COD REDUCTION AND DECOLORIZATION OF SWEET SORGHUM

DISTILLERY SLOPS BY SEQUENTIAL ANAEROBIC DIGESTION

AND PRELIMINARY ELECTROOXIDATION

AARON JAMES ESPIRITU PALACPAC

SUBMITTED TO THE FACULTY OF

DEPARTMENT OF CHEMICAL ENGINEERING
COLLEGE OF ENGINEERING AND AGRO-INDUSTRIAL TECHNOLOGY
UNIVERSITY OF THE PHILIPPINES LOS BAÑOS
IN PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR THE
DEGREE OF

BACHELOR OF SCIENCE IN CHEMICAL ENGINEERING

APRIL 2012

Permission is given to the following people to have access to this thesis:

Available to the general public No

Available only after consultation with author/thesis adviser Yes
Available only to those bound by confidentiality agreement Yes

Signature of Student: ___________________

Signature of Thesis Adviser: ___________________
 The thesis attached hereto, entitled “COD REDUCTION AND
DECOLORIZATION OF SWEET SORGHUM DISTILLERY SLOPS BY
SEQUENTIAL ANAEROBIC DIGESTION AND PRELIMINARY
ELECTROOXIDATION” prepared and submitted by AARON JAMES ESPIRITU
PALACPAC in partial fulfillment of the requirements for the degree of BACHELOR
OF SCIENCE IN CHEMICAL ENGINEERING is hereby accepted.

________________________________ ________________________________
DHAN LORD B. FORTELA MICHAEL VINCENT O. LAURIO
Member Member
Guidance Committee Guidance Committee
__________ __________
Date Signed Date Signed

________________________________ ________________________________
VERONICA P. MIGO CATALINO G. ALFAFARA
Co-Adviser Adviser and Chair
Guidance Committee Guidance Committee
__________ __________
Date Signed Date Signed

______________________________
MANOLITO E. BAMBASE JR.
Chair
Department of Chemical Engineering
__________
Date Signed

______________________________
ARNOLD R. ELEPAÑO
Dean
College of Engineering and Agro-Industrial Technology
__________
Date Signed
 BIOGRAPHICAL SKETCH

Born on September 19, 1988 in the town of Sta.Cruz, Laguna, the author is the

eldest child of Aldo B. Palacpac, a police officer, from Los Baños, Laguna and Julieta V.

Espiritu, a registered nurse, of Nagcarlan, Laguna. He started pre-school at Nagcarlan

Day Care Center and attended Kindergarten at Holy Angel School in Los Baños, Laguna.

He finished his basic and secondary education at Maquiling School Incorporated and at

Los Baños National High School, respectively.

In 2005, he entered the University of the Philippines Los Baños (UPLB) and took

up Bachelor of Science in Chemical Engineering. He also served as a UPLB ROTC cadet

officer and hailed as the Corps Commander from 2006 to 2008.

AARON JAMES E PALACPAC

iii
 ACKNOWLEDGMENT

With profound gratitude and sincere appreciation, I would like to thank the
following persons and institutions that helped me in the completion of this study.

Dr. Catalino G. Alfafara, my thesis adviser and chair of my guidance

committee, for his patience, guidance, suggestions and encouragement starting from the
thesis outline to the completion of this work. Your words of wisdom and encouragement
have given me a profound impact in conducting research work and served as an
inspiration in the Chemical Engineering Department.

Dr. Veronica P. Migo, my co-adviser, for accommodating me to conduct this

study in the Central Analytical Services Laboratory (CASL) of BIOTECH; for
accompanying me in the sleepless overnight experiments in the laboratory and for the
suggestions, encouragement and guidance in doing this thesis.

Engr. Michael Vincent O. Laurio and Engr. Dhan Lord B. Fortela, members
of my guidance committee, for offering their time, for listening and commenting for my
work to be better. Every suggestion you have made contributed to the improvement of
this manuscript.

Engr. Rhebner E. Arocena, my partner in the conduct of this study, for

accompanying me in the completion of this work and for the enthusiasm in working
together in the laboratory despite the number of runs we need to perform.

Engr. Maria Cristina V. Mahilum, my panelist in my thesis outline defense, for

the suggestions and comments in the conduct of this study and for being lab mates for
quite a short time in CASL.

Prof. Monet Concepcion C. Maguyon, my academic and previous thesis

adviser, for accepting me as your advisee and for making sure that you have transferred
me to your mentors. Your suggestions and comments on my previous study are very
much appreciated.

I would also like to thank the staff of CASL, for guiding me during my stay in the
laboratory. To Sir Lorenz Fabro, for guiding us the proper procedure on the equipments
in the laboratory; to Tita Resie Valderrama, for being a second mother in the lab, and
your influence in the other staffs made my work easier; to Tito Hermie Lapitan, for
helping me cleaning some of the lab equipments and apparatus during the conduct of my
study and also for the conversations during merienda breaks in the lab; to Dr. Lorele
Trinidad, for accompanying me, together with Dr. Migo, during my overnight stay in the
lab; to the Research Associates, Kuya Emil, Kuya Jeff, Kuya Carlo and Kuya Arn, for
the conversations and funny moments in the lab. I would not enjoy my stay in CASL
without the help and presence of these people.

iv
 To the faculty and staff of the Chemical Engineering Department, for giving
us, UPLB Chemical Engineering students, the best education we need. Without your
guidance and support, we will not succeed in our chosen field. Indeed, you are the best
mentors during our stay in this university.

I would also like to extend my appreciation to the Valderrama Family especially

to Tita Resie Valderrama, and to Engr. Arnulfo Valderrama, for accomodating me in
their home in times I need to stay out doing this study and other academic related
activities. I found my second home in your hands.

I would also like to share my deepest thanks to my buddies since high school, the
Butete Squad: Arvin Valderrama, Carlo Remo, Chancellor Molinyawe, Jastin
Revilleza, Garee Hernandez, Gebriel Remolacio, Patrick Garcia and Van Joseph
Rofiles. Without your encouragement, I would not be able to find strength in completing
this piece of work.

To my fraternal brothers in the UP Vanguard Incorporated-Los Baños

Chapter, I would like to thank and salute my mentors for developing in me courtesy,
discipline and stability under pressure and for accepting me to be part of the “Chosen
Few.” I would also extend my warmest gratitude to you Sirs, for indoctrinating me our
shibboleth of DUTY well performed, HONOR untarnished and COUNTRY above-self.

To Lt.Col. Vivian A. Gonzales, UPLB ROTC Commandant, and to the DMST

staffs, who encouraged me in the completion of this work and for the appreciation of my
endeavors for the UPLB ROTC unit and for the UPLB Corps of Cadets.

To my family: Tita Neng, who accompanied me since my childhood days until

my final days as an undergrad student, and tirelessly done her role as a second mother to
me; Tita Resy, who guided me the true essence of being a Christian and a second mother
as well and to the advises in writing this thesis manuscript; to my sister Ara, who have
accompanied me through all the years and encouraging me to finish my degree; and to
my dearest parents: Mommy and Daddy, who have supported me in my chosen field,
this work is dedicated to you.

Most of all to the Almighty Lord, who makes all things happen. To You be all
the glory.

Do not go where the path may lead. Go where there is no path and leave a trail.
-Ralph Waldo Emerson

v
 TABLE OF CONTENTS

PAGE

TITLE PAGE i

APPROVAL PAGE ii

BIOGRAPHICAL SKETCH iii

ACKNOWLEDGMENT iv

TABLE OF CONTENTS vi

LIST OF TABLES xi

LIST OF FIGURES xii

LIST OF APPENDICES xiv

LIST OF APPENDIX TABLES xv

ABSTRACT xvii

1. INTRODUCTION 1

1.1. Background and Significance of the Study 1

1.2. Objectives 4

1.3. Scope and Limitations of the Study 5

1.4. Time and Date of the Study 5

2. REVIEW OF RELATED LITERATURE 6

2.1. Bioethanol production from sweet sorghum 6

Sweet Sorghum 6

Conversion of sweet sorghum to ethanol 7

vi
 PAGE

Wastewater generation from the production of 8

sweet sorghum to ethanol

2.2. The Philippine Biofuels Industry 9

Republic Act 9367 or The Biofuels Law of 2006 9

Wastewater regulations 10

2.3 Wastewater Characteristics 11

Biochemical Oxygen Demand (BOD) 11

Chemical Oxygen Demand (COD) 12

Color 12

pH 12

Temperature 13

2.4. Distillery Wastewater Treatment Methods 13

Biological Treatment 13

Physico-chemical Treatment 14

2.5. Anaerobic Digestion Process 14

General Process Description 14

Conditions and Variables Influencing Anaerobic 16

Digestion

pH and buffering 17

Temperature 17

Nutrient and sludge addition 17

Hydraulic Retention Time (HRT) 18

Organic Loading Rate (OLR) 18

vii
 PAGE

Agitation 19

2.6. Electrochemical Processes 19

Principle of Electrolysis 19

Faraday’s Law of Electrolysis 20

Charge Dose 21

2.7. Ways of Electrochemical Treatment of Wastewater 21

Direct Electrolysis 22

Indirect Electrolysis 22

Electrofloatation 23

Electroflocculation 23

Electrocoagulation 24

Electrooxidation 24

3. METHODOLOGY 27

3.1. Preparation and Characterization of Sweet Sorghum 25

Distillery Effluent

3.2. Anaerobic Digestion Experiments 26

Preparation and characterization of inoculum 26

Batch Anaerobic Digestion (Acclimatization 26

Phase) and Semi-continuous Anaerobic Digestion
Set-up

Experimental Conditions for Anaerobic Digestion 28

Analytical Methods 29

pH 29

Gas Production 29

viii
 PAGE

Color Analysis 30

COD analysis 30

Data Analysis 31

3.3. Preliminary Electrooxidation Experiments 31

Batch Electrolysis Set-up 31

Experimental Conditions 32

Analytical Methods 33

Data Analysis 33

Scale-up and Operating Conditions 33

Charge Dose 33

Energy Requirement and Preliminary Cost 34

Estimation

4. RESULTS AND DISCUSSION 35

4.1. Wastewater characteristics 35

4.2. Time course profiles for wastewater parameters operated

at different organic loading rates (OLR) during
acclimatization 36

4.3. Anaerobic Digestion of Sweet Sorghum Distillery Slop at 37

different organic loading rates (OLR) in the Semi-
continuous experiments

pH profiles 38

Residual Chemical Oxygen Demand 39

(COD) profiles

Gas production profiles 42

ix
 PAGE

Residual Color profiles 44

4.4. Preliminary Electrooxidation of the Chloride- 45

Supplemented Sweet Sorghum Biodigester Effluent

pH and Temperature profiles after electrooxidation 45

Residual COD profiles 46

Decolorization Efficiency profiles 49

Charge Dose and Energy Requirements 51

Computations of the Electrooxidation Process

5. SUMMARY AND CONCLUSION 55

6. RECOMMENDATIONS 58

7. REFERENCES 60

8. APPENDICES 63

x
 LIST OF TABLES

TABLE PAGE

2-1 Comparison of cost for different ethanol feedstock in 6

the Philippines (Dar, 2007)
2-2 Characteristics of distillery stillage from molasses, 8
cane juice and mixed (molasses+juice)
(Demafelis, 2010)
2-3 Effluent standards for inland water Class C 11
(DENR, 1990)
2-4 Environmental and operating conditions for maximum 16
methane production (Malina and Pohland, 1992)
3-1 Schedule of Analysis for Batch and Semi-Continuous 28
Anaerobic Digestion Experiments
4-1 Characteristics of sweet sorghum distillery effluent 35
compared to the effluent standards of DAO No. 35
4-2 Energy requirements for color removal and COD 54
reduction for digested effluent

xi
 LIST OF FIGURES

FIGURE PAGE

2-1 Anaerobic Digestion Process 15

2-2 Mechanism of Direct and Indirect Electrolysis 22

2-3 Mechanism of Electrofloatation 23

2-4 Mechanism of Electrocoagulation 24

3-1 Simple distillation set-up for the generation of sweet 25

sorghum distillery effluent (slop)

3-2 Anaerobic Digestion Experimental Set-up 27

3-3 Actual Anaerobic Digestion Experimental Set-up 27

3-4 Orion pH meter 29

3-5 SHIMADZU UV-1601: UV-Vis spectrophotometer 30

3-6 Set-up for dichromate open-reflux method for COD 31

determination

3-7 Diagram of batch electrolysis set-up 32

4-1 Sweet sorghum distillery slop 35

4-2 Time course profiles for pH, residual COD, COD removal 37
efficiency, and gas production during acclimatization

4-3 pH profile of digesters at the two hydraulic retention time 38

in the semi-continuous experiments

4-4 Residual COD for the two HRT in the semi-continuous 40

experiments

4-5 Percent (%) COD removal for the setup of HRT = 60 41

days in the semi-continuous experiments

xii
FIGURE PAGE

4-6 Mean cumulative gas production for the two HRT in the 43
semi-continuous experiments

4-7 Flame test for gas production in the semi-continuous 43

experiment

4-8 Residual color for the two HRT in the semi-continuous 45

experiments

4-9 pH and temperature time courses during the electro- 46

oxidation of anaerobically digested sweet sorghum slops

4-10 COD time course in electro-oxidation 47

4-11 Foams formed during electrolysis 48

4-12 COD removal efficiency in electro-oxidation 48

4-13 Residual color time course in electro-oxidation 49

4-14 Color removal efficiency in electro-oxidation 50

4-15 Color removal of wastewater in electro-oxidation 50

4-16 Charge dose determination for COD removal in electro- 51

oxidation

4-17 Charge dose determination for color removal in electro- 52

oxidation

xiii
 LIST OF APPENDICES

APPENDIX PAGE

A. Raw Data 64

B. Derived Data 77

C. Sample Calculations 81

D. Chemical Oxygen Demand (COD) Determination 87

E. Material Safety Data Sheets 90

xiv
 LIST OF APPENDIX TABLES

APPENDIX PAGE
TABLE

A1 pH profile of digesters during acclimatization 64

phase
A2 Cumulative gas production of the digesters during 64
acclimatization phase
A3 Volume of standard ferrous ammonium sulfate 65
titrant used for 10-mL sample volume
(acclimatization phase)

A4 pH profile of digesters during semi-continuous 66

anaerobic digestion experiments
A5 Cumulative gas production of the digesters during 67
semi-continuous anaerobic digestion experiments
A6 Volume of standard ferrous ammonium sulfate 69
titrant used for 10-mL sample volume (semi-
continuous phase)
A7 Absorbance profile of sweet sorghum distillery 72
slop for the semi-continuous anaerobic digestion
experiments
A8 pH and temperature profile at an operating 74
current of 1.5A and 25 g/L NaCl supplementation
A9 Volume of standard ferrous ammonium sulfate 75
titrant used for 10-mL sample volume
(electrooxidation experiments)
A10 Absorbance profile digested effluent from 76
preliminary electro-oxidation
B1 COD profile of digesters during acclimatization 77
phase
B2 COD reduction profile of digesters during 77
acclimatization phase
B3 COD profile of digesters at each HRT for the 77
semi-continuous anaerobic digestion of sweet
sorghum distillery slop
B4 COD reduction profile of digester at HRT = 60 78
days for the semi-continuous anaerobic digestion
of sweet sorghum distillery slop
B5 COD profile of digested effluent before and after 79
electro-oxidation at 1.5A operating current and 25
g/L NaCl supplementation.

xv
APPENDIX PAGE
TABLE

B6 COD reduction profile of the digested effluent 79

before and after electro-oxidation treatment
B7 Color reduction profile of the digested effluent 79
before and after electro-oxidation treatment
B8 Charge dose determination for color removal and 80
COD reduction of the digested effluent at 1.5A
and 25 g/L NaCl supplementation
C1 Volume of standard ferrous ammonium sulfate 81
titrant used for 10-mL sample volume (sample
computation)
C2 COD reduction profile of the digested effluent at 83
1.5A operating current and 25 g/L NaCl
supplementation

xvi
 ABSTRACT

PALACPAC, AARON JAMES ESPIRITU, College of Engineering and Agro-

Industrial Technology, University of the Philippines Los Baños, April 2012. COD
Removal and Decolorization of Sweet Sorghum Distillery Effluent by Sequential
Anaerobic Digestion and Preliminary Electrooxidation.

