whi ch i s avai l ab l e atN T IS PB 95- 104766.G -5 O rgan i s m M et hodol ogy C i t at i on 1 T o ta l C o lif

o rm .... .... .... . 2

F e c a l C o lif

o rm s .... .... ... 2 H e te ro tro p h ic b a c te ria
.. 2 T u rb id ity
.... .... .... .... .... ... T o t al C o l i f o rm F erm ent at i o n T echni q ue 3, 4 , 5 T o t al C o l i f

o rm M em b rane Fi l t er T echni q ue 6 O N P G -M U G T e st 7 Fe cal C o l i f

o rm P roce d ure 8 Fe cal C o l i f

o rm F i l t er P roce d ure PourPl at e M eth od S im P la te 11 N eph el ometr i c M eth od N eph el ometr i c M eth od G re a t L a k e s In stru m e n ts H ach Fi l t er T r ak 9 2 2 1 A . B . C 9 2 2 2 A , B , C 9223 9221 E 9222 D 9215 B 2130 B 1 8 0 .1 9 M eth od 2 10 10133 12 T he procedur es s hal l be do ne i n acc ordanc e wi th the do cumen t s l i s t ed bel ow. T he i ncorpo r at i on byr ef er enc e o f th e fo ll o wi ng d o cum ent s l i st ed i n f o o tno t es 1 , 6 , 7 , 9
- 1 2 was ap p roved b y the D i rect o r o f the Fe d eral R egi ste r in acc ordanc e wi th 5 U
. S. C
. 5 52 ( a) and 1 C FR p art51. Co pi es of the do cumen t s maybe obt ai ned f r om the s our ces l i s t ed bel ow. I nf

ormat i on r ega r di ng obt ai ni ng thes e document s ca n be obt ai ned f r om the Sa f

e D rinking W at erH ot l i ne at 8 0 0
- 4 2 6
- 4 7 9 1
. D o c u m e n t s m a yb e i n s p e c t e d a tEPA ' s Dr i n k i n g W a t e rDo c k e t , 1 3 0 1 Co n s t i tu t i o n A v e n u e , NW
. , EPA W es t , Room B 102, W as h i n g t on , D C 20460 ( T el eph on e: 202- 566- 2426) ; ora tth e O f f i ce of the F ed eral R eg i s t er , 800 N or th Capi t ol Str ee t , N W , Su i t e 700, W as h i n g t on , D
. C. 20408. Exc ept wh er e n ot ed, al l me th ods r ef ert o Stan da rd Metho ds f or the Exam i na t i on of

W at erandW

astewa ter , 18 e di t i on 1 th ( 1 9 9 2 ) , 1 9 e d i t i o n ( 1 9 9 5 ) , o r 2 0 e d i t i o n ( 1 9 9 8 ) , A m e r i c a n P u b l i c H e a l th A s s o c i a t i o n , 1 0 1 5 F i f t e e n th S tr e e t , NW , th th W as h i n g t on , D
. C. 20005. T h e ci t ed meth ods publ i s h ed i n a n yof th es e th r ee e di t i on s ma ybe u s ed. T h e t i me f

r om s ampl e col l ec t i on t o i n i t i at i on of a n al y s i s ma yn ote x ce ed 8 h ou r s . Sy s t ems mu s th ol d s ampl es be l ow 10 2 d e g . C d u rin g tra n sit. La ct os e br oth , as commer ci al l yav ai l abl e, m aybe us ed i n l i euof l aury l trypt os e br oth , i f th e s y s t em con du ct s a tl ea s t25 3 par al l el t es t s betwe en th i s medi u m an d l aury l trypt os e br oth u s i n g th e wa t er n ormal l y t es t ed, and thi s compar i s on demon s tr at es th atth e f al s e- pos i t i v e r at e an d f al s e- n eg at i v e r at e f

ort ot alc ol i f

orm, u s i n g l ac t o se br oth , i s l es s th an10 p e rc e n t. M edi a s h ou l d c ov eri n v ert ed tu bes a tl ea s ton e- h al f t o two- th i r ds a f

t erth e s ampl e i s a dded. 4 N o req uirem en tex i st s t o run the co m p l eted p ha se o n 1 0 p erc en to f all t o t al co l i f

o rm - p o si t i ve co nfi rm ed tub es. 5 M I agar al so m ay b e used. P repa rat i o n a nd use o f M I agar i s setf

o rth i n the art i cl e, ` ` N ew m ed i um f o r the si m ul t aneo us 6 det ec t i on of

t ot alc ol i f

orm[ s ] a n d Escheri ch i a c o liin w a te r' ' b y B re n n e r, K .P ., et a l., 1 9 9 3 , A p p l. E n v iro n . M ic ro b io l. 5 9 :3 5 3 4 -3 5 4 4
. A lso a v a ilab le fro m the O ff ic e o f W a te r R e so u rc e C e n te r (R C -4 1 0 0 T ), 1 2 0 0 P e n n sylv a n ia A v e ., N W ., W as h i n g t on , D C 20460, EP A 600/ J- 99/ 225. V er i f i ca t i on of

c ol on i es i s n otr equ i r ed. T h e ON PG
- M U G T e s ti s a l s o kn ow n a s th e Au t oa n a l y s i s Col i l e rtS y s t e m. 7A- 1 Br oth ma ybe h e l d up t o th r e e mo n th s i n a t i g h t l yc l os e d s c r e w c a p tu be a t4 de g . C. 8

` ` M eth ods f

orth e D et ermi n at i on of I n or g an i c Subs t an ce s i n Env i r on men t alSa mpl es ' ' , EP A/ 600/R- 93/ 100, Au g u s t1993. 9 Ava i l abl e atN T I S, PB 94- 121811. G LI M ethod 2, ` ` T urbi di ty ' ' , N ov em b er 2 , 199 2, G r eatLak es I ns trument s , I nc. , 885 5 N orth 55th Str eet , M i l wau kee , 10 W i s con s i n 53223. A d es cr i pt i on of

the Si mP l at e method, “ I D E X X Si mP l at e T M H P C T es tM ethod f or H et er otr op hs i n W at er ”, N ovemb er 11 2000, ca n be obt ai n ed f r om I D EX X La bor at or i es , I n c. , O n e I D EX X D r i v e, W es tbr ook , M ai n e 04092, t el eph on e ( 800) 321- 0207. A de scr i p t i o n of the H ach F i l t erT rak M etho d 10 1 3 3 , “D et erm i nat i o n of T urbidi tyby Laser N ep hel o m etry ”, January 12 2000, Rev i s i on 2. 0, ca n be obt ai n ed f r om H ac h Co. , P. O
. B ox 389, Lov el an d, Col or ado 80539- 0389. Phon e: 800- 227- 4224. 3. Ground W at er R ul e ( t o be adde d af t er r ul e pr omul gat i on)