5310 TOTAL ORGANIC CARBON (TOC)

Abstract:5310 A.
Introduction

1. General Discussion

The organic carbon in water and wastewater consists of multiple organic compounds in
various oxidation states. Some of these compounds can be oxidized further via biological or
chemical processes. Biochemical oxygen demand (BOD), assimilable organic carbon (AOC),
and chemical oxygen demand (COD) methods may be used to characterize these fractions.
Total organic carbon (TOC) is a more convenient and direct expression of total organic
content than BOD, AOC, or COD but does not provide the same kind of information. TOC is
independent of the organic matter's oxidation state and does not measure other organically
bound elements (e.g., nitrogen and hydrogen) or inorganics that can contribute to the oxygen
demand measured by BOD and COD. The amount of TOC in water sources varies widely
from <0.1 to >25 mg/L in drinking water to >100 mg/L in wastewater. Although TOC is not a
direct substitute for BOD, AOC, or COD testing, several studies have demonstrated that
correlations can be developed and TOC can be used as a surrogate for some of these other
parameters in certain cases. This may be acceptable under some regulatory frameworks.
These relationships must be established independently for each set of matrix conditions (e.g.,
various points in a treatment process and water types). Correlations should be developed with
caution and validated seasonally.

Organic contaminants in water may degrade ion-exchange capacity, serve as a carbon or

energy source for undesired biological growth, react with disinfectants to form byproducts
that could harm human health or aquatic organisms, or be otherwise detrimental to the water's
purpose. Before organically bound carbon can be analyzed, the molecules must be converted
into carbon dioxide (CO2), a molecular form that can be measured quantitatively. TOC
methods use either high temperature (typically 680 to 950°C) with catalysts and oxygen or
air, or lower temperatures (<100°C) with ultraviolet irradiation and/or one or more chemical
oxidants and catalysts to convert organic carbon into carbon dioxide. The CO 2 may be purged
from the sample, dried, and transferred via carrier gas to a nondispersive infrared analyzer or
coulometric titrator. Alternatively, CO2 may be separated from the sample via a CO2-selective
membrane into high-purity water, where the increase in conductivity corresponds to the
amount of CO2 passing through the membrane.

2. Fractions of Total Carbon

The methods and instruments used to measure TOC analyze fractions of total carbon (TC)
and measure TOC via two or more determinations. These fractions of total carbon are defined
as:

• inorganic carbon (IC)—carbonate, bicarbonate, and dissolved CO2;

• total organic carbon (TOC)—all carbon atoms covalently bonded in organic molecules;

• dissolved organic carbon (DOC)—the fraction of TOC that passes through a 0.45-μm-pore-
diameter filter;
• suspended organic carbon (also called particulate organic carbon)—the fraction of TOC
retained by a 0.45-μm filter;

• purgeable organic carbon (also called volatile organic carbon)—the fraction of TOC

removed from an aqueous solution via gas stripping under specified conditions; and

• nonpurgeable organic carbon (NPOC)—the fraction of TOC not removed by gas stripping.
In most cases, inorganic carbon is purged and not “determined,” in which case only NPOC is
determined and purgeable organic carbon is assumed to be negligible.

In many water samples, the IC fraction is many times larger than the TOC fraction. IC can be
eliminated by adjusting samples to pH 4 or less to convert IC species to CO 2. Samples are
normally adjusted to pH 2 or less; however, this may maximize hydrophobic molecule loss to
the sample vessel walls. Adjusting to pH 4 minimizes such losses, but IC-removal efficiency
must be checked and carbonate minerals in unfiltered samples may not completely dissolve.
The CO2 is then removed by purging the sample with a purified gas (free of both CO 2 and
organic contaminants) or by vacuum degassing. Sample purging also removes purgeable
organic carbon, so the organic carbon measurement made after eliminating IC interferences is
actually an NPOC determination. In many surface and ground waters, the purgeable organic
carbon contribution to TOC is negligible, so in practice the NPOC determination is
commonly substituted for TOC.

Alternatively, TOC may be determined by separately measuring TC and IC, and calculating
the difference. If the sample contains far more IC than TOC, subtracting IC from TC is not
the preferred approach to determining TOC because of the relatively large error involved.
The purgeable fraction of TOC is a function of the specific conditions and equipment used.
Sample temperature and salinity, gas-flow rate, type of gas diffuser, purging-vessel
dimensions, volume purged, and purging time affect the division of TOC into purgeable and
nonpurgeable fractions. When separately measuring purgeable organic carbon and NPOC in
the same sample, use identical purging conditions for both fractions. Also, consider purging
conditions when comparing purgeable organic carbon or NPOC data from different
laboratories or instruments.