Major Adviser: Dr. Catalino G. Alfafara

Co-Adviser: Dr. Veronica P. Migo

Chemical oxygen demand (COD) reduction and decolorization of sweet sorghum

distillery effluent was investigated using anaerobic digestion followed by preliminary
electrooxidation. Anaerobic digestion was conducted in order to investigate the effects of
organic loading rates (OLR) in the anaerobic treatment of the wastewater. OLRs of 5.534
mg COD/L-d (HRT=30d) and 2.766mg COD/L-d (HRT=60d) were used in the study
under a semi-continuous mode of operation. The organic loading for the 30d HRT
digester was considered to be too high for the proper functioning of an anaerobic
digestion treatment system. The steady-state pH was acidic with no removal of COD.
On the other hand, the digester with 60d HRT showed a stable digester operation. A pH
range of 6.87 to 8.42 was achieved which was in the range of optimum anaerobic
digestion. The COD removal efficiency was 30.82%, and gas production was 1,819
mL/day. A second stage treatment of 12-hr electrooxidation at 1.5A and 2.5% chloride
supplementation resulted in further removal of 24% of COD and 41% of color. Increase
in both temperature and pH were observed during the electrochemical treatment, but
these eventually leveled-off to stable values. Charge dose for COD removal (a scale-up
operating factor) was found to be 11.632 Coulombs/mg COD removed. The computed
energy requirement was found to be 1.75x10 -5 kWh/mg pollutant removed while the
energy cost was PhP1,159.14/m3. The resulting effluent after anaerobic digestion and
preliminary electrooxidation were not yet compliant with DENR Class C standards.
Further studies are recommended to extend operation at 60d HRT (anaerobic digestion)
and to increase chloride supplementation or operating current in electrooxidation to
improve DENR compliance.

xvii
 1. INTRODUCTION

1.1. Background and Significance of the Study

Production of biofuels from agricultural materials, as well as organic waste

materials, for use as an alternative fuel has been attracting worldwide interest because of

the increasing demand for limited non-renewable energy resources and unpredictable

prices of oil and natural gas. Climate change mitigation is also considered as one of the

driving forces for the use of biofuels. The production and use of biofuels largely depend

on the potential available feedstock sources for which further studies are being conducted

in order to produce a renewable energy in an environmentally responsible way (Biofuel

Technology Handbook, 2008). Most of the countries worldwide are developing their

source for the production of biofuels in order to achieve sustainable energy security and a

less-polluting environmental landscape.

The Philippines is one the countries that depends on energy source from other

nations. But through the creation of Republic Act 9367, otherwise known as the Biofuels

Law of 2006, dependence on imported oil, such as diesel and ethanol, will be minimized.

The Biofuels Law ensures the availability of alternative and clean energy without any

detriment to the natural ecosystem, biodiversity and food reserves of the country. This

law also mandates biofuel-blending on gasoline and diesel fuels. Since its implementation

in 2007, five percent (5%) and one percent (1%) blending of bioethanol and biodiesel

were used for gasoline and diesel respectively. Four years after the implementation of this

law, blending for both fuels will be increased. Ten percent (10%) bioethanol blending

will be required for gasoline and two percent (2%) biodiesel for diesel fuels.
 2

Several feedstocks have been used for the production of bioethanol. Corn is the

primary used feedstock in the US while sugar cane is used in Brazil. These two countries

are the largest producer of ethanol in the world market (Kaparaju et al, 2010). In the

Philippines, corn and sugar cane are primarily consumed by Filipinos as food; thus, the

use of these crops for biofuels is discouraged, due to the “food versus fuel” debate. To

resolve this issue, several feedstocks have been eyed for use in the production of

bioethanol. One feedstock that is under consideration is sweet sorghum. India is presently

the largest producer of ethanol, using sweet sorghum as the feedstock (Pant and

Adholeya, 2006). Ethanol is produced by fermentation of the sweet sorghum juice

(extracted from the stalk), which has a sugar content of about 16% to 23% Brix (Dar,

2007).

There is presently no ethanol distillery in the Philippines that uses sweet sorghum

as its feedstock. With the implementation of the Biofuels Law, an increase in ethanol

distilleries is expected in order to cope up with the demand for ethanol in the market.

Sweet sorghum will be one of the priority feedstock since it will not be included in the

food versus fuel issue. In order to comply with the mandates of RA 9367 in producing

clean energy without any detriment to the natural ecosystem of the country, an

environmental component for the development of sweet sorghum production facility

should be considered. Production of ethanol produces wastewater in the form of slops.

Ten to 18 liters of slops are produced per liter of ethanol which will be equivalent to

75,000 mg/L to 120,000 mg/L COD (Demafelis, 2010). Aside from a high COD content,

slops have very dark brown color, high ash content and low pH which will be difficult to

dispose on the environment (Pant and Adholeya, 2006).

 3

In order to reduce the pollution that can be potentially caused by distillery slops

and to meet the wastewater effluent standards of in the country, biological and physico-

chemical treatment should be used. Most distilleries that use molasses for the production

of ethanol have used anaerobic digesters as part of their wastewater treatment facility

(Wilkie et al, 2000). Distillery slops from ethanol production using molasses spent wash

resulted 60 to 70% maximum COD removal using anaerobic digestion (Banerjee and

Biswas, 2004). The residual COD after anaerobic treatment is usually attributed to non-

biodegradable organic color compounds (brown to dark-brown Maillard reaction

products). In order to remove the remaining 30 to 40% non-biodegradable COD, a second

stage treatment involving advance oxidation is usually needed. Strong oxidizing

treatments are necessary to destroy the remaining organic color substances which resist

biological treatment. However, previous studies on the management of distillery are more

on the treatment of molasses-based slops, and little or no studies have so far been made

on sweet sorghum-based slops.

In this study, the treatability of sweet sorghum distillery effluent using anaerobic

digestion and preliminary electro-oxidation was investigated. A first-stage biological

treatment was first applied using anaerobic digestion for the purpose of removing most of

the biodegradable COD of the distillery slop. An acclimatization phase was first done in

order to have an active microbial population for the anaerobic digester. Effect of two

organic loading rates on the treatment performance in a semi-continuous system was

investigated. Parameters such as pH, gas production, color and COD were monitored. A

second-stage advance oxidation using electro-oxidation was also needed for the removal

of remaining non-biodegradable COD (in the form of color) was also investigated. Batch
 4

electrolysis was performed to investigate the effect of current on the decolorization

profile of the treated effluent as well as COD removal. Parameters such as color, pH and

COD were monitored. COD measurements were done for initial, middle and final

sampling interval for the operating current. The results were used to obtain engineering

relationships useful for operation and scale-up. These relationships were based on the

concept of charge dose.

1.2 Objectives of the Study

The general objective of this study is to assess the treatability of sweet sorghum

distillery effluent through anaerobic digestion and preliminary electro-oxidation

treatment. Specifically, this study aimed:

1. To assess wastewater characteristics in terms of pH, color, and organic matter

such as chemical oxygen demand (COD), and biochemical oxygen demand

(BOD);

2. To determine the effect of two organic loading rates (2,766 mg COD/L-day,

5,533.61 mg COD/L-day) on treatment performance in a semi-continuous system

in terms of pH, color, COD (and if possible, BOD), and biogas production, after

an initial start-up or acclimatization in the batch mode;

3. To determine which of the two organic loading rates would give a better

anaerobic digestion performance; and

 5

4. To use the digester slop for the second stage preliminary electro-oxidation

experiments, and make initial investigations on the effect of operating current and

chloride supplementation on decolorization and COD removal.

1.3. Scope and Limitations of the Study

This study focused on the anaerobic treatment of sweet sorghum distillery slop

primarily done for COD removal. A second stage treatment using electro-oxidation was

done in order to further decolorize the sweet sorghum slop after biological treatment. The

sweet sorghum distillery slop was produced using a laboratory-to-pilot scale fermentation

and distillation and could not reflect the properties of that in large-scale slop production.

Population of the anaerobic bacteria was not considered due to complexity and

constraints of time and resources. The effect of temperature to gas production was also

not considered. For the electro-oxidation part, the effect of different currents was not

considered but sample points were increased in order to have reliable data for the analysis

of the effect of current to color removal efficiency.

1.4. Time and Date of the Study

This study was conducted at the Central Analytical Services Laboratory (CASL)

of the National Institute of Molecular Biology and Biotechnology (BIOTECH),

University of the Philippines Los Baños (UPLB) from June 2011 to October 2011.
 2. REVIEW OF LITERATURE

2.1. Bioethanol Production from Sweet Sorghum

Sweet Sorghum

Sweet sorghum (Sorghum bicolor) belongs to the family of the Poaceae. The

genus of sorghum has many species of which sweet sorghum is one of the most grown

plants. Similar to sugarcane it is cane-like plant with a high sugar content of the stalk

(Rutz and Janssen, 2008). It grows under drier and warmer conditions than many other

crops. It only needs 1/3 water of sugarcane and is tolerant to drought, heat, water logging

and salt-alkali. But sweet sorghum can be grown in temperate areas as well. Additionally,

sweet sorghum has a relatively short growing season and can be harvests 1-3 times per

year. In contrast to sugarcane, sweet sorghum can be also sown very well and has not to

be propagated by stalk clippings.

In the Philippines, sweet sorghum is envisioned to be an effective

alternative feedstock to sugarcane for ethanol production (Dar, 2007). Table 2-1 shows

the comparison for different feedstock cost and their ethanol yield in the Philippines.

Table 2-1. Comparison of cost for different ethanol feedstock in the

Philippines
FEEDSTOCK
PRICE (Php)/MT
FEEDSTOCK Liter/ha/year COST (PhP)/Liter
Min Max Min Max
Sugarcane 1,000 1,100 6,120 13.89 15.28
Molasses 4,550 5,400 806 19.06 22.62
 7

Table 2-1 continued . . .

FEEDSTOCK
PRICE (Php)/MT
FEEDSTOCK Liter/ha/year COST (PhP)/Liter
Min Max Min Max
Cassava 1,500 5,800 5,549 8.38 32.4
Corn 8,500 10,000 5,282 20.92 24.61
Sweet
8,138 13.98 15.672
Sorghum
- Stalk 550 600 5,625 12.22 13.33
- Grain 6,000 7,000 2,513 17.91 20.90
Source: Dar, 2007

Conversion of Sweet Sorghum to Ethanol

Ethanol can be produced from either the grain or stalk juice of sweet sorghum.

Fermentation of sweet sorghum juice is done using the specific yeast Saccharomyces

cerevisiae, which is frequently used to ferment glucose to ethanol. Traditional

fermentation processes rely on yeasts that convert six carbon sugars to ethanol (Rutz and

Janssen, 2008). Theoretically, 100 grams of glucose will produce 51.4 g of ethanol and

48.8 g of carbon dioxide.

Ethanol produced by fermentation results in a solution of ethanol in water. For

ethanol to be purified, water must be removed. Distillation is used to separate water and

ethanol but purity of the latter will be around 95-96% only due to the formation of water-

ethanol azeotrope. In order to produce the required ethanol purity of 99.5 to 99.9%,

further dehydration is required using molecular sieves. The final product is called

anhydrous ethanol.
 8

Wastewater Generation from the Production of

Sweet Sorghum to Ethanol

Distillation is one of the unit operations involved in the production of ethanol

from sweet sorghum. Like other ethanol distilleries that use molasses, waste generation is

in the form of slops (stillage). Molasses spent wash (MSW) is one of the most difficult

waste to dispose because of high pH, high temperature, dark brown color, high ash

content and high percentage of dissolved organic and inorganic matter (Pant and

Adholeya, 2006).

In a study cited by Demafelis (2010) on waste management in the emerging

biofuels industry in the Philippines, 10 to 18 liters of slops are produced per liter of

ethanol which will be equivalent to 75,000 to 120,000 mg COD per liter of slop. In 2010,

the bioethanol requirement of the country reached around 3,000 MLi and that slop

produced was around 5,000 MLi. Characteristics of distillery slops using molasses and

juices from the presentation of Roxol Bioenergy Corporation were summarized in Table

2-2.

Table 2-2. Characteristics of distillery stillage from molasses, cane juice & mixed (molasses +
juice)

CHEMICAL DATA MOLASSES JUICE JUICE + MOLASSES

pH 4.2 - 5.0 3.7 - 4.6 4.4 - 4.6
Temperature (oC) 75 - 95 75 - 95 75 - 95
BOD (mg O2/L) 45,000 6,500 - 16,500 19,800
COD (mg O2/L) 120,000 15,000 - 33,000 45,000
Total Solids (mg/L) 90,500 23,700 52,700
Volatile Solids
65,000 20,000 40,000
(mg/L)
Fixed Solids (mg/L) 25,500 3,700 12,700
Nitrogen (mg N/L) 450 - 1,600 150 - 700 480 - 710
 9

Table 2-2 continued . . .

CHEMICAL DATA MOLASSES JUICE JUICE + MOLASSES

Phosporus (mg
100 - 290 10 - 210 9 - 200
P2O5/L)
Potassium (mg K2/L) 3,740 - 7,830 1,200 - 2,100 3,340 - 4,570
Calcium (mg Ca/L) 450 - 5,180 130 - 1,540 1,330 - 4,600
Magnesium (mg
420 - 1,520 200-490 580 - 700
MgO/L)
Sulphate (mg SO4/L) 6,400 600 - 700 3,700 - 3,730
Carbon (mg/L) 11,200 - 22,900 5,700 - 13,400 8,700 - 12,100
C/N Rate 16.0 - 16.27 19.7 - 21.07 16.4 - 16.43
Organic matter (mg/L) 63,400 19,500 38,000
Reducing matter
9,500 7,900 8,300
(mg/L)
Source: Demafelis, 2010

From Table 2-2, parameters of distillery slops such as pH, temperature, BOD,

COD, total solids and organic matter are indicators that wastewater from distilleries are

needed to be treated for it to pass standards set for discharge to the environment.

2.2. The Philippines Biofuels Industry

Republic Act 9367 or the Biofuels Law of 2006

The Philippines roadmap towards energy self-sufficiency can be clearly viewed

through the implementation of the RA 9367 also known as Biofuels Law. It mandates the

use of biofuels and aims to:

(a) develop and utilize indigenous renewable and sustainable clean energy

sources to reduce dependence on imported oil;

(b) mitigate toxic and greenhouse gas (GHG) emissions;

 10

(c) increase rural employment and income; and

(d) ensure availability of alternative and renewable clean energy without

any detriment to the natural ecosystem, biodiversity and food reserves

of the country.

Also, included in this law’s mandate, is the pre-blending of liquid fuels with

biofuels that are manufactured in the Philippines for use of motors and engines. There

required blending for both gasoline and diesel fuel is as follows:

(a) Gasoline: Five percent (5%) bioethanol by volume within two years of

enactment, rising to 10% two years thereafter, subject to the

determination of its feasibility by the government body created by the

Act, the National Biofuels Board (NBB).

(b) Diesel: a minimum of 1% biodiesel by volume within three months of

enactment, rising to 2% two years thereafter, also subject to NBB’s

determination of feasibility and availability of local biodiesel supply.

Wastewater Regulations

The Department of Environment and Natural Resources (DENR) is the primary

government agency that implements water protection in the country. In order to protect

water bodies in the country, standards are being monitored by DENR for wastewater

discharges for the residential, industrial and agricultural sectors. DENR Administrative

Order (DAO) Nos. 34 and 35 series of 1990 (Water Usage, Classification and Effluent

Regulations) and Republic Act 9275 (known as Clean Water Act of 2004) are two of the

guidelines in the proper implementation of wastewater discharges in the country.

