ASIA International Multidisciplinary Conference 2018

Abstract ID: AIMC-2018-STEM-64

Effects of Adding Oil, Fat and Grease (FOG) Waste from Grease Trap on Volatile Fatty Acids
(VFA) Accumulation in An Anaerobic Digester of Food Waste

Hanifah Khairiah *a and Cindy Rianti Priadi b

Departement of Civil Engineering, University of Indonesiaa,b
*Corresponding author Email: *hanifahkhairiah@gmail.com

Abstract
Volatile Fatty Acid (VFA) is an indicator process of an anaerobic digester that run well or not. This
research was aimed to determine the suitability of FOG wastes was operated using AD on VFA accumulation by
evaluating the performance of those AD waste based on the effluent quality. The conducted effluent tests were
pH, temperature, TS, VS, COD Reduction, ammonia, VFA, and alkalinity. This research used Continuous Stirred
Tank Reactor (CSTR) with area of work about 400 L that works mesophilic temperature. The steady state food
waste and cow mannure is gained when the averages of COD Reduction, %VSD, and pH, exceeding 54.24 ±
7.11 %, 75.812 ± 6.47%, and 6.0 ± 3.89%, respectively. Eventually, the AD Process was continued by adding
GTW that was mixed with food waste and cow mannure in ratio 1:7. Results of testing ratio 1:7 are acquired
averages of COD Reduction, VSD, and pH, are 57.60 ± 6.93 %; 88.89 ±6.18%, and 5.82 ± 5.63%, respectively.
Results of testing ratio 1:2 are acquired averages of COD Reduction, VSD, and pH, are 62.98 ± 6.74 %; 99.51
± 0.27%, and 6.27 ± 3.64%, respectively.

Keywords: Anaerobic Digester; Fat, Oil, and Grease; Volatile Fatty Acid; Long Chain Fatty Acid; Grease Trap
Waste; Food Waste

1. Introduction
Organic materials in Anaerobic Digester (AD) process may come from organic wastes, urban domestic
wastes, and fat, oil, and grease (FOG) wastes (Lee, et al., 2009). By a high organic compound (138 g/L), FOG
wastes are able to increase process efficiently in the anaerobic digester up to 65%, so that those wastes use can
become a co-substrate in the process of an anaerobic digester (Zhu, et al., 2011).
These wastes are recognized are able to increase efficiency in the AD process, however, the high-fat
compound in the FOG is going to become an inhibitor in that process. This inhibitor in the AD process is caused
by FOG waste is the Long Chain Fatty Acids (LCFA) that is released during the hydrolysis of oils and fats. The
LCFA tends to form a stable emulsion, to form surface, absorb the microbial cells that lead to foam formation,
sludge flotation, and washout, and temporary inhibiting microbes. This problem can be prevented by maintaining
a right balancing between LCFA accumulation and substrate biodegradation (A. J. Cavaleiro, et al., 2015).
Besides LCFA, one of the inhibitors that hamper works of methanogenic bacteria in FOG waste
treatments using the AD is caused by accumulating of Volatile Fatty Acids (VFA) in high quantities in the initial
process(Pereira, Sousa, Mota, & Alves, 2004). The accumulation of VFA is caused by hydrolysis/acidification of
bacteria that is produced from excessive substrates (excessive OLR) in the short time. As results, the pH will be
low and the capability of methanogenic bacteria to change acetic acid to be methane becoming low (Mata-
Alvarez, et al., 2011). Boe et al. (2010) explain that a production of excessive VFA accumulation during AD
production will exhibit un-balanced AD process, that the VFA accumulation can be an indicator of the AD
process run well or not.
According to Dai et al. (2013), the VFA can be an inhibitor if its concentration is more than 5.000 mg/L, and
Dong et al. (2015) its concentration is more than 2.000 mg/L (as Acetic Acids) and >6.000-8.000 mg/L (as Total
Volatile Acids) will interfere the AD process. In a research conducted by Martín-González et al. (2010) shows
that an optimum ratio of VFA/alkalinity is in the range of 0.3 to 0.6 for sludges of wastewater treatment plant
(WWTP) using AD. Whilst, during added FOG wastes, the VFA/alkalinity ration is around 0.4 (Neczaj, et al.,
2012). Hence, it is important to monitor the accumulation of VFA periodically in detecting problems and to
conduct necessary operational changes before occurring operational disruption of the AD reactor.
2. Literature Review
2.1. Food Waste

