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An easy uncertainty evaluation of the COD titrimetric analysis in correlation

with quality control and validation data. Method applicability region

Article  in  Analytical Methods · October 2012

DOI: 10.1039/C2AY25687H

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Analytical
Methods
PAPER

An easy uncertainty evaluation of the COD titrimetric

analysis in correlation with quality control and
Cite this: Anal. Methods, 2012, 4,
4204 validation data. Method applicability region
Elisavet Amanatidou,* Eleni Trikoilidou, Georgios Samiotis, Nikolas- P. Benetis
and Nikolaos Taousanidis

Received 12th April 2012
Accepted 25th October 2012
DOI: 10.1039/c2ay25687h
www.rsc.org/methods
Chemical Oxygen Demand (COD) estimates the amount of organic oxidizable matter in wastewaters. It is a
critical parameter for assessing the quality of effluents prior to discharge, in the design and operation of
wastewater treatment plants, and in environmental modelling and monitoring. In this study COD was
determined by a closed reflux titrimetric method according to 5220C, APHA, 21st edn, 2005. This
method is applicable to COD values between 40 and 400 mg L
1 and chloride concentrations up to
2000 mg L
1. Chloride interference is a serious problem in COD measurement for wastewaters
containing low organic matter concentrations. The reliability of the results depends on a proper
validation of the measurement procedure that includes the calculation and evaluation of the
measurement uncertainty. This study develops a practical, understandable and simple way for
calculating and assessing the Combined Standard Uncertainty (CSU) of COD measurement, mainly based
on already existing quality control and validation data. The CSU assessment is based on the existing
literature trueness and precision target limits, without any additional assumptions. Diagrams correlating
the trueness of the COD measurement mean values with different chloride concentrations and the
laboratory control charts’ limits offer a tool for improving the performance of the determination and
define the method application area. The validation data that were obtained in our laboratory
(accredited according to ISO 17025) and the proposed methodology for CSU calculation were verified in
the frame of intra and inter-laboratory measurements.

1 Introduction containing low organic matter and high chloride concentra-

Chemical Oxygen Demand (COD) is dened as the amount of a
specied oxidant that reacts with the sample under controlled
conditions. Both the organic and inorganic components of a
sample are subjected to oxidation, but in most cases the organic
components predominate and are of greater interest. COD is
one of the most widely used parameters in the eld of waste-
water analysis and is frequently used as an indicator of waste-
water pollution with organic compounds. The determination of
COD in wastewaters is of highest importance for operation
control of industrial and municipal wastewater treatment
plants, wastewater effluent monitoring, etc. It requires,
however, a detailed study of the presence of interfering species
that can signicantly affect the estimation of the target oxid-
ability of the sample.1
The most common interference in COD measurement is due
to coexisting chloride ions, especially for wastewaters
Technological Educational Institution of West Macedonia, Department of Pollution
Control Technologies, Environmental Chemistry and Wastewater Treatment
Laboratory, Koila, Kozani 50100, Greece. E-mail: eamanatidou@kozani.teikoz.gr;
Fax: +30 24610 39682; Tel: +30 24610 68015
tions. There are a number of analytical methods for the deter-
mination of COD. In this study the COD closed reux titrimetric
method according to 5220C, Standard Methods for the Exami-
nation of Water and Wastewater, APHA, 21st edn, 20052 was
used by the Laboratory of Environmental Chemistry and
Wastewater Treatment (accredited according to ISO 17025).
This method is applicable to COD values between 40 and
400 mg L
1 and chloride concentrations up to 2000 mg L
1.
The principle of the closed reux titrimetric COD measure-
ment method is the oxidation of the organic matter by a boiling
mixture of a known excess of 0.01667 M potassium dichromate
(K2Cr2O7) and concentrated sulfuric acid in the presence of a
catalyst (Ag+ salt). Aer digestion, the remaining unreduced
K2Cr2O7 is titrated with ferrous ammonium sulfate (FAS),
Fe(NH4)2(SO4)2$6H2O, to determine the amount of K2Cr2O7
consumed and the oxidizable matter is calculated in terms of
oxygen equivalents.
Oxidation of most organic compounds occurs to 95 to 100%
of the theoretical value. The most common interference is
coming from chloride ions. Chloride reacts with silver ions to
precipitate silver chloride and thus inhibits the catalytic activity
of silver. Such interferences are negative tending to restrict the

