440.0-1

Method 440.0

Determination of Carbon and Nitrogen in Sediments and Particulates

 of Estuarine/Coastal Waters Using Elemental Analysis

Carl F. Zimmermann

Carolyn W. Keefe

University of Maryland System

Center for Environmental Estuarine Studies

Chesapeake Biological Laboratory

Solomns, MD 20688-0038

and

Jerry Bashe

Technology Applications, Inc.

26 W. Martin Luther King Drive

Cincinnati, OH 45219

Revision 1.4

September 1997

Work Assignment Manager

Elizabeth J. Arar

National Exposure Research Laboratory

Office of Research and Development

U.S. Environmental Protection Agency

Cincinnati, Ohio 45268
Revision 1.4 September 1997 440.0-2

Chemical Abstracts Service

Analyte Registry Numbers (CASRN)

Carbon 7440-44-0

Nitrogen 1333-74-0

Method 440.0

Determination of Carbon and Nitrogen in Sediments and Particulates

of Estuarine/Coastal Waters Using Elemental Analysis

1.0 Scope and Application

1.1 Elemental analysis is used to determine particulate carbon (PC) and particulate nitrogen (PN) in estuarine and coastal waters and sediment. The method

measures the total carbon and nitrogen irrespective of

source (inorganic or organic).

1.2 The need to qualitatively or quantitatively deter- be passed through a number 10 sieve or a 2-mm mesh

mine the particulate organic fraction from the total sieve.

particulate carbon and nitrogen depends on the dataquality objectives of the study. Section 11.4 outlines 3.2 Material Safety Data Sheet (MSDS) -- Written

procedures to ascertain the organic/inorganic particulate information provided by vendors concerning a chemical’s

ratio. The method performance presented in the method toxicity, health hazards, physical properties, fire, and

was obtained on particulate samples with greater than reactivity data including storage, spill, and handling pre80% organic content. Performance on samples with a cautions.

greater proportion of particulate inorganic versus organic

carbon and nitrogen has not been investigated. 3.3 Instrument Detection Limit (IDL) -- The mini1.3 Method detection limits (MDLs) of 10.5 μg/L and which gives an analyte signal equal to three times the

1

62.3 μg/L for PN and PC, respectively, were obtained for standard deviation of the background signal at the sea 200-mL sample volume. Sediment MDLs of PN and lected wavelength, mass, retention time, absorbance line,

PC are 84 mg/kg and 1300 mg/kg, respectively, for a etc.

sediment sample weight of 10.00 mg. The method has

been determined to be linear to 4800 μg of C and 700 μg 3.4 Method Detection Limit (MDL) -- The minimum

of N in a sample. Multilaboratory study validation data are concentration of an analyte that can be identified, meain Section 13. sured, and reported with 99% confidence that the analyte

1.4 This method should be used by analysts experienced in the theory and application of elemental analysis. 3.5 Linear Dynamic Range (LDR) -- The absolute

A minimum of 6 months experience with an elemental quantity over which the instrument response to an analyte

analyzer is recommended. is linear.

1.5 Users of the method data should set the data- 3.6 Calibration Standard (CAL) -- An accurately

quality objectives prior to analysis. Users of the method weighed amount of a certified chemical used to calibrate

must document and have on file the required initial the instrument response with respect to analyte mass.

demonstration of performance data described in Section

9.2 prior to using the method for analysis. 3.7 Conditioner -- A standard chemical which is not

2.0 Summary of Method

2.1 An accurately measured amount of particulate

matter from an estuarine water sample or an accurately

weighed dried sediment sample is combusted at 975EC

using an elemental analyzer. The combustion products

are passed over a copper reduction tube to convert the 

oxides of N into molecular N. Carbon dioxide, water vapor

and N are homogeneously mixed at a known volume,

temperature and pressure. The mixture is released to a

series of thermal conductivity detectors/traps, measuring

in turn by difference, hydrogen (as water vapor), C (as

carbon dioxide) and N (as N2). Inorganic and organic C

may be determined by two methods which are also

presented.

