JOURNAL OF SCIENCE INNOVARE Volume 03, Number 02, September 2020, Page 35-38

http://journal.unpak.ac.id/index.php/jsi p-ISSN: 2615-3769 ; e-ISSN: 2615-3750

DEVELOPMENT OF COD (CHEMICAL OXYGEN DEMAND)

ANALYSIS METHOD IN WASTE WATER USING UV-VIS
SPECTROPHOTOMETER
Uswatun Hasanah a*), Ade Heri Mulyati a), Sutanto a), Diana Widiastuti a),
Siti Warnasih a), Yulian Syahputri a), Tri Panjia)

a)
Universitas Pakuan, Bogor, Indonesia
Corresponding Author: uswatun.hasanah@unpak.ac.id

Article history: received 08 August 2020; revised 17 August 2020; accepted 29 August 2020

Abstract. The COD (Chemical Oxygen Demand) analysis method based on the Indonesian National Standard (SNI) is a
revision of SNI 06-6989.2-2004, Water and wastewater - Part 2: Method of testing for chemical oxygen demand (COD)
with closed reflux spectrophotometrically. This SNI uses references from international standard methods, namely
Standard Methods for the Examination of Water and Wastewater, 21st Edition, editor of LS Clesceri, AE Greenberg, AD
Eaton, APHA, AWWA and WEF, Washington DC, 2005, Methods 5220 D (Closed Reflux, Colorimetric Methods). The
purpose of this study was to validate the total method for testing chemical oxygen demand (COD) in water and
wastewater with the reduction of Cr2O7 2- spectrophotometrically in the range of COD values of 100 mg / L to 900 mg /
L measurements were made at a wavelength of 600 nm. and COD values less than or equal to 90 mg / L measurements
were made at a wavelength of 420 nm. The results showed that the method SNI 06-6898.2- 2009 has good validation
results including the r results obtained from the calibration curve equation of 0.998, a precision of 1.82%, an accuracy of
98.25% and LOD and LOQ of 12.27 mg / L and 25, 61 mg / L.

Keywords: wastewater; COD analysis; UV-VIS spectometry; validation, LOD

I. INTRODUCTION The COD analysis method based on the Indonesian

Water is one of the natural resources which has a very National Standard (SNI) is a revision of SNI 06-6989.2-2004,
important function for human life, as well as for advancing Water and wastewater - Part 2: Method of testing for
the general welfare so that it is the basic capital and the main chemical oxygen demand (COD) with closed reflux
factor of development. The main need for the spectrophotometrically. This SNI uses references from
implementation of good health is the availability of adequate international standard methods, namely Standard Methods
water in terms of quantity and quality, namely meeting the for the Examination of Water and Wastewater, 21st Edition,
requirements for cleanliness and safety. This water must also editor of LS Clesceri, AE Greenberg, AD Eaton, APHA,
be available continuously, attractive and acceptable to the AWWA and WEF, Washington DC, 2005, Methods 5220 D
community in order to encourage people to use it [1]. (Closed Reflux, Colorimetric Methods) [4].
Increased human activities will cause various problems, one This SNI has been tested in testing laboratories in the
of which is water pollution in water sources because it framework of validating and verifying the method and
receives a pollution load that exceeds its carrying capacity. consensed by the 13-03-S1 Technical Sub-Committee,
Pollution that causes a decrease in water quality can come Water Quality from the 13-03 Technical Committee,
from point sources and non-point sources. Centralized waste Environmental Quality and Environmental Management
such as industrial waste, livestock business waste, hotel with related parties. This SNI has been agreed and approved
waste, and hospital waste. Meanwhile, the waste is scattered, in a consensus meeting with meeting participants
such as agricultural waste, plantation waste and domestic representing producers, consumers, scientists, technical
waste [2]. agencies and related governments on 29 October 2008 in
Domestic waste is waste originating from bathrooms, Serpong, Tangerang - Banten and has been through a poll on
latrines, kitchens, washing clothes, washing household 18 March 2009 to 18 June 2009, with the final RASNI result.
appliances, pharmacies, restaurants and so on quantitatively. With the stipulation of SNI 06-6898.2-2019, the application
This waste consists of organic substances in the form of of SNI 06-6989.2-2004 is declared invalid. SNI users should
solid or liquid, hazardous and toxic materials (B3), dissolved be able to research SNI validation related to this method, so
salts, and bacteria, especially the fecal coli, pathogenic they can always use the latest edition of SNI [5].
bodies, and parasites [3]. The purpose of this research is to validate the total
method that must be done for each method of analysis, either

