AFDC 15 (6770) P3/REV. TZS 615: 2017 
DRAFT TANZANIA STANDARD  
 
Pasta products  – Specification  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
TANZANIA BUREAU OF STANDARDS  
                                                                                                                                                                                                                 
 
                                                                                                                                         
1 
  
0  Foreword  
Macaroni, spaghetti , vermicelli and noodles  belong to a class of food products generally known as pasta 
products .  
This Tanzania Standard, therefore, has been developed in order to encourage the grow th of the industry 
in the country, and at the same time ensure safe and good quality products .  
In the preparation of this Tanzania Standard assistance was derived from :-  
 
a) IS:  1485 – 2015 Macaroni, Spaghetti and Vermicelli and Egg noodles - Specification; published  
     by the Bureau of Indian Standards.  
 
b) SLS 420 :1989 Specification for pasta products ; published by Sri Lanka Standards Institution .  
 
c) EAS 173:2004 pasta products  - Spec ification ; published by  East African Community.  
In reporting the results of a test or analysis made in accordance with provision of this Tanzania Standard , 
if the final value observed or calculated is to be rounded off , it shall be done in accordance with TZS 4 
(See clause 2)  
1.0 Scope   
This Tanzania Standard prescribes the requirements , methods of sampling and test for Pasta products  
which are macaroni , spaghetti, vermicelli , noodles  and similar commodities intended for human 
consumption . 
2.0 Normative  references  
The following referenced standards are indispensable for the application of this Tanzania Standard. For 
dated references, only the edition cited applies. For undated references, the latest edition of the 
referenced standard (including any amendme nts) applies :  
AFDC 15(6523) P 3 Bakery products - methods of sampling  
AFDC 15  (6522) P3 bakery products  - method of analysis  
AFDC15 (6524) P 3 Glossary of terms relating to bakery industry  
TZS 4 - Rounding off numerical values.  
TZS 59 - Water - Distilled quality - Specification  
TZS 76 Methods for determination of arsenic content.  
TZS 109 - Code of hygiene for food processing units - General . 
TZS 111 Bakery units – Code of hygiene  
TZS 122 -2/ISO/TS 6579 -2 Microbi ology of food and animal feed — horizontal method for the detection,  
enumeration a nd serotyping of Salmonella — Part 2: Enumeration by a miniaturized most probable 
number technique . 
TZS 132/ EAS 35 – Fortified food grade salt - Specification.  
TZS 268 General atomic absorption spectrophotometric me thod for determination of lead in food stuffs.  
TZS 439 – Wheat flour – Specification  
TZS 538/ EAS 38 – Packaging and labeling of food  
TZS 730 -2/ ISO 16649 -2: Microbiology of food and animal feeding stuffs – Horizontal method for the 
enumeration of -b-glucuronidase -positive Escherichia coli – Part 2 – Colony -count technique at 44 0C 
using 5 -bromo -4-chloro -3-indolyl -b-D-glucuronide  
TZS 2426 -2:/ISO 21527 -2: Microbiology of food and animal feeding stuffs - Horizontal method for the                                                                                                        
 
                                                                                                                                         
2 
 enumeration of yeasts and moulds - Part 2: Colony count technique in products with water activity le ss 
than or equal to 0,95  
 
 
3 Terms and definitions  
 
For the purpose of this Tanzania standard the terms in AFDC15 (6524) P2  and the following shall apply;  
3.1 pasta  
a type of food typically made from an unleavened dough of wheat flour mixed with water or eggs, and 
form into sheets or other shapes, then cooked by boiling or baking.  
 
Note 1:  pasta is a very generic term used to describe a commonly consumed food group which 
encompasses noodles, spaghetti, macaroni and similar commodities  
 
Note 2: rice flour,  or legumes such as beans or lentils, are sometimes used in place of wheat flour to yield 
different tastes and textures, or as gluten -free alternative.  
 
3.2 macaroni  
a variety of dry pasta traditionally shaped into narrow tubes produced in various shapes and sizes, made 
with durum wheat.  
 