 11

DAO No. 35 has set the effluent standards for Class C inland water (industrial

water supply for manufacturing after treatment) as classified by DAO No. 34 and is

summarized in Table 2-3.

Table 2-3. Effluent standards for inland water class C

PARAMETER UNIT VALUES
Color PCU 200
Temperature (maximum rise
in oC in receiving body of oC rise 3
water)
pH (range) --- 6.0 - 9.0
COD mg/L 150
BOD5 mg/L 80
Total Suspended Solids (TSS) mg/L 90
Source: DENR Administrative Order No. 35, 1990

2.3. Wastewater Characteristics

The quality of wastewater is determined by characterization of pollution

parameters such as pH, color, temperature, COD and BOD. These parameters should be

monitored and controlled due to the potential harm it may cause in the environment.

Biochemical Oxygen Demand (BOD)

Biochemical oxygen demand (BOD) measures the degree of microbially-mediated

oxygen consumption by contaminants in water. It is a major test made to determine the

organic polluting power of a waste because it measures the total amount and rate of
 12

oxygen, which will be needed to prevent nuisance odor formation and oxygen depletion

of streams. The standard period of test is 5 days using the Azide-modification method.

Chemical Oxygen Demand (COD)

Chemical oxygen demand (COD) measures the capacity of water to

consume oxygen during decomposition (Smith and Scott, 2005). It is also an indicator of

the amount of organic matter present in water. COD can be determined using the

dichromate open reflux method. The amount of unreacted dichromate is titrimetrically

determined to know the amount of chemical oxidizing agent (K2Cr2O7) required to

completely oxidize the organic material present (Rajeshwar and Ibanez, 1997).

Color

Color is a qualitative characteristic of water to assess its general condition. Dark

color wastewater hinders photosynthesis in bodies of water by blocking sunlight and can

cause eutrophication and therefore deleterious to aquatic life (Satyawali & Balakrishnan,

2008). It will also reduce the required oxygen needed by aquatic life. Color can also be an

indicator of the amount of organic matter in wastewaters. The standard method for the

measurement of color is the Platinum-Cobalt Method in terms of Platinum Cobalt Units

(PCU).

pH

pH measures the acid or base character of a liquid or solution. The measure is

based upon the hydrogen ion concentration of the liquid solution. The measure is
 13

expressed as a number from 0 to 14, where liquid solutions with pH < 6 are considered

acidic, pH > 8 are basic, and pH = 7 are neutral. Wastewaters should have pH values

between 6 to 9 in order to have minimum impact on the environment (Perry and Green,

1997). Wastewaters with pH values less than 6 will tend to be corrosive as a result of

excess hydrogen ions. On the other hand, wastewaters that have pH above 9 will cause

some of the metal ions to precipitate as carbonates and hydroxides.

Temperature

Temperature is also a considered parameter in wastewaters. High temperature of

wastewaters decreases amount of dissolved oxygen in water, which is deleterious to

aquatic life (Smith and Scott, 2005).

2.4. Distillery Wastewater Treatment Methods

According to Satyawali and Balakrishnan (2008), there are two major ways on

treating distillery wastewaters: biological and physico-chemical treatment.

Biological Treatment

Biological treatments for distillery wastewaters include anaerobic and aerobic

digestion. Since distillery effluents have high organic content, anaerobic treatment is

more favorable than direct aerobic treatment. Anaerobic treatment converts over half of

COD into biogas (Wilkie, 2000) while aerobic treatment just convert 50% of the COD to

sludge after treatment (Sennitt, 2005 as cited by Satyawali and Balakrishnan, 2008).

Anaerobic digestion is discussed in detail in section 2.5.

 14

Physico-chemical Treatment

Slops after biological treatment can still have a BOD of 250-500 mg/L (Satyawali

and Balakrishnan, 2008). Also, even biological treatment resulted in significant COD

removal, the effluent still remains dark in color. The color, due to melanoidin, is barely

affected by conventional biological treatment such as methane formation and activated

sludge process (Migo et al, 1993 as cited by Satyawali and Balakrishnan, 2008). Physico-

chemical treatment for distillery effluents includes adsorption, coagulation, flocculation

and oxidation. Coagulation, flocculation and oxidation will be discussed in detail in

section 2.6.

2.5. Anaerobic Digestion Process

Anaerobic biodegradation of organic material proceed in the absence of oxygen

and the presence of anaerobic microorganisms. Anaerobic digestion is the consequence of

a series of metabolic interactions among various groups of microorganisms (Verma,

2002). The first group of microorganisms secretes enzymes, which hydrolyzes polymeric

materials to monomers such as glucose and amino acids. These are subsequently

converted by acetogenic bacteria to higher volatile fatty acids, H2, and acetic acid.

Finally, the methanogenic bacteria convert H2, CO2, and acetate to CH4.

General Process Description

 15

According to Verma (2002), the anaerobic digestion process could be divided to

three processes: hydrolysis or liquefaction, acetogenesis and methanogenesis (Figure 2-

1).

Figure 2-1. Anaerobic Digestion Process

Source: http://www.wtert.eu/default.asp?Menue=13&ShowDok=12

The first stage of the process is the conversion of insoluble complex organic

matter, such as cellulose, into soluble molecules such as sugars, amino acids and fatty

acids by the fermentative bacteria. The complex polymeric matter is hydrolyzed to

monomer by hydrolytic enzymes secreted by microbes. In the second stage, anaerobic

bacteria, convert the product of the first phase to simple organic acids, carbon dioxide

and hydrogen. Generally, acids that will be produced are in the form of acetic acid

(CH3COOH), propionic acid (CH3CH2COOH) and butyric acid (CH3CH2CH2COOH).

The products formed in acetogenesis are due to a number of different microbes. Finally,
 16

in the third stage, methane is produced by methanogens in two ways: either by means of

cleavage of acetic acid molecules to generate carbon dioxide and methane, or by

reduction of carbon dioxide with hydrogen. Methane production is higher than reduction

of carbon dioxide but limited hydrogen concentration in digesters results in that the

acetate reaction is the primary producer of methane (Omstead et al, 1980 as cited by

Verma, 2002).

Conditions and Variables Influencing Anaerobic

Digestion

Several parameters should be monitored in order for an anaerobic process to

operate effectively and efficiently. Since one of the primary concerns of anaerobic

digestion is to convert the COD of the wastewater into biogas, environmental and

operating conditions for maximum gas production should be considered. Table 2-4 shows

the conditions and variables influencing biogas production in an anaerobic digestion.

Table 2-4. Environmental and operating conditions for

maximum methane production
CONDITIONS
VARIABLE
Optimum Extreme
pH 6.8 - 7.4
Temperature
Mesophilic 30 - 35 oC 20 - 40 oC
Thermophilic 50 - 56 oC 45 - 60 oC
Hydraulic Retention 10 - 15 days 7 - 30 days
Time (HRT)
Gas Composition
Methane 65 - 70 60 - 75
Carbon Dioxide 30 - 35 25 - 40
Source: Malina and Pohland, 1992
 17

pH and buffering. Most anaerobic processes operate at optimum pH near neutral.

Malina and Pohland (1992) mentioned that generally, the working pH value for

anaerobic bacteria usually ranges from 6.4 to 7.8. However, in most experimental

work, an optimum range of 6.8 to 7.4 shows good gas production.

Buffering system is also desirable and essential for anaerobic digestion

(Hadlocon, 2008). Some of the supplementary buffers may be in the form of

sodium bicarbonate (NaHCO3) and sodium hydroxide (NaOH).

Temperature. According to Malina and Pohland (1992), anaerobic digestion can

be operated in a wide range of temperature from 300C to 600C. This could be

divided into the mesophilic and thermophilic ranges.

Mesophilic bacteria could survive at an optimum temperature of 30 to 35

0
C. On the other hand, the optimum temperature for which thermophilic bacteria

will be efficient is at temperature range of 500C to 560C. An effective temperature

range must be established in order for the growth of sensitive methanogens will

not be disturbed with slight changes in temperature.

Nutrient and sludge addition. Anaerobic bacteria must be supplied with nutrients

for its optimum growth. Carbon, nitrogen and phosphorus must be with the

highest concentration of these nutrients for these are required in the reproduction

of anaerobic bacteria (Villa, 2009). Some wastes inherently have these

components, but for most industrial wastewater, supplementary nutrients are

added for the anaerobic process to be in optimum operation.

 18

Hydraulic Retention Time (HRT). The Hydraulic Retention Time (HRT) is a

measure of the average length of time that a soluble compound remains in a

constructed reactor. Retention time can be reduced by either increasing the rate of

feeding or by diluting the feed and keeping the loading rate constant. HRT can be

computed using Equation 2-1.

= (Equation 2-1)

where

Q is the volumetric flowrate of substrate (L/day)

V is the reactor volume (L)

Organic Loading Rate (OLR). Organic loading rate (OLR) is presented as

the amount of organic matter supplied daily, as expressed in mg COD/L-day. It is

also a measure of the biological conversion capacity of the anaerobic digester

system. Maximum OLR is dependent upon the nature of waste and can be

computed using Equation 2-2.

= (Equation 2-2)

where
Cso is the initial COD concentration (mg COD/L)
HRT is the hydraulic retention time (days)
 19

From Equation 2-2, OLR has an inverse relationship with HRT. Longer

HRT would mean lower OLR while shorter HRT would require higher OLR.

Loading rate for an anaerobic process is one of the important parameters

that could influence the performance of the digester. Excess mass rate of organic

solids that will be added to the digester will cause accumulation of volatile acids

that can inhibit the methanogenic activity and flow of reduction in an ideal control

action (Statfford et al, 1981 as cited by Villa, 2009).

Agitation. The purpose of mixing in a digester is to blend the fresh

material with digestate containing microbes. Furthermore, mixing prevents scum

formation and avoids temperature gradients within the digester (Verma, 2002).

However excessive mixing can disrupt the microbes so slow mixing is preferred.

The kind of mixing equipment and amount of mixing varies with the type of

reactor and the solids content in the digester.

2.6. Electrochemical Processes

Principle of Electrolysis

Electrolysis is the production of a chemical change by the passage of electricity

through a solution that contains ions or through a molten ionic compound (Maguyon,

2004).

Electrolytic cells are electrochemical cells used to cause a chemical change

through the application of electricity. It consists mainly of an electric source, electrodes

 20

(anode and cathode) and electrolytes. When the electrodes are placed in the electrolyte

and a direct current is passed, the electrons from the electric source flow toward one

electrode making it negatively-charged (cathode). The other electrode becomes

positively-charged (anode). The positive ions migrate toward the cathode and the

negative ions of the electrolyte move toward the anode and transfers electron.

Faraday’s Law of Electrolysis

The number of reactant molecules involved in an electrode reaction is related

stoichiometrically to the number of charges flowing in the circuit (Bagotsky, 2006). This

is the basic argument of the laws formulated by Michael Faraday between 1832 and

1833.

Faraday’s first law of electrolysis states that in electrolysis, the quantities of

substances involved in the chemical change are proportional to the quantity of electricity

which passes to the electrolyte. In equation form, Faraday’s first law illustrated as:

= (Equation 2-3)

On the other hand, Faraday’s second law of electrolysis states that the masses of

different substances set free or dissolved by a given amount of electricity are proportional

to their chemical equivalents. It can be shown as:

= ( ) (Equation 2-4)

where
 21

Q is the electrical charge (Coulumbs)

n(e) is the mole of electrons
I is the electrical current (ampere)
t is the time (seconds)
F is the Faraday’s constant (96,484 coulumbs/mol)

Charge Dose

Charge dose has been defined as the amount of electricity required to move a

target pollutant. It has been used as empirically derived factor for scale-up and operation

of electrolytic reactors (Alfafara et al, 2003). Charge dose is the amount of charge

required to remove a unit mass of pollutant with unit of coulumbs (Alfafara et al, 2002).

= (Equation 2-5)
( − )

where
Q is the electrical charge passed (Coulumbs)
I is the electrical current (ampere)
T is the time (seconds)
C0 is the initial concentration of pollutant (mg/L)
C is the final concentration of pollutant (mg/L)
V is the volume of reactor (L)

2.7. Ways of Electrochemical Treatment of Wastewater

Many electrochemical processes for environmental pollution control involve the

direct reaction of species at electrode surfaces, while others involve production of active

species at the electrode and further reaction with the targeted pollutants. Some of the

treatments of wastewater using electrochemical processes include direct electrolysis,

 22

indirect electrolysis, electroflotation, electroflocculation, electrocoagulation and electro-

oxidation.

Direct Electrolysis

Direct electrolysis involves processes in which electron transfer reaction to or

from the undesired pollutant occurs at the surface of an electrode (Rajeshwar and Ibanez,

1997). Figure 2-2.a shows the mechanism of direct electrolysis.

Indirect Electrolysis

Indirect electrolysis uses an electrochemically generated redox reagent as

chemical reactant (or catalyst) to convert pollutants to less harmful products. The redox

reagent acts as an intermediary for shuttling electrons between the pollutant substrate and

the electrode (Rajeshwar and Ibanez, 1997). Figure 2-2.b shows the mechanism of

indirect electrolysis.

Figure 2-2. Mechanism of Direct Electrolysis (a) and (b)

Indirect Electrolysis.
Source: Rajeshwar and Ibanez, 1997
 23

Electroflotation

Electroflotation, as shown in Figure 2-3, involves the electrolytic

production of gases that can be used to attach pollutants to gas bubbles and carry

them up to the top of the solution where they can be more easily collected and

removed (Rajeshwar and Ibanez, 1997).

Figure 2-3. Mechanism of Electrofloatation

Source: http://www.infomarine.gr/news/index.php?
mod=article&cat=newproducts&article=461&page_
order=1&act=print

Electroflocculation

Electroflocculation is an electrochemical production of agents that promote

particle bridging or coalescence (Rajeshwar and Ibanez, 1997). The process involves

application of an electric current to sacrificial electrodes, usually aluminum, inside a

processing tank.
 24

Electrocoagulation

Electrocoagulation refers to the electrochemical production of destabilization

agents that bring about charge neutralization for pollutant removal (Rajeshwar and

Ibanez, 1997). It involves the passage of an electric current, usually by using aluminum

electrodes. Aluminum goes into solution at the anode and hydrogen gas is released at the

cathode. Figure 2-4 illustrates the mechanism of eletrocoagulation process.

Figure 2-4. Mechanism of Electrocoagulation

Source: Rajeshwar and Ibanez, 1997

Electrooxidation

Electro-oxidation is a combination of direct and indirect electrolytic

processes that involves the oxidation and reduction of organic and inorganic pollutants

(Rajeshwar and Ibanez, 1997). It is similar to other electrochemical process that applies

electric current to electrodes.

 3. MATERIALS AND METHODS

3.1. Preparation and Characterization of Sweet Sorghum Distillery Slop

Since there are no currently available sweet sorghum-based distilleries in the

Philippines, the sweet sorghum distillery slop was produced from laboratory-to-pilot

scale fermentation followed by distillation. Sweet sorghum juice was collected from

Mariano Marcos State University (MMSU) and was evaporated to form syrup for storage

purposes.

The sweet sorghum syrup was subjected to anaerobic ethanol fermentation using

yeast cells. The fermented broth was then separated from the yeast cells and distilled

using the set-up shown in Figure 3-1.

Figure 3-1. Simple Distillation Set-up for the generation of

sweet sorghum distillery slops
 26

Distillation was done at about 78.1°C (the boiling point of ethanol) for three hours. After

distillation, the slops were collected in a container and stored in a cold room. The distillery slop

was characterized in terms of BOD, COD, color, pH, Nitrogen and Phosphorus contents and Total

Suspended Solids (TSS).

3.2. Anaerobic Digestion Experiments

Preparation and Characterization of Inoculum

The inoculum that was used was an acclimatized sludge obtained from an active

anaerobic digester of a local ethanol production plant. It was characterized in terms of

BOD, COD, color, pH, nitrogen and phosphorus content, and Total Suspended Solids

(TSS).

Batch Anaerobic Digestion (Acclimatization Phase)

and Semi-Continuous Anaerobic Digestion Set-up

The anaerobic digestion set-up used in this experiment is shown in Figure 3-2.