3.0 Definitions

3.1 Sediment Sample -- A fluvial, sand, or humic

sample matrix exposed to a marine, brackish or fresh

water environment. It is limited to that portion which may

mum quantity of analyte or the concentration equivalent

concentration is greater than zero.

necessarily accurately weighed that is used to coat the

surfaces of the instrument with the analytes (water vapor,

carbon dioxide, and nitrogen).

3.8 External Standards (ES) -- A pure analyte(s)

that is measured in an experiment separate from the

experiment used to measure the analyte(s) in the sample.

The signal observed for a known quantity of the pure 

external standard(s) is used to calibrate the instrument
440.0-3 Revision 1.4 September 1997

response for the corresponding analyte(s). The instru- which has been certified for specific analytes by a variety

ment response is used to calculate the concentrations of of analytical techniques and/or by numerous laboratories

the analyte(s) in the sample. using similar analytical techniques. These may consist of

3.9 Response Factor (RF) -- The ratio of the re- These materials are used as an indication of the accuracy

sponse of the instrument to a known amount of analyte. of a specific analytical technique.

3.10 Laboratory Reagent Blank (LRB) -- A blank 3.17 Quality Control Sample (QCS) -- A solution of

matrix (i.e., a precombusted filter or sediment capsule) method analytes of known concentrations which is used

that is treated exactly as a sample including exposure to to fortify an aliquot of LRB or sample matrix. The QCS is

all glassware, equipment, solvents, and reagents that are obtained from a source external to the laboratory and

used with other samples. The LRB is used to determine different from the source of calibration standards. It is

if method analytes or other interferences are present in used to check laboratory performance with externally

the laboratory environment, the reagents, or the appa- prepared test materials.

ratus.

3.11 Field Reagent Blank (FRB) -- An aliquot of

reagent water or other blank matrix that is placed in a

sample container in the laboratory and treated as a

sample in all respects, including shipment to the sampling

site, exposure to sampling site conditions, storage,

preservation, and all analytical procedures. The purpose

of the FRB is to determine if method analytes or other

interferences are present in the field environment.

3.12 Laboratory Duplicates (LD1 and LD2) -- Two

aliquots of the same sample taken in the laboratory and

analyzed separately with identical procedures. Analyses

of LD1 and LD2 indicate precision associated with laboratory procedures, but not with sample collection, preservation, or storage procedures.

3.13 Field Duplicates (FD1 and FD2) -- Two separate samples collected at the same time and place under

identical circumstances and treated exactly the same

throughout field and laboratory procedures. Analyses of

FD1 and FD2 give a measure of the precision associated

with sample collection, preservation and storage, as well

as with laboratory procedures.

3.14 Laboratory Fortified Blank (LFB) -- An aliquot

of reagent water or other blank matrices to which known

quantities of the method analytes are added in the

laboratory. The LFB is analyzed exactly like a sample,

and its purpose is to determine whether the method is in

control, and whether the laboratory is capable of making

accurate and precise measurements.

3.15 Laboratory Fortified Sample Matrix (LFM) -- An

aliquot of an environmental sample to which known

quantities of the method analytes are added in the

laboratory. The LFM is analyzed exactly like a sample,

and its purpose is to determine whether the sample matrix

contributes bias to the analytical results. The background

concentrations of the analytes in the sample matrix must

be determined in a separate aliquot and the measured

values in the LFM corrected for background concentrations.

3.16 Standard Reference Material (SRM) -- Material

pure chemicals, buffers or compositional standards.

4.0 Interferences

4.1 There are no known interferences for estuarine/coastal water or sediment samples. The presence of

C and N compounds on laboratory surfaces, on fingers,

in detergents and in dust necessitates the utilization of

careful techniques (i.e., the use of forceps and gloves) to

avoid contamination in every portion of this procedure.

5.0 Safety

5.1 The toxicity or carcinogenicity of each reagent

used in this method has not been fully established. Each

chemical should be regarded as a potential health hazard

and exposure to these compounds should be as low as

reasonably achievable. Each laboratory is responsible for

maintaining a current awareness file of OSHA regulations

regarding the safe handling of the chemicals specified in

this method. A reference file of material safety data

2-5

sheets (MSDS) should also be made available to all

personnel involved in the chemical analysis.