3.3 spaghetti  
A long, thin, cylindrical, solid pasta, made of milled wheat and water.  
 
3.4 vermicelli  
a traditional type of pasta round in section similar to spaghetti.  
 
3.5 noodles  
types of food made from unleavened dou gh which is rolled flat and cut, stretched or extruded into narrow 
long strips or strings  
 
3.6 egg noodles  
It is a narrow strip of pasta dough made with eggs.  
 
4. Requirements  
4.1 General requirements  
4.1.1 Form and dimensions  
The forms and dimensions of macaroni, spaghetti, vermicelli and egg noodles shall be as given below: - 
4.1.1.1 Macaroni  
They may be of two types,  namely long macaroni or cut macaroni.  
        a) Long macaroni – shall be in the form  of tubular rods, smooth or corrugated . These shall have the  
outer diameter ranging between 3mm and 5 mm and of wall thickness of about 1 mm.  
         b) Cut -macaroni - shall be obtained by extrusion and may be in the form of tubular elbows, shells, 
alphabets, numerals, stars, wheels, rings, rice, melon seeds etc.  
4.1.1.2 Spaghetti - shall be in the form of solid rods of diameter 1 mm.  
4.1.1.3 Vermicelli - shall be in t he form of solid rods or of slightly curved or bend product of diameter 
between 0.5 mm and 1 .25 mm.  
4.1.1.4 Egg noodles – The material shall be in the form of ribbons, and shall have not less than 5.5 
percent by mass of egg solids or egg yolk solids when tested by the method given in Annex  E. 
                                                                                                        
 
                                                                                                                                         
3 
 4.1.2 Other physical characteristics  
Macaroni, spaghetti, vermicelli and noodles shall be  
- of good characteristic colours, flavour and odour , 
- Free from rancidity, bitterness, mustiness or any other undesirable taste o r odour.  
- Free from impurities, any foreign matter, cracks, flaws, mould, insect infestation or other spoilage.  
- Of the materials that retain  its shape and show no signs of disintegration and swell appreciably 
when plunged into vigorously – boiling water an d boiled for 10 minutes.  
NOTE - In case of instant or quick cooking noodles, the material shall be boiled for the time declared in the cooking 
instructions. The material shall be smooth to the touch (not stick together) and shall not contain any added colo uring 
matter.  
4.1.3 Enrichment  
The product may be fortified with one or more of the following:  
     a) Minerals – calcium, phosphorus and iron.  
     b)  Vitamins – A, B complex and D.  
4.2 Ingredients  
4.2.1 Essential ingredients  
The following materials shall be used in the manufacture of macaroni, spaghetti, vermicelli and noodles 
and shall be of food grade quality and conform to the relevant Tanzania standard specifications, where 
available  and other materials shall be free from foreign matter, insect infestation and objectionable 
flavours and odours.  
The materials are wheat flour, water and Egg (only in the case of egg noodles).  
4.2.2 Optional Ingredients  
In addition to the essential ingred ients specified under 4.2.1, any of the following ingredients may be used 
in the manufacturer of macaroni, spaghetti, vermicelli and egg noodles:  
a) wheat durum semolina  
 b) Milk 
c) Soya flour  
d) Vegetable or vegetable products  
e) Spices  
f) Gluten  
g) Edible o ilseeds flour  
h) Edible common salt                                                                                                        
 
                                                                                                                                         
4 
 i) Fruit or fruit products (preserved, dehydrated or pulp).  
j) rice  
4.3 Specific requirements  
4.3.1 The product shall also comply with the requirements given in Table 1.   
Table 1  - Requirements for pasta products  
                         
S/N Characteristics  Requirements  Method of Test  
(Refer to annex)  
1 Moisture, per cent by mass, max  12  AFDC 15 (6522) P3  
2 Total ash (on dry basis),per cent by mass 
max.  1.0 AFDC 15 (6522) P3  
3 Acid insoluble ash (on dry basis) per cent 
by mass max.  0.05  AFDC 15 (6522) P3  
4 Total protein (Nx 5.7) (on dry basis), per 
cent by mass, min  10  A 
5 Cooking test: Total solids in  
gruel, per cent by mass, max  
 10  B 
6 Free acidity (ml of 1N, NaOH solution per 
100g of product), max.               4 C 
 
4.4 Food additives    
Only the food additives permitted by CODEX STAN 192 in the manufacture of pasta products  may be 
used without exceeding the limits.  
 