Food waste is the largest component of urban waste, either in developed or developing countries,
ranging from 12-62% of total waste. Food waste means the amount of waste generated during the process of
making food and after eating activities related to the behavior of sellers and consumers. One third of all food
produced for human consumption is wasted globally, as much as 1.2 billion tons of cubic meters per year. The
problem of global food waste is of particular concern to be addressed both in reducing hunger and in improving
environmental sustainability by reducing carbon emissions and landfill use (Zhang et al., 2017).

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2.2. Fat, Oil, And Grease Waste

The disposal of oil and fat waste into sewerage channels has a disastrous effect of being able to release
high concentrations of pathogens, nutrients, and solid foods for water bodies that are at risk to public health and
polluting the environment (Li, Champagne, & Anderson, 2011). Waste oil and fats are comparatively complex
and heterogeneous wastes (Martín-González, et al., 2010)so they can not be easily biologically described, but
they are not easily processed by other conventional processing due to their consistency. The transformation of
fatty acids contained in oils and fats from unsaturated states into saturated forms and the influence factors of
water hardness can affect the physical characteristics of oils and fats so as to form precipitates (Williams, et al.,
2012).
2.3. Volatile Fatty Acid (VFA)

VFA is an important intermediate produced in the acidogenesis stage and the stage of acetogenesis. The
main products of VFA during acidogenesis process are acetic acid, propionate, and butyrate (Jiang et al., 2013).
The concentration of VFA has an effect on the pH value, if the concentration of VFA increases then the pH value
will decrease. This decrease in pH may be due to increased volatile fatty acid concentration (VFA) which is a
consequence of the rapid degradation of organic macromolecules (Seadi et al., 2008). The presence of VFA, then
leads to a decrease in alkalinity characterized by a decrease in pH (Wu, et al., 2016). It can affect the continuity
of AD so that the need to control the concentration of VFA. Control can be performed by feeding the buffer to
stabilize the pH value in the reactor (Yang et al., 2015). During the fermentation stage in the AD process, having
a VFA concentration greater than 5,000 mg / l may interfere with the AD process.
An increase in the amount of volatile fatty acid (VFA), produced from the degradation of large amounts
of organic macromolecules such as carbohydrates, lipids, and proteins. An increase in VFA in the early stages of
digestion may also inhibit the reduction of volatile solids (VS) (Neczaj et al., 2012). The addition of 100 percent
oil and fat waste in the AD process did not result in an increase in methane gas, indicating the presence of
inhibitors that inhibit the activity of methanogenic bacteria in forming VFA. Research on anaerobic co-digestion
between the organic fraction of urban solid waste and oil and fat waste is stated that when added oil and fat
waste, in the early stages of the co-digestion process there will be an increase in VFA value and then gradually
drop to a very small value ignored (Martín-González, et al., 2010).

3. Methodology/Materials
3.1. Reactor Specification

This research used a pilot-scale AD reactor, clylindrical shape with its volume 500 L and 400 L in an
active volume, so as there was 20 cm high of headspace. There was an inlet channel on the top of the reactor that
was made from Stainless Steel with 3’’ in diameter. It had a valve that served to enter feedstocks into the reactor.
This inlet channel was linked by circulation pipes with 1’’ in diameter to circulate the reactor content. On the
reactor’s side, there was an effluent channel with 1’’ in diameter that was equipped with a valve functioning as
drainage channel of reactor’s effluent. Furthermore, there was a sludge valve with 2’’ in diameter in the bottom
of the reactor that was used to remove the reactor’s contents. The reactor was connected to control, a stirring
system, and gas and re-circulation systems. On the outside of the reactor was coated with an insulator namely an
aluminum foil to keep the temperature in the system to be a mesophilic condition (Tassakka, 2016). The
following figure is a whole picture of reactor AD.