5.2 The acidification of samples containing reactive

materials may result in the release of toxic gases, such as

cyanides or sulfides. Acidification of samples should be

done in a fume hood.

5.3 All personnel handling environmental samples

known to contain or to have been in contact with human

waste should be immunized against known disease

causative agents.

5.4 Although most instruments are adequately

shielded, it should be remembered that the oven temperatures are extremely high and that care should be

taken when working near the instrument to prevent

possible burns.

5.5 It is the responsibility of the user of this method to

comply with relevant disposal and waste regulations. For

guidance see Sections 14.0 and 15.0.
Revision 1.4 September 1997 440.0-4

6.0 Apparatus and Equipment

6.1 Elemental Analyzer

6.1.1 An elemental analyzer capable of maintaining a

combustion temperature of 975EC and analyzing particulate samples and sediment samples for elemental C and

N. The Leeman Labs Model 240 XA Elemental Analyzer

was used to produce the data presented in this method.

6.2 A gravity convection drying oven. Capable of

maintaining 103-105EC for extended periods of time.

6.3 Muffle furnace. Capable of maintaining 875EC ±

15EC.

6.4 Ultra-micro balance. Capable of accurately

weighing to 0.1 μg. Desiccant should be kept in the

weighing chamber to prevent hygroscopic effects.

6.5 Vacuum pump or source capable of maintaining

up to 10 in. Hg of vacuum.

6.6 Mortar and pestle.

6.7 Desiccator, glass.

6.8 Freezer, capable of maintaining -20EC ± 5EC.

6.9 47-mm or 25-mm vacuum filter apparatus made

up of a glass filter tower, fritted glass disk base and 2-L

vacuum flask.

6.10 13-mm Swinlok filter holder.

6.11 Teflon-tipped, flat blade forceps.

6.12 Labware -- All reusable labware (glass, quartz,

polyethylene, PTFE, FEP, etc.) should be sufficiently

clean for the task objectives. Several procedures found

to provide clean labware include washing with a detergent

solution, rinsing with tap water, soaking for 4 hr or more

in 20% (v/v) HCI, rinsing with reagent water and storing

clean. All traces of organic material must be removed to

prevent C-N contamination.

6.12.1 Glassware -- Volumetric flasks, graduated

cylinders, vials and beakers.

6.12.2 Vacuum filter flasks -- 250 mL and 2 L, glass.

6.12.3 Funnel, 6.4 mm i.d., polyethylene.

6.12.4 Syringes, 60-mL, glass.

7.0 Reagents and Standards

7.1 Reagents may contain elemental impurities which

affect analytical data. High-purity reagents that conform

to the American Chemical Society specifications should 6

be used whenever possible. If the purity of a reagent is in

question, analyze for contamination. The acid used for

this method must be of reagent grade purity or equivalent.

A suitable acid is available from a number of manufacturers.

7.2 Hydrochloric acid, concentrated (sp. gr. 1.19)-

HCI.

7.3 Acetanilide, 99.9% + purity, C8H19NO (CASRN

103-84-4).

7.4 Blanks -- Three blanks are used for the analysis.

Two blanks are instrument related. The instrument zero

response (ZN) is the background response of the instrument without sample holding devices such as capsules

and sleeves. The instrument blank response (BN) is the

response of the instrument when the sample capsule,

sleeve and ladle are inserted for analysis without standard

or sample. The BN is also the laboratory reagent blank

(LRB) for sediment samples. The LRB for water samples

includes the capsule, sleeve, ladle and a precombusted

filter without standard or sample. These blanks are

subtracted from the uncorrected instrument response

used to calculate concentration in Sections 12.3 and 12.4.

7.4.1 Laboratory fortified blank (LFB) -- The third blank

is the laboratory fortified blank. For sediment analysis,

add a weighed amount of acetanilide in an aluminum

capsule and analyze for PC and PN (Section 9.3.2). For

aqueous samples, place a weighed amount of acetanilide

on a glass fiber filter the same size as used for the

sample filtration. Analyze the fortified filter for PC and PN

(Section 9.3.2)

7.5 Quality Control Sample (QCS) -- For this method, the QCS can be any assayed and certified sediment

or particulate sample which is obtained from an external

source. The Canadian Reference Material, BCSS-1, is

just such a material and was used in this capacity for the

data presented in this method. The percent PC has been

certified in this material but percent PN has not.

