Accepted Manuscript

Determination of chemical oxygen demand (COD) using an alternative wet chemical

method free of mercury and dichromate

Marit Kolb, Müfit Bahadir, Burkhard Teichgräber

PII: S0043-1354(17)30510-9
DOI: 10.1016/j.watres.2017.06.034
Reference: WR 12989

To appear in: Water Research

Received Date: 30 January 2017

Revised Date: 30 May 2017
Accepted Date: 11 June 2017

Please cite this article as: Kolb, M., Bahadir, Mü., Teichgräber, B., Determination of chemical oxygen
demand (COD) using an alternative wet chemical method free of mercury and dichromate, Water
Research (2017), doi: 10.1016/j.watres.2017.06.034.

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 ACCEPTED MANUSCRIPT

Sample Preparation

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turbid sample
colloid /

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suspension
Results of WW-Samples
+ Al2(SO4)3 3000

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+ NaOH

COD-Mn(III)/Ag (mg/L)
2500

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filtration

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2000
clear filtrate +H2SO4
1500

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to pH 1 y = 0.7999x - 1.9515
+ AgNO3 R² = 0.9935
1000

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discard AgCl 500
filter residue
of Al(OH)3
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0
filtrate 0 1000 2000 3000 4000
digestion COD-Cr(VI)/Hg (mg/L)
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with Mn(III)
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CODFR CODFL

COD = CODFR + 2,4 x CODFL

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1 Determination of Chemical Oxygen Demand (COD)

2 using an alternative wet chemical method free of mercury and dichromate

3 Marit Kolba, Müfit Bahadira, Burkhard Teichgräberb

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a
5 Institute of Environmental and Sustainable Chemistry, Technische Universität

6 Braunschweig, Hagenring 30, 38106 Braunschweig, Germany

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b
7 Emschergenossenschaft/Lippeverband, Kronprinzenstraße 24, 45128 Essen,

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8 Germany

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9 m.bahadir@tu-braunschweig.de
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10 teichgraeber.burkhard@eglv.de
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11 corresponding author: m.kolb@tu-braunschweig.de

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13 Abstract

14 Worldwide, the standard methods for the determination of the important wastewater
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15 parameter chemical oxygen demand (COD) are still based on the use of the

16 hazardous chemicals, mercury sulfate and chromium(VI). However, due to their

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17 properties they are meanwhile classified as “priority pollutants” and shall be phased
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18 out or banned in the frame of REACH (current European Chemical Law: Registration,

19 Evaluation, Authorization and restriction of Chemicals) by the European Union.

20 Hence, a new wet-chemical method free of mercury and chromium(VI) was

21 developed. Manganese(III) was used as oxidant and silver nitrate for the removal of

22 chloride ions. The quantification was performed by back titration of manganese(III)

23 with iron(II) as done in the standard method. In order to minimize losses of organic
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24 substances during the precipitation of silver chloride, suspended and colloid organic

25 matter had to be separated by precipitation of aluminum hydroxide in a first step. In

26 these cases, two fractions, one of the suspended and colloid matters and a second of

27 the dissolved organic substances, are prepared and oxidized separately. The method

28 was tested with potassium hydrogen phthalate (KHP) as conventional COD reference

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29 substance and different types of wastewater samples. The oxidation of KHP was

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30 reproducible in a COD range of 20 to 500 mg/L with a mean recovery rate of 88.7%

31 in comparison to the standard COD method (DIN 38409-41). Also in presence of

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32 1000 mg/L chloride a recovery rate of 84.1% was reached. For a series of industrial

33 and municipal wastewater samples a high correlation (R2= 0.9935) to the standard

34
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method with a mean recovery rate of 78.1% (± 5.2%) was determined. Even though
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35 the results of the new method are not 100% of the standard method, its high
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36 correlation to the standard method and reproducibility offers an environmentally

37 benign alternative method with no need to purchase new laboratory equipment.

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38
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39 Keywords
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40 chemical oxygen demand, COD, wastewater, chloride interference, mercury, silver,

41 chromium(VI), manganese(III)
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42

43 Abbreviations

COD chemical oxygen demand

FL filtrate

FR filter residue

HP hydroxide precipitate

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KHP potassium hydrogen phthalate

PFA perfluoro alkoxy polymer

REACH Current European Chemical Law:

Registration, Evaluation, Authorization and restriction of CHemicals
RSD relative standard deviation

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SS suspended solids

TOC total organic carbon

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UV/VIS ultraviolet/visible

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WW wastewater

WWTP wastewater treatment plant

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45 Introduction

46 The chemical oxygen demand (COD) is an important parameter for the determination

47 of the organic load in water. In particular, for operation of wastewater treatment

48 plants (WWTP) and taxation of wastewater (WW) polluters as well as for the

49 characterization of water quality, this parameter is used worldwide and is part of

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50 many directives dealing with water quality.

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51 It is based on the almost complete oxidation of the organic load in WW to carbon

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52 dioxide (CO2) and water (H2O). The oxidation of the organic load with potassium

53 dichromate (K2Cr2O7) is the internationally used standard method since the end of

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54 the 1970s. Many water samples, e.g., municipal and industrial wastewater, and
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55 seawater contain high amounts of chloride in the range of g/L. Since chloride will be

56 oxidized by Cr(VI) as well, this leads to non-reproducible elevated COD values.

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57 Hence, mercury sulfate (HgSO4) is used in order to mask chloride ions by complex
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58 formation. Furthermore, silver sulfate (Ag2SO4) in sulfuric acid is added as catalyst

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59 for the oxidation of relatively recalcitrant compounds (Moore, 1951). These wet

60 chemical COD procedures are regulated by very similar standard methods (DIN
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61 38409-H41 (1980), ISO 6060 (1989), ASTM D1252-06 (2012)). The principle of the

62 DIN 38409-H41 method is shown in Figure 1.

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63 Figure 1
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64 While measuring an important environmental parameter, these methods produce

65 relatively large volumes of hazardous wastes containing Hg(II) and Cr(VI). In the

66 meantime, mercury and its compounds are classified by the European Union as a

67 “priority pollutant” and should be “phased out” (EU Directive 2008/105/EC (2008))

68 due to their high toxicity for human beings and the ecosystem (Drasch et al. 2004).

69 Cr(VI) is a strong carcinogen (Stoecker 2004) and should be banned from 2017
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70 according to REACH (2006). Therefore, there is a high demand for the development

71 of an alternative standard method for COD measurement free of hazardous

72 chemicals. E.g., the use of mercury is already banned in Sweden and the standard

73 COD determination method is just used with a time limit exception. For this reason,

74 Sweden performed a pre-commercial procurement of mercury free COD analyses

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75 methods for wastewater and waste products (Arnell et al. 2016).

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76 Alternative to the standard COD, the total organic carbon (TOC) is already in

77 discussion since several years and was also introduced as an alternative parameter

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78 to COD in the wastewater directive of Germany (AbwV 2004) using a factor of 4 for

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79 the TOC content. However, the acceptance has been so far low, as TOC and COD
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80 can differ from each other up to a factor of 8 due to different oxidation stages of

81 organic carbon in different chemical compounds (e.g., ethane versus oxalate) leading
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82 to a completely different oxygen consumption. The parameter needed for successful

83 operation of wastewater treatment plants is the oxygen consumption.