Figure 1. Specification of AD Reactor

Source: Tassaka, 2016

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3.2. Research Materials

3.2.1. Inoculum

The used inoculum for start-up process came from an unit AD effluent of Pasar Kopro Grogol
Petamburan, West Jakarta as much as 130 L, and an unit AD effluent of Engeneering Faculty University of
Indonesia as much as 170 L. Moreover, an additional inoculum in form of cow rumen came from slaughterhouse
(Rumah Potong Hewan, RPH) as much as 40 L.
3.2.2. Substrates

The used substrates in this research were FOG wastes coming from grease trap. These wastes consisted
of three layers such as oil on the top, water on the middle, and sludge on the bottom of layers. Before deciding
which layer of grease trap wastes to be used more efficiently, there was an initial characteristic test of fatty acid
profiles or LCFA. Through this test, there would be produced waste data with the highest compound and had a
potential in inhibiting methanogenesis. The selected FOG waste with the lowest LCFA compound was used to
restrain inhibitors in the AD process.
The next substrate was food waste. The used food waste was derived from selecting waste, due to the
AD food waste substrate was not intended any skeletoned wastes, hard fruit skins. Those two kinds of rubbish
would inhibit the AD operational process. Those sorted substrates then were put in the container and were stored
in the temperature of -200C.
3.2.3. Co-Substrate

The co-substrates that was used in this research was cow mannure with ratio 9:1 (b:b) which referred to
the research conducting by Siregar (2015) that have the optimum result for producing methane gas. According to
Seadi (2013), animal mannure has a high C/N value ratio that exceeds 25/1 and rich in nutrients are able to
support microbial development.

3.3. Research Steps/Operating Reactor AD

3.3.1. Start-up Process

In operating the reactor, it was started by a start-up process in ±6 months of works. It was conducted on
July 2017 to February 2018 by supplying feedstock of food waste. During the start-up process, monitoring pH
and temperature was conducted to keep the stability of the process in optimum situation. When pH decreased
below 7, replenishing buffer NaHCO3 solution is necessarily carried out. After adding feedstock of food waste
exceeded steady state condition, co-substrate of cow mannure was added to increase biogas volume until the
steady state was reached again.
3.3.2. Input Process

The used substrates were FOG wastes, and food waste coming from University of Indonesia. During the
research period, the used OLR were 10 kgVS/m3. Those substrates were stored in a cooler with -200C of
temperature before use. They were put into the cooler three times in a week (Monday, Wednesday, and Friday).
Before inputting, those substrates were weighed to measure their quantities based on the determined OLR.
Furthermore, those food waste were inputted into the reactor via inlet channel manually. After exceeding steady
state, feedstock variation was conducted by adding FOG wastes.
Variation in adding FOG wastes into food waste was in ratio 1:7 and 1:2, respectively. Variation change
of FOG waste was taken until the steady state exceeded namely COD reduction exceeded 50% and parameters of
pH and temperature were stable, and biogas production was consistent with time duration about 20 days (Lopez,
et al., 2014). The reactor was equipped by mechanic-stirring paddles periodically as 4 hours in a day and 30 rpm
velocity. Steering as long as 4 hours was carried out for 2 hours before and after adding substrates. Below is a
flowchart of operating AD.
3.3.3. Sampling Test

Taken samples from the reactor were leachate and gas. The sampling of leachates was carried out every
day before inputting process occurred that represented parameters of the process. The input process was taken
through an effluent port that was located in the center of the reactor. A 100 mL of leachates were sampled using
sampling bottle to measure its pH and temperature. A representative sample was gained by removing leachates
about 200 – 300 mL before sampling was taken. During samples were measured for their characteristics, they
were stored in a room in 4oC of temperature. An effluent sample was taken out three times in a day at the time
before feeding.

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3.4. Data Analysis

Collecting data that had been derived by measuring pH and temperature was conducted directly using a
pH meter and a thermometer before inputing substrates. Measuring parameters such as Total Solid (TS), Volatile
Solid (VS), ammonia, Chemical Oxigen Dissolved (COD), alkalinity, and VFA, was taken during effluent AD.
After those testing data were acquired, therefore statistical analysis for the data as follows.
1. Student test, T-Test, to measure the difference between specific values and average population group. This
T-Test was carried out for comparing three variables such as 1) food waste + cow mannure with a ratio of
FOG wastes + food waste was 1:7; 2) food waste + cow mannure with a ratio of FOG wastes + food waste
was 1:2: 3) The Variation of 1:7 and 1:2
2. Correlation test, to determine the strength or level of linear correlation all data. The more linear a
relationship (straight line), the more strong or high level of the both variables or more. If r value = -1 meant
the perfectly negative correlation; r = 0 meant there was no correlation; and r = 1 meant the correlation was
very strong. Correlation test was conducted among testing parameters.