8.0 Sample Collection, Preservation and

Storage

8.1 Water Sample Collection -- Samples collected

for PC and PN analyses from estuarine/coastal waters

are normally collected from a ship using one of two

methods; hydrocast or submersible pump systems. Follow the recommended sampling protocols associated

with the method used. Whenever possible, immediately

filter the samples as described in Section 11.1.1. Store

the filtered sample pads by freezing at -20EC or storing in

a desiccator after drying at 103-105E C for 24 hr. No

significant difference has been noted in comparing the

two storage procedures for a time period of up to 100

days. If storage of the water sample is necessary, place
440.0-5 Revision 1.4 September 1997

MDL = (t) X (S)

where, S = Standard deviation of the replicate analyses.

t = Student’s t value for n-1 

degrees of freedom at the 

99% confidence limit; t = 3.143 

for six degrees of freedom.

the sample into a clean amber bottle and store at 4EC

until filtration is done. When beginning the use of this method, on a quarterly

8.1.1 The volume of water sample collected will vary calibration standards and acceptable instrument perforwith the type of sample being analyzed. Table 1 provides

a guide for a number of matrices of interest. If the matrix

cannot be classified by this guide, collect 2 x 1L of water

from each site. A minimum filtration volume of 200 mL is

recommended.

8.2 Sediment Sample Collection -- Estuarine/coastal sediment samples are collected with benthic

samplers. The type of sampler used will depend on the be established for PC and PN using a low level estuarine

type of sample needed by the data-quality objectives. water sample, typically three to five times higher than the 7

Store the wet sediment in a clean jar and freeze at -20EC estimated MDL. The same procedure should be followed

until ready for analysis. for sediments. To determine MDL values, analyze seven

8.2.1 The amount of sediment collected will depend on through the entire analytical procedure (Section 11).

the sample matrix and the elemental analyzer used. A

minimum of 10 g is recommended.

9.0 Quality Control

9.1 Each laboratory using this method is required to

operate a formal quality control (QC) program. The

minimum requirements of this program consist of an initial

demonstration of laboratory capability and the continued

analysis of laboratory reagent blanks, laboratory duplicates, field duplicates and calibration standards analyzed

as samples as a continuing check on performance. The

laboratory is required to maintain performance records

that define the quality of data thus generated.

9.2 Initial Demonstration of Performance 

(Mandatory)

9.2.1 The initial demonstration of performance is used

to characterize instrument performance (MDLs, linear dynamic range) and laboratory performance (analysis of QC

samples) prior to the analyses conducted by this method.

9.2.2 Linear dynamic range (LDR) -- The upper limit of

the LDR must be established by determining the signal

responses from a minimum of three different concentration standards across the range, one of which is close to

the upper limit of the LDR. Determined LDRs must be

documented and kept on file. The LDR which may be

used for the analysis of samples should be judged by the

analyst from the resulting data. The upper LDR limit

should be an observed signal no more than 10% below

the level extrapolated from the lower standards. Determined sample analyte concentrations that are 90% and

above the upper LDR must be reduced in mass and

reanalyzed. New LDRs should be determined whenever

there is a significant change in instrument response and

for those analytes that periodically approach the upper

LDR limit, every 6 months or whenever there is a change

in instrument analytical hardware or operating conditions.

9.2.3 Quality control sample (QCS) (Section 7.5) --

basis or as required to meet data quality needs, verify the

mance with the analyses of a QCS. If the determined

concentrations are not within ± 5% of the stated values,

performance of the determinative step of the method is

unacceptable. The source of the problem must be identified and corrected before either proceeding with the

initial determination of MDLs or continuing with analyses.

9.2.4 Method detection limits (MDLs) -- MDLs should

replicate aliquots of water or sediment and process

These replicates should be randomly distributed throughout a group of typical analyses. Perform all calculations

defined in the method (Section 12) and report the concentration values in the appropriate units. Calculate the

MDL as follows:1

MDLs should be determined whenever a significant

change in instrumental response, change of operator, or

a new matrix is encountered.