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84 In the past, different instrumental methods for COD determination were developed,

85 based on physical-chemical methods like thermal, electrochemical, and

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86 photocatalytic oxidation of WW (ASTM, D 6238 – 98 2011, Silva et al. 2009, Lee et

87 al. 1999, Kim et al. 2000, Zhang and Zhao, 2008, Li and Song 2009). Some
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88 instruments are on the market especially for online monitoring of WWTPs. However,
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89 these methods have not been introduced into monitoring laboratories so far. One

90 reason might be their relatively high prices. Another disadvantage is their disability to

91 analyze suspended solids (SS) in water samples, which are an important part of the

92 total COD content. SS can cause problems in the operation of the instruments by

93 clogging of tubes and other instrumental parts.

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94 The aim of the investigation presented in the following was to develop a wet chemical

95 method that can be performed also in small remote laboratories worldwide, even in

96 developing countries with common inexpensive laboratory equipment and without the

97 use of hazardous chemicals.

98 Three main working pathways were followed for the method development: First, the

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99 substitution of mercury using an alternative procedure for eliminating the chloride

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100 interferences; second, the substitution of Cr(VI) by using an alternative non-

101 hazardous chemical oxidant; and third, combining the two steps of chloride

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102 elimination and alternative oxidation to one unified reaction scheme.

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103 In order to avoid the use of HgSO4 for the elimination of chloride, different methods
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104 were reported in the past. Ballinger et al. (1982) added a high surplus of AgNO3

105 instead of HgSO4. The authors used 50% more Ag+ than given in the standard COD
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106 method, before they performed the oxidation with Cr(VI) and sulfuric acid. Also
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107 Geerdink et al. (2009) suggested to precipitate chloride ions as AgCl, however, by
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108 addition of Ag2SO4 solution instead of HgSO4 already before addition of Cr(VI)-

109 oxidant, in contrast to the standard method (compare Figure 1). In order to minimize
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110 the oxidation of chloride a reduced reaction temperature of 120 °C and an Ag+/Cl-

111 ratio of 1.7 were employed, and the oxidation was performed in a closed tube.
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112 However, Ballinger et al. (1982) and Geerdink et al. (2009) still used Cr(VI) as
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113 oxidant. Miller and Bryton (1995) developed a procedure using Bi(V) in cartridges for

114 a selective oxidation of Cl- to Cl2 gas. An acidified WW sample is sucked under slight

115 vacuum through a small filter plate and the Bi(V) cartridge. Particles are retained on

116 the filter and the filter is combined afterwards with the filtrate for oxidation. The

117 authors stated that chloride removal was effective up to 1000 mg/L, and organic

118 compounds are prevented from oxidation if the sample is passed not too slowly

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119 (Miller et al. 2001). The disadvantage of this method is the very low sample volume of

120 0.6 mL, which makes it difficult to pass a representative aliquot of a SS containing

121 WW. Further trials were made to oxidize chloride preliminary before adding Ag2SO4

122 as catalyst and to consider its consumption of oxidant (1 mg Cl = 0.226 mg COD) just

123 by calculation (Dobbs and Williams, 1963). However, the authors recognized that

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124 especially in case of high concentrations of nitrogenous matter an increased

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125 consumption of Cr(VI) was obtained. They suggested that it might occur a reaction

126 series of cyclic changes from chlorine back to chloride by an intermediate reaction to

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127 chloramine. Mathematical calculation of chloride oxidation was re-adopted by Saral

128 and Goncaloglu (2008). However, they still used the standard Cr(VI)/Hg method and

129
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applied the mathematical correction only for highly chlorinated WW samples (500 –
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130 14,000 mg Cl-/L), which contain chloride levels above the complexing capacity of
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131 HgSO4. The authors did not report about an influence of nitrogen containing

132 compounds, but they reported for real samples percentage errors in the range
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133 of -8.95% to 12.32%.

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134 In order to substitute Cr(VI), oxidants should have at least the same or a higher redox

135 potential than Cr(VI) (E0 = 1,36 V) and they should be less hazardous. From this
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136 point of view, MnO4- (E0 =1,51 V), Mn(III) (E0 = 1,54 V), Ce(IV) (E0 = 1,72 V), S2O82-
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137 (E0 = 2,1 V), and H2O2/(Fe2+) (E0 = 2,8 V) are potential candidates. However, kinetic
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138 limitations are in general a much more important criterion for the suitability of an

139 oxidant, because the redox potential is just an equilibrium parameter. Due to this

140 aspect, MnO4- was already excluded for the determination of COD in the past

141 because the oxidation of many organic substances was very low in practice due to

142 kinetic limitations (Cameron and Moore 1957). E.g., Phthalic acid, which is used as

143 reference substance for the standard Cr(VI)-COD, is 100% oxidized by Cr(VI),

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144 whereas its permanganat-COD reaches only 1% of the theory (Janicke 1983).

145 Pohling (2015) reported that in domestic WW only 25% of constituents are oxidized

146 by permanganate and thus is not in use for domestic and industrial WW. Oxidants

147 like Fenton’s reagent (H2O2/(Fe2+)) and peroxodisulfate (S2O82-) are very strong, and

148 hence are in use in order to completely destroy organic substances in WW (e.g.,

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149 Rivas et al., 2001, Kilic et al. 2014). However, the reagents are very instable. For this

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150 reason, the oxidant concentrations are similarly reduced by blanks as by samples,

151 and a quantitative COD determination becomes impossible. In case of Ce(IV), the

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152 oxidation of organic substances is also relatively low. For instance, Korenaga et al.

153 (1993) used it for online oxidation of glutamic acid and lactose. The authors reported

154
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results that were comparable with the incomplete oxidation by MnO4-.
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155 Miller et al. (2001) suggested the use of Mn(III) for COD determination, which was
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156 obtained by a comproportionation of Mn(VII) and Mn(II) in sulfuric acid. The

157 preparation of Mn(III) was based on an early paper of Ubbelohde (1935). According
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158 to Miller et al. (2001), Mn(III) was stabilized by a surplus of Mn(II) of 3 : 1 in the
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159 reagent solution. They developed a spectrophotometric COD test using potassium

160 hydrogen phthalate (KHP) for calibration. For chloride elimination, they used Bi(V) as
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161 mentioned above. They described good correlations with the conventional Cr(VI)-

162 COD for some, however, not for all tested wastewater samples. However, their test
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procedure was based on a cuvette test with only 0.5 mL sample volume. Handling SS
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164 containing WW samples with such small sample volumes appears highly problematic.

165 Considering the results reported in the literature and of own pre-tests, AgNO3 for

166 chloride elimination through precipitation and Mn(III) reagent for the oxidation of the

167 organic load promised to be obviously the most suitable reagents in order to

168 substitute HgSO4 and K2Cr2O7 in an alternative wet chemical method. In

169 consequence, our method development was based on the use of these two reagents.
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170 1 Materials and Methods

171 1.1 Reagents and reference standards

172 All reagents used were of analytical grade. Water was of ultrapure quality (deionized,

173 conductivity < 0.05 µS/cm, membrane filtrated).

174 2.1.1 Preparation of Mn(III) reagent

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175 150 mL MnSO4 solution (0.378 mol/L in water) was diluted with 480 mL water in a

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176 2,000 mL wide-mouthed Erlenmeyer flask and filled up with 720 mL sulfuric acid (ρ =

177 1.84 g/mL, c = 18 mol/L) in an ice bath. At the beginning, the sulfuric acid was added

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178 in small portions in order to avoid evaporation losses. Since the sulfuric acid

179 concentration in the reagent mixture significantly influences its boiling point, the final

180
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composition of the Mn(III) reagent solution was controlled after cooling to 20 °C with
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181 an areometer (ρ20°C = 1.554 g/mL ± 0.002 g/mL).
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182 1.330 g KMnO4 was dissolved in water, filled up to 500 mL and boiled under reflux for

183 2 h. After cooling to 60 °C, the solution was filtr ated through a glass fiber filter disk
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184 into an amber glass bottle for the removal of the manganese dioxide (MnO2) formed.
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185 The filtrate was stored at room temperature in the dark.