4. Results and Discussion

4.1. Acclimatization

Acclimatization process was carried out for 120 days using inoculum coming from AD process of food
waste and cow rumens. Only checking pH and temperatures are taken in in this process. The result of monitored
pH and temperature is shown in Figure 2 below.

Figure 2. Result of pH and Temperatures in Acclimatization Process

Based on Figure 2 above, pH of initial inoculum is started from 6.8 which is suitable with optimum pH in
the methanogenesis namely in the range of 6.5-8.2 (Lee et al., 2009). Afterwards, the pH tends to fluctuate and
has decreased to 5.7 as a result of a very low pH of the substrate that is dominated by food waste, namely ±5,0
(Zarkadas, et al, 2015), and forms accumulated organic acids as a result of a high organic compound reaction
(Yang et al., 2015).In case of a decrease in AD operating process, adding buffer NaHCO 3 solution is taken to
increase pH. The gained average pH during acclimatization process is 6,8 ± 0,79. In Figure 2 above delineates
operating AD using mesophilic temperature with its average about 28,5 ± 0,90 that is suitable for optimum
mesophilic temperature for AD process namely 25-45 oC (Buoaziz, 2014).
4.2. Characteristics of Substrates

The used substrates in this research consist of three steps such as an addition of food waste, adding food
waste and co-substrate cow mannure and addition of food waste, cow mannure, and FOG waste from grease trap
waste (GTW). Moreover, substrate mixing is aimed to exceed optimum condition of operating AD e.g. through
pH parameter and comparing C/N ratio is able to be improved by adding co-substrates of cow mannure (Divya,
Gopinath, & Merlin Christy, 2015). Table 1 below lists the characteristic of used substrates.
Table 1. Characteristics of Substrates
a b c
Parameters Unit
TS % 26,65 24,58 22,86
VS %TS 92,18 90,59 92,30
COD g/L 396 399 380,05
C mg/L 52,09 41,18 87,36
N mg/L 2,5 1,8 3,42
C/N - 20,74 22,63 25,56

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Annotation:
a
: Food waste
b
: Food waste + Cow mannure
c
: Food waste + Cow mannure + Wastes of FOG
Based on the Table above, the TS value has met the processing criteria using AD dry system namely in
the range of 20 – 50% (Kwietniewska & Tys, 2014). The gained VS value of substrate is quite high namely in
the range of 90-92 %TS. The high value of VS is going to influence the amount of Volatile Solid Destruction
(VSD) value. The VSD used a stable AD for food waste can reach 74-81% (Babaee & Shayegan, 2011). This
indicates that food waste can be degraded significantly. The high VSD is able to decrease volumes and mass of
waste generation. Further, the gained concentrates on COD substrates are classified high namely in the range of
380-399 g/L. The high organic content makes those substrates are potentially processed using AD(Elsamadony
& Tawfik, 2015). For CN value that is an important indicator in biological processing system using AD has
values in the range of 20 – 25 which corresponds to the recommended comparison namely 20:1 – 30:1 with an
optimum condition in 25:1 for anaerobic bacteria growth (Lin et al., 2011). Based on the substrate’s
characteristics in Table 1, all variables are compatible to become substrates using AD.
The next step is a selection of substrates for FOG wastes coming from grease trap waste (GTW). Those
waste in the GTW consist of three layers such as FOG on the top layer, mixing of oil, fat, water and sludge in the
middle layer, and sludge in the bottom layer. This selection is conducted in LCFA test where the substrate from
FOG waste will be chosen based on the lowest value of LCFA in preventing inhibitors in the AD process. Here
in Table 2 below is the result of LCDA test.
Table 2. Testing Result of LCFA of FOG wastes from Grease Trap
Saturated Fatty Acids Deposition Oil Mixing
% (mg/100 g)
Caprylic Acids (C8:0) 39,09 215,09 3,22
Capric Acid (C10:0) 29,97 154,95 1,32
Lauric Acid (C12:0) 183,72 1004,69 8,54
Myristic Acid (C14:0) 107,33 575,79 4,69
Palmitic Acid (C16:0) 2085,25 9201,98 74,42
Stearic Acid (C18:0) 390,49 1346,57 15,01
Arachidic Acid (C20:0) 103,42 136,54 1,13
Behenic Acid (C22:0) 822,38 515,96 15,99
Sub Total 3761,65 13151,58 124,32