3. Method Selection

The high-temperature combustion method (5310B) is suitable for samples with higher TOC
levels, which otherwise would have to be diluted for some of the persulfate methods (5310C).
Generally, 5310B determines organic carbon levels in compounds that are chemically
refractory. These compounds cannot be determined by 5310C. High-temperature combustion
may be desirable for samples containing high levels of suspended organic carbon or more
than 500 mg/L of chloride or other halides, where carbon may not be oxidized efficiently by
persulfate and/or ultraviolet (UV) irradiation methods. When analyzing saline waters, care
must be taken when using the combustion method. Manufacturers may offer different catalyst
packing schemes that will increase the life of the column and minimize the formation of salt
cakes. Interlaboratory studies have shown biases on the order of 1 mg C/L using older high-
temperature instruments. With newer instruments, bias has been reduced and detection limits
as low as 0.010 mg C/L have been reported. Some high-temperature combustion instruments
are not designed for levels <1 mg C/L. Method 5310B accumulates nonvolatile residues in
the analyzer, while in 5310C residuals are drained from the analyzer after each analysis.
Method 5310C generally provides better sensitivity for lower-level samples (<1 mg C/L); it is
useful for TOC as low as 0.010 mg C/L.

Because the sensitivity range of the methods overlaps, other factors may dictate method
choice. A method may be chosen based on desired precision, ease of use, cost, or other
factors.

Samples high in particulate organic carbon are problematic for any TOC technique. Even if
the particulates can be homogenously suspended, they may selectively adhere to sample
introduction portions of the analyzer. The best approach is to filter the samples using carbon-
free filters and analyze the filtrate for DOC, then analyze the filters for particulate organic
carbon using an appropriate carbon analyzer.

To qualify a particular instrument for use, demonstrate that the single-user precision and bias
given in each method can be reproduced. Preferably, demonstrate overall precision by
conducting in-house studies with more than one operator.

Evaluate the selected method to ensure that data quality objectives are attained. Evaluate the
method detection level in a matrix as similar as possible to the unknowns. Be aware that TOC
analyzers handle instrument blanks in a variety of ways and that the true magnitude of the
blank may not be readily apparent to the analyst. Some instruments “zero out” much of the
blank by adjusting the detector's baseline reading. Some enter blank values in units (e.g., mv
responses) rather than absolute concentrations, and others accumulate the total blank in the
system during a blank run. Carefully observe the variability of low-level measurements, and
both the method detection level and the blank whenever reagents or instrument operations are
changed.

Methods 5310B and C note that when a water blank is analyzed, the carbon level in the blank
water contributes to the observed blank value. However, an instrument blank should only
represent the contribution of reagents or system upset (injection blank). Organic carbon-free
water exposed to the atmosphere can rapidly gain TOC. Modern instrumentation provides
schemes to recycle previously combusted water or allows reagent blanks to be run in the
absence of injected water. Some instruments have on-line capability or sample-introduction
procedures so low-TOC water can be injected without exposure to the atmosphere. Blank
values should be based on internally recycled water or reagent blanks, and external blank
water injections should not be subtracted from determined sample values. Determine the
background TOC level in the reagent water used to prepare standards. Handle this water
similarly to the standards and subtract the background TOC level from the TOC level of the
standards. Do not subtract the background TOC level from the samples unless they are
diluted; in which case, subtract the dilution water's contribution to the TOC concentration.
Evaluate each instrument and/or external sparging system for IC removal. IC removal
efficiency can be significantly altered by minor changes in the sparging system or by changes
in sample temperature. Whenever the sparging system is altered, re-evaluate IC-removal
efficiency. The methods show expected single-operator and multiple-laboratory precision.
These equations are based on referenced interlaboratory studies that, in some cases, were
performed on older equipment. Observe the testing range because the error and bias generally
will be some significant fraction of the low standard. Consult references to determine the type
of equipment and conditions of the interlaboratory study. Determine instrument performance
by analyzing waters with matrices similar to those of the unknowns.
4. Filtering Samples for DOC Determinations