186 150 mL of the prepared KMnO4 solution (0.0168 mol/L) was continuously added
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187 under stirring to the total volume of the prepared MnSO4/H2SO4 solution within 3 min.

188 Thereby, manganese(III) sulfate is formed through the comproportionation of Mn(VII)

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and Mn(II). The extinctions of the reagents were measured with a UV-VIS spectral
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190 photometer (DR 6000, Hach Lange GmbH, Düsseldorf, Germany). Quartz glass cells

191 with 1 mm and 10 mm optical pathlength were used for the measurements. Due to a

192 high absorbance of the Mn(III) reagent in the UV range (200 – 400 nm), 1 mm quartz

193 cells were needed to reduce the optical pathlength and to resolve the spectrum also

194 in this range.

195
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196 2.1.2 Reference standards

197 0.170 g Potassium hydrogen phthalate (KHP) (dried for 2 h at 105 °C) was dissolved

198 in water. After adding 5 mL H2SO4 (ρ = 1.84 g/mL, c = 18 mol/L), the resulting

199 solution was filled up with water to 1,000 mL final solution. The KHP solution had a

200 theoretical COD of 200 mg/L. A reference solution of chloride (Cl-) (1000 mg/L) was

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201 produced by solving NaCl in water.

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202 1.2 Preparation of glass ware

203 All glassware was cleaned carefully before use (completely free of fat and dust). After

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204 rinsing with water and detergent first, an additional treatment with hydrogen peroxide

205 and with Mn(III) reagent was performed in order to completely remove oxidizable

206 impurities.
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208 2.3 Sample preparation

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209 Turbid aqueous samples were homogenized first with a laboratory blender (Vortex
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210 Genie 2, Scientific Industries, New York USA) for 3 min at 9,500-13,000 rpm. If
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211 needed, the samples were diluted with water and blended again at 5,000 rpm. An

212 aliquot of 10 mL sample was transferred into a test tube. An overview of the whole
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213 procedure is shown in Figure 2.

214 Figure 2
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215
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216 2.3.1 Step 1 – Separation of SS and colloids

217 1 mL Al2(SO4)3 solution (0.10 mol/L) was added to all aqueous samples in the test

218 tubes that contained particulate matter and colloidal constituents (e.g. wastewater

219 samples and leachates) in order to coagulate colloids and separate them together

220 with particle matters. After addition of Al2(SO4)3 the sample was mixed well with a

221 vibration shaker. If no Al(OH)3 was precipitated, 1-2 droplets of NaOH (11 mol/L)
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222 were added to adjust the pH slightly alkaline. After agglomeration of the precipitate,

223 the sample was filtrated through a glass fiber filter disk (25 mm diameter, GF 50 025,

224 Hahnemuehle, Dassel, Germany) in a filter holder made of perfluoro alkoxy polymer

225 (AHF Analysentechnik GmbH, Tübingen, Germany) connected to a vacuum

226 apparatus (Chromabond, Macherey & Nagel, Düren, Germany) containing a second

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227 test tube. The increase of sample volume through adding 1.00 mL Al2(SO4)3 solution

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228 from 10 to 11 mL was considered by means of calculation (see 2.7). Afterwards, a

229 third test tube was placed under the filtration unit and residues of Al(OH)3 precipitate

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230 were transferred with 1 mL water onto the filter in order to quantitatively recover the

231 Al(OH)3 precipitate. Filter holder and filter were washed twice with 1 mL water each.

232
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The filtrate from the third tube with the rinsing water was discarded. The filter carrying
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233 the residues of Al(OH)3 were used for oxidation (see 2.4).
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234

235 2.3.2 Step 2 – Precipitation of chloride

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236 10 mL clear original sample or the filtrate of Al(OH)3 precipitation were acidified in
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237 the test tube with 1-2 droplets of H2SO4 (ρ = 1.84 g/mL, c = 18 mol/L) to pH 1.

238 Samples with chloride contents ≤ 1,000 mg Cl-/L were treated with 1.00 mL AgNO3
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239 solution (0,60 mol/L) and mixed with the test tube shaker. At chloride contents of

240 1,000 – 2,000 mg Cl-/L, 2.00 mL AgNO3 solution were added. The increase of sample
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241 volume through 1 or 2 mL AgNO3 solution was considered by means of calculation

242 (see 2.7). In order to agglomerate the AgCl precipitate and to improve its filterability

243 the sample was allowed to stand for 30-60 min. Afterwards, the sample was filtered

244 through a second glass fiber filter disk into another test tube as described above. The

245 filter residue of AgCl was not rinsed. 5 mL aliquot of the AgCl filtrate was used for

246 oxidation with Mn(III) reagent.

247
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248 2.4 Oxidation Procedure
249 50 mL Mn(III) reagent each was transferred with a dispenser or a volumetric pipet

250 into the digestion vessels containing stirring bars that were treated previously with the

251 Mn(III) reagent.

252 For the determination of blank values, 5 mL water was given to the Mn(III) reagent

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253 under stirring. Through mixing while adding water, a too high local dilution of Mn(III)

254 reagent was avoided.

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255 For the digestion of the filter residues, the loaded filter disks were given into the

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256 prepared vessels containing 50 mL Mn(III) reagent. Further 5 mL water was added

257 into the digestion vessels. This was necessary in order to adjust the reagent

258
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concentration for the digestion of filter residues the same as for the oxidation of 5 mL
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259 of the filtrates.

260 For the digestion of the filtrates, aliquots of each 5 mL were pipetted to 50 mL
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261 reagent in the digestion vessels.

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262 After connecting an air-cooled condenser on the digestion vessel the reaction mixture
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263 was heated up to the boiling point of 135 °C ± 2 °C within 15 min and remained

264 boiling for 120 min under reflux. The open end of condenser was covered with a
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265 small beaker or with glass wool in order to prevent foreign material from entering the

refluxing mixture.
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266
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267

268 2.5 Titration

269 After cooling the reaction mixture to room temperature in an ice bath, 4 droplets of

270 the Ferroin indicator solution (Merck KGaA, Darmstadt, Germany) were added and

271 the remaining unreduced Mn(III) was titrated back with a standard volumetric

272 ammonium iron(II) sulfate solution (0.012 mol/L) (1:10 dilution of 0.12 mol/L

273 (NH4)2Fe(SO4)3, Grüssing GmbH, Filsum, Germany) until the indicator changed from
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274 blue-green to red-orange. The titer of the ammonium iron(II) sulfate solution was

275 determined at the beginning of the method development using 10 mL of K2Cr2O7

276 (0.002 mol/L) as it is the primary standard for a volumetric solution of ammonium

277 iron(II) sulfate. In order to avoid the use of any Cr(VI), an indirect titer determination

278 was applied with sodium oxalate as primary standard. Dried sodium oxalate was

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279 weighed and titrated with the 0.0168 mol/L KMnO4 solution, which was also used for

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280 the preparation of the Mn(III) reagent. Afterwards, the titer-known KMnO4 was used

281 for the determination of the titer of the ammonium iron(II) sulfate volumetric solution.