Based on Table 2 above, C12 – C22 are grouped into LCFA. The palmitic acid (C16) has the highest value
than other. This happens because the food waste come mostly from processed foods using vegetable oils which
are palmitic acids as their main constituent components. Of three layers of FOG wastes of the GTW, there is
conducted a substrate selection of the lowest contents of its LCFA in order to be inhibited in the AD process.
Based on the result of LCFA content test, the mixing of oil, fat, water, and sludge, is the lowest content.
4.3. Influence of Adding FOG wastes on VFA Accumulation

The concentration of VFA is affected to pH value where the VFA concentration increase, the pH value will
decrease. The decreasing pH is able to be caused by increasing of VFA concentration that is a consequence of a
very fast degradation of organic macromolecule (Seadi et al., 2008). The existence of VFA then triggers the
occurrence of decreasing alkalinity characterizing by decreasing pH (Wu et al., 2016). This matter can affect the
sustainability of AD, so that is needed a control of VFA concentration. This control is able to be carried out by
delivering a buffer material for stabilizing the pH in the reactor (Yang et al., 2015).

Testing VFA is conducted on substrates in this research in order to seek an initial VFA before
conducting the AD process and effluent to know VFA degradation in the AD process. Figure 3 below depictures
the VFA results on substrates and effluents.

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Figure 3. Result test of VFA on Substrates and Effluents

The VFA consists of five acids such as acetic acid (C2), propionic acid (C3), isobutyric acid (iC4), n
butyric acid (nC4), isovaleric acid (iC5), and n valeric acid (nC5). The primary product of VFA during
acidogenesis process are acetic acid, propionic acid, and butyric acid (Jiang et al., 2013). Based on Figure 3
above, there has been a decrease concentration of VFA substrate’s food waste and GTW which can be seen in
result effluent of AD process. This decrease of VFA is about 94.5% indicates that acidogenesis process has run
well because of the organic acids such as VFA. This VFA has been changed to become acetate, carbon dioxide,
and/or hydrocarbon that can be used for substrates to produce methane (Jain, Jain, Wolf, Lee, & Tong, 2015)
resulting in a decrease of VFA content.
VFA/Total Alkalinity (TA) test is a better control of AD process than just pH monitoring. The ratio of
VFA/TA will give more indications of problems on the future AD process, whilst pH monitoring would not
indicate any problems, so the AD process is late to be handled (Feng et al., 2013). Therefore, the VFA and TA
tests are conducted for 24 days by using substrates of food waste and oil wastes in the variation of (1:7) and 22
day in the variation of (1:2) to co-substrate of cow mannure.
Here down is the ratio VFA/TA on effluents of adding FOG waste from the grease trap.

Figure 4. Ratio of VFA/TA of Effluent AD

In a research about anaerobic co-digestion organic fractions between municipal solid wastes and FOG
wastes are avowed that during adding FOG wastes, in the initial co-digestion process, the VFA value will
increase and then it decreases gradually until to the lowest value, so it can be ignored later (Martín-González et
al., 2010). Figure 4 points out that an initial process with a high VFA/TA ratio as high as 1.58 that indicates
instability in the AD process because this ratio is above of 0.8 (Callaghan, Wase, Thayanithy, & Forster, 2002).
This instability is able to be caused by adding the GTW that is not degraded yet by microorganisms, and quantity
of acidogenic and acetogenic bacteria exceeds the quantity of methanogenic microorganism (Gray, 2008).
Furthermore, an addition of the GTW also results increasing of organic loads where the VFA is high and the
alkalinity is low. Hereafter, until the ratio of day 17 is in the range of 0.4 – 0.8 that signifies some the AD
process occurring some instability (Callaghan et al., 2002). This is able to be rendered instability of
microorganism activity in conversing suspended organic contents as a result of accumulating VFA. The AD