The reactor vessel (Tank 1, the anaerobic digestion reactor) is a 4-L glass container

which contained the inoculum (anaerobic sludge) and the substrate (sweet sorghum

distillery slop) in a 4:1 ratio (by volume). The total volume of the reaction mixture

(considering the above proportions for wastewater and inoculum) will be 3.1 liters. The

biogas produced from the anaerobic digestion reactor in Tank 1 was channeled to Tank 2

(the gas collection tank), where it displaced and equivalent volume of water in Tank 3

(displaced-water collection tank). The volume of water displaced is a measure of the

volume of biogas produced.

The rubber connector tubing was clamped when gas production was measured in

a graduated cylinder. Two (2) set-ups were prepared, with duplicates, to obtain various

treatments. The reactor vessels were gas-tight and were sealed by rubber stoppers with

two openings, as shown in Figure 3-2: (1) sampling port 5, and (2) biogas exit port 6.
 27

Sampling for the analyses was done by connecting a tygon tubing at the sampling

port 5, a syringe was then attached to the free end of this tubing to extract the sample.

Figure 3-3 shows the actual anaerobic digestion set-up.

Figure 3-2. Anaerobic Digestion Experimental Set-Up

Figure 3-3. Actual Anaerobic Digestion Experimental Set-Up

 28

Experimental Conditions for Anaerobic Digestion

Four set-ups were used in this first phase of acclimatization with a twenty-day

(20d) hydraulic retention time (HRT) that was considered for monitoring, sampling and

analyses. Using the experimental set-up in Figure 3-2, 10 ml samples were collected

every day except weekends using the syringe attached to the reactor. Gas volume was

also measured. Monitoring schedule is given in Table 3-1.

Table 3-1. Schedule of Analysis for Batch Anaerobic Digestion and Semi-continuous
Experiment
TYPES OF SCHEDULE OF ANALYSIS
ANALYSIS Acclimatization Semi-Continuous
1. pH Daily Daily
2. Color Daily Daily
3. Gas Volume Daily Daily
Every 5 days for HRT =
4. COD Every 5 days 30d Every 10 days for
HRT = 60d

For the semi-continuous anaerobic digestion experiment, two (2) HRT’s (30 days

and 60 days) were considered. The corresponding feeding rates of the sweet sorghum

distillery slop at the above HRT’s are 66.67 mL/day (OLR = 5.534 mg COD/L-day) and

33.33 mL/day (OLR = 2.766 mg COD/L-day), respectively.

Two set-ups (30day and 60day HRT’s) were used with duplicate for each for

monitoring, sampling and analyses. Feeding of the distillery slops was done daily during

weekdays. On weekends, the feed rates were adjusted on Fridays to compensate the

feeding for Saturdays and Sundays. Before loading a fresh feed of slops, an equivalent

volume of spent slop was withdrawn from the reactor. The feeding system was similar to

a “draw-fill” system. The withdrawn samples were taken for analysis of pH, color and

COD. All the spent digested slops were collected and accumulated for use in the second
 29

step preliminary electro-oxidation experiment. Agitation was done before and after

adding the fresh slops to the reactor for the purpose of homogenous mixing. Monitoring

schedule for the semi-continuous anaerobic digestion experiment is shown in Table 3-1.

Analytical Methods

pH. The acidity and basicity of the collected samples was analyzed using the

Orion pH meter (Thermo Fisher Scientific, USA) as shown in Figure 3-4.

Figure 3-4. Orion pH meter (Thermo Fisher Scientific, USA)

Gas production. Gas production was measured by water displacement method

through a gas trap (Figure 3-2). The displaced water in a water receiver (500-ml for batch

experiment and 6-L for semi-continuous experiment) was transferred in a graduated

cylinder for measurement. The volume of the displaced water is approximately equal to

the volume of the biogas produced inside the reactors.

To qualitatively assess gas quality, a flame test was done at the end of each HRT.
 30

Color Analysis. Color was analyzed by measuring the absorbance at 462nm

using the SHIMADZU UV-1601: UV- Vis spectrophotometer (Figure 3-5). This

wavelength corresponds to the maximum absorbance of Pt-Co standards. The absorbance

readings were converted to color in terms of TCU (total color units) using a calibration

curve.

Figure 3-5. SHIMADZU UV-1601: UV-Vis spectrophotometer

COD Analysis. The organic matter in the wastewater was measured as COD

using the Standard Methods for the Examination of Water and Wastewater (1992).

Dichromate open reflux method (set-up shown in Figure 3-6) was used for determining

COD.
 31

Figure 3-6. Set-up for dichromate open-reflux

method for COD determination.

Data Analysis. Graphs of color, COD, gas production and pH with respect to

time were constructed. Removal efficiency of COD (as well as compliance with DENR

standards) was also evaluated as index of performance.

3.3. Preliminary Electrooxidation Experiments

Batch Electrolysis Set-up

The electrolysis set-up shown in Figure 3-7 includes a 1-L beaker reactor (with a

working volume of 800 mL), a pair of electrodes and a power supply. Uniform mixing

was done using a magnetic stirrer system.

The electrodes used were composed of a sintered platinum anode and a stainless

steel cathode. The dimensions of the electrodes were 5 cm x 15 cm. Two rubber strips

(approximately 0.5 cm thick) were placed in between the electrodes using a Teflon tape

to ensure spacing and to avoid short circuiting. The electrodes were submerged in the

treated effluent exposing the largest possible area. Wires with alligator clips served as

connectors to the electrodes with direct current regulated power supply. A multimeter
 32

was used to monitor the voltage and the operating current throughout the electro-

oxidation experiment.

Figure 3-7. Diagram of batch electrolysis set-up

Source: Santos, 2005.

Experimental Conditions

Electro-oxidation experiment of treated effluent from anaerobic digestion was

conducted using the set-up shown in Figure 3-7. In a study conducted by Beltran (2005),

chloride ion supplementation is necessary for treating distillery effluent to achieve higher

color and COD removal at a much shorter time.

Sodium chloride (NaCl) crystals were used for chloride supplementation, and

were added to the biodigester effluent at a concentration of 2.5% (w/v), to increase its

conductivity as well as to provide additional chloride ions which served as raw materials

for electrochemical oxidant formation. Operating current was at 1.5 A. Samples were
 33

taken at hourly intervals time and were analyzed for absorbance (color), COD, pH and

temperature.

Analytical Methods

The analytical methods for the electro-oxidation experiments were similar to

those for anaerobic digestion (except for gas production).

Data Analysis

Time-course for the COD, color, pH and temperature, evaluated at the operating

current (1.5A), were graphically constructed. Using these graphs, the pollutant

parameters (COD and color) removal efficiencies were evaluated. Results of these graphs

determined whether complete color and COD removal is possible of the treated sweet

sorghum distillery effluent from anaerobic digestion using electro-oxidation. Also, these

graphs determined the effect of operating current on COD removal efficiency.

3.4. Scale-up and Operational Conditions

Charge Dose

Charge dose is proposed as a scale up parameter for electrolytic reactors

(Alfafara, et al. 2002). It is defined as the amount of charge required in removing a unit

mass of target pollutant (expressed as Coulumbs per mg pollutant removed). Linearizing

equation 2-5, charge dose could be evaluated for the electro-oxidation of treated distillery

effluent.

=
( − ) (Equation 2-5)
 34

3.5. Energy Requirement and Preliminary Cost Estimation

Energy requirement for the electrolytic treatment of sweet sorghum distillery slop using

the treated effluent from the anaerobic digestion experiments was assessed using Equation 3-1.

Charge dose value obtained was used in evaluating this criterion.

= (Equation 3-1)
3.6 10

where

W is the energy requirement (kWh/mg pollutant removed)

Q is the charge dose (coulomb per mg pollutant removed)
E is the operating voltage (V)

Energy cost was computed by multiplying the prevailing electric cost rate with the energy

requirement.
 4. RESULTS AND DISCUSSION

4.1. Wastewater characteristics

The distillery slop which was obtained from the distillation of the fermented

sweet sorghum juice using the set-up in Figure 3-1 is shown in Figure 4-1. It had a dark-

brown color which was similar to the color of molasses-based distillery slops. The

wastewater characteristics of the sweet sorghum distillery slop, in comparison to the

DENR Class C inland water effluent standards are shown in Table 4-1.

Figure 4-1. Sweet sorghum distillery slop

Table 4-1. Characteristics of sweet sorghum distillery slop and acclimatized sludge
compared to the effluent standards of DAO No. 35
SWEET
DENR CLASS C
SORGHUM ACCLIMATIZED
PARAMETER STANDARD (Inland
DISTILLERY SLUDGE
Water)
EFFLUENT
COD, mg/L 166,000 136,457 150
BOD5, mg/L 78,361 13,460 80
Color, TCU 196,390 224,961 200
pH 5.05 7.43 6.0 to 9.0
 36

The pollutant parameters of the sweet sorghum distillery slop such as COD, BOD,

color and pH were much higher than the standard set by the DENR for inland water class C.

Before disposal, the wastewater must undergo a treatment process. For this wastewater

(which contains an extremely high organic matter content), anaerobic biological treatment

(i.e. anaerobic digestion) is usually the recommended choice of treatment. The reasons are:

(1) anaerobic digestion is effectively a controlled and enclosed version of the anaerobic

breakdown of organic waste which converts organic substances to methane; (2) biomass

acclimatization allows more organic compounds to be transformed; (3) less biological sludge

produced; (4) lower nutrient demand; and (5) less energy required.

4.2. Time Course Profiles for Wastewater Parameters Operated at Different

Organic Loading Rates (OLR) During Acclimatization

Parameters such as pH, COD and gas production were investigated to assess the

performance of the anaerobic digestion system in the acclimatization phase of the

experiment, for organic loading rates of 5.534 g/L-d COD (hydraulic retention time, HRT

= 30 days) and 2.766g/L-d COD (HRT = 60 days). Figure 4-2 shows the time profiles of

the above-mentioned parameters for both HRTs during the start-up phase.

Figure 4-2a shows the change in pH of the system for the 20-day acclimatization

period. The pH of the system at both organic loading rates fluctuated but do not change

significantly. Also, both organic loading rates did not decrease beyond pH = 7 which was

an indication of good performance of the system in the acclimatization phase. Figure 4-2b

shows the residual COD behavior during start-up phase at the two organic loading rates.

The residual COD for both organic loading rates decreased in the 20-day start-up period.

The COD removal efficiency, shown in Figure 4-2c, increased by 44.63% and 43.86%
 37

for HRT = 30 days and HRT = 60 days, respectively. Decrease in the residual COD and

increase in removal efficiency also indicated good performance of the anaerobic

digestion system during start-up/acclimatization. Gas production, with simultaneous

COD reduction (shown in Figure 4-2d), also increased, indicating the conversion of COD

into biogas. These observations were observed at both organic loading rates tested. The

time profiles of the indicated wastewater parameters indicated a reasonably good

acclimatization.

Figure 4-2. Time course profiles for (a) pH, (b) residual COD, (c) COD removal efficiency, and (d) gas
production during acclimatization

4.3. Anaerobic Digestion of Sweet Sorghum Distillery Slop at Different Organic

Loading Rates (OLR) in the Semi - Continuous Experiments
 38

pH profiles

The pH profiles of the (sweet sorghum slops) anaerobic digestion set ups for

the two organic loading rates (2.766 g/L-d or 60d HRT and 5.534 g/L-d or 30d HRT)

are shown in Figure 4-3.

Figure 4-3. pH profile of digested effluent at the two hydraulic retention time in the semi-
continuous experiments.

For a 60d HRT, the pH decreased initially during the first 10 days from an

initial pH of 7.5 to a pH of 6.87. Eventually, the pH increased to a pH of 8.42 and

achieved steady state at 7.5. The pH profile for the 60d HRT anaerobic digester was

within the pH range for an optimally functioning anaerobic digester (Malina and

Pohland, 1992).

On the other hand, for the 30d HRT digester, the pH continuously decreased

to more acidic conditions at pH = 5.50, where it achieved its steady-state pH value in

the acidic range. The decrease in steady state pH at higher OLR can be attributed to

the higher fatty acid production by acidogenic bacteria in the system at higher organic

loads. Undesirable fatty acid production leading to acidic pH is called “souring”.

 39

Low pH and excessive acid production and accumulation could also displace the

more neutral pH bicarbonate buffer system (Pohland and Suidan, 1987) and these

conditions would be inhibitory to methanogens (Malina and Pohland, 1992). Since

the pH of the system at the higher OLR of 5.534 g/L-d (30d HRT) was below 6.0,

methane forming bacteria may have been inhibited, since excessively high amounts

of volatile acids (substrate for methane-forming bacteria) accumulated in the digester

(Vermi, 2002). Stoppage of feeding was done in an attempt to revive the souring

digester. However, pH values still decreased and COD values increased. Gas

production also leveled-off even after stoppage of feeding was done. The reactor may

have already in a state of extreme high organic overload, so that recovery would no

longer be possible.

Based on the pH values, the anaerobic digestion setup with a HRT of 60 days

(OLR = 2.766 g/L-d) can be considered suitable for functional gas production and

COD reduction as compared to the setup with a HRT of 30 days (OLR = 5.534 g/L-

d).

Residual Chemical Oxygen Demand (COD) profiles

One of the key parameters for assessing anaerobic digestion performance is

the ability of the microorganisms to degrade the organic carbon compounds that

cause water pollution. This can be determined by monitoring the decrease in the

Chemical Oxygen Demand (COD), which is a measure of organic pollutants in the

wastewater. The COD profiles of the anaerobic digestion setups at the 30 day HRT

and 60 day HRT are shown in Figure 4-4.

 40

Figure 4-4. Residual COD for the two HRT in the semi-continuous
experiments

Figure 4-4 shows that only the set-up for the 60d HRT showed a decrease in

residual COD after about 60 days of operation. The COD profiles for the 30d HRT

operation indicated that the system indicated signs of “souring” under this organic

loading rate. The 30d HRT digester can be considered as “overloaded” with COD.

Under these conditions, acidogenic bacteria grow faster than the methanogenic

bacteria, as a result of higher COD introduced into the system. Overgrowth of

acidogenic bacteria results in the overproduction of volatile fatty acids in the reactor,

causing the pH to go acidic. When the pH goes acidic, methanogenic bacteria can

become inhibited and may also die. Inhibition or death of methanogenic bacteria

will stop the conversion of the fatty acids into CO2, thus no more reduction in COD

occurs. Consequently, biogas production also stops. In Figure 4-4, there was no

observed decrease in the residual COD concentrations after long time operation of the

30d HRT set up. The increase in the residual COD over time for the 30d HRT

digesters may be attributed to possible COD exerted by some acidogenic biomass

which may have not been completely removed during centrifugation of the sample
 41

during COD analysis. The effluent samples from the 30d HRT operation also had a

strong organic acid smell.

The COD values of an organic loading rate of 2.766 g/L-d (HRT = 60days),

continuously decreased after day 25. This could be attributed to good pH values

which promoted methane gas production by methanogenic bacteria. Since the pH was

within the optimum range needed for methanogens to survive, higher gas production

ensured conversion of organic matter to methane and carbon dioxide, which have

decreased COD values. Figure 4-5 shows the percent reduction of COD for HRT of

60 days (OLR = 2.766 g/L-d).

The highest COD reduction was 33.05% corresponding to a COD value of

32,547 mg/L. However, the COD value did not still pass the DENR Class C effluent

standards and should be further treated to conform to these standards. A second-stage

treatment was done in order to further reduce the organic matter content of the

wastewater (by electrooxidation) and this is discussed in Section 4.4.

Figure 4-5. Percent (%) COD removal for the setup of HRT = 60 days in the
semi-continuous experiments
 42

Gas production profiles

To assess the performance of the anaerobic digestion system (based on

organic loading rates), gas production was monitored. Figure 4-6 shows the mean

cumulative gas production throughout the time course of the semi-continuous

experiment. An increase in the organic loading rate generally corresponds to a higher

gas production. The mean cumulative gas volume for the two set-ups increased daily.