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282

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283 2.6 Reference method AN
284 The results obtained with the new method were compared with the German standard

285 method (DIN 38409-H41, 1980). For this purpose, either original samples (with or
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286 without dilution, depending on the concentration level) were oxidized by Cr(VI) with

287 addition of HgSO4 and Ag2SO4 according to the DIN method or parts of the samples
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288 (filtration residues, filtrates) after sample preparation according to the developed
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289 method. In order to check the efficiency of the chloride elimination of the developed

290 method, no HgSO4 was added before oxidation in some cases.

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291
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292 2.7 Calculations

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293 CODtot (mg/L) = CODFR + CODFL

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c (FeII ) f
295 CODFR = • (VB - VE ) • F
VP
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c (FeII ) f V VP + VAl + VAg

297 CODFL = • (VB - VE ) • P • •F
VP VA VP

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298 CODtot: COD total amount in the original sample
299 CODFR: COD of the filter residue, if sample preparation was applied
300 CODFL: COD of the filtrate if sample preparation was applied
301 c(FeII): concentration of ammonium iron(II)-sulfate-solution in mol/L

M(O2 ) g • 1000mg
302 f: equivalence factor (here f = 8,000 mg/mol, f = )
4 • mol • g

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303 VP: sample volume for the sample preparation (here VP = 10 mL)
304 VB: volume of the ammonium iron(II) sulfate solution in mL spent for blank

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305 sample.

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306 VE: volume of the ammonium iron(II) sulfate solution in mL spent for the
307 digested aliquot of the filtrate or of filter residue

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308 VA: volume of the digested aliquot of the filtrate (here VA = 5 mL)
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309 F: dilution factor of the sample

310 VAl: volume of added Al2(SO4)3 solution in mL (here VAl = 1)

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311 VAg: volume of added AgNO3 solution in mL (here VAg = 1 or 2)

312
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313 2 Results and Discussion

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314 3.1 Elimination of chloride

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315 In order to avoid the use of mercury for masking chloride, precipitation with AgNO3

316 was used. It was known from literature, that precipitation of chloride as AgCl will
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317 reduce the COD content of wastewater samples by co-precipitation of suspended

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318 particles and colloids that will be adsorbed onto the surface of the precipitate

319 (Cameron and Moor 1957, Wagner 1976). Therefore, some authors like Ballinger et

320 al. (1982) and Geerdink et al. (2009) suggested to precipitate chloride with Ag+

321 directly as AgCl in the sample before oxidation with Cr(VI) and not to separate the

322 precipitate. Following the method of Ballinger, we spiked municipal wastewater

323 (COD = 624 mg/L) diluted 1:2 with 1000 mg Cl-/L. Chloride was precipitated with

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324 AgNO3 and oxidized with Cr(VI) and sulfuric acid. However, compared with the

325 standard method an enhanced COD value of 114% (± 6%, n=4) was found. This

326 indicated that despite of a surplus of Ag+, chloride was still subject to oxidation.

327 Hence, in our method the suspended particles were separated first by filtration with a

328 glass fiber filter using a filter holder of perfluoro alkoxy polymer (PFA). The ultrapure

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329 polymer that is commonly in use for ultra-trace analysis has an extremely smooth and

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330 hydrophobic surface, which causes less interfering physical-chemical interaction like

331 adsorption. Figure 3 shows the results of Cr(VI)-COD determination of the different

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332 fractions after preparation of a municipal WW, spiked with 1000 mg/L Cl-, according

333 to step 1 (separation of particles without addition of Al2(SO4)3) and step 2

334
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(precipitation of AgCl) of the sample preparation procedure. Identical COD values of
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335 93 mg/L were obtained for the filter residues (FR) after oxidation with Cr(VI) with and
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336 without addition of HgSO4 (Figure 3 A). This shows that no chloride ions were

337 retained in the fraction FR after the filtration procedure, otherwise the COD would be
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338 higher in case of not being masked by Hg. The COD results of the filtrates (FL) also
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339 did not differ with and without addition of Hg. Thus, the filtrates did not contain COD-

340 interfering amounts of chloride in the filtrate after separation of the AgCl precipitate
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341 according to preparation step 2. The total COD, calculated as sum of CODFR and

342 CODFL, amounted to 251 mg/L, which was 98% of the COD of the original sample
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determined according the standard Cr(VI)/Hg-method. This shows that no

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344 considerable organic was lost during the precipitation and filtration procedure by

345 sorption processes on the AgCl precipitate or on the surface of the filtration

346 equipment. The Cr(VI)-COD after filtration and precipitation of AgCl was also

347 determined for 3 replicates of another municipal WW sample (spiked with 1000 mg

348 Cl-/L) and compared with the standard Cr(VI)/Hg-COD. A mean recovery rate of

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349 101% and a variation coefficient of 4.7% showed the repeatability and accuracy of

350 the alternative chloride elimination procedure (Figure 3 B).

351 Figure 3 A and B

352 Furthermore, the Cr(VI)-COD of the AgCl precipitate was also determined without

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353 adding Hg. However, the obtained COD of 60 mg/L (Figure 3 A) clearly showed that

354 an important part of Cl- from the precipitate was oxidized by Cr(VI) and thus the

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355 precipitate had to be separated obligatory by filtration from the wastewater before

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356 oxidation.

357 In case of a landfill leachate (about 1,300 mg Cl-/L), which was free of SS, the

358
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precipitation of AgCl was directly performed in a 1:5 diluted sample as no filtration of
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359 particles was needed. However, after separation of the AgCl precipitate, the Cr(VI)-

360 COD recovery was obviously reduced to 46%. The yellow-brownish coloring of the
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361 normally white AgCl precipitate also indicated that parts of the organic content of the
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362 sample were co-precipitated and thus discarded together with the AgCl precipitate. In
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363 order to avoid these COD-losses, a co-precipitation with Al(OH)3 was introduced in

364 place of only filtration of SS in the first preparation step. In a pre-test, the precipitate
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365 and the filtrate were oxidized with the standard method (Cr(VI)/Hg) and the COD-

366 values were compared with the COD-values obtained without any pre-treatment in
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367 order to check the recovery rates of this preparation step. Good recovery rates of
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368 95% (landfill leachate) and 97% (municipal WW) were found for the sum of CODFR

369 and CODFL (Figure 4). In case of the landfill leachate, the Cr(VI)-COD of the Al(OH)3

370 precipitate (CODFR) was 70% in comparison to the standard method. This showed

371 that a high portion of colloids was present in the leachate that had to be separated

372 before precipitation of AgCl. In order to remove chloride totally from the Al(OH)3

373 precipitate, a rinsing step with pure water was introduced. The rinsed precipitate was
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374 checked for purity by solving it in HNO3 and adding AgNO3. AgCl did not precipitate,

375 indicating that the Al(OH)3 precipitate was free of chloride.

376 Hence, the addition of Al2(SO4)3 was introduced into the first preparation step as a

377 well working procedure to analyze turbid and colloidal samples by the method. This

378 step is not needed for the oxidation of real solutions of organic compounds in water.

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379 In such cases, the sample preparation with 10 mL clear sample can directly start with

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380 step 2. Then, only the filtrate of the AgCl precipitation has to be digested (Figure 2).

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381 Figure 4

382 3.2 Alternative oxidation reagent for COD

383
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Mn(III) was used for the development of an eco-friendly alternative oxidation agent
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384 instead of Cr(VI). It was obtained by the comproportionation of Mn(VII) and Mn(II)
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385 according to the following chemical equation:

386 2 KMnO4 + 8 MnSO4 + 8 H2SO4 5 Mn2(SO4)3 + K2SO4 + 8 H2O

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387 The development of the reagent solution was based on the work of Miller et al. (2001)

388 and Domini et al. (2009).