5.0 Hygiene  
5.1 The pasta products  shall be prepared and packaged under hygienic condition in acc ordance with TZS 
109 and TZS 111 (see clause 2)  
 
5.2 pasta products shall be free from pathogenic organisms and shall comply with microbiological limits in 
Table 2.  
 
 
    Table 2 - Microbiological limits for pasta products  
 
Types of micro -organism       Limits   Test methods  
 
Yeast and mould count (cfu per g,  max)  
            102          
TZS 2426 -2  
 
 E.coli   (cfu per g )  
      absent  TZS 730 -2  
 
Salmonella  in 25g   
     absent  TZS 122 -2  
 
6.0 Contaminants  
The pasta products  shall comply with the heavy metal contaminants limits stipulated in Table 3  
 
 
 
                                                                                                        
 
                                                                                                                                         
5 
 Table 3  Requirements for Heavy metal in pasta products  
 
S/N Contaminant  Limits (max ) mg/kg   Test me thod  
 
1        Lead (Pb)  0.5 TZS 268  
 
2         Arsenic(A s) 1.0 TZS 76  
 
7 .0 Sampling and method of test  
 
7.1 Sampling      
Representative samples of the pasta product shall be drawn as prescribed in annex D. 
 
7.2 Te st methods          
Test shall be c arried out as specified in Tables 1, 2 and 3.  
 
7.3 Criteria for conformity  
Criteria for conformity - The criteria ascertaining conformity to the requirements specified in this Tanzania 
Standard shall be as given in annex D.G 
 
8.0 Pack ing, marking  and labelling  
 
8.1 Pack ing 
 
The pasta products shall be packaged in food grade materials that will ensure the safety and integrity of 
the product throughout the shelf life.   
 
8.2 Marking and l abelling  
 
8.2.1  Labelling of  pasta products  shall be done in accordance with the requirements of the TZS 538(EAS 
38) and the following particular shall be clearly and indelibl y marked on each pack of  pasta products on 
the outer side of the wrapper; printing ink if any, shall be non -toxic and non -transferable:  
a) Name of the product;  macaroni, spaghetti, vermicelli, noodles and egg noodles  
b) If enriched with vitamins; (i)  the word ‘vitaminized’ placed in brackets, below (a); and (ii) the 
details of enrichment and quantities added;  
c) If spices have been added, the word ‘spiced’ to be given on the label  
d) If egg has been added, it shall be declared on the label;  
e) All ingredients shall be declared on the label  in descending order of proportions ; 
f) Name and address of the manufacture r; 
g) Batch or code number;  
h) Net mass;  
i) Date of manufacture;  
j) Expiry date;  
k) Cooking instructions; and  
l) Instruction for the disposal of packaging materials  
 
 
8.2.2 Each pack may also be marked with the TBS Certification mark.  
  
NOTE – The use of TBS Certification mark is gover ned by the provisions of the  standards Act, 2009. Details of 
conditions under which a licence for use of the TBS Certification mark may be granted to manufacturers, may be 
obtained from TBS.  
 