9.3 Assessing Laboratory Performance 

(Mandatory)

9.3.1 Laboratory reagent blank (LRB) -- The laboratory

must analyze at least one LRB (Section 3.10) with each

batch of 20 or fewer samples of the same matrix. LRB

data are used to assess contamination from the laboratory environment. LRB values that exceed the MDL

indicate laboratory or reagent contamination. When LRB

values constitute 10% or more of the analyte level determined for a sample, fresh samples or field duplicates of

the samples must be prepared and analyzed again after

the source of contamination has been corrected and

acceptable LRB values have been obtained. For aqueous samples the LRB is a precombusted filter of the

same type and size used for samples.
Revision 1.4 September 1997 440.0-6

Response factor (μv/μg) = RN-ZN-BN

 WTN

where, RN = Average instrument response to 

standard (μv)

ZN = Instrument zero response (μv)

BN = Instrument blank response (μv)

and, WTN = (M)(N )(AW/MW) a

where, M = The mass of standard material in 

μg

N = Number of atoms of C, N or H, in a

a molecule of standard material

AW = Atomic weight of C (12.01), N 

(14.01) or H (1.01)

MW = Molecular weight of standard 

material (135.2 for acetanilide)

9.3.2 Laboratory fortified blank (LFB) -- The laboratory

must analyze at least one LFB (Section 7.4.1) with each

batch of samples. Calculate accuracy as percent recovery. If the recovery of any analyte falls outside the

required control limits of 85-115%, that analyte is judged

out of control, and the source of the problem should be

identified and resolved before continuing analyses.

9.3.3 The laboratory must use LFB analyses data to

assess laboratory performance against the required control limits of 85-115% (Section 9.3.2). When sufficient

internal performance data become available (usually a

minimum of 20-30 analyses), optional control limits can

be developed from the percent mean recovery (x) and the

standard deviation (S) of the mean recovery. These data

can be used to establish the upper and lower control

limits as follows:

Upper Control Limit = x + 3S

Lower Control Limit = x - 3S

The optional control limits must be equal to or better than

the required control limits of 85-115%. After each five to

ten new recovery measurements, new control limits can

be calculated using only the most recent 20-30 data

points. Also the standard deviation (S) data should be

used to establish an ongoing precision statement for the

level of concentrations included in the LFB. These data

must be kept on file and be available for review.

9.4 Assessing Analyte Recovery and Data

Quality

9.4.1 Percent recoveries cannot be readily obtained

from particulate samples. Consequently, accuracy can

only be assessed by analyzing check standards as

samples and quality control samples (QCS). The use of

laboratory fortified matrix samples has not been assessed.

10.0 Calibration and Standardization

10.1 Calibration -- After following manufacturer’s

installation and temperature stabilization procedures,

daily calibration procedures must be performed and

evaluated before sample analysis may begin. Single

point or standard curve calibrations are possible, depending on instrumentation.

10.1.1 Establish single response factors (RF) for each

element (C,H, and N) by analyzing three weighed portions

of calibration standard (acetanilide). The mass of

calibration standard should provide a response within

20% of the response expected for the samples being

analyzed. Calculate the (RF) for each element using the

following formula:

If instrument response is in units other than μv, then

change the formula accordingly.

10.1.2 For standard curve preparation, the range of

calibration standard masses used should be such that the

low concentration approaches but is above the MDL and

the high concentration is above the level of the highest

sample, but no more than 90% of the linear dynamic

range. A minimum of three concentrations should be

used in constructing the curve. Measure response versus

mass of element in the standard and perform a

regression on the data to obtain the calibration curve.

11.0 Procedure

11.1 Aqueous Sample Preparation

11.1.1 Water Sample Filtration -- Precombust GF/F

glass fiber filters at 500EC for 1.5 hr. The diameter of filter

used will depend on the sample composition and instrument capabilities (Section 8.1.1). Store filters covered if

not immediately used. Place a precombusted filter on

fritted filter base of the filtration apparatus and attach the

filtration tower. Thoroughly shake the sample container

to suspend the particulate matter. Measure and record

the required sample volume using a graduated cylinder.