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389 The reaction of KMnO4 to Mn(III) was documented by the UV/VIS-spectra in Figure 5.

The spectrum of the obtained reagent was obviously different from the spectra of
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390
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391 unreacted KMnO4 and MnSO4 in the concentrations that were used for the

392 preparation of Mn(III) reagent. The spectrum of the Mn(III) reagent showed one broad

393 maximum at the VIS range λ = 514 nm and a shoulder in the UV range at λ = 290

394 nm. Miller et al. (2001) reported only the VIS-spectrum of the prepared Mn(III)

395 reagent in a range of 400 – 700 nm with a broad maximum at 510 nm and an

396 extinction of about 1.1 (10 mm optical pathlength). The shape of our reagent

397 spectrum was identical with the spectrum given by Miller et al. (2001) in the VIS
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398 range. In order to show that a comproportionation of Mn(VII) and Mn(II) occurred

399 under the preparation conditions, only KMnO4 was mixed with H2SO4 at the same

400 concentration as for the reagent preparation but without addition of MnSO4, and

401 heated like the Mn(III) reagent for COD determination. The marked change of the

402 spectrum showed that KMnO4 was degraded (Figure 5, dotted line). The spectrum of

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403 the reaction product was similar to the Mn(III) reagent spectrum, however, with

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404 maxima in a much lower intensity. The shoulder at 290 nm showed an extinction of

405 only 0.36, and at 514 nm a very broad low maximum with an extinction of 0.0012 was

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406 measured. The spectrum was an indication that a small amount of Mn(III) was formed

407 by disproportionation during the degradation of KMnO4. However, KMnO4 was not

408
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stable under the harsh conditions, that were used for the oxidation, and thus the sole
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409 use of KMnO4 would not be an alternative to the preparation of the Mn(III) reagent.
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410

411 Figure 5
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412 The reagent composition and preparation were optimized by changing the

413 concentrations of KMnO4, MnSO4 and H2SO4 in different directions based on the
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414 limited parameters given by Miller et al. (2001) and comparing the COD recovery

415 rates of a KHP standard (data not given here). The best results were found at a
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416 Mn(III) concentration of 0.0084 mol/L. This value was calculated without considering
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417 the volume contraction, which is caused while adding H2SO4. A real concentration of

418 0.009 mol/L was determined by titration with Fe(II). This concentration was in the

419 range of the solubility limit of Mn(III) (0.01 mol /L) (Miller et al 2001). The ratio of

420 Mn(II)/Mn(III) of the optimized reagent was 3.7/1, which was slightly higher than the

421 ratio of 3/1 given by Miller et al. (2001) and much higher than the ratio of 0.24/1

422 which was obtained using the procedure given by Domini et al. (2009). With regard to

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423 the H2SO4 concentration, the highest recovery rates were obtained with 8.64 mol/L

424 H2SO4, calculated without considering the volume contraction. Such a high

425 concentration of H2SO4 was not only needed to solve Mn(III), but also to enhance the

426 boiling point of the reagent in order to reach the needed high reaction temperature for

427 the oxidation. The boiling point of the optimized reagent was 135 °C ± 2 °C. At higher

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428 and lower concentrations of H2SO4 the recovery rates of KHP were reduced. With the

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429 optimized reagent, mean recovery rates of 87.2% ± 4.0% (n=73) were reached for

430 KHP with a theoretical COD of 200 mg/L. In comparison, the recovery rate for KHP

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431 with the standard Cr(VI)/Hg method was 99.6% ± 2.4% (n=15). This means that a

432 small part of averaged 12.8% of KHP was not covered by the oxidation reaction with

433 Mn(III).
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434 In order to check this hypothesis, a KHP-standard, which was already digested with
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435 the Mn(III) reagent, was post-oxidized with the standard Cr(VI)/Hg method. However,

436 the added amount of Cr(VI) was not reduced, showing that no further KHP or
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437 oxidizable degradation products were remained in the solution. This finding
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438 suggested, that parts of the KHP were oxidized incompletely by Mn(III), e.g., to CO

439 instead of CO2. A release of CO is well known for the reaction of formic acid with
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440 H2SO4 (Schierz 1923). We could also prove the partly oxidation of formic acid to CO
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441 with the Mn(III) reagent in a 10-fold preparation volume and a cascade of two
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442 cumulative preparations through the detection of CO in a gas trap filled with Tollens

443 reagent. However, in case of KHP, the detection of CO was not possible. The amount

444 of CO that could be formed from KHP was calculated on the base of the mean loss of

445 13% KHP and the maximum concentration of KHP (600 mg/L) that could be oxidized

446 by the method. The calculated formation of CO from KHP (about 1.6 mg C per

447 digestion) was below the detection limit of the Tollens reagent (about 8 mg C per

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448 digestion). Hence, the reason for the reduced recovery rate using Mn(III) reagent in

449 comparison to Cr(VI)/Hg could not be clarified so far.

450 In order to enhance the recovery rates further experiments were performed. The

451 prolongation of the digestion time from 2h to 4h, a reduction of the volume ratio of

452 sample to reagent from 1:10 to 1:20 and the digestion in a microwave at

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453 temperatures above the boiling point were tested without any success. In the

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454 standard method Ag+ is used as catalyst (e.g. DIN 38409-41, 1980), thus also

455 catalysts were tested. Anyway, the addition of Ag+ to the filtrate happens in step 1 of

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456 the sample preparation (CODFL). A surplus of AgNO3 was added for chloride

precipitation. However, the addition of Ag+ to a KHP standard did not increase the

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457 AN
458 recovery rates. The addition of CuSO4, which is used as Missouri catalyst for

459 digestion of samples with concentrated sulfuric acid for the determination of Kjeldahl
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460 nitrogen, and further the addition of TiO2, were tried without improvement of the

461 recovery rates. These results agree with the findings of Miller et al. (2001). They
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462 reported without giving details that all tested catalysts did not improve the oxidation of
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463 organics with Mn(III).

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464

465 Nevertheless, the oxidation of KHP with Mn(III) was reproducible and also linear (R2=
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466 0.9996) in a COD range of 20 to 500 mg/L (Figure 5). The relative standard deviation
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467 (RSD) for nine replicates in the concentration range of 50 to 500 mg COD/L was ≤

468 3.5% with a mean recovery rate of 88.7%. For the lowest standard concentration of

469 20 mg COD/L, the relative standard deviation was a bit higher (8.4%). In order to

470 check the influence of the chloride elimination step, the oxidation with Mn(III) was

471 also performed with KHP standard solutions that were additionally spiked with NaCl

472 in a high concentration of 1000 mg Cl-/L (Figure 6). Also under these conditions, a
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473 linear correlation of the data was received (R2= 0.9999, n=5) with a good

474 reproducibility. The RSD for five replicates in the range of 50 to 500 mg COD/L was

475 ≤2.1% with a little lower mean recovery rate of 84.1% compared with Mn(III)-

476 oxidation without any sample preparation step. For the lowest standard of 20 mg

477 COD/L the RSD was still 7.7%.

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478 Figure 6

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479 The concentration of the reagent was determined through titration with ammonium

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480 iron (II) sulfate solution (0.012 mol/L) (see 2.5). The mean concentration was 0.0090

481 mol/L ± 0.00013 mol/L measured at 93 days. The decrease of the Mn(III)

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482 concentration was 0.00017 mol/L in 5 days and 0.00029 mol/L in 7 days. The mean
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483 extinctions of the reagent, which was prepared at 18 different days were 1.270 ±