Ideally, a 0.45-μm pore-size filter is required to determine DOC; however in practice, glass
fiber filters with larger nominal pore sizes are often used.

a. Filter apparatus: Glass, tetrafluoroethylene (TFE), stainless steel, acetyl resin, or other

material that neither adsorbs nor leaches organic substances and is capable of holding the
selected filters may be used. Polypropylene syringes with syringe-type filter holders or
peristaltic pumps with in-line filter holders may be used as an alternative to vacuum filtration.

b. Filters: Use a glass fiber filter without organic binder or else filters made of TFE,
polycarbonate, silver, polyethersulfone, or hydrophilic polypropylene. Other filters that
neither adsorb nor leach organic carbon into water may be used, especially if colloidal matter
must be removed. The specific filter used must be documented along with the data. Filters
commonly used are 2.2 to 4.7 cm in diameter, although this may vary. Syringe filters are also
common. Alternatively, centrifugation may be used to remove colloidal material.

Pre-rinse the filter with organic-free water to remove soluble impurities. Glass fiber filters
may be combusted at high temperatures (450 to 500°C for at least 1 h) before rinsing. If
alternate separation techniques, filters, or filter preparations are used, demonstrate that
equivalent results are produced. Filter pore size may influence test results, especially in raw
waters. When filtering samples, check applicable regulations to see if the filter must have an
absolute pore size of 0.45 μm because glass fiber filters do not meet this requirement. For
highly turbid samples, pre-filters (including glass fiber) may be used to remove interferences
and minimize clogging of 0.45-μm filters. Studies have shown that hydrophilic
polyethersulfone or polypropylene filters are the best options.1

c. Filter blanks: Filters can both release and adsorb DOC. Before use, a filter must be rinsed
to remove any potential DOC. Also, for each filter type and manufacturer, determine and
document the volume required so the resulting filtrate analysis yields a TOC result <1/2
MRL. Begin with three successive rinses of 25 mL each, and analyze the third rinse for TOC.
If analysis of the third aliquot yields a result <1/2 MRL, then the volume used is acceptable.
If the value exceeds 1/2 MRL, continue with another 25 mL rinse and analyze. Continue the
process until a sufficient rinsing volume has been determined.

d. Filter to waste: If possible, pass a portion of the sample through the filter to waste before
beginning to collect filtrate. For the recommended filters, 25 mL was generally found to be
an adequate amount to discard before collecting sample. The filter-to-waste volume can be
determined on a low suspended solids sample by collecting aliquots of filtrate and
determining DOC on the aliquots until the difference in DOC values between sample and
filtrate is insignificant.

e. Filter plugging: Filter plugging is a common problem. A glass prefilter that has been
heated to 500°C for 1 hour in a muffle furnace or an organic-free glass fiber filter that has
been demonstrated to be free of interferences and pre-rinsed immediately before use has been
shown to be effective in reducing plugging.

5. Quality Control Applicable to All Methods

a. Initial demonstration of performance: This should be done whenever a new instrument is
set up, a method or instrument is modified, or a new operator is trained.

1) Initial demonstration of calibration and precision—Analyze at least 4 replicates of a mid-

range calibration check sample prepared from a different source than that used for the
calibration. The average obtained value should be within 15% of the theoretical, and the
relative standard deviation (RSD) should be <10%.

2) Method detection limit—Determine the method detection limit (MDL) by preparing a

standard at 2 to 5 times the expected MDL in reagent water. (NOTE: MDLs determined using
standards prepared in reagent water may not be representative of the MDL for complex
sample matrices, so consider developing matrix-specific MDLs when matrix effects indicate
that a reagent-water-based MDL is inappropriate.) Perform at least seven non-consecutive
injections of the standard over a minimum of 3 d. The MDL is calculated as follows:

Where:

S = standard deviation of replicate analyses.

t(n – 1,1 – α = 0.99) = Student's t value for the 99% confidence level with n – 1 degrees of
freedom (t = 3.14 for 7 replicates), and

n = number of replicates.

3) Demonstration of IC-removal efficiency—Purged samples may absorb organics or

CO2 from the atmosphere, so analyze samples immediately after purging or cap them tightly
until an aliquot is injected. Use the procedure in 5310A.5b10) to demonstrate IC removal
efficiency. The necessary sparging time may be determined by sparging for various times and
selecting the time required to obtain a value <1/2 of the MRL.