Pour the measured sample into the filtration tower, no

more than 50 mL at a time. Filter the sample using a

vacuum no greater than 10 in. of Hg. Vacuum levels

greater than 10 in. of Hg can cause filter rupture. If less

than the measured volume of sample can be practically

filtered due to clogging, measure and record the actual

volume filtered. Do not rinse the filter following filtration.

It has been demonstrated that sample loss occurs when

the filter is rinsed with an isotonic solution or the filtrate.8

Air dry the filter after the sample has passed through by

continuing the vacuum for 30 sec. Using Teflon-coated

flat-tipped forceps, fold the filters in half while still on the

fritted glass base of the filter apparatus. Store filters as

described in Section 8.
440.0-7 Revision 1.4 September 1997

Corrected

Concentration (μg/L) = sample response (μv)

Sample volume (L) x RF (μv/μg)

where, RF = Response Factor (Section 10.1.1)

Corrected Sample Response (Section 

7.4)

Corrected

Concentration (mg/kg) = sample response (μv)

Sample weight (g) x RF (μv/μg)

where, RF = Response Factor (Section 10.1.1) 

Corrected Sample Response (Section 

7.4)

Note: Units of μg/g = mg/kg

11.1.2 If the sample has been stored frozen, place the Analyze the samples for particulate C. The resultant data

sample in a drying oven at 103-105E C for 24 hr before are particulate organic carbon.

analysis and dry to a constant weight. Precombust one

nickel sleeve at 875E C for 1 hr for each sample.

11.1.3 Remove the filter pads containing the particulate

material from the drying oven and insert into a precombusted nickel sleeve using Teflon-coated flat-tipped

forceps. Tap the filter pad using a stainless steel rod.

The sample is ready for analysis.

11.2 Sediment Samples Preparation

11.2.1 Thaw the frozen sediment sample in a 102-

105EC drying oven for at least 24 hr before analysis and

dry to a constant weight. After drying, homogenize the

dry sediment with a mortar and pestle. Store in a desiccator until analysis. Precombust aluminum capsules at

550EC in a muffle furnace for 1.5 hr for each sediment

sample being analyzed. Precombust one nickel sleeve

at 875EC for 1 hr for each sediment sample.

11.2.2 Weigh 10 mg of the homogenized sediment to

the nearest 0.001 mg with an ultra-micro balance into a

precombusted aluminum capsule. Crimp the top of the

aluminum capsule with the Teflon-coated flat-tipped forceps and place into a precombusted nickel sleeve. The

sample is ready for analysis.

11.3 Sample Analysis

11.3.1 Measure instrument zero response (Section 7.4)

and instrument blank response (Section 7.4) and record

values. Condition the instrument by analyzing a conditioner. Calibrate the instrument according to Section 10

and analyze all preliminary QC samples as required by

Section 9. When satisfactory control has been established, analyze samples according to the instrument

manufacturer’s recommendations. Record all response

data.

11.3.2 Report data as directed in Section 12.

11.4 Determination of Particulate Organic and

Inorganic Carbon

11.4.1 Method 1: Thermal Partitioning -- The difference

found between replicate samples, one of which has been

analyzed for total PC and PN and the other which was

muffled at 550EC and analyzed is the particulate organic

component of that sample. This method of thermally

partitioning organic and inorganic PC may underestimate

slightly the carbonate minerals’ contribution in the

inorganic fraction since some carbonate minerals

decompose below 500EC, although CaCO does not. 3

9

11.4.2 Method 2: Fuming HCI -- Allow samples to dry

overnight at 103-105EC and then place in a desiccator

containing concentrated HCI, cover and fume for 24 hr in

a hood. The fuming HCI converts inorganic carbonate in

the samples to water vapor, CO and calcium chloride. 2

10

12.0 Data Analysis and Calculations

12.1 Sample data should be reported in units of μg/L

for aqueous samples and mg/kg dry weight for sediment

samples. 

12.2 Report analyte concentrations up to three significant figures for both aqueous and sediment samples.

12.3 For aqueous samples, calculate the sample concentration using the following formula:

12.4 For sediment samples, calculate the sample concentration using the following formula:

12.5 The QC data obtained during the analyses

provide an indication of the quality of the sample data and

should be provided with the sample results.