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JOURNAL OF SCIENCE INNOVARE Volume 03, Number 02, September 2020, Page 35-38
http://journal.unpak.ac.id/index.php/jsi p-ISSN: 2615-3769 ; e-ISSN: 2615-3750

for new methods or just repetition based on literature. sulfuric acid reagent solution into the tube or ampoule, as
Method validation has the main objective of demonstrating stated in the following table:
the reliability of a particular method (European Medicines
Table 1. Sample test and reagent solutions for various
Agency, 2011). This COD validation method is used for
digestion vessels
testing chemical oxygen demand (COD) in water and
wastewater with spectrophotometric reduction of Cr 2O7 2- in
the COD value range of 100. mg / L up to 900 mg / L
measurements were made at a wavelength of 600 nm and
COD values less than or equal to 90 mg / L measurements
were made at a wavelength of 420 nm [6].

II. RESEARCH METHODS

Curve Calibrartion
Calibration The calibration curve is made in the
Tools and Materials following steps:
The tools used in this study were DO bottles, a) turn on the instrument and optimize the
incubation cabinets or water coolers, temperature 20 ° C ± 1 ° spectrophotometer test instrument according to the
C, dark; bottles from glass 5 L - 10 L; volumetric pipettes instructions for using the tool for COD testing. Set the
1.0 mL and 10.0 mL; 100.0 mL volumetric flask; 200.0 mL wavelength at 600 nm or 420 nm;
and 1000.0 mL; PH meter; DO meter is calibrated; shaker; b) measure the absorption of each working solution then
blender; oven; and analytical scales. For COD, visible record and plot the levels of COD; create a calibration
spectrophotometer (400 nm to 700 nm), cuvette; digestion curve from the data above and determine the equation for
vessel, burette; volumetric flask 50.0 mL; 100.0 mL; 250.0 the straight line; b) if the linear regeneration correlation
mL; 500.0 mL and 1000.0 mL; 5.0 mL volumetric pipette; coefficient (r) <0.995, check the condition of the tool and
10.0 mL; 15.0 mL; 20.0 mL and 25.0 mL; beaker glass; repeat steps a) to c) until the coefficient r ≥ 0.995 is
magnetic stirrer. The materials used in this study were obtained. Measurement of the test sample
K2Cr2O7, H2SO4, HgSO4, Ag2SO4 powder, NH2SO3H For COD test samples 100 mg / L up to 900 mg / L a)
(Sulfamic Acid), demineralized water. cool the refluxed sample slowly to room temperature to
prevent the formation of sediment. If necessary,
Analysis of Cod in Wastewater Using a Uv-Vis occasionally open the sample cap when cooling to
Spectrophotometer prevent gas pressure; b) let the suspension settle and
The research method includes the preparation of make sure the part to be measured is really clear;
reagents, stock standard solutions and working standard c) measure the absorption of the test sample at a
solutions, optimization of detection parameters on the UV- predetermined wavelength (600 nm); d) calculate COD
vis spectrometer and validation of the analysis method. levels based on the linear equation of the calibration
Validation of the analysis method includes parameters of the curve; e) do a duplo analysis. - For samples of COD less
calibration curve, precision, accuracy, and recovery test.The than or equal to 90 mg / L a) cool the refluxed sample
validation of the COD analysis method carried out is the slowly to room temperature to prevent the formation of
total validation which refers to SNI 6989.2: 2009. Validation sediment. If necessary, occasionally open the sample cap
parameters include linearity, accuracy, precision, LOD and when cooling to prevent gas pressure; b) let the
LOQ. suspension settle and make sure the part to be measured
is really clear; c) use water reagent as reference solution;
1) Taking COD test samples d) measure the absorption of the test sample at a
Preparation of test samples predetermined wavelength (420 nm); e) calculate COD
Who is the test sample, namely by a) homogeneous levels based on the linear equation of the calibration
the test sample; b) wash digestion vessel and cover with 20% curve; f) do a duplo analysis.
H2SO4 before use; Preservation of test samples If the test
sample cannot be tested immediately, then the test sample is
preserved by adding concentrated H2SO4 until the pH is less III. RESULTS AND DISCUSSION
than 2 and stored in a cooler at a temperature of 4 ° C ± 2 °
C with a maximum recommended shelf life of 7 days. 3.5 In this study, the COD method validation refers to
Preparation of working solutions Create a work solution SNI 06-6898.2-2019. The sample used was sago waste water.
series from the KHP main solution with 1 (one) blank and at Preparation of test samples by washing 16x100mm size
least 3 proportionally different levels that are in the culture tubes using 20% H2SO4. Preservation of the test
measurement range. samples using concentrated H2SO4 until the pH is less than 2,
Digestion procedure then stored in a cooler at 4˚C ± 2˚C with a storage time of 1
pipette the volume of the test sample or working week.
solution, add the digestion solution and add a sufficient