However, the set-up with an OLR of 5.534g/L-d (HRT = 30 days) leveled off on the

52nd day. This level off in the cumulative gas production means that there is no more

net production of biogas after about 25 days of operation. The digester for the 30d

HRT set up already failed after 25 days. This is also supported by the pH (Fig 4-3)

and residual COD (Fig 4-4) profiles which was previously shown. As shown in Fig

4.3, the pH fell to values below 6, indicating the accumulation of organic fatty acids

and inhibiting the methane-forming bacteria to convert organic matter to methane

which hindered gas production. Since there was no gas production, there was no

conversion of organic matter to methane.

On the other hand, the setup with an OLR of 2.766g/L-day (HRT = 60 days)

continuously increased until the 120th day at a rate of 1,818.9 mL/day. Since the setup

with an HRT of 60 days has a pH range within the optimum range of 6.8 to 7.4 (Fig.

4-3), cumulative gas production was continuously increased (Fig. 4-5), which

resulted to conversion of organic matter to methane and carbon dioxide, residual

COD of the system decreased (Fig 4-4).

Flame test was also done to check for the quality of gas that was produced by

the digester. Figure 4-7 shows the quality of gas through the color of the flame. The

picture shows a yellow color of the flame. Methane gas when burned has a blue

flame. However, flame colors with yellow, orange, green and red that are mixed in
 43

the blue flame may indicate incomplete combustion of organic compounds such as

methane. Oxygen needed for complete combustion may not have been enough.

Nevertheless, the presence of the flame is a reasonable test which can verify the

presence of methane in the gas. To investigate the composition of the gas produced

by the digester, methods for determining the composition of biogas produced would

be ideal. However, this could not be done to unavailability of measuring facilities in

the laboratory.

Figure 4-6. Mean cumulative gas production for the two HRT in the semi-continuous
experiments
 44

Figure 4-7.Flame test for gas production in the semi-continuous

experiment

Residual color profiles

Figure 4-8 shows the residual color profiles for each setup in the semi-

continuous phase in the anaerobic digestion of sweet sorghum distillery slop. Both

set-ups exhibited fluctuating values of color. No significant change in color can be

seen observed from the graph. Color values for both HRT reached as high as 250,000

TCU. Insignificant change in color can be attributed to repolymerization of

melanoidin (dark-brown colored substance generated by Maillard reaction of amino

acids and carbohydrates) for which the brown color of the wastewater retained or

even increased (Agarwal et al, 2010). Studies on the conventional biological

treatment (aerobic and anaerobic treatment) of distillery slops showed that

degradation of melanoidin could be up to 6 to 7% only (Agarwal et al, 2010).

Physicochemical methods for decolorization of distillery effluents after biological

treatment such as adsorption, coagulation and flocculation and oxidation process,

may be required. For this study, electro-oxidation was used as a second stage

treatment for further decolorization of the digested effluent and is discussed in

Section 4.4.
 45

Figure 4-8.Residual color for the two HRT in the semi-continuous

experiments

4.4. Preliminary Electrooxidation of the Chloride-Supplemented Sweet Sorghum

Biodigester Effluent

pH and Temperature profiles after electrooxidation

Figure 4-9 shows the pH and temperature profile during the electrooxidation

of anaeraboically digested sweet sorghum slops at 1.5A and 2.5% chloride

supplementation. The use of 2.5% chloride supplementation was based on the study

of Beltran (2005).

From an initial pH of 7.59, the pH increased to 8.03 after 1-hr electrolysis

time. After the 12-hr electrolysis, the pH was increased to 8.6 as shown in Figure 4-

9a. The pH-buffering elements which could be present in the system could have a

role in pH stabilization even if there was a continuous production of hydroxide ions

at the cathode (Maguyon, 2004).

On the other hand, there was a 12 degree rise in temperature (in Celsius units)

during the progress of electrolysis, as shown in Figure 4-9b. The rate of temperature
 46

rise was slow and it stabilized to about 39oC, from the 8th to the 11th hour of the

electro-oxidation.

Figure 4-9. pH and temperature time courses during the electro-

oxidation of the digested effluent

Residual COD profiles

The COD time profile for the electro-oxidation experiment is shown in Figure

4-10. The graph shows that at the operating current of 1.5A and chloride

supplementation of 2.5%, there was a decrease in COD in the 12hr electrolysis

period. However, it is evident in the graph that there was only a slight decrease of

COD for the duration of electrolysis. This condition may be due to lower oxidizing
 47

power generated at 1.5A. The lower oxidizing power may also be due to insufficient

(chlorine-based) oxidants generated at the anode (i.e. chloride supplementation may

not be enough). Also, another possibility was that an operating current of 1.5A could

only oxidize easily biodegradable materials and the oxidants produced were not

enough to treat the remaining organic matter in the digested slop.

Excessive foaming, as shown in Figure 4-11, also occurred during electrolysis

causing a decrease in reaction when the foams were removed. To recover the amount

of wastewater that was loss due to foaming, the liquid after the settling of the foam

was returned back into the reactor. However, this condition may have compromised

the homogeneity of the wastewater treated in the reactor. Further study to solve this

problem is recommended.

Figure 4-10. COD time course in electro-oxidation

 48

Figure 4-11. Foams formed during electrolysis

Figure 4-12 shows the removal efficiency of COD with time. The graph

shows that the removal efficiency at the operating current of 1.5A and 2.5% chloride

supplementation achieved 24.14% COD removal after 12-hr electrolysis. The residual

value of COD was 28,253 mg/L after 12 hours and it is still higher than the effluent

standard. Thus, longer electrolysis time (and possibly a higher operating current by

increasing the power source, additional electrodes, or additional chloride

supplementation) may be needed to meet the standards for effluent imposed by the

DENR.

25

20
% COD Removal

15

10

0
0 2 4 6 8 10 12
Time (hr)

Figure 4-12. COD removal efficiency in electro-oxidation

 49

Decolorization Efficiency profiles

Figure 4-13 shows the residual color time course during the electrolysis of

(sweet sorghum) anaerobic digester effluent at an operating current of 1.5A and

chloride supplementation of 2.5%. The graph shows that color was removed during

the 12-hr electrolysis period, but the final residual color of 69,231 TCU was still

higher than the effluent standard. The maximum decolorization removal efficiency

(as shown in Figure 4-14) was 41%. The results indicate that higher operating current

and more electrolysis time should be studied in order to achieve complete color

reduction.

Figure 4-13. Residual color time course in electro-oxidation

Figure 4-15 shows a comparative visual observation of the effect of

12-hr electro-oxidation at 1.5A and 2.5% chloride supplementation on the color of the

digester effluent. The picture shows a discernable, but only slight reduction in color.
 50

Charge Dose and Energy Requirements

Computations of the Electrooxidation Process

Charge dose has been used as an operating and scale up factor to relate the

operating current on electrolysis time of an electrochemical wastewater treatment

 51

process at any scale (Alfafara et al, 2003). Charge dose can be determined using

equation 2-5.

= (Equation 2-5)
( − )

By simply plotting linearized curve of It vs (Co-C)V, the charge dose, Q,

which is equal to the slope of the line, can be determined. With the charge dose

known (from the slope of the regression line in Figure 4-16), the operating conditions

of electrolysis (electrolysis time given the operating current or vice versa) for any

reactor scale could be easily obtained.

Figure 4-16 and Figure 4-17 show the linearized plot for the charge dose

computations for COD removal and decolorization, respectively.

70000

60000

50000
y = 11.632x + 5137.3
It, Coulombs

40000 R² = 0.9636

30000

20000

10000

0
0 1000 2000 3000 4000 5000 6000
(Co-C)V, mg

Figure 4-16. Charge dose determination for COD removal in electro-oxidation

 52

80000
y = 2.3032x + 10083
70000
R² = 0.8931
60000

It, Coulombs
50000
40000
30000
20000
10000
0
0 5000 10000 15000 20000 25000 30000
(Co-C)V, TCU-L

Figure 4-17. Charge dose determination for color removal in electrooxidation

The charge dose for COD removal was found to be 11.632 C/mg pollutant

removed with a regression coefficient (R2) equal to 0.9636 while charge dose for

color removal was found to be 2.3032 C/Abs-L removed with a regression

coefficient (R2) equal to 0.8931.

For practical applications, operation of electro-oxidation based on

maximum charge dose obtained ensures the removal of the pollutants with lower

charge dose values. Among the two parameters considered (COD and color),

COD consumed the highest amount of charge. Thus, charge dose for COD maybe

used as the operating and scale-up factor for this process. Other operating

conditions, most especially operating current and electrolysis time can be derived

from the charge dose taking into consideration other parameters such as reactor

volume, operating current and concentration of pollutant to be removed.

The charge dose can also be used to explore different operating conditions

(for example, operating current or electrolysis time) to achieve maximum or

 53

complete decolorization. As a sample calculation, if the reactor scale is set at 100

L, and the electrolysis time is set at 12 hours, the required operating current for

complete decolorization can be computed from equation 4-1 (and the determined

value for the decolorization charge dose) as 626 A.

On the other hand, energy requirements can also determine to estimate the

energy cost of the process and economic feasibility. Energy required in kWh per

milligram of pollutant removed can be computed using equation 3-1. The

equation describes a direct relationship between charge dose and specific energy

requirement.

= (Equation 3-1)
3.6 10

Where, W is the specific energy requirement (kwH/mg pollutant

removed), Q is the charge dose (Coulomb/mg pollutant removed) and E is the

operating voltage in volts.

From the computed energy requirement, the energy cost of the process can

be determined using equation 4-1.

= ( − ) (Equation4-1)

Where, P is the estimated energy cost (Php/mg pollutant removed), W is

the specific energy requirement (kWh/mg pollutant removed), Co is the initial

 54

pollutant concentration (mg/L pollutant), C is the final pollutant concentration

(mg/L pollutant) and R is the prevailing industrial power cost.

The average voltage required in the operation of the process was 5.4V.

The charge dose computed for COD and color removed in the process and its

corresponding energy requirement, as well as the cost for the 12-hr treatment and

removal of each pollutant are summarized in Table 4-2 .

Table 4.2. Energy requirements for color removal and COD reduction for digested effluent

ENERGY
AVERAGE
PARAMETER CHARGE DOSE REQUIREMENT
VOLTAGE COST
(Pollutant) (C/mg pollutant) (kWh/mg
(Volts)
pollutant)
3.31E-6 PhP
Color 5.4 2.2032
kWh/TCU-L 0.000024/TCU-L
COD 5.4 11.632 1.74E-5 kWh/mg PhP 0.000129/mg

The specific energy requirement was obtained by multiplying the charge

dose for pollutant removed with the average operating voltage (5.4V) during

electrolysis. Then, the energy cost per pollutant removed was determined by

multiplying the specific energy requirement with the prevailing industrial power

cost (PhP7.39/kWh)(Meralco, 2011).

The computed energy costs in terms of pollutant removed were

PhP0.000017/TCU-L and PhP0.000090/mg, for color removed and COD reduced,

respectively. In terms of volume of slop treated, the costs of treatment were PhP1,

176.90/m3 for color removal and PhP1, 159.14/m3 for COD reduction. Since

initial color and COD values were high, a high treatment cost is expected.
 5. SUMMARY AND CONCLUSIONS

Treatment for the sweet sorghum distillery slop was investigated using two-stage

treatment: anaerobic digestion followed by preliminary electro-oxidation. Both wastewater

treatments were evaluated for the removal of pollutants from the sweet sorghum distillery slop

such as Chemical Oxygen Demand (COD) and color.

For the first stage anaerobic digestion, sweet sorghum distillery slop was produced by

fermentation of sweet sorghum juice followed by distillation process to separate ethanol and the

slops. After collecting the slops, two (2) replicated set-ups were prepared. Hydraulic retention

times (HRT) of 30 days and 60 days were used for each setup. Acclimatization was done for the

anaerobic sludge for 20 days to develop reasonably acclimatized microorganisms for the

biodigestion of sweet sorghum distillery slop. Parameters such as pH, COD and gas production

were monitored for the acclimatization phase, and results showed that the micoorganisms in the

sludge were reasonably acclimatized.

The effect of two organic loading rates (OLRs) (corresponding to two hydraulic retention

times, or HRTs) was evaluated using semi-continuous (“draw-fill”) experiments. The OLRs

tested were 5.534 g COD/L-d (30 d HRT) and 2.766 g COD/L-d (60 d HRT). Profiles for pH,

COD, and gas production until the approach of a reasonable steady state was obtained.

Results showed that the digester operated at the 30d HRT had signs of a failing or “sour”

anaerobic digestion. The pH values reached to as low as 5.50 which is beyond the optimum

condition for good anaerobic digestion. The COD values did not decrease, indicating COD

removal, gas production also leveled-off at a mean cumulative value of 30,074 mL. Based on the

poor performance in terms of COD removal and biogas production, the organic loading at 30 days
 56

HRT (5.534g/L-d) can be considered too high for the proper functioning of anaerobic digestion of

sweet sorghum distillery slops. Overloading can lead to overproduction of organic acids,

resulting in a decrease in reactor pH, and death or inhibition of methanogenic bacteria.

On the other hand, the digester operated at 60d HRT, showed a stable digester operation

with good steady state pH, good COD removal and good biogas production. pH values of 6.87 to

8.42 were in the range of optimum condition for anaerobic digestion. Residual COD decreased

with an initial value of 48, 721 mg COD/L to a value of 33,704 mg COD/L which corresponds to

a removal efficiency of 30.82%. Gas production continuously increased and has a production rate

of 1,818.9 mL/day. Color values fluctuated and showed no significant decolorization through

anaerobic digestion. With respect to compliance with the DENR Class C effluent (inland water),

only the pH passed the standard. The COD and color values were still much higher than the

standard. However, compared to the digester operated at 30d HRT, the digester operated at 60d

HRT has better operating condition as shown in the results of the pollution parameters. An

attempt to have a second stage treatment by application of electrochemical methods was made in

order to further improve pollution reduction performance, in terms of COD removal and

decolorization.

The digester effluent was then subjected to a preliminary batch electro-oxidation

experiment using a sintered platinum anode, which oxidizes the pollutants by generating a

chlorine-based oxidizing agent that reacts with the pollutant. Removal efficiencies of COD and

color were measured using a chloride supplementation of 2.5% operating at 1.5A in a period of

12 hours. Preliminary evaluation of electro-oxidation with higher currents was also tried.

The 12-hr preliminary electro-oxidation at 1.5A with a 2.5% chloride supplementation

resulted in 24.14% COD removal and 41.04% color removal. COD and color values were still
 57

higher than the effluent standards based on DAO No. 35 requirements as cited from Table 4-1.

Low operating current, as well as insufficient chloride supplementation for the generation of

enough oxidants could cause the low COD removal and decolorization efficiency of

electrooxidation treatment. Another possible cause could be the resistance of organic compounds

of the wastewater to electrooxidation.

Charge dose for the pollutants removed were also evaluated. In this system, it was

defined as the amount of charge needed to remove a unit of pollutant. For practical applications,

operation of electro-oxidation based on maximum charge dose obtained ensures the removal of

the pollutants with lower charge dose values. Among the two parameters considered (COD and

color), COD consumed the highest amount of charge. Thus, charge dose for COD maybe used as

the operating and scale-up factor for this process. Charge dose was estimated to be about 11.632

C/mg COD removed. Corresponding energy requirement and maximum energy cost were also

computed from the charge dose. An energy requirement of approximately 1.75 x 10-5 kWh/mg

pollutant removed and a maximum energy cost of 0.000129 Php/mg COD reduction. Considering

the very high initial color and the very high initial COD of the sweet sorghum distillery slop, this

would translate to a per volume treatment cost of 1,159.14 Php/m3.

Under the present operating conditions tested, the anaerobic digestion treatment followed

by preliminary electro-oxidation was not yet sufficient to treat sweet sorghum distillery slops,

conforming to the DENR effluent Class C standards. However, further extending the anaerobic

digestion at the loading rate of 2.766 g COD/L-d (60d HRT), might give some improvement.