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process is able to be categorized stable in the day 20 (0.36 – 0.34) because the ratio value of VFA/TA is under
0.4 (Neczaj et al., 2012) .
The ammonia content tends to be low namely in averaged 1.18 ± 9.23% that is not an inhibitor in the
AD process. Ammonia is inhibitors in the AD process if its concentration reaches 3.00 g/L (Tian, Duan, Lin, Li,
& Zhong, 2015). The ammonia formed in the AD are results of degrading protein, fatty acids, and nucleic acid.
While substrates of food wasteand FOG wastes contain a lot of carbohydrates and lipids inflicting in low
ammonia value.

4.4. Effects of Adding FOG wastes on AD Performance of Food waste

Before operating the reactor, the bacteria are stabilized firstly in the AD process. This stability process
is conducted after the acclimatization process over by administering food waste using OLR 10 Kg VS/m3. It is
purposed to keep bacteria activity in order to be still alive and growing. Further, the reactor is operated by adding
co-substrates such as cow mannure. According to Seadi (2013), animal mannure has a high C/C that exceeds
25/1 and rich in nutrition which is able to support the growth of microbes. In another side, the cow mannure has
a high buffer capacity to stabilize AD process that tends to keep a low pH through an easy access and cheap.
Furthermore, in the AD process, adding GTW as substrates functionates to know its effects on VFA
accumulation. An addition of this GTW is caused by it has a high COD that FOG wastes use is able to be created
substrates in the anaerobic digester. Figure 5 below exhibits result of effluent tests by some parameters such as
VSD, COD, pH, and ammonia.

Figure 5. Results of Effluents Testing

Figure 5 above explains that other indicators that show an addition of FOG from GTW that is more
efficient in keeping the stability of AD process, are %VSD and COD reduction. Values of %VSD and % COD
reduction until the end of the process are getting an increase that indicates the higher effectivity of AD
processing unit in untangling organic contents of substrates.
Furthermore, results of temperature and pH testing tend to fluctuate, however, it still points a stable
condition of AD process. The optimum pH in the hydrolysis step and acidification is in the range of 5.5 –
6.5(Kothari, Pandey, Kumar, Tyagi, & Tyagi, 2014) and in the methanogenesis is in the range of 6.5 – 8.2(Lee et
al., 2009), whilst in this research is gained average pH is about 5.8 ±5.29%. Statistical data analysis that has been
conducted is T-Test to know two data groups resulting in a significant difference and data stability. Those
statistical data are able to be seen in Table 3 below.

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Table 3. Results of Statistical Data Testing

Parameter t stat t P Test Statistical
Critical Results
TS % 7,9838 1,7459 2,85E-07 √
VS % 5,4207 1,8331 0,000211 √
VSD % - 1,7291 1,41E-09 √
10,3945
COD % 4,7178 1,7056 3,54E-05 √
Reduction
Ammonia mg/L 0,2925 1,7959 0,387687 X
Abbreviation √: t stat > t Critical and P < 5 % (data is stable and there is a significant difference); X: t stat < t
Critical and P > 5 % (data is stable and there is a significant difference)
Based on Table 3 above, testing parameters such as TS, VS, VSD, and COD reduction, have a significant
difference among the AD processes. This significant difference of the processes using substrates of food waste
and co-substrate of cow mannure with additional FOG waste in the variation is marked by t stat > t critical with
its level of confidence is about 95%. There is no significant difference for Ammonia due to its data is not stable
and its concentration tends to be small and fluctuated.

5. Conclusion
Results show that variation 1:2 can be an effective method to reduct a FOG waste by AD process, we can
conclude that the best variation cause of can reduction COD, increase %VSD and biogas volume. Whilst effects
of those waste on VFA accumulation are happened decreasing VFA values that are marked by value ratio of
VFA/TA is below 0.8 in the end process that figures the AD process running in stable condition cause of VFA
Concentrate has below 6000 mg/L. However, AD Process were able to reduction COD above 50 % and increase
% VSD 69 – 99%.

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