4204 | Anal. Methods, 2012, 4, 4204–4212 This journal is ª The Royal Society of Chemistry 2012
Paper
oxidizing action of the dichromate ion. Under the digestion
procedure applied in COD analysis, chlorides react with
dichromate ions to produce chlorine and chromic ions, causing
a positive interference, eqn (1).
6Cl
+ Cr2O72
+ 14H+ / 2Cr3+ + 3Cl2 + 7H2O (1)
The most oen used standard methods for COD determina-
tion are suitable for samples containing less than 2000 mg L
1
Cl
ions. This limitation renders the COD determination inap-
plicable for brackish and sea water which have a concentration of
chloride ions around 0.55–1.37 g kg
1 and 19.2 g kg
1 respec-
tively. More complicated, expensive and time consuming tech-
niques designed to measure COD in saline waters are available.
Difficulties that are caused by the presence of chloride ions can
be overcome largely by complexing with mercuric sulfate (1 g
HgSO4 per 50 mL sample), in the Hg : Cl
ratio of 10 : 1, up to
2000 mg L
1 Cl
,2 before the reuxing procedure (closed reux
titrimetric method) that leads to formation of a soluble salt or a
soluble unreactive complex (chloride masking).2,3
HgSO4 reacts with Cl
and forms the easily soluble HgCl2 or
complexes of type HgCl42
are formed (eqn (2) and (3)) in the
sample. The extent to which HgCl2 in aqueous solutions is ionized
is small. The Hg2+ concentration in saturated HgCl2 solution is
10
8 g ion per L, which is less than the H+ concentration in water
(covalent bond). Thus the presence of sulfuric acid in the solution
does not give replacement reaction of chloride.
The reactions of chloride masking are shown in eqn (2) and (3).
HgSO4 + 2Cl
/ HgCl2 + SO42
(2)
HgSO4 + 4Cl
/ HgCl42
+ SO42
(3)
Many other approaches that attempt to manage the problem
of chloride ion interference including addition of silver salts to
mask chloride ions are utilized. For example, addition of
chromium(III) to lower the oxidation potential of the reaction in
eqn (1), determination of the amount of chloride ions oxidized
by iodometric titration followed by a subsequent correction for
chloride ions, oxidation with permanganate, etc.3,4 In the
oxidation of the organic load by potassium permanganate
KMnO4 for the COD measurement (Japanese Mn-COD method),
the COD is known as the permanganate value. Compared to the
Mn-COD, the Cr-COD method that is used as the standard in
USA and Europe has a higher detection limit and severe chlo-
ride ion interference because of the stronger oxidant and the
longer digestion time.4
The reliability of the results depends on a proper validation
of the measurement procedure that includes the calculation
and evaluation of the measurement uncertainty.
There have been reported many different approaches of how
to calculate and assess the measurement uncertainty. Drolc
et al. (2003) identied the major uncertainty sources and
compared the combined uncertainty with the experimentally
determined variation. The results showed that the major sour-
ces of uncertainty arose from repeatability at a high concen-
tration level and volumetric steps at a low concentration level.1
This journal is ª The Royal Society of Chemistry 2012
Analytical Methods
Viana da Silva et al. (2011) evaluated the magnitude of the
estimated measurement uncertainty by comparing the relative
or absolute expanded measurement uncertainty with dened
target values.5
The main objective of the present study is to develop a
practical, understandable and simple way for calculating and
assessing the measurement uncertainty of COD, mainly based
on the already existing quality control and validation data. The
absence of established measurement uncertainty target limits
led this laboratory to develop a methodology for the Combined
Standard Uncertainty assessment based on the existing true-
ness and precision target limits, without any additional
assumptions. Moreover, the laboratory identied the method
application area of COD measurement, by correlating uncer-
tainty, trueness, precision and laboratory applied control limits,
with and without chloride interference. The proposed meth-
odology is an alternative way for ensuring the application of the
method under controlled conditions.
2 Material and methods
The aim of this study is to (a) develop an easily applied method
for the uncertainty evaluation and assessment of COD titri-
metric measurement useful for both the academic and the
industrial sector and (b) develop a way for determining safe
control chart limits taking into account the interferences under
the condition of acceptable accuracy and precision. This
approach was examined by method validation and verication
and both trueness and precision as well as repeatability and
reproducibility (from intra and inter-laboratory comparisons)
estimation.6
Trueness is the closeness of agreement between the average
of an innite number of replicate measured quantity values and
a reference quantity value.7 The terminology is very similar to
that used in accuracy, but trueness applies to the average value
of a large number of measurements.8
Precision is the closeness of agreement between indications
or measured quantity values obtained by replicate measure-
ments on the same or similar objects under specied condi-
tions. The ‘specied conditions’ can be, for example,
repeatability conditions of measurement, intermediate preci-
sion conditions of measurement, or reproducibility conditions
of measurement.7 Precision increases as random variations
decrease. It is possible to have results, which are precise but not
accurate. They may be all close to a mean or average value but
because of some systematic effect there is a bias rendering them