484 0.006 at λ = 514 nm (10 mm quartz cuvette) and 2.618 ± 0.027 at λ = 290 nm (1 mm
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485 quartz cuvette) showing the high reproducibility of the preparation. The Mn(III)
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486 reagent was stable for at least 5 days at room temperature under light exclusion. The
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487 decrease of the extinction after 5 days was about 0.04 at λ = 514 nm and 0.05 at λ =

488 290 nm, and after 7 days 0.09 at λ = 514 nm and 0.13 at λ = 290 nm.
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489
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490 3.3 COD determination of WW samples

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491 Different WW samples were analyzed with the developed method. In the first step,

492 the two fractions CODFR and CODFL of the sample preparation (Figure 2) were

493 digested with the Mn(III) reagent as well as with Cr(VI)/Hg. The results were

494 compared with the results, which were obtained when the samples were prepared

495 following the DIN 38409-41 standard procedure without further pretreatment (Table

496 1). In the range of analytical variability, the proportions of the two fractions were

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497 equal for both oxidation methods. In case of the municipal WW, around 75% of the

498 COD was found in the filter residue (CODFR) and 25% in the filtrate (CODFL), whereas

499 in case of the industrial WW the distribution was opposite, 90% of the COD was

500 found in the filtrate (CODFL) and only 10% in the filter residue (CODFR). Using the

501 Cr(VI)/Hg reagent for oxidation, the recovery rates of the total COD (CODFR+FL) was

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502 100% for the municipal WW and 102% for the industrial WW showing that the

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503 developed sample preparation for the elimination of chloride did not cause any losses

504 of COD in comparison with the DIN method. In case of oxidation with Mn(III), the

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505 recovery rates were lower compared with the DIN-method. A recovery rate of 77%

506 was obtained for the municipal WW and of 82% for the industrial WW. These rates

507
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were a little lower as the mean recovery rate of 87% for the oxidation of KHP.
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508 Different reasons might be responsible for the lower recovery rates of the COD
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509 values of WW samples compared to the recovery rates for KHP-standard solution.

510 First, in case of KHP, the recovery rate of 87% for the Mn(III)-COD was determined
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511 as CODFL without any sample preparation. Experiments that were performed with a
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512 NaCl spiked KHP standard and thus included the precipitation of AgCl showed little

513 reduced recovery rates of 84% (see above). The first step of the sample preparation
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514 with precipitation of Al(OH)3 and separated determination of CODFR might cause

515 small additional losses. Second, the multitude of different constituents in WW

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samples are supposed to have different recovery rates in case of being oxidized by
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516

517 Mn(III) compared to being oxidized by Cr(VI). E.g., for the Cr(VI)-COD of the amino

518 acid glycine a recovery rate of 96% (n=3) and for the Mn(III)-COD of 73% ± 1.25%

519 (n=9) were found compared to the theoretic COD. For the amino acid glycine, these

520 results showed that the Mn(III)-COD was 75% of the Cr(VI)-COD which was lower as

521 the recovery rate of the Mn(III)-COD for KHP (87%). Also for the oxidation with Cr(VI)

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522 it is known that not all organic compounds are oxidized to 100% and that the Cr(VI)-

523 COD recovery rates are different (Janicke 1983).

524 Table 1

525 However, a series of different WW samples (4 municipal WWs, 3 industrial WW, and

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526 2 landfill leachates) showed a high correlation (R2 = 0.9935) between the COD

527 obtained with the Mn(III)-method including the developed sample preparation and the

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528 standard method (DIN 38409-41 (1980)) (Figure 7). The mean recovery rate of all

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529 WW samples was 78.1% (± 5.2%), which shows that the lower COD values in

530 comparison with the DIN method were reproducible for the WW samples tested.

531
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Figure 7
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532 Also in case of analyzing different dilutions of one WW sample, the results for an
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533 industrial WW sample showed a high linearity with R2 of 0.9992 and also for an

534 municipal WW with R2 of 0.9851 (Figure 8).

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535 Figure 8
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536 Furthermore, the repeatability of the Mn(III)-method was tested analyzing five
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537 samples of different WW (2 municipal WW, 1 landfill leachate) in parallel, respectively

538 (Table 2). The COD values of the filter residues (CODFR, RSD 1.5 – 2.8%) had an
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539 obviously higher repeatability than the values of the filtrates (CODFL, RSD 4.5 –
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540 21.0%). Overall, the RSD of total COD (CODFR+FL) of the three samples were in a

541 range of 3.6 – 5.1% and thus showed a high repeatability of the developed method.

542 Table 2

543

544 4 Conclusions

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545 Even though the recovery rates for COD of different wastewater samples with the

546 developed Mn(III)-COD-method are not 100% compared with standard method (DIN

547 38409-41), the results showed a high correlation with the standard method and also

548 a high reproducibility among each other. Therefore, it might be possible that the

549 Mn(III)/Ag-COD could be beneficially used as conventional parameter with a

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550 correction factor in order to enable the comparison to all data that were obtained with

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551 the standard Cr(VI)/Hg-method. However, this has to be validated by further analyses

552 also through further laboratories since the number of samples is still too small.

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553 The preparation of the Mn(III) reagent and the sample preparation is so far relatively

554 time consuming compared to the standard method and has to be performed very

555
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precisely and needs some training, however automation of some steps (e.g.
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556 automatic mixing of pre reagents to Mn(III) reagent) should be possible in order to
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557 reduce the working effort.

558 The main advantage that the novel method substitutes the neurotoxic Hg and
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559 carcinogenic Cr(VI) might compensate some of its disadvantages. Hence, it offers an
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560 environmentally benign alternative method in the frame of the ongoing processes in

561 REACH and the development of international environmental quality standards.

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562 Already the application of the developed sample preparation steps for chloride

563 elimination, in combination with Cr(VI) oxidation instead of Mn(III) oxidation would be
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environmentally benign due to the absence of mercury.

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564

565 Furthermore, the wet-chemical method developed makes use of the same COD

566 equipment as for the current standard method in use. With this, the laboratories

567 worldwide do not need to purchase new equipment and detectors for the endpoint

568 determination.

569 Overall, after further validation steps, the method might be ready to be introduced

570 into a national and/or international standardization process.

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571

572 Acknowledgements

573 The authors gratefully acknowledge that this work was funded by the German

574 Federal Environmental Foundation, grant numbers: Az. 32317/01 and Az. 32317/02.

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575

576 References

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577 AbwV, 2004. German Wastewater Regulation. Regulation about the requirements for discharge of

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578 wastewater into surface water bodies, in the version published on 17. June 2004 (BGBl. I p. 1108,

579 2625), last amended by article 6 of the regulation on 2. May 2013 (BGBl. I S. 973), in German.

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580 Arnell, M., Lopez, M., Palmgren, T., 2016. Per-commercial procurement of a mercury free COD
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581 analysis method for wastewater and waste products. SP Technical Research Institute of

582 Schweden, SPRapport 2016:39, Boras, Schweden.

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583 ASTM D6238-98, 2011. Standard test method for total oxygen demand in water. West Conshohocken,

584 PA, USA.

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585 ASTM D1252-06, 2012. Standard test methods for chemical oxygen demand (dichromate oxygen
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586 demand) of water, West Conshohocken, PA, USA.

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587 Ballinger, D., Lloyd, A., Morrish, A., 1982. Determination of chemical oxygen demand of wastewaters

588 without the use of mercury salt. Analyst, 107, 1047-1053.