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JOURNAL OF SCIENCE INNOVARE Volume 03, Number 02, September 2020, Page 35-38
http://journal.unpak.ac.id/index.php/jsi p-ISSN: 2615-3769 ; e-ISSN: 2615-3750

Validation Results of Cod Analysis Method in the linear regression equation for the calibration curve are y
Wastewater Using a Uv-Vis Spectrophotometer = 0.0004x + 0.0051 with a value of r = 0.999. Following the
recovery results are presented in table IV.
A. Linearity
The result of the determination of the KHP Table 4. COD accuracy test results
(Potassium Hydrogen Phthalate) calibration curve is a linear
line equation that can be used to calculate COD levels in Sample % Recovery
wastewater samples. The linear regression line equation 1 96.39
illustrates the relationship between the KHP concentration 2 98.06
and the uptake of the KHP reaction compound with its 3 99.66
reagents. The determination of the calibration curve in this 4 98.03
study used 5 series of concentrations, namely 100 mg / L, 5 96.33
200 mg / L, 300 mg / L, 400 mg / L, 500 mg / L with 2 6 99.65
repetitions each. The wavelength used for absorption is 7 99.66
600nm. The results of the KHP standard curve determination Average 98.25
are presented in table II.
Accuracy is expressed as a percentage of recovery (%
Table 2. Result of Determination of KHP Calibration
recovery). The test results showed that the average%
Curve to Determine recovery in COD levels gave good results and met the
KHP
concentration Repetition 1 Repetition 2 accuracy test requirements of 98.25%. The% recovery result
(mg / L) is said to meet the requirements if it shows a percentage
100 0.045 0.041 value between 85% -115%.
200 0.096 0.085
300 0.128 0.123
400 0.171 0.174
D. Detection Limit and Quantitation Limit Test (LOD
500 0.213 0.205 and LOQ)
r 0.998 0.998 The determination of LOD and LOQ in this study
Linear Regression y = 0.0004x +0.0073 y = 0.0004 + 0.0005 was carried out by making a blank for 11 replications and a
Line Equations R2 = 0.9964 R2 = 0.996
sample of 5 replications. Based on the results of LOD and
LOQ measurements obtained, respectively, are 12.27 mg / L
The calibration curve has a good correlation
and 25.61 mg / L. The results of LOD and LOQ show that
coefficient (r) with a value of 0.998 each. This shows that
the sensitivity of the method is quite good.
the calibration curve is linear because r> 0.995 is in
accordance with SNI 06-6898.2-2009. The value of r that
approaches 1 indicates a relationship between concentration
IV. CONCLUSION
(x) and response (y) which is interpreted as linearity.
Based on the research results, it can be concluded that
B. Accuracy / Precision
the method SNI 06-6898.2-2009 has good validation results
Determination of the precision value is done in 7
including the r results obtained from the calibration curve
repetitions. precision is determined based on the standard
equation of 0.998, a precision of 1.82%, an accuracy of
deviation (SD). Based on the test results, it can be seen that
98.25% and LOD and LOQ of 12.27 mg / L and 25, 61 mg/L.
the UV-VIS spectrophotometric method used for
determining COD levels has a precision of 1.82% <2%.
Acknowledgements
Presented in table III.
We want to thank Fakultas Matematika dan Ilmu
Table 3. COD Level Precision Pengetahuan Alam Universitas Pakuan for funding this
No. COD levels % research and to Threcia Valentina for helping research in the
Average SD
Sample (mg / L) RSD laboratory.
1 146.75
2 151.75
3 146.75 REFERENCES
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JOURNAL OF SCIENCE INNOVARE Volume 03, Number 02, September 2020, Page 35-38
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[4] Apha. 1989. Standard methods for the examination of

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