13.0 Method Performance

13.1 Single Laboratory Performance

13.1.1 Single laboratory performance data for aqueous

samples from the Chesapeake Bay are provided in Table

2.

13.1.2 Single-laboratory precision and accuracy data for

the marine sediment reference material, BCSS-1, are

listed in Table 3.

13.2 Multilaboratory Performance

13.2.1 In a multilab study, 13 participants analyzed

sediment and filtered estuarine water samples for

particulate carbon and nitrogen. The data were analyzed
Revision 1.4 September 1997 440.0-8

using the statistical procedures recommended in ASTM letter and spirit of any sewer discharge permits and

D2777-86 for replicate designs. See Table 4 for regulations, and by complying with all solid and hazardsummary statistics. ous waste regulations, particularly the hazardous waste

13.2.2 Accuracy as mean recovery was estimated from further information on waste management consult The

the analyses of the NRC of Canada Marine Sediment

Reference Material, BCSS-1. Mean recovery was 98.2% available from the American Chemical Society at the

of the certified reference carbon value and 100% of the address listed in Section 14.2.

noncertified nitrogen value.

13.2.3 Overall precision for analyses of carbon and

nitrogen in sediments was 1-11% RSD, while the

analyses of both particulate carbon and nitrogen in

estuarine water samples was 9-14% RSD.

13.2.4 Single analyst precision for carbon and nitrogen

in sediment samples was 1-8% RSD and 4-9% for water

samples.

13.2.5 Pooled method detection limits (p-MDLs) were

calculated using the pooled single analyst standard

deviations. The p-MDLs for particulate nitrogen and

carbon in estuarine waters were 0.014 mg N/L and 0.064

mg C/L , respectively. The p-MDLs for percent carbon

and nitrogen in estuarine sediments were not estimated

because the lowest concentration sediment used in the

study was still 20 times higher than the estimated MDLs.

Estimates of p-MDLs from these data would be

unrealistically high.

14.0 Pollution Prevention

14.1 Pollution prevention encompasses any technique

that reduces or eliminates the quantity or toxicity of waste

at the point of generation. Numerous opportunities for

pollution prevention exist in laboratory operation. The

EPA has established a preferred hierarchy of environmental management techniques that places pollution

prevention as the management option of first choice.

Whenever feasible, laboratory personnel should use

pollution prevention techniques to address their waste

generation. When wastes cannot be feasibly reduced at

the source, the Agency recommends recycling as the next

best option.

14.2 For information about pollution prevention that

may be applicable to laboratories and research institutions, consult Less is Better: Laboratory Chemical Management for Waste Reduction, available from the American Chemical Society’s Department of Government Relations and Science Policy, 1155 16th Street N.W.,

Washington D.C. 20036, (202) 872-4477.

15.0 Waste Management

15.1 The Environmental Protection Agency requires

that laboratory waste management practices be conducted consistent with all applicable rules and regulations. The Agency urges laboratories to protect the air,

water and land by minimizing and controlling all releases

from hoods and bench operations, complying with the

identification rules and land disposal restrictions. For

Waste Management Manual for Laboratory Personnel,

16.0 References

1. 40 CFR, Part 136, Appendix B. Definition and

Procedure for the Determination of the Method

Detection Limit. Revision 1.11.

2. Carcinogens - Working With Carcinogens,

Department of Health, Education, and Welfare,

Public Health Service, Center for Disease Control,

National Institute for Occupational Safety and

Health, Publication No. 77-206, Aug. 1977.

3. OSHA Safety and Health Standards, General

Industry, (29 CFR 1910), Occupational Safety and

Health Administration, OSHA 2206, (Revised,

January 1976).

4. Safety in Academic Chemistry Laboratories,

American Chemical Society Publication, Committee

on Chemical Safety, 3rd Edition, 1979.

5. Proposed OSHA Safety and Health Standards,

Laboratories, Occupational Safety and Health

Administration, Federal Register, July 24, 1986.

6. Rohrdough, W.G. et al. Reagent Chemicals,

American Chemical Society Specifications, 7th

Edition. American Chemical Society, Washington,

DC, 1986.

7. Holme, N.A. and A.D. McIntyre (eds). 1971.

Methods for the Study of Marine Benthos.