 
 
                                                                                                        
 
                                                                                                                                         
6 
 ANNEX A 
DETERMINA TION OF TOTA L PROTEIN  
A.1 APPAR ATUS  
A -1.1 Kjelda hl appara tus  
A-1.2 Kjeldahl fla sk - of capac ity 500 ml.  
A -2 REAG ENTS  
A -2.1 anhydrous sodium sulph ates  
A -2 .2 Copper sulphate  
A -2.3 Concentrated sulphuric ac id - sp. gr. 1.84  
A -2.4 Sodium hydroxide sol ution  - Dissolve about 225 g of sodium hydroxide in 50 0 ml of water .  
A -2 .5 Standard sulphuric acid  - 0.1 M.  
A -2 .6 Methyl red indicator solution  - dissolve one gram of methyl red in 200 ml of rectif ied 
spirit (95 per cent by volume)  
A-2.7 Standard so dium hydroxide solution - 0.1 M.  
A.3 PROCEDURE  
A.3.1 Transfer carefully about one gram of the prepared sample , accurately weighed , into the Kjeldahl 
flask, taking precaution to see that particles of the material do not stick on the neck of the flask. Add  
about 10g of anhydrous sodium sulphate, about 0 .5 g of copper sulphate and 25 ml concentrated 
sulphuric ac id.  
Place the flask in an inclined position . Heat below the boi ling point of the acid until frothing ceases . 
Increase heat until ac id boils vigorous ly and digest for 30 minutes after the mixture becomes clear and 
pale green or colour less. Cool the contents of the f lask. Transfer quantitatively to the round bottom flask , 
with water , the total quant ity of water used be ing about 200 ml . Add with shaking a few pieces of p umice 
stones to prevent  bump ing. Add about 30 ml of the sodium hydrox ide solution (enough to make the 
solution alkal ine) caref ully through th e side of the flask so that it does not mix at onc e with the ac id 
solution but f orms a layer below  the a cid layer .  
Assemble the apparatus taking ca re that the tube extends be low the surface of the standard sulphuric 
acid con tained in the beaker . Mix the co ntents of the flask b y shake until all am moni a has passe d over 
into the standard su lphuric acid. Shut off the burner and immediately detach the flask from the condenser . 
Rinse the condenser thoroughly with water into the beaker . Wash the dip tube careful ly so that a ll traces 
of the c ondensate a re transferred to the beaker . When all t he washing has dra ined into the beaker  add 
two or three drops of methyl red indicator solution and titrate with the standard sodium h ydroxide solution.  
 
A.3.2 Carry out a blank determinat ion using all  reagents in the same quantities but without th e material to 
be tested.  
 
 
                                                                                                        
 
                                                                                                                                         
7 
  
A.4 CALCULATION  
      798 (A - B) N  
Total protein (on dry basis ), per cent by mass     __  
          M1 (100 - M)  
Where  
                     B = volume in ml of the standard sodium hydroxide solution u sed to n eutralize the acid in the 
blank determin ation,  
A = volume in ml of the sta ndard sodium hydroxide solution used to   n eutralize the 
excess of acid in the test with the materia l,  
N = normality of the standard sodium hydroxide solution,  
M1 = mass in g of the prepar ed mate rial taken for the test, 
and  
M = moisture , percent by mass of the material.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
                                                                                                        
 
                                                                                                                                         
8 
 ANNE X B 
DETERMINA TION OF T OTAL SOLIDS IN GRUE L 
B -1 APPARATUS  
Lipless beaker - tall-form, of capacity 500 ml .  
B-2 PROCEDURE  
Take 250 ml of water in the lipless bea ker and heat over hot -plate or any suitable bu rner til l the water 
boils. Introduce 25 g of the material (previously broken into about 10 mm lengths in case of long goods) 
and stir thoroughly with a glass rod . Cook for 10 minutes with occasional stirr ing. At the end of 10 
minutes allow the material to drain for five minutes. Measure the volume of gruel collected . Pipette out 20 
ml of the gruel ; after  
stirring well to give an even distribution of the solids con tent, into a tared petri dish and evaporate to 
dryness on a water -bath. Transfer the petri dish to a hot air -oven mainta ined at 105 ± 2°C and dry to 
constant mass .  
 