all much higher (or lower) than the true value.8
Trueness and precision alone may omit some uncertainty
sources. For this reason it is necessary to compare trueness and
precision computation (results) with target limits.
For minimization of the error propagation, a controlled
experimental procedure was applied, proposed recently by this
laboratory as “Errors Propagation Break-up (ERB)”.9 The
Combined Standard Uncertainty (CSU) was calculated accord-
ing to the error propagation law.10 The controlled experimental
procedure can be applied by considering a measurement
Anal. Methods, 2012, 4, 4204–4212 | 4205
Analytical Methods
procedure as a series of discrete operations each of which may
be assessed separately.
The method validation and verication was achieved
considering the repeated analysis data of in-house standards, as
well as inter-laboratory comparison programmes, both being
useful in establishing method precision.
The results of the inter-laboratory measurements conrmed
the assumptions and veried the calculations of the Combined
Standard Uncertainty at the 95% condence level.
The procedure steps involved during the present investiga-
tion were:
(i) Evaluation of Combined Standard Uncertainty (CSU)
mathematical expression.
(ii) Calculation of trueness and precision.
(iii) Comparison of calculated trueness and precision with
standard target limits.
(iv) Calculation of type A uncertainty if trueness and preci-
sion are acceptable.
(v) CSU calculation.
(vi) Method application area determination, with the
contribution of trueness, precision and control limits in one
composite control chart and in correlation with the inuence of
the interfering substance.
The key point in this procedure is to bring trueness and
precision within the target limits. Only under this condition the
other steps can be applied. In this way the laboratory uses
international literature accepted target limits to assess CSU and
at the same time to ensure the measurement accuracy.
2.1 Uncertainty calculation of COD titrimetric
determination
The evaluation of the Combined Standard Uncertainty (CSU) is
a statistical approach of measurement, where the different
uncertainty sources are estimated and combined into a single
value. The basis for the estimation of measurement uncertainty
to be proposed is derived from the existing data; no special
investigation is required from the laboratories.
For the uncertainty estimation the steps involved are: (a)
measurand specication, (b) uncertainty source identication,
(c) uncertainty component quantication, and (d) CSU
calculation.
Table 1 Uncertainty components in titrimetric COD measurements
Uncertainty components Uncertainty error sources
1. Sample volume - Filling
- Temperature
- Tolerance
- Reading
2. Random phenomena (type A uncertainty) - Repeatability
- Intermediate precision
3. Titration - Burette technical data
4. Weighing - Balance technical data (included in standard solution preparation uncertainty)
5. Standard solutions preparation - Weight, dilutions (included in type A uncertainty and in bias)
6. Reagents purity - Included in standard solution preparation uncertainty
7. Method and laboratory total bias - Standard solutions
4206 | Anal. Methods, 2012, 4, 4204–4212
Paper
2.1.1 IDENTIFICATION OF UNCERTAINTY COMPONENTS IN COD
TITRIMETRIC DETERMINATION. In the closed reux titrimetric
COD measurement method, the possible sources of error that
constitute the Combined Standard Uncertainty (CSU) compo-
nents are presented in Table 1. Some of them were considered
signicant and were taken into account in the CSU
calculation.
Because the in-house or eld sampling stages do not form
part of the specied procedure, the components of uncertainty
related to the sampling procedure were not included in this
calculation procedure.
2.1.2 CALCULATION OF UNCERTAINTY COMPONENTS. The
measurement procedure is considered as a series of discrete
operations. Each of them may be assessed separately to obtain
estimates of the uncertainty associated with it.
Accordingly, as is described below, the separate uncer-
tainties for individual or groups of components were estimated
and expressed as standard uncertainty that contributed to the
CSU calculation.
The digestion and the standard solution preparation
uncertainty were included in the repeatability (r)/intermediate
precision7 (within laboratory reproducibility, Rw)11 and it
was not taken into account in calculating the Combined
Standard Uncertainty (CSU). Under controlled conditions of
the method application, the total bias is insignicant
compared to the combined uncertainty and it can be
neglected in the total uncertainty calculation. In the
following is presented the uncertainty calculation of the main
components as well as the CSU of the closed reux titri-
metric COD determination.9
All the assumptions were conrmed by repeated intra- and
inter-laboratory measurements.9–12
Sample volume uncertainty. The volume of the solution con-
tained in the volumetric ask is subjected to three major
sources of uncertainty9,10,13–15
(a) the volumetric glassware uncertainty (tolerance);
(b) the repeatability (glassware lling) uncertainty, u(R); and,
(c) the uncertainty caused by the volume difference between
the glassware calibration temperature (20  C) and the solution
temperature,
u(t) ¼ DV/O3 and DV ¼ V20  aw  DT
Data sources
- Glassware calibration certicate
- Measurements
- Literature data
- Measurements
- Burette calibration certicate
- Balance calibration certicate
- Measurements
- Reagent certicates of analysis
- Certicates of analysis
- Measurements
This journal is ª The Royal Society of Chemistry 2012
Paper Analytical Methods

sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
where V20: glassware volume at 20  C, aw: volume expansion
Sr 2
coefficient, DT: temperature variation. uðbiasÞ ¼ uCRM 2 þ (7)
n
By a, b, and c, the combined sample volume uncertainty,
u(V), is calculated in eqn (4): where uCRM ¼ a/2 (a ¼ certicate value), Sr ¼ standard
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
 2  2 deviation.
tol DV
uðV Þ ¼ pffiffiffi þ½uðRÞ þ pffiffiffi 2
(4) If the contribution of the method and laboratory total bias
3 3
uncertainty is insignicant, it is not taken into account in the
However, the glassware lling uncertainty13 is included in Combined Standard Uncertainty calculation and eqn (6) is
the measurement of the repeatability or/and the intermediate reformed to eqn (8):
precision (type A uncertainty), as well as in the standard solu- sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
    2ffi
u Vsample 2 uðV titration Þ
tion preparation uncertainty and it cannot be taken into uðcÞ ¼ c  þðRSDrÞ2 þ (8)
Vsample Vtitration
account in the volume uncertainty estimation. So the nal
relation is expressed in eqn (5): This equation can be applied safely in all titrimetric
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
 2  2 determinations.
tol DV
uðV Þ ¼ pffiffiffi þ pffiffiffi (5) The contribution of the sensitivity coefficient has been
3 3
included in the estimation of component uncertainties and was
This way of calculation can be used also for automatic considered equal to unity.6
pipettes and dispensers. For instruments of variable volume, The expanded uncertainty, for a coverage factor k, is evalu-
the determination of the tolerance and the lling DV in the ated from the relationship: U ¼ ku(c). For an approximate level
closer volume of the measurement are necessary. of condence of 95%, k ¼ 2 and U ¼ 2u(c).
Titration uncertainty. Similarly, in the titration uncertainty are
involved the above three steps. 3 Results and discussion
Type A uncertainty. The estimation of type A uncertainty,10 was
For the purpose of this study, solution standards of 20, 40, 200
performed under repeatability and intermediate precision
and 400 mg L
1 COD with and without the presence of chloride
conditions and expressed as RSDr or RSDRw. Because of the
were measured by the closed reux titrimetric method, as
Errors Propagation Break-up (ERB) procedure applied by this
control samples. The COD standard solutions were prepared by
laboratory, the use of one of the above type A uncertainty
suitable dilutions of potassium hydrogen phthalate stock
expressions did not cause any important differentiation from
solution (KHP). The presence of chloride was ensured by addi-
the total uncertainty calculation, as was conrmed by the rele-
vant intra- and inter-laboratory measurements. The ERB tion of NaCl in the solution stock. The inuence of the dilution
procedure considers that the measurement is a series of from the solution stock was controlled by repeated COD
measurements of diluted standards and comparison with the
discrete operations whose uncertainties can be assessed sepa-
laboratory control charts limits. The concentrations of the COD
rately. Furthermore the ERB imposes specic limitations to
standard solutions used for this study are given in Table 2.
some of these operations depending on their contribution in
For the measurements, a high purity reagents calibrated
the total uncertainty. This is a controlled experimental proce-
Brand automatic burette, a calibrated Hamilton soop
dure in the frame of the bias source elimination and was
dispenser, a calibrated Brand Transferpette S pipette, a cali-
recently presented by this laboratory.9
Combined Standard Uncertainty calculation. Taking into brated WTW CR2200 digester as well as a calibrated Mettler
account the uncertainty components mentioned, the Combined Toledo XS205 balance and glassware were used. The prepara-
tion of the standard solutions had signicant contribution in
Standard Uncertainty (CSU) of closed reux titrimetric COD
the estimation of the control sample concentration uncertainty.
determination is expressed by eqn (6).1,8,9
According to the ERB procedure, this laboratory calculated
uðcÞ ¼ c
separately the concentration uncertainties of the standard
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
    2  2ffi solutions and only standard solutions with uncertainty #1–2%
u Vsample 2 2 uðVtitration Þ uðbiasÞ were used for achieving the CSU management in the framework
 þðRSDrÞ þ þ (6)
Vsample Vtitration c0 of the internal quality control applied.8 The use of such
where u(Vsample), sample volume uncertainty; RSDr, type A
uncertainty expressed by the repeatability or the intermediate
Table 2 Concentrations of standard COD solutions
precision; u(Vtitration), burette volume uncertainty and u(bias),
method and laboratory total bias.9 The method and laboratory Concentrations of
total bias9 is estimated by the analysis of the Certicated standard solutions
Reference Material (CRM) under repeatability conditions. It is mg L
1 COD Chloride concentrations mg L
1 Cl