Furthermore, additional electrooxidation tests at higher chloride supplementation, higher current

or extended electrolysis time might also improve treatment performance that might hopefully lead

to DENR compliance. Further studies may be useful for improvement of the treatment strategy.
 6. RECOMMENDATIONS

The anaerobic treatment and preliminary electro-oxidation used in this study were

shown to have a small effect on the removal of pollutants such as COD, BOD and color

in sweet sorghum distillery slop. In order for both treatment to be completely applicable,

further evaluation of the following is recommended:

6.1. Anaerobic Digestion Experiments

Effect of organic loading rate in the performance of the system

The study focused on two (2) organic loading rates (for 2 hydraulic

retention times) only for which both OLRs are more than 1 g/L-d. Additional

loading rates lower than the OLRs used in this study must be further studied in

order to evaluate the performance of other OLRs and to recommend an optimum

OLR for the treatment of sweet sorghum distillery slops. Longer HRTs would be

evaluated as well to check the performance of the anaerobic digester.

Quality of gas produced

Composition of gas should be evaluated to verify the quality of methane

and other gas included in the production of gas by the system. Good gas

production does not always ensure good gas composition. Percentage of methane

and carbon dioxide as well as other gases in the gas produced by the digester

should be evaluated to check for the quality of the gas.

 59

6.2. Electro-oxidation experiments

Effect of chloride supplementation on the removal efficiency of the electro-oxidation

process

Chloride supplementation used in this study was based on the optimum chloride

supplementation used in other studies on the electrochemical treatment of digester

effluent. Further studies using different chloride supplementation is recommended to

evaluate the performance of electro-oxidation using various chloride supplementations.

Measurement at higher operating currents

The study evaluated treatment performance of electro-oxidation at 1.5A only.

Higher operating currents may be tried in order to know if further increase in current

would result in a significant change in the removal efficiencies of the electro-oxidation

process.

Foaming management

Preliminary experiments on electro-oxidation at higher currents were conducted

to determine the effect on removal efficiencies. However, higher currents resulted to

massive foaming and thus, decreasing the reactor volume of the electro-oxidation

process. Further studies on minimizing foaming without affecting COD content of the

wastewater should be done.

 LIST OF REFERENCES

AGARWAL, R., LATA, S., GUPTA, M. & SINGH, P. (2010). Removal of melanoidin
present in distillery effluent: A review. Journal of Environmental Biology July
2010, 521-528. Retrieved December 13, 2011 from
http://www.jeb.co.in/journal_issues/ 201007_jul10/paper_22.pdf.

ALFAFARA, C.G., KAWAMORI, T.K., NOMURA, N., KIUCHI, M. &

MATSUMURA, M. (2003). Electrolytic removal of ammonia from brine
wastewater: scale-up operation and pilot-scale evaluation. Journal of Chemical
Technology and Biotechnology 79, 291-298. doi: 10.1002/jctb.986

ALFAFARA, C.G., NAKANO, K., NOMURA, N., IGARASHI, T. & MATSUMURA,

M. (2002). Operating and scale-up factors for the electrolytic removal of algae
from eutrophied lakewater. Journal of Chemical Technology and Biotechnology
77, 871-876. doi: 10.1002/jctb.649

BAGOTSKY, V.S. (2006). Fundamentals of Electrochemistry (2nd ed). New Jersey: John
Wiley & Sons Inc.

BANERJEE, S. & BISWAS, G.K. (2004). Studies on biomethanation of distillery wastes

and its mathematical analysis. Chemical Engineering Journal 102, 193-201. doi:
10.1016/j.cej.2004.05.006.

BELTRAN, A.B. (2005). Electrolytic decolorization of Sequential Batch Reactor (SBR)

distillery effluent from an alcohol distillery plant. Unpublished undergraduate
thesis. College of Engineering and Agro-Industrial Technology. University of the
Philippines Los Baños, College, Laguna.

DAR, W.D. (2007). Sweet sorghum bioethanol technology. International Crops Research
Institute for the Semi-Arid Tropics. Retrieved March 20, 2011 from
http://www.icrisat.org/Biopower/philippines-event/DarSweetSorghumMay07.pdf.

DEMAFELIS, R.B. (2010). Waste management in the emerging Philippine biofuel

industry. Lecture Seminar. Drilon Hall, Southeast Asian Regional Center for
Graduate Study and Research in Agriculture (SEARCA). University of the
Philippines Los Baños, College, Laguna.

DENR Administrative Order No. 34, 1990

DENR Administrative Order No. 35, 1990

Hadlocon, L.J.S. (2008). Inoculum development for the anaerobic degradability of wash
water from sodium hydroxide – methanol transesterification of Jatropha curcas L.
Unpublished undergraduate thesis. College of Engineering and Agro-Industrial
Technology. University of the Philippines Los Baños, College, Laguna.
 61

International Society for Southeast Asian Agricultural Sciences. (March 2007).

Feasibility study for an integrated anhydrous alcohol production plant using sweet
sorghum as feedstock. Final Report. Retrieved March 20, 2011 from
http://www.bar.gov.ph/biofuelsinfo/downloads/FS%
20of%20Sweet%20Sorghum.pdf

KAPARAJU, P., SERRANO, M. & ANGELIDAKI, I. (2010). Optimization of biogas

production from wheat straw stillage in an UASB reactor. Elsevier jounal of
applied energy 87, 3779-3783. doi: 10.1016/j.apenergy.2010.06.005.

MAGUYON, M.C.C. (2004). Treatability study for the electrochemical treatment of

piggery wastewater. Unpublished undergraduate thesis. College of Engineering
and Agro-Industrial Technology. University of the Philippines Los Baños,
College, Laguna.

MALINA, J.F. & POHLAND, F.G. (1992). Design of anaerobic processes for the
treatment of industrial and municipal wastes. USA: Technocomic Publishing Co.,
Inc.

MERALCO. (November 2011). How do you light up a nation? Meralco report. Retrieved
on January 2, 2012 from http://gpcci.org/home/wp-
content/uploads/2011/11/MERALCO-Presentation-Nov.-2011.pdf.

PANT, D. & ADHOLEYA, A. (2007). Biological approaches for treatment of distillery

wastewater: A review. Bioresource Technology Journal 98, 2321-2324. doi:
10.1016/j.biortech.2006.09.027.

POHLAND, F.G. & SUIDAN, M.T. (1987). Prediction of pH stability in biological

treatment systems. Chemistry of Wastewater Technology, A.J. Rubin Ed: 441-
463.

RAJESHWAR, K. & IBANEZ, J.G. (1997). Environmental electrochemistry:

fundamentals and applications in pollution sensors and abatement. London:
Academic Press.

RUTZ, D. & JANSSEN, R. (2008). Biofuel technology handbook (2nd Ed). Munchen,
Germany: WIP Renewable Energies Inc.,

SAMUDRO, G. & MANGKOEDIHARDJO, S. (2010). Review on BOD, COD and

BOD/COD ratio: A triangle zone for toxic, biodegradable and stable levels.
International Journal of Academic Research 2(4), 235-239. Retrieved December
15, 2011 from http://its.ac.id/personal/files/pub/3298-sarwoko-sarwoko_ijar24-
2010(4-29).pdf
 62

SANTOS, R.B. (2005). Electrolytic treatment of simulated textile wastewater containing

azo-dispersed dye. Unpublished undergraduate thesis. College of Engineering and
Agro-Industrial Technology. University of the Philippines Los Baños, College,
Laguna.

SATYAWALI, Y. & BALAKRISHNAN, M. (2008). Wastewater treatment in molasses-

based alcohol distilleries for COD and color removal: a review. Journal of
Environmental Management 86, 481-497. doi: 10.1016/j.jenvman.2006.12.024.

SMITH, P.G. & SCOTT, J.S. (2005). Dictionary of Water and Waste Management. (2nd
ed). London: IWA Publishing Inc.

VERMA, S. (2002). Anaerobic digestion of biodegradable organics in municipal solid

wastes. Unpublished undergraduate thesis. Fu Foundation of Engineering and
Applied Science, Columbia University, New York.

VILLA, B.C. (2009). Anaerobic treatability of wastewater from the production of ethyl
ester via sodium hydroxide-catalyzed transesterification of Jatropha curcas L. oil.
Unpublished undergraduate thesis. College of Engineering and Agro-Industrial
Technology. University of the Philippines Los Baños, College, Laguna.

WILKIE, A., RIEDIESEL, K.J. & OWENS, J.M. (2000). Stillage characterization and
anaerobic treatment of ethanol stillage from conventional and cellulosic
feedstocks. Biomass and Bioenergy Journal 19, 63-102. Retrieved March 25,
2011 from http://www.sciencedirect.com/science/article/pii/S0961953400000179.
APPENDICES
 APPENDIX A

RAW DATA

I. ANAEROBIC DIGESTION EXPERIMENTS

A. Acclimatization Phase

1. Daily pH monitoring

Appendix Table A1. pH profile of digesters during

acclimatization phase
DAYS GAS VOLUME, mL
30A 30B 60A 60B
0 0 0 0 0
1 0 0 0 0
2 0 0 0 0
3 0 0 0 0
4 43.0 40.0 43.5 39.0
7 156.5 143.5 116.5 154.0
8 212.0 211.0 220.0 222.0
11 354.0 378 378.5 365.0
12 409.0 432.0 435.5 435.5
14 514.5 518.5 529.0 531.0
15 574.5 587.0 598.5 607.5
18 764.5 800.5 787.0 802.5
20 910.0 940.0 917.0 942.0

2. Cumulative gas production

Appendix Table A2. Cumulative gas production of the

digesters during acclimatization
phase
DAYS pH
30A 30B 60A 60B
0 8.48 8.51 8.44 8.5
5 8.14 8.2 8.06 8.24
11 8.11 8.22 8.16 8.01
12 8.21 8.21 8.15 8.13
13 8.1 7.99 7.83 7.81
14 8.17 8.06 8.1 7.87
 65

Appendix Table A2 continued . . .

DAYS pH
30A 30B 60A 60B
15 8.16 8.04 8.01 8.06
18 7.9 7.93 7.73 7.86
19 7.83 7.94 7.86 7.92
20 7.88 8.05 8.02 8.01

3. Chemical Oxygen Demand (COD) Determination

Appendix Table A3. Volume of standard ferrous ammonium sulfate titrant

used for 10-mL sample volume

SAMPLING VOLUME OF Fe(NH4)2(SO4)2 (mL)

PERIOD SOLUTION
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Trial 1 Trial 2
Standard
25.25 25.20 25.25 25.20
Solution
1 Blank 24.70 24.70 24.70 24.70
QAS 23.40 23.40 23.40 23.40
Sample 22.15 22.05 21.90 22.10
Standard
25.25 25.25 25.25 25.25
Solution
5 Blank 24.90 24.90 24.90 24.90
QAS 23.90 23.90 23.90 23.90
Sample 22.60 22.60 22.50 22.50
Standard
25.30 25.50 25.10 25.15
Solution
11 Blank 22.60 22.60 24.35 24.35
QAS 21.90 21.90 23.20 23.20
Sample 20.50 20.60 22.15 22.30
Standard
25.30 25.50 25.10 25.15
Solution
15 Blank 22.60 22.60 24.35 24.35
QAS 21.90 21.90 23.20 23.20
Sample 20.80 20.70 22.40 22.60
Standard
25.30 25.50 25.30 25.50
Solution
20 Blank 24.75 24.75 24.75 24.75
QAS 23.55 23.55 23.55 23.55
Sample 23.35 23.25 23.20 23.25
 66

B. Semi-continuous Anaerobic Digestion Experiments

1. Daily pH monitoring

Appendix Table A4. pH profile of digesters during semi-continuous anaerobic digestion experiments

pH pH
DAYS DAYS
30A 30B 60A 60B 30A 30B 60A 60B
0 7.41 7.36 7.49 7.50 49 5.95 5.83 7.66 7.59
1 7.58 7.51 7.59 7.60 50 5.90 5.79 7.55 7.62
2 7.33 7.26 7.41 7.32 51 5.91 5.79 7.51 7.56
3 7.20 7.11 7.21 7.19 52 5.94 5.83 7.56 7.59
4 6.97 7.04 7.08 7.11 53 5.87 5.80 7.55 7.65
7 7.05 7.12 7.19 7.27 56 5.82 5.79 7.68 7.75
8 6.84 6.83 7.02 7.07 57 5.77 5.75 7.58 7.59
9 6.81 6.70 7.01 7.04 58 5.75 5.73 7.57 7.58
10 6.80 6.75 6.82 6.92 59 5.73 5.75 7.56 7.54
14 6.58 6.49 7.12 7.11 60 5.72 5.71 7.61 7.60
15 6.47 6.42 7.02 7.11 65 5.68 5.73 7.71 7.66
16 6.53 6.45 7.07 7.08 66 5.74 5.72 7.72 7.70
17 6.51 6.38 7.01 7.00 67 5.68 5.65 7.80 7.76
18 6.49 6.42 7.18 7.14 70 5.66 5.61 7.72 7.74
21 6.52 6.43 7.26 7.34 71 5.68 5.62 7.78 7.78
22 6.45 6.39 7.40 7.49 72 5.65 5.62 7.69 7.68
23 6.40 6.32 7.42 7.46 73 5.58 5.63 7.79 7.75
24 6.34 6.26 7.28 7.32 74 5.64 5.58 7.75 7.72
25 6.30 6.16 7.25 7.30 77 5.63 5.57 7.74 7.75
28 6.10 6.05 7.64 7.79 79 5.63 5.56 8.39 8.44
29 5.98 6.03 7.46 7.67 80 5.61 5.56 8.02 7.86
30 6.03 5.99 7.47 7.62 84 5.62 5.54 7.87 7.80
31 5.97 5.96 7.43 7.41 85 5.50 5.53 7.81 7.80
32 5.92 5.92 7.33 7.38 86 5.62 5.55 7.84 7.97
35 6.00 5.99 7.56 7.63 87 5.60 5.50 7.98 8.16
36 5.98 5.95 7.61 7.57 88 5.61 5.54 7.94 8.20
37 5.99 5.99 7.54 7.64 91 5.62 5.55 7.75 7.78
38 5.95 5.93 7.48 7.47 92 7.74 7.78
39 5.93 5.88 7.51 7.49 93 7.78 7.79
42 5.89 5.81 7.64 7.61 94 7.66 7.65
43 5.87 5.79 7.54 7.52 95 7.65 7.69
44 5.88 5.81 7.52 7.54 98 7.63 7.64
45 5.84 5.76 7.51 7.49 99 7.62 7.62
46 5.86 5.77 7.51 7.50 100 7.58 7.59
 67

Appendix Table A4 continued . . .

pH
DAYS
30A 30B 60A 60B
101 7.59 7.61
102 7.54 7.57
105 7.58 7.56
106 7.51 7.50
107 7.52 7.49
108 7.58 7.56
109 7.54 7.53
112 7.47 7.49
113 7.57 7.59
114 7.56 7.57
115 7.53 7.54
116 7.58 7.57
119 7.61 7.60
120 7.57 7.57

2. Daily Gas Production

Appendix Table A5. Cumulative gas production of the digesters during semi-continuous anaerobic
digestion experiments
DAILY GAS PRODUCTION, mL DAILY GAS PRODUCTION, mL
DAYS DAYS
30A 30B 60A 60B 30A 30B 60A 60B
0 0 0 0 0 23 1100 797 1662 2158
1 217 216 214 185 24 1003 -56 2026 2080
2 824 842 455 490 25 453 1058 2338 2494
3 897 978 597 573 28 314 212 1806 1809
4 1143 1156 650 600 29 560 944 2192 2246
7 1020 1016 604 603 30 293 633 2411 2407
8 1025 1122 736 862 31 49 264 2369 2384
9 1543 1572 1086 1119 32 0 95 2300 2341
10 1063 1065 941 872 35 75 51 1785 1773
11 1393 1496 1129 1359 36 42 132 2528 2295
14 1244 1355 1004 1065 37 0 49 2343 2200
15 1111 1333 1340 1449 38 0 4 2141 2121
16 939 1261 1399 1487 39 99 0 2496 2423
17 1060 1132 1349 1357 42 73 195 1325 1201
18 640 1169 1361 1524 43 111 132 1953 1661
21 676 717 1406 1514 44 109 138 1940 1815
22 1064 1026 1885 1959 45 174 226 1947 2045
 68

Appendix Table A5 continued . . .

