66 Clean 2008, 36 (1), 66 – 69

Ayrton F. Martins1
Lucas E. W. de Jesus1
Research Article
N
dia Vendruscolo1
Tibiri
G. Vasconcelos1 Semi-Micro Reflux Procedure for Minimization of
Marcelo L. Wilde1
Chloride Interference by COD Determination
1
Departamento de Qu
mica, Universidade
Federal de Santa Maria, Brasil. In this study, a semi-micro procedure for the COD (Chemical Oxygen Demand) was
undertaken to allow the measurement of effluent samples with a high chloride con-
tent. It was found that the COD values for standard and effluent samples, corre-
sponded closely to those obtained using the standard method of Burns and Marshall.
When the method was applied to samples of industrial wastewater, with a very high
chloride content, the recommended procedure proved to be accurate and precise,
with RSDs of 6% and 4.3% for the standard and effluent solutions, respectively. The
required amount of hazardous reagents was similar to that found in the standard
closed reflux method. It was also possible to determine the total amount of oxidized
halides. The use of this low-cost, laboratory-made system made it unnecessary to
acquire an expensive, imported analytical system, while maintaining a high level of
accuracy in the determinations.
Keywords: Semi-micro Analysis; Chemical Oxygen Demand; Reflux Method; Chloride Interference;
Received: October 15, 2007; revised: November 12, 2007; accepted: November 17, 2007
DOI: 10.1002/clen.200700154

1 Introduction
The COD is an important parameter for the evaluation of the
organic load of waters and effluents. The mixing of dichromate
with concentrated sulphuric acid, using silver sulphate as a catalyst,
under an open or closed reflux, with a heating time of 2 h (at
1508C), can oxidize 85 to 95% of the existing organic load (CxHyOz) to
water and carbon dioxide gas, reactions (1) and (2) [1, 2]:
CxHyOz + (2x – z)H2O q xCO2 + (y + 4x – 2z)H+ + (y + 4x – 2z)e – (1)
6 Cl – + Cr2O72 – + 14H+ fi 2 Cr3+ + 3 Cl2 + 7H2O (2)
One of the great difficulties of this kind of determination is the
serious interference caused by halides, in particular, chlorides.
Since the standard method is based on the use of a strong oxidizing
acid medium, a consumption of oxidant by chlorides often occurs
and this is accompanied by precipitation of the silver catalyst [1].
For this reason, the presence of excessive amounts of chlorides
causes a positive deviation from the results of the COD determina-
tion, following reaction (2).
This interference can be reduced by the addition of mercury sul-
phate, resulting in the formation of a complex compound (HgCl2),
reaction (3):
HgSO4 + 2 Cl – fi HgCl2 + SO42 – (3)
As a consequence, it is possible to determine COD when dealing
with concentrations of up to 2,000 mg L – 1 of chlorides. However,
Correspondence: Prof. Dr. A. F. Martins, Departamento de Qu
mica, Uni-
versidade Federal de Santa Maria, 97105-900 Santa Maria, RS – Brasil.
E-mail: martins@quimica.ufsm.br
the analytical procedure can become unreliable when the organic
load of the samples contains as low as 100 to 200 mg L – 1 of O2, and
the chloride concentrations are as high as those observed in sea-
water [3, 4].
Other authors also suggest different methods and techniques for
the improvement of the standard method for the COD determina-
tion. Domini et al. [5] proposed three different digestion methods to
determine the COD using closed microwave-assisted, open micro-
wave-assisted and ultrasound-assisted systems, all allowing the
reagents to be consumed in the same amounts as occurs in the clas-
sical reference method.
LaPara et al. [6] suggest improving the standard method of COD
determination by reducing the use and disposal of hazardous
reagents, employing a miniaturized closed reflux method and col-
orimetric analysis, and describing the application to synthetic and
real samples, which are not subject to chloride interference.
In this study, a laboratory-made semi-micro reflux procedure is
proposed to determine the COD of samples with high chloride con-
tent. The procedure is applicable to samples of environmental and
industrial significance. The developed semi-micro procedure
requires similar amounts of hazardous reagents as the standard
closed reflux method.
2 Experimental
2.1 Samples
The most widely used standard solutions which can be employed in
oxidative methodologies are aqueous solutions of D-glucose and
potassium hydrogen phthalate (KHP). The concentrations of these
chemicals can be correlated with theoretical COD values, i. e., 1.176
mg COD/mg KHP and 1.067 mg COD/mg D-glucose [1].