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589 Cameron, W.M., Moore, T.B., 1957. The influence of chloride on the dichromate-value test. Analyst,
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590 82, 677-682.

591 DIN 38409-H41, 1980. German standard methods for the examination of water, wastewater and

592 sludge, summary effect and material parameters (group H), determination of the chemical oxygen

593 demand (COD), in the range above 15 mg/L, Beuth Verlag, Berlin, in German.

594 Dobbs, R. A., Williams, R.T., 1963. Elimination of chloride interference in the chemical oxygen

595 demand test. Analytical Chemistry, 35 (8), 1064-1067.

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596 Domini, C.E., Vidal, L., Canals, A.,2009.: Trivalent manganese as an environmentally friendly oxidizing

597 reagent for microwave- and ultrasound-assisted chemical oxygen demand determination.

598 Ultrasonics Sonochemistry. http://dx.doi.org/10.1016/j.ultsonch.2009.01.008.

599 Drasch, G., Horvat, M., Stoeppler, M., 2004. Mercury. In (eds) Anke, M., Ihna,t M., Stoeppler, M.

600 Elements and their compounds in the environment. 2nd Edition. Wiley-VCH Verlag, Weinheim, pp.

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601 931.

602 EU (2008) Directive 2008/105/EC of the Europeans Parliament and of the Council of 16 December

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603 2008, on environmental quality standards in the field of water policy.

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604 Geerdink, R.B., Brouwer, J., and Epema, O.J., 2009. A reliable mercury free chemical oxygen demand

605 (COD) method. Analytical Methods, 1, 108-114.

606 ISO 6060, 1989. Water quality - Determination of the chemical oxygen demand, Genf, Schweiz.

607
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Janicke, W., 1983. Chemical oxidability of water constituents. WaBoLu Berichte 1/1983, Dietrich
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608 Reimer Verlag GmbH, Berlin, in German.

609 Kilic, M.Y., Yona, T., Mert, B.K., 2014. Landfill leachate treatment by Fenton and Fenton-like oxidation
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610 processes, Clean, http://dx.doi.org/10.1002/clen.201200714.

611 Kim, Y-C, Sasaki, S., Yano, K., Ikebukuro, K., Hashimoto, K., Karube, I., 2000. Relationship between
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612 theoretical chemical oxygen demand for specific classes of organic chemicals. The Analyst,
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613 http://dx.doi.org/10.1039/b007005j.

614 Korenaga, T., Zhou, X., Okada, K., Moriwake, T., Shinoda, S. 1993. Determination of chemical oxygen
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615 demand by a flow-injection method using cerium (IV) sulfphate as oxidizing agent. Analytica

616 Chimica Acta, 272, 237-244.

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617 Lee, K-H, Ishikawa, T., McNiven, S.J., Nomura, Y., Hiratsuka, A., Sasaki, S., Arikawa, Y., Karube, I.
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618 1999. Evaluation of chemical oxygen demand (COD) based on coulometric determination of

619 electrochemical oxygen demand (EOD) using a surface oxidized copper electrode. Analytica

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621 Li, C., Song, G. 2009. Photocatalytic degradation of organic pollutants and detection of chemical

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623 http://dx.doi.org/10.1016/j.2009.01055.

624 Miller, D.G., Brayton, S.V. 1995. US005683914A patent. Method for chloride ion removal prior to

625 chemical oxygen demand analysis, Filed 25.09.1995.

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626 Miller, D.G., Brayton, S.V., Boyles, W.T. 2001. Chemical oxygen demand analysis of wastewater using

627 trivalent manganese oxidant with chloride removal by sodium bismuthate pretreatment. Water

628 Environment Research. 73 (1), 63-71.

629 Moore, W. A., Ludzack, F.J., Ruchhoft, C. C. 1951, Determination of oxygen-consumed values of

630 organic wastes. A comparative study of methods. Analytical Chemistry 23, 1297-1300.

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631 REACH 2006. Regulation (EC) No 1907/2006 of the European Parliament and of the Council of 18.

632 December 2006 concerning the Registration, Evaluation, Authorisation and Restriction of

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633 Chemicals (REACH) establishing a European Chemicals Agency, amending Directive 1999/45/EC

634 and repealing Council Regulation (EEC) No 793/93 and Commission Regulation (EC) No 1488/94

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635 as well as Council Directive 76/769/EEC and Commission Directive 91/155/EEC, 93/67/EEC,

636 93/105/EC and 2000/21/EC.

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637 Pohling, R. (2015): Permanganate index. Determination of the oxygen consume with potassium
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638 permanganate. In: Chemical reactions in water analysis, Springer Verlag, Berlin Heidelberg (in

639 German), 227-230. DOI 10.1007/978-3-642-36354-2_29

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640 Rivas, F.J., Beltran, F.J., Gimeno, O., Frades, J. 2001. Treatment of olive oil mill wastewater by

641 Fenton’s Reagent. Journal of Agriculture and Food Chemistry, 49 (4), 1873-1880.
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642 Saral, A., Goncaloglu, B. I. 2008. Determination of real COD in highly chlorinated wastewaters. Clean,
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643 http://dx.doi.org/10.1002/clen.200800073
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644 Schierz, E.R. 1923. The decomposition of formic acid. Journal of the American Chemical Society,

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646 Silva, C.R., Conceicao, C.D.C., Bonifacio, V.G., Filho, O.F., Teixeira, M.F.S. 2009. Determination of
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647 the chemical oxygen demand (COD) using a copper electrode: a clean alternative method. Journal

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651 Ubbelohde, A.R.J.P. 1935. 380. Oxidation mechanisms in aqueous solution. Journal of the Chemical

652 Society (Resumed), 1605-1607

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653 Wagner, R. 1976. The COD method in Wasterwater Charges Act (AbwAG). Vom Wasser, 76, 139-

654 159. In German.

655 Zhang, S. Zhao, H. 2008. A new approach prevailing over chloride interference in the photo

656 electrochemical determination of chemical oxygen demand. The Analyst,

657 http://dx.doi.org/10.1039/B805095C.

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Table 1: COD-Determination of the two fractions CODFR and CODFL after sample preparation of municipal wastewater and industrial
wastewater with the developed method. Oxidation with Mn(III) (n=2) and Cr(VI)/Hg (n=1) reagents in comparison to the
standard method DIN 38409-41 (1980).

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Municipal Wastewater Industrial Wastewater
Cr/Hg Mn/Ag Cr/Hg Mn/Ag
ഥ
X DR RC ഥ
X RSD DR RCDIN ഥ
X DR RCDIN Xഥ RSD DR RCDIN

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DIN
(mg/L) (%) (%) (mg/L) (%) (%) (%) (mg/L) (%) (%) (mg/L) (%) (%) (%)
CODFR 535 74.7 74.7 405 1.1 73.6 256 9.3 10.6 224 2.88 10.1

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CODFL 181 25.3 25.3 146 8.4 26.4 2510 90.7 92.6 1992 1.17 89.9
CODFR + FL 716 100 100 550 3.0 100 76.9 2766 100 102 2216 0.76 100 81.8

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CODDIN 715 100 100 2710 100 100

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DR: Distribution ratio of the total COD (CODFR+FL) in the two fractions CODFR and CODFL
RCDIN: Recovery rate of the COD in the different fractions in relation to the CODDIN

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Table 2: Repeatability of COD-determination with the Mn(III)-method,
including sample preparation steps 1 and 2.
Municipal Municipal Landfill
Wastewater Wastewater leachate
A B
N=4 N=5 N=5