the combination of the standard deviation of the mean for n
20 1000 2000 3000 — — —
determinations and of the standard uncertainty of the certied 40 1000 2000 2500 3000 — —
value of the CRM, (at a level of condence of 95%). The total 200 1000 2500 3000 4000 5000 6000
bias7 is expressed in eqn (7): 400 2000 2500 3000 4000 5000 6000 7000 8000

This journal is ª The Royal Society of Chemistry 2012 Anal. Methods, 2012, 4, 4204–4212 | 4207
Analytical Methods Paper

standard solutions contributed in reducing the type A uncer-
tainty of this study in which standard solutions were used as
control samples. Control charts for the internal quality control
were used.16
The titration was performed with 0.025 M ferrous ammo-
nium sulfate (FAS) solution, suitable for low COD samples,
instead of 0.1 M FAS proposed in Standard Methods for the
Examination of Water and Wastewater for most samples.
The FAS solution was standardized before each set of
measurements. If the FAS concentration was #0.024 M, a new
reagent was prepared and standardized, in the frame of the ERB
process applied.
The trueness, the precision and the uncertainty of the COD
measurements in the above four concentration levels, with and
without the presence of chloride, were calculated and compared
with the relative target limits (Tables 3 and 4).
To obtain means and standard deviations, 16 samples were
processed for each COD concentration level. It should be noted
that all the experimental data, more than 2000 COD measure-
ments were made over a period longer than four years.
Calculating trueness, the agreement among the average
values that are obtained by a number of measurements is esti-
mated, as a parameter of the method systematic errors. True-
ness is expressed by eqn (9):
Trueness ¼ 100  (Xmean
m)/m (9)
where m is the true value.
Depending on the measurand concentration, the trueness is
acceptable in comparison with the limits from Table 3.17
If trueness is acceptable, precision, which is a parameter of
the method random errors, is calculated as RSDr, and
compared with RSDrmax as given in Table 4 (ref. 18), for the level
of concentration m.
Precision is expressed by eqn (10).18
RSDr ¼ Sr/Xmean (10)