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Clean 2008, 36 (1), 66 – 69 Semi-Micro COD Determination 67

Synthetic samples with a theoretical COD of 200 mg L – 1 and a

chloride content of 1.2% (w/v) were used for the standard measure-
ments. An effluent stream from the production of the herbicide Tri-
fluraline, i. e., amination water (AW), was used as a source of real
samples. This effluent had a chloride concentration of 121,000 mg
L – 1 (12.1%) and a relatively low organic load. As a result, the interfer-
ence in the free determination of COD is limited to a range of up to
2,000 mg L – 1 of chloride [3, 4]. It is not possible to use a simple dilu-
tion to ensure conformity within the working range (a 2% Cl – ,
(w/v)), since the organic load falls below the quantification limit
(LOQ, see Sec. 3.2.1).

2.2 Apparatus and Procedure

The COD determinations were carried out in accordance with the
Standard Methods [1], as recommended by Baumann [4]. This proce-
dure was modified by using a smaller sample (2 mL), sulphuric
Ag2SO4 (3 mL) and 0.04167 M of K2Cr2O7 (1 mL) solution volumes. The
amount of HgSO4 added was proportional to the chloride, using a
predicted Hg:Cl – ratio of 10:1.
The proposed system, is demonstrated in Fig. 1 and is composed
of digestion tubes (height, h = 19 cm and inner diameter, di = 2.0 cm)
and a condenser (h = 17.5 cm and di = 1.5 cm), both made of boronsi-
licate glass. During the digestion process at 1508C for 2 h, a flow
rate of 3 L min – 1 of nitrogen was added to the digestion tubes
through PTFEm tubing (di = 0.8 mm), by means of a gas distributor, in Figure 1. Proposed semi-micro system for COD determination: (1) N2
order to remove the evolving chlorine. The reception glass tubes (h = cylinder, (2) Peristaltic pump, (3) Digestion block, (4) Digestion tubes, (5)
48 cm and di = 1.1 cm) trapped the chlorine, which was carried Cooling system, (6) Gas distributor, (7) PTFE tubing, (8) Trapping tube
down through Tygon tubing (di = 0.8 mm), by merging the current for halides, and (9) Tygon tubing.
in 40 mL of iodide solution (4 g of KI/300 mL water) acidified with 15
mL of acetic acid [4]. The inlet temperature of the cooling water was
maintained at 10 – 158C during the process.
When the digestion was completed, the tubes were withdrawn
and the N2 flow rate was increased to 5 L min – 1 to cool down the
process and prevent back-siphoning of the KI solution. The calcula-
tions were worked out by means of Eqs. (4 – 6) [4]:
ðBr
AÞ N N N 8000
CODK2 Cr2 O7 ¼ ð4Þ
mLSample

8000 N D N E
CODCl
¼ ð5Þ
mLSample

CODTotal ¼ CODK2 Cr2 O7
CODCl
ð6Þ

where Br is the volume of ferrous ammonium sulphate (FAS) used

for titrating the blank, A is the volume of FAS spent titrating the
samples and N is the FAS normality used as the titrant. D is the vol-
ume of sodium thiosulphate spent titrating the iodine, the excess of Figure 2. Influence of the chloride concentration on the COD value, (KHP
which generates the triiodide ion (I3 – ) [7], and E is the normality of standard solution of 200 mg L – 1 O2).
the sodium thiosulphate.

observed from Fig. 2, an increase of the chloride concentration leads

3 Results and Discussion
3.1 Effect of the Chloride Ion Concentration
A search was undertaken to gauge the degree of interference by
keeping the COD content at 200 mg L – 1 of O2 (A LOQ) and varying
the chloride concentrations from 0.6 to 1.8% (w/v), below the upper
limit of 2,000 mg L – 1. The results are shown in Fig. 2. As can be
to a proportional increase of COD, which can be correlated to an
inefficient gas-liquid transfer by high concentrations of chloride. In
contrast, the pattern that emerges from the standard method is a
decrease of the COD values, which can be attributed to an incom-
plete complexation by the mercury chloride [3]. As a result, it was
decided to deal with chloride concentrations close to 1200 mg L – 1,
in which the relative error is ca. 5%.

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68 A. F. Martins et al. Clean 2008, 36 (1), 66 – 69

Table 1. COD values for synthetic samples of D-glucose and AW effluent samples with a correction procedure for the interference of chlorides.