B- 3 CA LCULATION  
 
 
Total solids in gruel, percent b y mass  
5  
Where  
m2 = mass in g of petri dish with total solids,  
m1 = mass in g of empty pe tri dish, and  
V = volume of grue l in ml.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
                                                                                                        
 
                                                                                                                                         
9 
 ANNEX C 
 
DETERMINATION OF ACIDITY  
C-1 REAGENTS  
C-1.1 Sodium Hydroxide – 0.1N solution.  
C-1.2 Phenolphthalein Indicator Solution  
C-2 PROCEDURE  
C-2.1 Grind 10grams of the product an d add 100 millilitres of distilled water (start the test from within an 
hour from the grinding). Leave for one hour, stirring 3 times for two minutes each time, at approximately 
equal intervals. Titrate with 0.1N solution of NaOH using phenolphthalein as a n indicator.  
 
C-3 CALCULATION  
C-3.1 Free acidity (ml of 1N NaOH) =   10,000 x V x N  
                                                                      M 
Where:    V = volume, in ml, of standard sodium hydroxide solution used in the titration;  
                N = normality of sodium hydroxide solution; and  
                M = mass, in g, of the sample taken.  
                                                               
 
 
 
 
 
 
 
 
                                                                                                        
 
                                                                                                                                         
10 
                                                                ANNE X D 
                                                              SAMPLING  
                             D-1 GENERAL REQU IREMENTS FOR S AMPLING  
In drawing, prepa ring, storing and hand ling samples , the followin g precaution s shall be obser ved.  
D.1.1 Samples sha ll be taken in a p rotected place not exposed to d amp air, dust or soot.  
D.1.2 The sampling instrument shall be c lean and dry when used .  
D.1.3 Precautions shal l be taken to protect the samples, t he material being samp led, the sampling 
instrument and the contai ners for samp les fro m adventitio us conta mination.  
D.1.4 The samp les sha ll be placed in clean, odou rless and dry c ontainers. The sample containers shall 
be of such a size that they a re almost comp letely fi lled by the samp le.  
D.1.5 Each container s hall be sealed air -tight after fill ing and marked with full details of s ampling, batch or 
code number , name o f the manufac turer and othe r important pa rticulars of the consignm ent.  
D.1.6 Samples sha ll be s tored in such a manner that the temperatur e of the material  does not vary 
undul y from the normal tempe rature.  
D .1.7 Sampling shall be done by a person agreed to between t he purchaser and the ven dor.  
D.2 SCALE OF SAMPLING  
D.2.1 Lot - All the packages in a single consignment belonging to the same batch of manufa cture shall 
constitute a lot .  
Samples sha ll be tested from each lot for ascertaining confo rmity of the mate rial to the requirements of 
the spec ification .  
D .2.2 The number of conta iners to be tested from a lot shall depend on the size of the lot and shal l be in 
accordance with table 2.  
TABLE 2 - Number of con tainers to be selected for sa mpling  
Lot size ( number of  Sample size (number of  
packages in the lot), N  packages to be selected) n  
Up to 50  3  
51 to 150  5  
151 to 500  8  
501 to 3000  13  
Above 3000  20  
D.2.3 The pac kages t o be selec ted fo r sampling shall be chosen a t random from th e lot. In orde r to 
ensure r andomness of selec tion, r ando m numb er tables shall be used .  
D.3 TEST FOR CONFORMITY  
D.3.1 Each of the selected packages shall be opened and the co ntents examined separately for                                                                                                         
 
                                                                                                                                         
11 
 appearance and dimension (see 4 .1.1).   
D.3.2 The contents of the packages shall then be mixed to obtain a compos ite sample. The sample shall 
be tested for chem ical requirements (see 4.3 .1).  
D.4 CRITERIA FOR  CONFORMI TY  
D.4.1 The lot shall be considered satisfa ctory in respect of t he requirements of G .3.1 if eac h individual 
sample satisfies all the requiremen ts.  
D.4.2 The lot shall be considered satisfacto ry in respec t of the requiremen ts of D.3.2 if the tes t results on 
the composite sample satisfy the corresponding requirements.  '  
D.4.3 The lot shall be declared to be in conformity with all the req uirement s of this specification, if it has 
been found s atisfactory in accordance with D.4.1 and D.4.2.  
 