where Sr is the standard deviation under repeatability

Table 3 Trueness target limits 17 conditions.
If RSDr # RSDrmax, the method precision is acceptable in the
Concentration of analyte 100  (Xmean
m)/m concentration level examined with reference materials (in-
house standards).
m < 1 mg kg
1
50 to +20%
1 mg kg
1 < m < 10 mg kg
1
30 to +10%
In Tables 5 and 6 are presented the calculation of trueness,
m > 10 mg kg
1
20 to +10% precision and standard deviation results of COD under repeat-
ability conditions for a large number of measurements in four
concentration levels, with and without the presence of chloride.
Table 4 Relationship of concentration and relative standard deviation (RSDr) Trueness and precision of COD control sample measurements
under repeatability conditions18 without the presence of chloride are acceptable for all the
concentration levels (Table 5).
Concentration of analyte
The variation of the trueness relative to chloride concentra-
Average fraction of Corresponding tion, for all the COD concentration levels, is shown in Fig. 1a–d.
analyte in the sample concentration of the analyte RSDr maximuma In this gure are clearly shown the experimentally determined
limits of chloride concentrations in which the trueness is
10
9 to 10
8 1–10 mg kg
1 30%
acceptable (trueness limits:
20 up to +10%).
10
8 to 10
7 10–100 mg kg
1 20%
10
7 to 10
6 100 mg kg
1–1 mg kg
1 15%
Under the presence of chloride ions, trueness and precision
10
6 to 10
5 kg
1–10 mg kg
1 10% of COD low concentration control samples, measurements are
10
5 to 10
4 10 mg kg
1–100 mg kg
1 7.5% acceptable. For high COD concentrations, trueness and preci-
10
4 to 10
3 100 mg kg
1–1 g kg
1 5.0% sion are acceptable for higher chloride ion concentrations than
10
3 to 10
2 0.1–1% 3.5%
the original method prescription implies (Table 6).
10
2 to 10
1 1–10% 2.5%
10
1 to 10 10–100% 2.0%
More specically, for 200 mg L
1 COD and chloride concen-
trations greater than 3300 mg L
1 Cl
, the trueness is not
a
These values of relative standard deviation RSDr were obtained from acceptable because of the increasing systematic errors, while the
large experimental calculations of HORWITZ and based on these
results and the related graph. (Horwitz trumpet–funnel of Horwitz). precision is acceptable because the random phenomena are under
They are independent of the analyte’s nature, the substrate (matrix) of control. For 400 mg L
1 COD, the trueness and the precision are
the analyzed material and the method of analysis. acceptable for chloride concentrations up to <8000 mg L
1 Cl
.

Table 5 Trueness, precision and standard deviation of COD measurements in four concentration levels, without the presence of chloride

Trueness
COD mg L
1 Measurement average Standard deviation (SD) (
20 up to +10%) Precision Observations

20 21.70 2.7 8.5% 3.1% (up to 7.5%) Trueness and precision: A

40 39.3 4 1.75% 2.65% (up to 7.5%) Trueness and precision: A
200 202.25 4.2 1.12% 2% (up to 5%) Trueness and precision: A
400 393 7.5
1.85% 1.92% (up to 5%) Trueness and precision: A

4208 | Anal. Methods, 2012, 4, 4204–4212 This journal is ª The Royal Society of Chemistry 2012
Paper Analytical Methods

Table 6 Trueness, precision and standard deviation of COD measurements in four concentration levels, with the presence of chloridea

Measurement Trueness
COD mg L
1 Cl
mg L
1 average (
20 up to +10%) Precision

20 1000 21.9 9.50 A 7.53 A (up to 7.5%)

2000 33.33 66.63 NA 5.39 A (up to 7.5%)
3000 56.39 181.94 NA 18.32 NA (up to 7.5%)
40 1000 40.26 0.65 A 3.1 A (up to 7.5%)
2000 43.41 8.52 A 2.44 A (up to 7.5%)
2500 51.8 29.5 NA 13.83 NA (up to 7.5%)
3000 61.07 52.7 NA 16.39 NA (up to 7.5%)
200 1000 202.02 1.01 A 1.97 A (up to 5%)
2500 209.025 4.98 A 4.97 A (up to 5%)
3000 214.64 7.32 A 4.57 A (up to 5%)
4000 230.44 15.22 NA 2.30 A (up to 5%)
5000 271.99 35.99 NA 1.35 A (up to 5%)
6000 288.72 44.36 NA 1.35 A (up to 5%)
400 2000 395.50
1.125 A 0.93 A (up to 5%)
2500 399.31
0.17 A 1.3 A (up to 5%)
3000 399.39
0.15 A 0.9 A (up to 5%)
4000 382.30
4.42 A 2.77 A (up to 5%)
5000 391.89
2.03 A 0.68 A (up to 5%)
6000 399.35
0.16 A 1.84 A (up to 5%)
7000 426.16 6.54 A 0.53 A (up to 5%)
8000 446.13 11.53 NA 0.22 A (up to 5%)
a
A: acceptable, NA: not acceptable.

Fig. 1 Trueness variation relative to chloride concentration for four different COD concentrations (a) 20 mg L
1 COD, (b) 40 mg L
1 COD, (c) 200 mg L
1 COD and (d)
400 mg L
1 COD.