Predicted CODa) Predicted CODb) Predicted CODc) AW CODd)

(200 mg L – 1 of O2) (200 mg L – 1 of O2) (300 mg L – 1 of O2)
1.2% Cl – (w/v) 1.2% Cl – (w/v) 1.2% Cl – (w/v)

Observed COD (mg L – 1 of O2) 222.0 191.7 312.8 198.4

s (mg L – 1 of O2) l 13.4 l 16.3 l 19.0 l 8.5
RSD (%)e) 6 8.5 6.1 4.3
Er (%)f) 6.7 8.12 6.3 4.2
n 7 7 7 7
a)
D-glucose standard sample.
b)
KHP standard sample.
c)
AW effluent diluted 10 times + D-glucose (100 mg O2 L – 1).
d)
AW effluent diluted 10 times.
e)
Percentage relative standard deviation of seven determinations.
f)
Percentage relative error.

used with the same content of chloride as that used in the standard
and real tests.

3.4 Limits of Detection and Quantification

According to IUPAC [8], the detection limit is defined as LOD = 3S/b,
where S is the standard deviation of the response for blank repli-
cates and b is the angular coefficient of the calibration curve.
Accordingly, the LOD obtained was 25.4 mg L – 1 of O2 for the devel-
oped procedure. The LOQ, defined as 10S/b, was 84.7 mg L – 1 of O2.

3.5 Precision and Accuracy

Figure 3. Standard COD curves for the proposed procedure, (50 – 200
mg L – 1 of O2, KHP standard solutions).
3.2 Linearity of the Analytical Response
The analytical linearity was calculated by means of a variance analy-
sis. The significant Linear Regression and the nonsignificant Linear-
ity Deviation (P a 0.01) were checked, with KHP concentrations
between 50 and 200 mg L – 1 of O2, in triplicate.
A correlation coefficient of 0.9827 was found using a calibration
curve, Fig. 3. The results obtained showed that there was a good cor-
relation despite a relative error of 25%, which could be attributed to
the fact that the work was carried out close to the Limit of Detection
(LOD) (a 50 mg L – 1 of O2). Hence, it is recommended that the COD
contents of the samples should be above 100 mg L – 1 of O2 for use of
this method.
3.3 Validation of the Proposed Procedure
Both the real and standard samples were all analyzed seven times.
The COD data analysis was carried out by means of the t-test and var-
iance analysis (ANOVA), which ensured the reliability of the results.
The values of the recovery tests lay in the range of 95 to 111%. In the
case of the blank test, samples of distilled-deionised water were
Precision, as a relative standard deviation RSD (%), and accuracy, as
a relative percentage error Er (%), of the proposed procedure, were
measured by means of solutions containing two different concen-
trations of standards and AW samples, which were prepared and
analyzed seven times, i. e., n = 7. The data obtained are summarized
in Tab. 1. As can be observed from Tab. 1, the value found for the AW
effluent by the proposed procedure of 1,984.0 l 8.5 mg L – 1 of O2
with an RSD of 4.3% corresponds to the value of 2,032 l 400 mg L – 1
of O2 with an RSD of 19.7% found in previous studies [9, 10], using
the Burns and Marshall method of COD determination [3].
3.6 COD Determination od Standard and AW Effluent
Samples
The AW effluent with a concentration of 121,000 mg L – 1 of Cl – was
diluted 10 fold to obtain a COD of ca. 200 mg L – 1 of O2 in the pres-
ence of 1.21% of chloride, and thus in this case, remained within
the working range of the developed procedure. The results obtained
were compared to those of the standard solutions of KHP and D-glu-
cose. The results are shown as a Box-and-Whisker plot for the stand-
ard and AW effluent samples, with and without standard additions,
see Fig. 4.
However, there are few minor drawbacks to the proposed semi-
micro procedure compared to the standard method. These concern
the requirement for a N2 flow and the recirculation of cooling water
at a controlled temperature.
The main benefits of the proposed procedure include a reduced
consumption of the reagents and a higher reproducibility when
compared to the recommended standard method for COD determi-

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Clean 2008, 36 (1), 66 – 69 Semi-Micro COD Determination 69

tory has been solved, with the added benefit that this relatively inex-
pensive system means that there is no need to purchase an
imported, expensive commercial system.

Acknowledgement
The present study was carried out with the support of the National
Counsel of Scientific and Technological Development, CNPq, Brazil.

References
Figure 4. Box and Whisker Plot for standard and AW effluent samples for
the proposed procedure. (1) AW effluent sample, (2) D-glucose, (3) AW
effluent sample + D-glucose (100 mg L – 1 of O2), (4) KHP (200 mg L – 1 of
O2). SD: Standard deviation, and SE: Standard error.
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4 Conclusions
The low cost, laboratory-made, semi-micro reflux system for COD
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In the case of the real samples, where there was an industrial
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