DAILY GAS PRODUCTION, mL DAILY GAS PRODUCTION, mL
DAYS DAYS
30A 30B 60A 60B 30A 30B 60A 60B
46 49 183 1867 1930 84 0 0 1374 1212
49 346 128 1349 1690 85 0 0 1919 1890
50 147 401 2258 2103 86 0 0 2102 2133
51 266 150 1945 2047 87 0 0 2033 1896
52 97 0 2184 2138 88 0 0 1869 1592
53 0 0 2002 1905 91 0 0 1568 1389
56 0 0 1748 1709 92 2006 1777
57 0 0 1919 1687 93 2117 2150
58 0 0 1906 1845 94 2274 2288
59 0 0 2239 1988 95 2185 2217
60 0 0 2216 2438 98 1649 1478
63 0 0 1559 1576 99 2008 2025
64 0 0 1837 1840 100 2057 2247
65 0 0 2083 2246 101 2269 2340
66 0 0 2212 2350 102 2131 2115
67 0 0 1928 2036 105 1676 1572
70 0 0 1637 1686 106 1992 1993
71 0 0 1917 2042 107 2474 2450
72 0 0 2108 2175 108 2111 2094
73 0 0 1831 1880 109 2220 2036
74 0 0 2042 2058 112 1648 1641
77 0 0 1576 1612 113 2508 2559
78 0 0 1759 1795 114 2336 2413
79 0 0 2167 2336 115 2514 2521
80 0 0 2226 2328 116 2246 2249
81 0 0 2112 1595 119 1632 1679
120 2056 2342

3. Chemical Oxygen Demand (COD) Determination

 69

Appendix Table A6. Volume of standard ferrous ammonium sulfate titrant

used for 10-mL sample volume

SAMPLING VOLUME OF Fe(NH4)2(SO4)2 (mL)

PERIOD SOLUTION
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Trial 1 Trial 2
Standard
25.50 25.30 25.50 25.30
Solution
0 Blank 25.00 25.00 25.00 25.00
QAS 24.05 24.05 24.05 24.05
Sample 22.6 22.6 22.45 22.60
Standard
25.40 25.40
Solution
4 Blank 25.20 25.20
QAS 24.05 24.05
Sample 22.60 22.60
Standard
25.40 25.40 25.40 25.40
Solution
10 Blank 25.20 25.20 25.20 25.20
QAS 24.05 24.05 24.05 24.05
Sample 22.20 22.55 22.50 22.70
Standard
26.10 26.10
Solution
15 Blank 25.50 25.50
QAS 24.40 24.40
Sample 22.60 22.55
Standard
26.70 26.30 26.70 26.30
Solution
21 Blank 24.75 24.75 24.75 24.75
QAS 24.95 24.95 24.95 24.95
Sample 22.50 22.50 23.00 23.20
Standard
27.00 26.75
Solution
25 Blank 24.75 24.75
QAS 24.95 24.95
Sample 22.40 22.20
Standard
26.50 26.65 25.25 25.20
Solution
30 Blank 25.85 25.85 25.00 25.00
QAS 25.20 25.20 24.10 24.10
Sample 22.60 22.80 22.20 22.90
 70

Appendix Table A6 continued . . .

SAMPLING VOLUME OF Fe(NH4)2(SO4)2 (mL)

PERIOD SOLUTION
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Trial 1 Trial 2
Standard
26.50 26.65
Solution
35 Blank 25.85 25.85
QAS 25.20 25.20
Sample 22.10 22.80
Standard
26.75 26.60 26.70 26.30
Solution
39 Blank 26.00 26.00 25.00 25.00
QAS 25.25 25.25 24.95 24.95
Sample 21.85 21.90 23.00 23.20
Standard
27.20 27.25
Solution
45 Blank 26.65 26.65
QAS 24.80 24.80
Sample 21.60 22.00
Standard
25.70 25.80 25.25 25.20
Solution
50 Blank 25.30 25.30 25.00 25.00
QAS 24.50 24.50 24.10 24.10
Sample 20.90 20.75 22.65 23.00
Standard
26.35 26.30 25.25 25.20
Solution
60 Blank 24.75 24.75 25.00 25.00
QAS 24.95 24.95 24.10 24.10
Sample 21.55 21.65 22.70 22.65
Standard
26.10 26.10
Solution
65 Blank 25.65 25.65
QAS 25.10 25.10
Sample 21.40 23.80
Standard
26.10 26.10 25.00 25.15
Solution
70 Blank 25.65 25.65 24.60 24.60
QAS 25.10 25.10 24.60 24.60
Sample 21.80 21.75 23.00 22.95
 71

Appendix Table A6 continued . . .

SAMPLING VOLUME OF Fe(NH4)2(SO4)2 (mL)

PERIOD SOLUTION
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Trial 1 Trial 2
Standard
26.10 26.10
Solution
74 Blank 24.75 24.75
QAS 24.85 24.95
Sample 21.80 21.70
Standard
26.10 26.10 26.05 26.05
Solution
80 Blank 25.65 25.65 25.55 25.55
QAS 24.85 25.10 25.10 25.10
Sample 21.90 21.40 23.75 23.70
Standard
26.85 26.75
Solution
85 Blank 25.70 25.70
QAS 25.60 25.60
Sample 22.05 21.85
Standard
26.50 26.50 26.50 26.50
Solution
91 Blank 25.70 25.70 25.70 25.70
QAS 25.60 25.60 25.60 25.60
Sample 21.85 21.85 24.15 23.90
Standard
25.35 25.40
Solution
100 Blank 25.05 25.05
QAS 23.60 23.60
Sample 21.25 21.25
Standard
26.50 26.50
Solution
109 Blank 24.70 24.70
QAS 25.60 25.60
Sample 22.90 23.10
Standard
25.30 25.20
Solution
120 Blank 25.05 25.05
QAS 23.60 23.60
Sample 21.60 21.65

4. Color time course for the semi-continuous anaerobic digestion of sweet sorghum
distillery slop.
Appendix Table A7. Absorbance profile of sweet sorghum distillery slop for the
semi-continuous anaerobic digestion experiments
ABSORBANCE
TIME
HRT = 30 days
(Days)
Trial 1 Trial 2 Average
0 218.4960 229.8604 224.1782
1 196.1689 240.8611 218.5150
2 248.9869 208.3577 228.6723
3 216.4835 240.8611 228.6723
4 224.6094 208.3577 216.4835
7 241.0752 240.7081 240.8917
8 168.5245 153.1057 160.8151
9 160.8151 160.8151 160.8151
10 160.8151 183.9433 172.3792
11 153.1057 153.1057 153.1057
17 156.7865 156.7865 156.7865
18 156.7865 168.0077 162.3971
21 168.0077 160.5269 164.2673
23 141.9327 141.9327 141.9327
24 166.6356 154.2842 160.4599
25 154.2842 166.6356 160.4599
28 192.3317 152.2668 172.2993
29 156.2733 140.2474 148.2603
30 152.2668 156.2733 154.2701
31 150.0986 142.5792 146.3389
32 131.3001 153.8583 142.5792
35 176.4166 161.3778 168.8972
36 158.4021 166.2371 162.3196
37 185.8248 177.9897 181.9073
38 185.8248 205.4125 195.6186
39 152.7453 152.7453 152.7453
42 187.5472 199.1479 193.3476
43 175.9466 179.8135 177.8800
44 164.3966 172.1768 168.2867
45 191.6273 168.2867 179.9570
46 168.2867 179.9570 174.1219
49 141.6654 156.9513 149.3084
72
HRT = 60 days
Trial 1 Trial 2 Average
161.6736 157.8854 159.7795
236.7982 248.9869 242.8925
248.9869 179.9173 214.4521
212.4206 216.4835 214.4521
220.5465 257.1128 238.8296
253.9236 240.3410 247.1323
118.4134 129.9775 124.1954
83.7211 160.8151 122.2681
176.2339 203.2168 189.7253
160.8151 168.5245 164.6698
190.4501 164.2673 177.3587
168.0077 168.0077 168.0077
179.2289 156.7865 168.0077
162.5184 162.5184 162.5184
166.6356 158.4013 162.5184
183.1041 187.2213 185.1627
164.2863 156.2733 160.2798
160.2798 160.2798 160.2798
172.2993 172.2993 172.2993
161.3778 161.3778 161.3778
150.0986 168.8972 159.4979
172.6569 191.4555 182.0562
189.7423 177.9897 183.8660
201.4949 177.9897 189.7423
213.2475 209.3300 211.2888
195.2810 199.1479 197.2144
199.1479 226.2161 212.6820
222.3492 226.2161 224.2826
183.8471 203.2976 193.5724
195.5174 199.4075 197.4625
226.6383 211.0779 218.8581
202.8089 198.9874 200.8982
 73

Appendix Table A7 continued . . .

ABSORBANCE
TIME
HRT = 30 days HRT = 60 days
(Days)
Trial 1 Trial 2 Average Trial 1 Trial 2 Average
50 164.5942 160.7728 162.6835 210.4518 214.2733 212.3626
51 153.1298 156.9513 155.0406 176.0586 202.8089 189.4338
52 147.9226 187.8550 167.8888 199.8347 215.8077 173.8787
53 187.8550 159.9023 173.8787 167.8888 191.8482 179.8685
56 135.9429 139.9361 137.9395 195.8415 195.8415 195.8415
57 139.5166 151.5111 145.5139 167.5038 151.5111 159.5074
58 167.5038 151.5111 159.5074 175.5001 183.4964 179.4983
59 179.4983 175.5001 177.4992 179.4983 191.4928 185.4955
60 163.5056 143.5148 153.5102 167.5038 183.4964 175.5001
63 131.5203 119.5258 125.5230 147.5129 139.5166 143.5148
64 151.5998 159.5333 155.5665 191.2672 183.3337 187.3005
65 191.2672 183.3337 187.3005 195.2340 175.4002 185.3171
66 155.5665 163.5000 159.5333 191.2672 183.3337 187.3005
67 163.5000 171.4335 167.4667 179.3670 191.2672 185.3171
70 163.5000 155.5665 159.5333 230.9347 223.0012 226.9680
71 167.4667 183.3337 175.4002 195.2340 234.9015 215.0677
72 179.3670 223.0012 201.1841 183.3337 195.2340 189.2839
73 240.2361 177.6795 208.9578 181.8499 215.2134 198.5317
74 198.5317 198.5317 198.5317 194.3612 211.0430 202.7021
77 160.9977 160.9977 160.9977 190.1908 206.8726 198.5317
78 156.8273 190.1908 173.5091 223.5543 219.3839 221.4691
79 145.1123 165.0989 155.1056 201.0748 201.0748 201.0748
80 201.0748 201.0748 201.0748 177.0909 177.0909 177.0909
81 141.1150 153.1070 147.1110 181.0882 181.0882 181.0882
84 145.1123 157.1043 151.1083 205.0721 169.0962 187.0842
85 145.1123 165.0989 155.1056 197.0775 189.0828 193.0802
86 132.8957 140.1285 136.5121 190.7580 208.8399 199.7989
87 183.5252 183.5252 183.5252 205.2235 194.3744 199.7989
88 140.1285 158.2104 149.1695 197.9907 183.5252 190.7580
91 161.8268 165.4432 163.6350 197.9907 201.6071 199.7989
92 168.5066 179.1926 173.8496
93 193.4406 186.3166 189.8786
94 182.7546 189.8786 186.3166
95 200.5646 214.8126 207.6886
99 167.6087 171.1836 169.3961
100 214.0822 199.7826 206.9324
 74

Appendix Table A7 continued . . .

ABSORBANCE
TIME HRT = 30 days HRT = 60 days
(Days) Trial Trial
Average Trial 1 Trial 2 Average
1 2
101 185.4831 174.7585 180.1208
102 178.3334 181.9082 180.1208
105 181.9082 181.9082 181.9082
106 214.0822 214.0822 214.0822
107 206.9324 199.7826 203.3575
108 214.0822 199.7826 206.9324
109 221.2319 256.9808 239.1063
112 149.7343 153.3092 151.5217
113 135.4348 131.8599 133.6473
114 153.3092 146.1594 149.7343
115 164.0338 139.0096 151.5217
116 174.7585 153.3092 164.0338
119 174.7585 164.0338 169.3961
120 221.2319 221.2319 221.2319

II. PRELIMININARY ELECTRO-OXIDATION EXPERIMENTS

A. Initial operating conditions, pH and temperature time courses

Table A8. pH and temperature profile at an operating current of 1.5A and 25

g/L NaCl supplementation
TIME VOLUME VOLTAGE CURRENT TEMPERATURE
pH
(hr) (mL) (V) (A) (oC)
0 800 4.1 1.5 29 7.59
1 785 5.0 1.5 35 8.03
2 767 5.0 1.5 36 8.24
3 750 5.0 1.5 36 8.39
4 730 5.0 1.5 36 8.40
5 690 5.0 1.5 37 8.41
6 675 5.0 1.5 37 8.44
7 650 5.0 1.5 38 8.45
8 625 6.0 1.5 39 8.50
 75

Appendix Table A8 continued . . .

TIME VOLUME VOLTAGE CURRENT TEMPERATURE
pH
(hr) (mL) (V) (A) (oC)
9 600 6.9 1.5 39 8.50
10 600 7.0 1.5 39 8.55
11 590 6.0 1.5 39 8.60
12 550 5.2 1.5 41 8.60

B. Chemical Oxygen Demand Determination

Appendix Table A9. Volume of standard ferrous ammonium sulfate titrant used for 10-mL sample volume

VOLUME OF
VOLUME OF
TIME Fe(NH4)2(SO4)2 TIME
SOLUTION SOLUTION Fe(NH4)2(SO4)2 (mL)
(hr) (mL) (hr)
Trial 1 Trial 2 Trial 1 Trial 2
Standard Standard
29.20 29.20 29.20 29.20
Solution Solution
0 Blank 28.00 28.00 9 Blank 28.00 28.00
QAS 26.50 26.50 QAS 26.50 26.50
Sample 25.80 25.85 Sample 26.00 26.10
Standard Standard
29.20 29.20 29.20 29.20
Solution Solution
3 Blank 28.00 28.00 12 Blank 28.00 28.00
QAS 26.50 26.50 QAS 26.50 26.50
Sample 25.85 25.85 Sample 26.30 26.40
Standard
29.20 29.20
Solution
6 Blank 28.00 28.00
QAS 26.50 26.50
Sample 26.20 26.20
 76

C. Color time course for the preliminary electro-oxidation of the digested effluent

Appendix Table A10. Absorbance profile digested effluent from

preliminary electro-oxidation
TIME ABSORBANCE (462 nm)
(hr) Trial 1 Trial 2 Average
0 117.42 117.42 117.42
3 113.9780 113.9780 113.978
6 107.094 107.094 107.094
9 96.7675 96.7675 96.7675
12 69.2307 69.2307 69.2307
 APPENDIX B

DERIVED DATA

I. ANAEROBIC DIGESTION EXPERIMENTS

C. Acclimatization Phase

Appendix Table B1. COD profile of digesters during acclimatization phase

CHEMICAL OXYGEN DEMAND (mg/L)
TIME
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Mean Trial 1 Trial 2 Mean
0 50545.14 52527.31 51536.23 55500.55 51536.23 53518.39
5 47000.19 45544.55 46272.37 47524.75 47524.75 47524.75
11 41339.22 39370.69 40354.96 43781.14 40796.06 42288.60
15 35433.62 37402.15 36417.89 38806.01 34825.91 36815.96
20 27559.48 29528.02 28543.75 30512.28 29528.02 30020.15

Appendix Table B2. COD reduction profile of digesters during acclimatization

phase
MEAN COD (mg/L)
TIME
(Days) HRT = 30 HRT = 60
% Reduction % Reduction
Days Days
0 51536.23 53518.39
44.61420098 43.90685146
20 28543.75 30020.15

D. Semi-continuous Anaerobic Digestion Experiments

Appendix Table B3. COD profile of digesters at each HRT for the semi-continuous
anaerobic digestion of sweet sorghum distillery slop
CHEMICAL OXYGEN DEMAND (mg/L)
TIME
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Mean Trial 1 Trial 2 Mean
0 47,224.83 47,224.83 47,224.83 50,197.63 47,244.83 48,721.23
4 43,307.09 53,149.61 48,228.35
10 59,055.12 52,165.32 55,610.22 53,149.61 49,212.60 51,181.11
15 55,555.56 56,513.41 56,034.49
21 42,455.25 42,455.25 42,455.25 59,437.35 55,663.55 57,550.45
 78

Appendix Table B3 continued . . .