This journal is ª The Royal Society of Chemistry 2012 Anal. Methods, 2012, 4, 4204–4212 | 4209
Analytical Methods Paper

Table 7 Uncertainty of COD measurements in four concentration levels without the presence of chloridea

COD mg L
1  U

1
1
COD mg L  U COD mg L  U (calculated by using repeatability as
(calculated by using repeatability as (calculated by using intermediate type A uncertainty, including total
COD mg L
1 type A uncertainty) precision as type A uncertainty) laboratory bias)

20 21.70  5.4 21.5  6.4 21.7  5.6

40 39.30  8 — 39.3  8.3
200 202.25  8.5 199.3  8.6 202.25  8.7
400 393  15.2 394.1  18 393  15.3
a
U ¼ ku(c). For condence level 95%, k ¼ 2 and U ¼ 2u(c).

Table 8 Uncertainty of COD measurements in four concentration levels with the presence of chloride

COD concentrations (mg L
1) 20 mg L
1 Cl
1000 2000 3000

COD mean value mg L
1 20.75 21.9 56.39
U 6.63 2.4 20.66
40 mg L
1 Cl
1000 2000 2500 3000
COD mean value mg L
1 40.26 43.41 51.8 61.07
U 7 2.2 27.95 20
200 mg L
1 Cl
1000 2500 3000 4000 5000 6000
COD mean value mg L
1 201.85 209.025 214.64 230.44 271.99 288.72
U 7.52 16.77 20 5.33 8.90 8.00
400 mg L
1 Cl
2000 2500 3000 4000 5000 6000 7000 8000
COD mean value mg L
1 395.50 399.31 399.39 382.30 391.89 399.35 426.16 446.13
U 7.49 10.93 7.64 21.30 5.60 14.80 4.85 2.6

In Tables 7 and 8 are presented the Combined Standard
Uncertainty (CSU) results for a large number of COD measure-
ments at four concentration levels, with and without the pres-
ence of chloride. This laboratory examined the contribution of
the repeatability and intermediate precision in the CSU, by
calculating the type A uncertainty with repeatability and intra-
laboratory intermediate precision data, at the selected concen-
trations. Normally, the repeatability standard deviation, Sr,
excludes major uncertainty contributions and is not suitable for
the uncertainty estimation. Therefore, for the estimation of the
measurement uncertainty, the intermediate precision standard
deviation SRw is used most oen. However, by applying a
controlled experimental procedure and a strict internal quality
control for CSU management, i.e. the Error Propagation
Breakup (ERB) method,9 this laboratory contributed to reducing
the type A uncertainty even under repeatability conditions. In
applying the ERB methodology the consideration of either
repeatability or intermediate precision in the total uncertainty
calculation did not alter signicantly the results, as presented in
Table 7.
For example, in 200 mg L
1 COD the uncertainty calculated
by using repeatability as type A uncertainty is 8.5 mg L
1 and by
using intermediate precision as type A uncertainty is 8.6 mg
L
1. Because of the analytic accuracy improvement of the COD
testing by this laboratory through the use of rigorous quality
control and CSU management programs, the method and
laboratory total bias is not signicant and is not specied as a
separate performance characteristic in CSU calculation. Its

Fig. 2 Laboratory control charts (mean value  2S) at 200 mg L
1 COD (a) without the presence of chloride and (b) with 2500 mg L
1 Cl
.

4210 | Anal. Methods, 2012, 4, 4204–4212 This journal is ª The Royal Society of Chemistry 2012
Paper Analytical Methods