 
                                                            
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
                                                                                                        
 
                                                                                                                                         
12 
                                                                 ANNEX E 
 
                                                            (Clause 3.1.1.4 ) 
 
                     DETERMINATION OF EGG SOLIDS IN EGG NOODLES  
 
E-O PRINCIPLE    
Egg solid content of egg noodles is estimate d closely from cholesterol content of unsapon ifiable matter. 
When analyzing egg -free products or material containing less than 0·23 percent uns aponifiable matter, 
add 10 mg cholesterol before analyzing and correct results accordingly. The added Cholesterol should 
have melting point not less than 147°, and its purity should be checked by submitting 20 mg to the 
determination as given below.  
 
E.1 APPARATUS  
 
E.1.1 Cold Baths  
Prepare one bath with crushed ice and one with salt -ice mixture.  
 
E.1.2. Mohr Pipettes .  
One graduated to 0·01 ml and one to 0.1 ml.  
 
E.1.3 Filteration Bell Jar  
Size to accommodate 300 -ml Erlenmeyer flask connected to vacuu m source by 2 -way stopcock  
 
E-1.4 Filtering Device  
For filtering at O0 C, prepare filter tube of Knorr type. Tube should be about 20 mm  
internal diameter, body about 11cm long, and stem about 10cm long, provided with usual nickel or Monel 
metal disk. Ins ert this tube through bottom of container of such size that crushed ice can be packed 
around body of tube to height of 7 -8 cm. Fit tube into 500 -ml suction flask. In tube prepare mat of 
asbestos 6 -8 mm thick, packed fairly tight, and cover with layer of sa nd about 12 mm deep. Digest 
asbestos with acid and alkali. Purify sand by passing through 60 -mesh sieve, then treat with warm 
hydrochloric acid until extract is practically colour less. Wash. dry, and ignite sand.  
 
E-1.5 Separating Funnels  
Separating fun nels, one 250 -ml and one 500 -ml. Wash free of grease. Funnels must be ether -  
tight, with stopcock lubricated only with water.  
 
E-1.6 Sintered Glass Filter - 30-50 ml capacity.  
 
E-2 REAGENTS  
 
E-2.1 Bromine Solution  
Weigh to 0·1g a narrow -mouthed, glass - stoppered 25 -ml flask containing 5ml carbon  
tetrachloride. Add 4 -5 g bromine, weigh again and dilute with carbon tetra chloride to calcula  
ted final concentration of 0.22± 0.02 g bromine per ml. (This reagent should not be more than  
2 days old.) Store under refrigeration.   
 
E-2.2 Acetic Acid Solution  
Pipette or burette 200 ml glacial acetic acid into 250 -ml glass -stoppered volumetric flask;  
dilute to mark with water  and mix cautiously,  
 
E-2.3 Sodium Hypochlorite Solution  
Dissolve 88g reagent grade s odium hydroxide in 200 ml water in a wide -mouthed 3 -litre flask.  
Add about 1500 g crushed ice and pass in chlorine until 71 g is absorbed. Dilute to 2 litre and then store 
in dark bottles under refrigeration. Solution should be alkaline to phenolphthalein . Before using, check                                                                                                        
 
                                                                                                                                         
13 
 concentration of available chlorine as follows: Pipette 5ml solution into 100 ml water containing 2 g 
potassium iodide. Add 5 ml hydrochloric acid (1: 1) and titrate with 0·1 N Na 2S203 Solution should be 
equivalent in available chlorin e to 0·95 - 1·05 N NaOCI.  
 
E-2.4 Sodium Formate Solution  
Dissolve 50 g reagent grade sodium formate and dilute 100 ml.  
 
 E-2.5 Hydrochloric Acid  
Approximately 6 N. Mix 500 ml concentrated hydrochloric acid with water and dilute to 1 litre.  
 
E-2.6 Pota ssium Iodide Solution  
20 percent. Dissolve 20 g potassium iodide and dilute to 1 00 ml. This solution must be colorless when 
acidified with hydrochloric acid.  
 