4) TOC carryover check—Immediately following the analysis of the highest calibration

standard, analyze a blank. The concentration of the blank must be <1/2 of the MRL. If the
blank has a value >1/2 of the MRL, then the highest calibration point must be lowered until
the blank immediately following the point has a value <1/2 of the MRL. Analyze the high
calibration point/blank pair at the beginning of every analytical day to verify that carryover is
not occurring.

b. Ongoing quality control:

1) Holding time—For DOC, an unpreserved sample must be filtered and then preserved
within 48 hours of collection. For NPOC analysis, adjust the original sample or filtrate to pH
≤2. Samples must be stored at ≤6°C. Preserve samples with HCl, H 2SO4, or H3PO4,
depending on instrument recommendations. High chloride content in a sample may cause a
bias to the reported results on some instruments. Analyze all samples within 28 d of
collection.

When determining IC and then TOC via difference and high-temperature combustion,
refrigerate sample, do NOT adjust its pH, and analyze it as soon as possible because TOC
may biodegrade significantly before the regulatory 28-d holding time expires. Alternatively,
determine IC on an unacidified sample and then determine TOC by difference using a
second, acidified sample. Keep in mind that the IC result for the acidified sample used to
determine TOC does not represent the IC concentration of the unacidified sample.

2) Collect samples in glass bottles—Amber glass is preferred, but clear glass may be used if
the sample is protected from light. Alternative containers may be used if it is demonstrated
that the container does not remove or add TOC at a level greater than 1/2 MRL.

3) Calibration check sample—At least once per analytical day, analyze a mid-range
calibration check sample prepared from a different source than that used for the initial
calibration. The results should be within 10% of the theoretical.

4) Initial instrument blank—Analyze a blank consisting of recycled water or low TOC water.
The purpose is to determine if there is any TOC present in the instrument that may
contaminate the system. Instrument blank results should be <1/2 MRL and not affect the
linearity of the calibration curve. If the result is higher, analyze several blanks to clear the
system.

5) Method blank—After the initial calibration or after an existing calibration has been
verified, and prior to sample analysis, a method blank must be analyzed. The method blank
consists of low TOC water as well as any reagents that have been added to samples. The
value of the blank must be <1/2 of the MRL.

6) Continuing blank check—After every 10 samples, analyze a blank preferably consisting of

recycled combusted water or low-TOC water. The blank must contain any reagents that have
been added to the samples. With some instruments, this is difficult to do regularly. If so,
monitor a given lot of reagent water throughout the day's run. The results for subsequent
reagent-water blanks must be <1/2 MRL.

7) Continuing calibration check—After every 10 samples, analyze one of three calibration

check samples—low-range, mid-range, and high-range standard concentrations—on a
rotating basis. The low-range standard should be at or below the minimum reporting level.
The low-range sample should agree within 50% of the true value, and the mid- and high-
range samples should agree within 15%.

8) Duplicates—Perform a duplicate analysis for every ten samples (or part thereof) analyzed.
The RPD (relative percent difference) should be less than 15%. The duplicate analysis can be
a duplicated fortified sample.

9) Fortified sample—Spike one sample per every ten samples analyzed or part thereof. Spike
at 50 to 200% of the expected concentration. The spike level should be greater than 5 times
the MRL and generally within 50 to 200% of the expected concentration. The recovery
should be between 85 and 115%.

10) Inorganic carbon check—Confirm on a frequency that corresponds to the laboratories'

routine periodic maintenance schedule, or when maintenance has been conducted on an
instrument that may affect the tool/technique for inorganic carbon removal, that inorganic
carbon is sufficiently removed from the samples. Prepare a 102.5 mg/L IC test solution
(based on bicarbonate calculations and impurities) and analyze the solution to confirm that
the result is <1/2 MRL. The IC removal check may be performed on a different matrix, but
the IC level in the check sample should be higher than those in the unknown samples.

c. Corrective actions:

1) High blank values—Consult instrument manual. Check for contamination of reagents,

sample containers, and equipment. Both high inorganic-matrix samples and high-TOC
samples may cause an ongoing elevated blank. Diluting samples may help. Higher MRLs
may be required.

2) Bad calibration check samples—Consult instrument manual. The instrument may need to
be recalibrated.

3) Poor spike recovery or poor duplicate precision—This suggests a matrix problem, or it

may be caused by non-homogenous suspended particulates in the sample. Consult instruction
manual. Diluting samples may help.