CHEMICAL OXYGEN DEMAND (mg/L)

TIME
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Mean Trial 1 Trial 2 Mean
25 76,280.72 80,001.73 78,141.23
30 61,618.55 78,551.90 70,085.23 55,500.55 41,625.41 48,562.98
35 75,984.99 70,754.72 73,369.85
39 79,663.24 78,726.02 79,194.63 49,203.76 46,860.73 48,032.25
45 92,745.72 85,399.52 89,072.62
50 85,437.22 88,349.85 86,893.54 48,543.87 45,631.24 47,087.56
56 73,572.74 77,444.99 75,508.87
60 83,573.83 81,671.49 82,622.66 45,589.74 46,580.68 46,085.21
65 81,147.62 86,754.06 83,950.84
70 73,754.79 74,712.64 74,233.72 36,239.02 34,280.15 35,259.59
74 73,754.79 75,670.50 74,712.65
80 71,839.08 81,417.62 76,628.35 34,548.94 35,508.64 35,028.79
85 82,556.26 69,339.62 75,947.94
91 72,641.51 72,641.51 72,641.51 31,132.08 33,962.36 32,547.22
100 31,840.83 36,453.24 34,147.04
109 33,663.50 33,168.45 33,415.98
120 33,743.88 33,663.50 33,703.69

Appendix Table B4. COD reduction profile of digester at

HRT = 60 days for the semi-continuous anaerobic
digestion of sweet sorghum distillery slop

TIME
COD (mg/L) % REDUCTION
(Days)
0 48,721.23
30.8234008
120 33,703.69
 79

II. PRELIMININARY ELECTRO-OXIDATION EXPERIMENTS

A. Chemical Oxygen Demand Determination and Removal Efficiency

Appendix Table B5. COD profile of digested effluent before and after
electro-oxidation at 1.5A operating current and 25 g/L NaCl
supplementation.
TIME COD (mg/L)
(hr) Trial 1 Trial 2 Mean
0 37,671.23 36,815.07 37,243.15
3 36,815.07 36,815.07 36,815.07
6 34,246.58 32,534.25 33,390.41
9 30,821.92 30,821.92 30,821.92
12 29,109.59 27,397.26 28,253.42

Appendix Table B6. COD reduction profile of the

digested effluent before and after electro-oxidation
treatment
TIME MEAN COD %
(hr) (mg/L) REDUCTION
0 37,243.15
24.13793103
12 28,253.42

B. Color Removal Efficiency

Appendix Table B7. Color reduction profile of the

digested effluent before and after electro-oxidation
treatment
MEAN
TIME %
ABSORBANCE
(hr) REDUCTION
(462 nm)
0 117.4201357
41.04017027
12 69.23071206
 80

C. Charge Dose
Appendix Table B8. Charge dose determination for color removal and COD
reduction of the digested effluent at 1.5A and 25 g/L NaCl supplementation.

TIME COLOR REMOVED COD REDUCED

TIME (s)
(hr) (Co-C)V It mg It
0 0.00 0.00 0.00 0.00 0.00
3 10,800.00 2,581.58 16,200.00 321.06 16,200.00
6 21,600.00 6,970.26 32,400.00 2,600.60 32,400.00
9 32,400.00 12,391.57 48,600.00 3,852.74 48,600.00
12 43,200.00 26,504.18 64,800.00 4,944.35 64,800.00
 APPENDIX C

SAMPLE CALCULATIONS

I. ANAEROBIC DIGESTION EXPERIMENTS

A. Chemical Oxygen Demand Determination

Dichromate open reflux method was used to determine the COD of the sweet sorghum

distillery slop as shown in the following equation:

( − )(8000)( )
=

Where a = mL Fe(NH4)2(SO4)2 used for blank

b = mL Fe(NH4)2(SO4)2 used for sample

N = normality of Fe(NH4)2(SO4)2 used

( )( )
=
( ( ) ( ) )

Consider the following data:

Appendix Table C1. Volume of standard ferrous ammonium sulfate titrant

used for 10-mL sample volume

SAMPLING VOLUME OF Fe(NH4)2(SO4)2 (mL)

PERIOD SAMPLE
(Days) HRT = 30 Days HRT = 60 days
Trial 1 Trial 2 Trial 1 Trial 2
Standard
26.65 26.50
Solution
35 Blank 25.85 25.85
QAS 25.20 25.20
Sample 22.80 22.10

Considering trial 1 of the sample at HRT = 30 days,

( − )(8000)( )
= ( )

( . )( )
(25.85 − 22.80) (8000)
.
= (500)
10
 82

= 75,984.99

Considering trial 2 of the sample at HRT = 30 days,

( − )(8000)( )
= ( )

( . )( )
(25.85 − 22.10) (8000)
.
= (500)
10

= 70,754.72

(75,984.99 + 70,754.72)
= = 73,369.85
2

B. COD Removal Efficiency

Calculation for the removal efficiency of organic matter in the digester will be

evaluated using the equation below:

−
% = 100

Where Co = initial concentration of pollutant

C = final concentration of pollutant

Consider Table B of Appendix B:

(48,721.23 − 33,703.69)
% = 100
48,721.23

% = 30.82%
 83

II. PRELIMININARY ELECTRO-OXIDATION EXPERIMENTS

A. Chemical Oxygen Demand and BOD5 to COD Ratio Determination and Removal

Efficiencies

Calculations for COD and BOD5/COD, as well as removal efficiency, are similar

to the sample calculations of anaerobic digestion experiments of this appendix.

B. Charge Dose

Charge dose was determined by plotting (Co-C)V values with It as slope

of the curve passing through the origin as shown in the following equation:

=
( − )

where Q = electrical charge passed (Coulomb)

I = electrical current (Ampere)

t = electrolysis time (second)

Co = initial concentration of pollutant

C = final concentration of pollutant

V = operating volume (Liter)

Considering the chemical oxygen demand profile for charge dose determination of the

electro-oxidation experiment from Table B5 and Table B10 of Appendix B:

Appendix Table C2. COD reduction profile of the digested effluent at 1.5A operating current and
25 g/L NaCl supplementation.
TIME TIME VOLUME CURRENT
C (mg/L) Co (mg/L)
(hr) (s) (L) (A)
0 0 0.8 1.5 37243.1507 37243.15068
3 10800 0.75 1.5 36815.0685 37243.15068
6 21600 0.675 1.5 33390.411 37243.15068
9 32400 0.6 1.5 30821.9178 37243.15068
12 43200 0.55 1.5 28253.4247 37243.15068
 84

Consider t = 3 hrs (10,800 s)

= (1.5 )(10,800 ) = 16,200 −

( − ) = 37,243.15068 − 36,815.0685 (0.75 ) = 321.0616

For each time interval, the values of (Co-C)V and It were calculated shown in Table C3.

Appendix Table B8. Charge dose determination for the electro-oxidation of digested effluent with
1.5A and 25 g/L NaCl supplementation.
TIME TIME
It (Co-C)V
(hr) (s)
0 0 0 0
3 10800 16200 321.0616
6 21600 32400 2600.599
9 32400 48600 3852.74
12 43200 64800 4944.349

The values of It versus (Co-C)V were plotted and the slope of the linearized plot is the

charge dose as shown in the figure below.

70000

60000

50000
y = 11.632x + 5137.3
40000 R² = 0.9636
It

30000

20000

10000

0
0 1000 2000 3000 4000 5000 6000
(Co-C)V

The computed charge dose was 11.632 Coulombs/mg. Charge dose values for color

removal was also determined.

 85

C. Energy Requirement

Specific energy requirement for the electrochemical decolorization and COD

reduction of the digested effluent can be computed using the following equation:

=
3.6 10

Where W = specific energy requirement (kWh/mg pollutant

removed or kWh/Abs-L color removed)

Q = charge dose (Coulombs per mg pollutant removed or

Coulombs/Abs-L color removed)

E = operating voltage (V)

Considering the charge dose (11.632 Coulombs/mg pollutant removed) for COD

removed at 25 g/L NaCl supplementation and 1.5A operating current,

− 1 1 1ℎ
= = 11.632 (5.4 )
1 − 1000 3600

= 1.7448 10

Considering the calculated specific energy requirement and the prevailing

industrial power cost of PhP7.39/kWh (Meralco, November 2011), the energy cost on COD

removal of the digested effluent with an initial COD of 37,243.15 mg/L and final COD of

28,253.42 is calculated as follows:

ℎ ℎ 7.39 1000
= 1.7448 10 (37,243.15 − 28,253.42)
ℎ 1

ℎ 1,159.14
=
 86

D. Color Removal Efficiency

Color removal for the electro-oxidation treatment of the digested effluent was

calculated using the equation below.

−
% = 100

Where = absorbance at maximum wavelength at initial

time
= absorbance at maximum wavelength at time t

Consider Table A10 of Appendix A,

Appendix Table A10. Absorbance profile digested

effluent from preliminary electro-oxidation
TIME ABSORBANCE (462 nm)
(hr) Trial 1 Trial 2 Average
0 117.4201 117.4201 117.4201
3 113.9780 113.9780 113.978
6 107.0938 107.0938 107.0938
9 96.7675 96.7675 96.76753
12 69.2307 69.2307 69.23071

117.4201 − 69.2307
% = 100
117.4201

% = 41.04%
 APPENDIX D

CHEMICAL OXYGEN DEMAND (COD) DETERMINATION

(APHA Standard Methods for the Examination of Water and Wastewater, 1998)

Apparatus

a. Reflux apparatus, consisting of 500- or 250-mL Erlenmeyer flasks with ground-glass

24/40 neck and 300-mm jacket Liebig, West, or equivalent condenser with 24/40 ground-
glass joint, and hot plate having sufficient power to produce at least 1.4 W/cm2 of heating
surface, or equivalent.

b. Blender.

c. Pipets, Class A and wide-bore.

Reagents

a. Standard potassium dichromate solution, 0.04167M:

Dissolve 12.259 g K2Cr2O7, primary standard grade, previously dried at 150oC for 2h, in
distilled water and dilute to 1000 mL. This reagent undergoes a six-electron reduction
reaction; the equivalent concentration is 6 x 0.04167 M or 0.2500 N.

b. Sulfuric acid reagent:

Add Ag2SO4, reagent or technical grade, crystals or powder, to concentrated H2SO4 at the
rate of 5.5 g Ag2SO4/kg H2SO4. Let stand 1 to 2 days to dissolve. Mix

c. Ferroin indicator solution:

Dissolve 1.485 g 1,10-phenanthroline monohydrate and 695 mg FeSO4 · 7H2O in
distilled water and dilute to 100 mL.

d. Standard ferrous ammonium sulfate (FAS) titrant, approximately 0.25M:

Dissolve 98 g Fe(NH4)2(SO4)2 · 6H2O in distilled water. Add 20 mL concentrated H2SO4,
cool, and dilute to 1000 mL. Standardize this solution daily against K2Cr2O7 solution as
follows:

Dilute 25.00 mL standard K2Cr2O7 to about 100 mL. Add 30 mL concentrated H2SO4 and
cool. Titrate with FAS titrant using 0.10 to 0.15 mL (2 to 3 drops) ferroin indicator.

0.250 K Cr O ,
= 0.2500
,

e. Mercuric Sulfate, HgSO4, crystals or powder.

 88

f. Sulfanamic acid:
Required only if the interference of nitrites is to be eliminated.

g. Potassium hydrogen phthalate (KHP) standard, HOOC6H4COOK:

Lightly crush and then dry KHP to constant weight at 110oC. Dissolve 425 mg in
distilled water and dilute to 1000 mL. KHP has a theoretical COD of 1.176 mg O2/mg
and this solution has a theoretical COD of 500 μg O2/mL. This solution is stable when
refrigerated, but not indefinitely. Be alert to development of visible biological growth. If
practical, prepare and transfer solution under sterile conditions. Weekly preparation is
satisfactory.

Procedure

a. Treatment of samples with COD of >50 mg O2/L:

Blend sample if necessary and pipet 50.00 mL into a 500-mL refluxing flask. For
samples with a COD of >900 mg O2/L, use a smaller portion diluted to 50.00 mL.
Add 1 g HgSO4, several glass beads, and slowly add 5.0 mL sulfuric acid reagent,
with mixing to dissolve HgSO4. Cool while mixing to avoid possible loss of volatile
materials. Add 25.00 mL 0.04167M K2Cr2O7solution and mix. Attach flask to
condenser and turn on cooling water. Add remaining sulfuric acid reagent (70 mL)
through open end of condenser. Continue swirling and mixing while adding sulfuric
acid reagent. CAUTION: Mix reflux mixture thoroughly before applying heat to
prevent local heating of flask bottom and a possible blowout of flask contents.

Cover open end of condenser with a small beaker to prevent foreign material from
entering refluxing mixture and reflux for 2 h. Cool and wash down condenser with
distilled water. Disconnect reflux condenser and dilute mixture to about twice its
volume with distilled water. Cool to room temperature and titrate excess K2Cr2O7
with FAS, using 0.10 to 0.15 mL (2 to 3 drops) ferroin indicator. Although the
quantity of ferroin indicator is not critical, use the same volume for all titrations. Take
as the end point of the titration the first sharp color change from blue-green to reddish
brown that persists for 1 min or longer. Duplicate determinations should agree with
5% of their average. Samples with suspended solids or components that are low to
oxidize may require additional determinations. The blue-green may reappear. In the
same manner, reflux and titrate a blank containing the reagents and a volume of
distilled water equal to that of the sample.

b. Alternate procedure for low-COD samples:

Follow procedure of 4a, with two exceptions: (i) use standard 0.004167 M K2Cr2O7,
and (ii) titrate with standardized 0.025M FAS. Exercise extreme care with this
procedure because even a trace of organic matter on the glassware or from the
atmosphere may cause gross errors. If a further increase in sensitivity is required,
concentrate a larger volume of sample before digesting under reflux as follows: Add
all reagents to a sample than 50 mL and reduce total volume of 150 mL by boiling in
 89

the refluxing flask open to the atmosphere without the condenser attached. Compute
amount of HgSO4 to be added (before concentration) on the basis of a weight ratio of
10:1, HgSO4:Cl-, using the amount of Cl- present in the original volume of sample.
Carry a blank reagent through the same procedure. This technique has the advantage
of concentrating the sample without significant losses of easily digested volatile
materials. Hard-to-digest volatile materials such as volatile acids are lost, but an
improvement is gained over ordinary evaporative concentration methods. Duplicate
determinations are not expected to be as precise in 4a.

c. Determination of standard solution:

Evaluate the technique and quality of reagents by conducting the test on a standard
potassium hydrogen phthalate solution.

Computations

( − )(8000)( )
=

where a = mL Fe(NH4)2(SO4)2 used for blank

b = mL Fe(NH4)2(SO4)2 used for sample

N = normality of Fe(NH4)2(SO4)2 used

( )( )
=
( ( ) ( ) )
APPENDIX E
91
92
93
94
95
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118