E-2.7 Soluble Starch Solution  - 1 percent.  
 
E-2.8 Sodium Thiosulphate Solution  - 0·02 N.  
 
E-2.9 Sodium Sulphate Anhydrous   
Powdered to pass through 60 -mesh sieve.  
 
E-2.10 Potassium Hydroxide Solution   
a) 100 percent - Dissolve 10 g potassium hydroxide in 10ml water  
b) 1 percent  - Dissolve 10 g potassium hydroxide in water and make up to 1 litre.  
 
 E-2.11 Ammonium Molybdate Solution  
Dissolve 5 g ammonium molybdate and make to 100 ml.  
 
E-2.12 Ether  
a) Anhydrous, reagent grade   
 
b) Dry and peroxide free - (check for presence of peroxides, just prior to use).  
Rinse glass -stoppered cylinder with e ther, place 10ml ether in rinse d cylinder, and add 1mI freshly 
prepared 10 percent solution of cadmium potassium iodide.  Let stand protected from light for one hour, 
shaking occasionally, no color should develop in either liquid layer. To dry ether, shake  with excess of 
anhydrous calcium chloride and filter.   
 
E-3 PROCEDURE  
 
E-3.1 Determination of Unsaponifiable Matter   
Weigh 10 -g sample into 500 -ml Erlenmeyer flask and add, with shaking, 30 ml hydrochloric  
acid (1: 1). Heat on a steam bath for 30 min utes with occasional shaking. Cool under cold -water tap and 
add carefully with shaking, 30 g, potassium hydroxide pellets.  Add pellets at such rate that liquid may 
boil, but not so fast as to cause splattering. While flask is still hot, place on steam bat h, cover with small 
watch -glass and heat for 3 hours with occasional swirling. Cool, add 30 ml of 95 percent alcohol and 500 
ml water and mix well. (Samples  may be allowed to stand overnight at this point).  
     
Add 100ml ether, swirl mixture vigorously f or one minute, and  transfer to 500 -ml separating funnel, 
washing flask with 50 -ml and 25 -ml portions of ether . Wash flask with 50 -ml dilute potassium  hydroxide 
solution. Pour washings slowly into funnel while gently swirling liquid and continue swirling fo r 10 to 15 
seconds.   
 
Let liquid separate (about 10 minutes) and a slowly draw off soap solution into 250 -ml separating funnel, 
but do not draw off any emulsion or insoluble matter at interface. If emulsion forms and does not break to 
give  sharp interfac e within 10 minutes, pour 5 ml  alcohol into funnel and let stand until emulsion breaks.                                                                                                        
 
                                                                                                                                         
14 
 Rinse down sides of 500 -ml funnel with 10 ml dilute potassium solution  and draw this off into smaller 
funnel. Add 50 ml ether to smaller funnel and shake vigorously:  
 
After liquids have separated, discard lower layer. Add eth er layer to solution in larger funnel  and rinse 
smaller funnel with 10 ml ether. Wash ether solution as before with100 ml  dilute  potassium hydroxide 
solution, still retaining any insoluble matter o r emulsion in funnel.  In the same manner wash ether 
solution with two additional100 -ml portions of dilute potassium  hydroxide solution, testing portions of last 
washings for soap by acidifying with dilute hydrochloric  acid (1: 4). Acidified washings shoul d be clear or 
only faintly turbid. If necessary , repeat washings with dilute potassium hydroxide solution until acidified 
washing are clear.  
 
Wash ether solution by swirling with successive portions: 50 ml water, 50 ml water containing 0·5 ml 0·1 
N hydroc hloric acid, and two more 50 -ml portions water. Draw off as much water as possible without loss 
of ether solution.  
 
Filter ether solution into dry 500 -ml suction flask through 50 -g layer of sodium sulphate on sintered glass 
filter, using no suction for fir st several millilitres and then gentle suction for remainder.  
 