contribution in CSU is within the range 0.02% to 0.45% and it is

negligible as shown in Table 7;19–21 (0.045%, 0.33%, 0.045%,
0.022% for COD concentrations of 20, 40, 200 and 400 mg L
1
respectively). For 200 mg L
1 COD the uncertainty calculated by
using repeatability as type A uncertainty, including the total
laboratory bias, is 8.7 mg L
1.
From Tables 7 and 8 is also observed that by increasing the
chloride ion concentration the COD measurement mean
values also increase, especially for the COD concentrations
that can tolerate lower chloride concentrations. At the same
time, the trueness and precision is also increasing (Table 6).
When trueness and precision (or one of them) becomes
unacceptable, the measurement is imprecise, although the
uncertainty may not change signicantly. The uncertainty in
most cases, specied solely as an isolated value can be
considered acceptable, but arriving at a decision only from
this assessment is wrong.22 For this reason, it is necessary to
correlate the trueness with the laboratory control charts’ limits
under the condition of acceptable precision. The laboratory
control charts, which demonstrate that the measurement Fig. 4 Method application area (gray shadow) for 400 mg L
1 COD relative to
chloride concentration, trueness and control limits from the control chart con-
procedure performs within given limits, can be constructed
structed with chloride-free solution standard. The % values are the trueness
with solution standard in the presence of interfering corresponding to different COD mean values.
substances. In Fig. 2a are presented the laboratory control
charts at 200 mg L
1 COD without chlorides and with control
For control samples of 200 mg L
1 COD, chloride-free, the
limits 190–208 mg L
1 COD. In Fig. 2b are presented the
method is applicable for samples with chloride concentrations
laboratory control charts at 200 mg L
1 COD, with 2500 mg
<2500 mg L
1 Cl
(Fig. 3a) instead of 2000 mg L
1 Cl
proposed
L
1 Cl
and control limits 192.4–225.65 mg L
1 COD; in this
from the standard method. Also, for control samples of 200 mg
case the laboratory control limits are expanded. This control
L
1 COD with 2500 mg L
1 Cl
, the method is applicable for
charts can be applicable under the constraints of an accept-
able trueness and precision.
The correlations of the trueness of the COD measurement
Table 9 Results of COD inter-laboratory measurements
mean values with different chloride concentrations and the
laboratory control charts’ limits are presented graphically in Reference Laboratory
Fig. 3 and 4. Such diagrams, which can become useful tools for Date value Measurement z-score code
any laboratory, dene the method application area (gray
June–July 2008 96.2 97.2
0.1 026
region). The application area is limited from the lower and
April–May 2009 135 133
0.4 035
upper laboratory control limits and the acceptable trueness of October–November 2009 147 141 0.1 063
the COD measurement mean values in different chloride Dec–January 2010 144 151 0.5 046
concentrations. It can be implemented by laboratories in the Feb–March 2010 295 280
0.5 021
frame of analytical methods validation and internal quality June–July 2010 114 110
0.4 014
June–July 2010 34.9 35.57 0.8 061
control programs.

Fig. 3 Method application region (gray shadow) for control samples of 200 mg L
1 COD relative to chloride concentration, trueness and control limits; (a) control
limits from the control chart constructed with chloride-free solution standard; (b) control limits from the control chart constructed with 2500 mg L
1 Cl
solution
standard. The % values are the trueness corresponding to different COD mean values.

This journal is ª The Royal Society of Chemistry 2012 Anal. Methods, 2012, 4, 4204–4212 | 4211
Analytical Methods
samples with chloride concentrations <3300 mg L
1 Cl

(Fig. 3b). For control samples of 400 mg L
1 COD, chloride-free
and control limits 376–411 mg L
1 COD, the method is appli-
cable for samples with chloride concentrations <6400 mg L
1
(Fig. 4). In the case of using control samples with chlorides the
control limits will be expanded and the method can be appli-
cable for samples with chloride concentrations <8000 mg L
1
because trueness and precision are acceptable up to this
concentration (Table 6).
Therefore, COD measurements of urban and most industrial
wastewater, with such chloride concentration levels,23 can be
measured with satisfactory accuracy. Samples of higher
chloride concentrations can be measured by making suitable
dilutions which minimize the interference.
The lower COD quantitation limit, that can be determined
with acceptable trueness and precision as well as acceptable
level of uncertainty, is 20 mg L
1 COD for relative clean waters
with chloride ion concentration up to 1000 mg L
1 Cl
. This
COD concentration is lower than the method relative limit
(Tables 5–8).
The results of the inter-laboratory measurements conrmed
the assumptions and veried the calculations of the Combined
Standard Uncertainty.24
The inter-laboratory measurements were implemented
according to LEAP Scheme Chemistry for wastewater. The
results, as z-score, for the examined parameter are presented in
Table 9. The z-score is considered satisfactory for values
|z| # 2.
4 Conclusions
An easily applied method for the uncertainty evaluation of
closed reux titrimetric COD measurement, applicable in all
titrimetric determinations, useful for both the academic and
the industrial sector, was developed in this study.
Laboratory control charts, demonstrating the measurement
procedure within specic limits can be constructed with solu-
tion standards in the presence of interfering substances, having
a matrix similar to the samples. In this case the laboratory
control limits are expanded. Diagrams correlating the trueness
of the COD measurement mean values with different chloride
concentrations and the laboratory control charts’ limits offer a
tool for improving the performance of the determination and
denes the method application area. It can be applied by
laboratories as a way to achieve a good internal quality control
(QC) involved in this type of chemical determinations.
4212 | Anal. Methods, 2012, 4, 4204–4212
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