(CODFR) (mg/L)
X 339 391 355
SD (mg/L) 8.40 5.86 9.96

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RSD (%) 2.48 1.50 2.81

(CODFL) (mg/L)
X 238 101 566
SD (mg/L) 32.3 21.2 25.5

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RSD (%) 13.6 21.0 4.50

(CODFR+FL)
X 577 493 921
(mg/L)

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SD (mg/L) 29.5 20.8 33.3
RSD (%) 5.12 4.22 3.61

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20 mL WW sample

Add 10 mL oxidant
0.02 Mol (5.9 g) K2Cr2O7/L

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+ 80 g HgSO4/L

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Add 30 mL
Ag2SO4 (10 g/L)

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in conc. H2SO4

Reflux for 2 h at 148 °C

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Back-titrate
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non-consumed
Cr(VI) with Fe(II)
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Figure 1: Flow chart of standard COD method according DIN 38409-H41 (1980).
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turbid water sample clear
colloid / suspension or dilution real solution

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10 mL
sample 1 mL Al2(SO4)3
Step 1
1-2 dpl. NaOH

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separation of particles
und colloids

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filtration

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11 mL clear filtrate 10 mL clear sample

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acidification with H2SO4 to pH 1

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1 mL AgNO3
Step 2

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precipitation of AgCl
AgCl filter residue, to discard

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filter residue of
Al(OH)3 precip.
EP
5 mL aliquot of the filtrate
digestion with 50 mL
C

Mn(III) Step 3
AC

oxidation
CODFR CODFL

CODturbid = CODFR + 2.4 x CODFL CODclear = 2.2 x CODFL

Figure 2: Flow chart of sample preparation for turbid and clear water samples and oxidation with Mn(III) for COD determination.
 ACCEPTED MANUSCRIPT
300
Cr (VI)-COD (mg/L)
250
A

200

150

100

PT
50

RI
AgCl FR FR FL FL FR + FR + WW
- Hg - Hg + Hg - Hg + Hg FL FL + Hg
- Hg + Hg

SC
450
400 B
U
AN
350
Cr(VI)-COD (mg/L)

300
M

250
200
D

150
TE

100
50
EP

FR + FL of wastewater with Cr(VI)/AgCl

C

total wastewater with Cr(VI)/Hg

AC

Figure 3: Cr(VI)-COD values of municipal WW samples (dilution of 1:3, spiked with

1000 mg Cl-/L, and acidified with (H2SO4) to pH 1) after different sample
preparation steps,
A: Cr(VI)-COD of single fractions of one municipal WW sample, in
comparison to standard (Cr(VI)/Hg)-COD, n=1
B: Cr(VI)-COD of FR + FL of one municipal WW in comparison to standard
(Cr(VI)/Hg)-COD, n= 3.
FR: filter residue with particles (step 1 of sample preparation, without
using Al2(SO4)3)
 ACCEPTED MANUSCRIPT
FL: filtrate after separation of particles and AgCl precipitate (step 2 of
sample preparation)
WW: wastewater
- Hg: without addition of HgSO4
+ Hg: with addition of HgSO4

PT
RI
U SC
AN
M
D
TE
C EP
AC
 ACCEPTED MANUSCRIPT
1200
Landfill leachate
1000
Cr (VI)-COD (mg/L)

800

600

400

PT
200

RI
0
FR (HP) FL (HP) FR +FL (HP) WWtot

SC
1200

U
Municipal WW
1000
Cr(VI)-COD (mg/L)

AN
800

600
M

400
D

200
TE

0
FR (HP) FL (HP) FR + FL WWtot
(HP)
EP

Figure 4: Cr(VI)/Hg-COD values of landfill leachate and of municipal WW, (n=3)

C

after sample preparation with addition of Al2(SO4)3 (step 1).

AC

FR (HP): filter residue of Al (OH)3 precipitation and particles (step 1

of sample preparation)

FL (HP): filtrate after separation of Al(OH)3 precipitate and particles

(step 1 of sample preparation)

HP: hydroxide precipitate

WWtot: sample of wastewater (leachate) without sample

preparation
 ACCEPTED MANUSCRIPT

3.5

2.5
extinction

PT
1.5

RI
0.5

SC
0
200 300 400 500 600 700 800 900
wave length (nm)

Mn(III) reagent MnSO4 in RS

U KMnO4 in RS KMnO4+H2SO4
AN
Figure 5: UV/VIS-spectra of Mn(III) reagent (0.009 mol/L), KMnO4 (0.0017 mol/L)
M

and MnSO4 (0.038 mol/L) in the concentrations used in the Mn(III)

D

reagent mixture, and KMnO4 (0.0017 mol/L) mixed with H2SO4 in the
TE

concentration used for the Mn(III) reagent without addition of MnSO4 in

1 mm quartz cells.
EP

RS: reagent strength

C
AC
 ACCEPTED MANUSCRIPT

600
550
500
450
y = 0.8894x + 0.3429
Mn(III)-COD (mg/L)

400
R² = 0.9996
350

PT
300
y = 0.8486x - 1.8457
250
R² = 0.9999

RI
200
150

SC
100
50
0

U
0 50 100 150 200 250 300 350 400 450 500 550 600
AN
COD theoretical (mg/L)
M

Figure 6: Linearity of KHP oxidation with Mn(III);

continuous line: Mn(III) oxidation without sample preparation of KHP

D

standard solutions of 20, 50, 75, 100, 200 and 500 mg COD/L, n=9 (each
TE

replicate is represented by an extra symbol + , ,….., x….),

dashed line: Mn(III) oxidation after sample preparation step 2 of KHP

EP

standard solutions of 20, 50, 75, 100, 200 and 500 mg COD/L with 10 mg
C

Cl-/L respectively, n=5 (each replicate is represented by an extra symbol

AC

+, ,… .., x….).
 ACCEPTED MANUSCRIPT

3000

2500

PT
Mn(III)-COD (mg/L)

2000

RI
1500
y = 0.7999x - 1.9515
R² = 0.9935

SC
1000

500

U
AN
0
0 1000 2000 3000 4000
DIN-COD (mg/L)
M

Figure 7: Correlation of COD results of different water samples; Mn(III)-method

D

including sample preparation versus DIN 38409-41-method.

TE

Municipal WW A, B, C and D

Industrial WW A, B, C and D (food industry, textile cleaning, coke

EP

oven plant)

Landfill leachates A and B

C
AC
 ACCEPTED MANUSCRIPT

900
800
700 y = 6.1895x + 150.57
Mn(III)-COD (mg/L)

600 R² = 0.9992
500
400
300

PT
200 y = 4.156x + 34.174
100 R² = 0.9851
0

RI
0 20 40 60 80 100
Ratio of WW sample in the dilution (%)

SC
Industrial WW Municipal WW

Figure 8: Linearity of COD-values of a dilution series of an industrial and a

U
municipal WW sample, determined with Mn(III)-method including sample
AN
preparation.
M
D
TE
C EP
AC
 ACCEPTED MANUSCRIPT
Highlights

• Precipitation and separation of AgCl can substitute the use of neurotoxic Hg in

COD determination.

• Mn(III) proved to be an alternative oxidant for carcinogenic Cr(VI).

PT
• AgCl precipitation and Mn(III) oxidation can be successfully combined as

Cr(VI)/Hg free COD Method.

RI
• High correlation (R2= 0.9935) to the standard method was reached for

SC
wastewater samples with mean recovery rates of 78%

U
• Mn(III) COD might be able to be compared with existing Cr(VI) data by a
AN
correction factor.
M
D
TE
C EP
AC