Rinse separate funnel and filter with successive 10 -, 5-, 5-, and 5 -ml portions dry ether.  Rinse stem of 
funnel, add glass beads to flask, and evaporate on steam bath to about 20 -ml volume  (current of air 
directed across neck of flask will increase rate of evaporation). Heat 50 -ml glass -stoppered Erlenmeyer 
flasks at 100 - 105°C for 1 hour, cool in air for 30 minutes, and weigh. Transfer ether solution in suction 
flask quantitatively to weighe d Erlenmeyer flask. Evaporate ether and dry flask at 100°c. Weigh flask 
again and determine unsaponifiable matter by difference.  
 
Carry out blank determination using all reagents in above method without sample, and deduct this value 
from weight of unsapon ifiable matter. Calculate unsaponifiable matter.  
 
E-3.2 Determination of Egg Solids  
Wash down sides of flask containing unsaponifiable matter with 2·0 ml anhydrous ether delivered from 
Mohr pipette. Pack flask in ice bath up to neck and let stand for at least 10 minutes. From another 'Mohr 
pipette add 0.20 ml cold bromine solution, swirl, stopper replace  in ice bath for 10 minutes.  
 
During bromination, cool acetic acid solution in ice -salt bath until temperature is -5°C and in same bath 
cool a 25 -ml cylin der. Measure, 15 ml acetic acid solution into cold cylinder and add to reaction flask. 
Swirl reaction mixture for 3 minutes while holding in ice bath, and let stand in ice bath for 10 minutes. 
With slight suction pour mixture down stirring rod into cold fi lter tube, leaving rod in tube. Wash down 
sides of flask with 5 ml cold acetic acid  solution and replace in ice bath. When liquid in funnel recedes 
below surface of sand, add acetic acid from react ion flask. Again repeat washing with 5 ml cold acetic 
acid solution and suck free of excess liquid. Wash flask and tube with ice water, filling filter tube about five 
times. Drain flask and apply suction to filter to remove all excess water, discarding filtrate and  
washings.  
 
Remove crushed ice from around filt er tube and transfer tube to top of filtration bell jar. Place 300-ml 
Erlenmeyer flask under filter tube so that stem projects well into the neck of the flask. Wash filter with 10 
m l alcohol, 10 -, 5-, and 5 -ml portions of ether, and finally with 10 ml alc ohol, stirring sand gently with 
each portion of solvent, and let mixture stand for about 1 minute before applying suction. Wash stem of 
filter with few ml ether, remove 300 -ml flask, and to filtrate add 1 ml potassium hydroxide solution. Mix 
and wash down sides of flask with 5 ml ether. Evaporate on steam bath, using stream of clean air to 
remove all vapors.  
 
Add 40 ml hot water to residue, mix and neutralize with 6 N hydrochloric acid, using 1 drop methyl red 
indicator. Add 10 g NaCI, 3 g Na H 2P04H20, and  20ml N 2OCL solution. Bring to vigorous boil, remove 
from heat, and at once, with care, add 5ml'sodium formate solution. Cool and dilute to about 150ml. Add 5 
ml potassium iodide solution, 1 -2 drops ammonium molybdate solution and 25 ml 6 N hydrochloric ac id.                                                                                                        
 
                                                                                                                                         
15 
 Immediately titrate rapidly with thiosulfate, using starch indicator. Correct titre by blank value obtained on 
reagents, starting with addition of potassium hydroxide to alcohol ether solution.  
 
E-4 CALCULATIONS  
H-4.1 Cholesterol = 0. 55 + 0·688  
 (mg)                (ml 0.02 N Na 2 S203 )  
 
E-4.2 Percent sterol calculated as cholesterol in sample  
 (Moisture -free basis) = mg cholesterol  = C  
                                           (100-percent moisture)  
 
E·4.3 Percent commercial egg -yolk solids  
 (Moisture -free basis) = (C-0.024)100  = 35 (C - 0.024)  
                                                         (2.88 -0.024)  
E-4.4 Percent commercial whole -egg solids  
 (Moisture -free basis) = (C – 0.024) 100 = 48 (C – 0.024)  
                                           (2.11 – 0.024)