International Biome Program. IBP Handbook #16.

F.A. Davis Co., Philadelphia, PA.

8. Hurd, D.C. and D.W. Spencer (eds). 1991. Marine

Particles: Analysis and Characterization.

Geophysical Monograph: 63, American Geophysical

Union, Washington, DC 472p.

9. Hirota, J. and J.P. Szyper. 1975. Separation of total

particulate carbon into inorganic and organic

components. Limnol. and Oceanogr. 20: 896-900.

10. Grasshoff, K., M. Ehrhardt and K. Kremling (eds).

1983. Methods of Seawater Analysis. Verlag

Chemie.
440.0-9 Revision 1.4 September 1997

 Filter diameter

Sample matrix 47mm 25mm 13mm

Sample matrix volume

Open ocean 2000 mL 500 mL 100 mL

Coastal 1000 mL 400-500 mL 100 mL

Estuarine 500-700 mL 250-400 mL 50 mL

 (low particulate)

Estuarine 100-400 mL 75-200 mL 25 mL

 (high particulate)

Measured Measured

nitrogen carbon

concentration S.D. concentration S.D. A A

Sample (μg/L) (μg/L) (μg/L) (μg/L)

1 147 ± 4 1210 ± 49

2 148 ± 11 1240 ± 179

3 379 ± 51 3950 ± 269

4 122 ± 9 1010 ± 63

 Standard deviation based on 7 replicates. A

Mean

measured

Element T.V. value (%) %RSD %Recovery A B C

Carbon 2.19% 2.18 ± 3.3 99.5

Nitrogen 0.195% 0.194 ± 3.9 99.5

 True value. Carbon value is certified; nitrogen value is listed but A

 not certified

 Percent relative standard deviation based on 10 replicates. B

 As calculated from T.V. C

17.0 Tables, Diagrams, Flowcharts, and

Validation Data

Table 1. Filter Diameter Selection Guide

Table 2. Performance Data--Chesapeake Bay Aqueous

 Samples

Table 3. Precision and Accuracy Data - Canadian

 Sediment Reference Material BCSS-1
Revision 1.4 September 1997 440.0-10

Table 4. Overall and Single Analyst Precision Estimates from Collaborative Study

Analyte Sample N Mean Overall Overall Analyst Analyst (1) (2)

Conc. Std. Dev. %RSD Std. Dev. %RSD

Particulate A 11 0.0655 0.0081 12.4% 0.0050 7.6%

Nitrogen

(as N) in 

Estuarine 

Waters

B 12 0.0730 0.0076 10.3% 0.0057 7.7%

C 12 0.0849 0.0110 12.9% 0.0060 7.1%

D 12 0.126 0.0138 11.0% 0.0071 5.6%

E 11 0.182 0.0245 13.5% 0.0157 8.6%

Nitrogen 1 10 0.178 0.0190 10.7% 0.0131 7.3%

(as %N) in

Estuarine 

Water

2 10 0.295 0.0114 3.9% 0.0046 1.6%

3 10 0.436 0.0178 4.1% 0.0104 2.4%

4 10 0.497 0.0183 3.7% 0.0082 1.6%

5 10 0.580 0.0207 3.6% 0.0150 2.6%

Particulate B 12 0.369 0.0505 13.7% 0.0222 6.0%

Carbon 

(as C) in 

Estuarine

Waters

A 12 0.417 0.0490 11.8% 0.0230 5.5%

D 12 0.619 0..0707 11.4% 0.0226 3.6%

C 12 0.710 0.0633 8.9% 0.0367 5.2%

E 12 0.896 0.1192 13.3% 0.0569 6.4%

Carbon 1 13 1.78 0.1517 8.5% 0.1346 7.6%

(as %C) in

Estuarine

Sediments

2 13 2.55 0.0372 1.5% 0.0204 0.8%

3 13 3.18 0.0435 1.4% 0.0348 1.1%

4 13 4.92 0.1201 2.4% 0.0779 1.6%

5 13 5.92 0.0621 1.1% 0.0547 0.9%

(1) N = Number of participants whose data was used.

(2) Concentration in mg/L or percent, as indicated.