Environ. Eng. Res.

2022
Research Article
https://doi.org/10.4491/eer.2020.704
pISSN 1226-1025 eISSN 2005-968X
In Press, Uncorrected Proof

Biodiesel production from the black soldier fly larvae

grown on food waste and its fuel property
characterization as a potential transportation fuel
Jo-Yong Park1*, Sungyup Jung2*, Yong-Gyu Na1, Cheol-Hwan Jeon1, Hwa-Yeon Cheon1, Eun-
Young Yoon3, Sang-hoon Lee4, Eilhann E. Kwon2†, Jae-Kon Kim1†

1
Research Institute of Petroleum Technology, Korea Petroleum Quality & Distribution Authority, Cheongju 28115, Republic of
Korea
2
Department of Environment and Energy, Sejong University, Seoul 05006, Republic of Korea
3
Department of Integrative Bio-industrial Engineering, Sejong University, Seoul 05006, Republic of Korea
4
GreenTeko,Inc., Gyeonggi-do 15655, Republic of Korea
*
These authors contributed equally to this work.

Abstract
Insects have a great potential to be used as feedstocks for biodiesel production due to their intrinsic high lipid
contents. In particular, biodiesel production from black soldier fly larvae (BSFL) is promising because of its
advantages in engineering aspects such as a rapid proliferation, more lipid storage due to long-development time in
larva stage, self-separation from organic waste. In this study, biodiesel was synthesized through (trans)esterification
of BSFL grown on food waste to achieve both biofuel production from organic waste materials. In addition, all fuel
properties of the biodiesel were examined to evaluate its feasibility as a potential transportation fuel in Korea. The
highest yield of biodiesel obtained in this study was 86.51%. All the parameters of biodiesel produced from BSFL
satisfied the Korea fuel standard (KS M 2965) except oxidation stability. The quality criterion of oxidation stability
met the fuel standard with an addition of small quantity of tert-butylhydroquinone.

Keywords: Biodiesel, Black soldier fly larvae, Fatty acid methyl esters, Fatty acids, Fuel properties, Quality standards

This is an Open Access article distributed under the terms Received December 28, 2020 Accepted March 23, 2021
of the Creative Commons Attribution Non-Commercial Li- †
cense (http://creativecommons.org/licenses/by-nc/3.0/) Corresponding Author
which permits unrestricted non-commercial use, distribution, and repro- E-mail: ekwon74@sejong.ac.kr; jkkim@kpetro.or.kr
duction in any medium, provided the original work is properly cited. Tel: +82-2-3408-4166; +82-43-240-7931 Fax: +82-2-3408-4320
ORCID: 0000-0001-7438-7920 (E.E.K); 0000-0002-3458-8661 (J.K.K)

Copyright © 2022 Korean Society of Environmental Engineers http://eeer.org

 1 1. Introduction

2 Substantial quantity of food waste is generated in the world, and the sources of food waste

3 generation include household, restaurants, agricultural and food industries [1]. The European

4 Union (EU) reported that the annual production of food waste from 28 countries in the EU has

5 continuously increased, and the estimation of food waste production in 2020 is 116 million tons

6 [2]. In Korea, the annual production of food waste was approximately 5 million tons [3].

7 Currently, the food waste is being disposed composting or landfill methods. Composting is a

8 broadly used and well-established way to valorize food waste for its recycling as ingredients in

9 the agricultural industry. However, composting process requires continuous and proper

10 managements for carbon to nitrogen ratio, moderate temperature (50 to 70˚C), and aerobic

11 conditions with long reaction time to convert food waste into desired composts [4]. Landfill

12 methods produce biogas, composed of CO2 and CH4 from biological reactions in the oxygen-

13 limited conditions. It releases leachate and greenhouse gases, and the completion of the

14 anaerobic reactions takes a few months [5]. In addition, the installation of landfill plant requires

15 high capital costs and large spaces [6]. Thus, it is beneficial to find an innovative and

16 environmentally benign way to dispose and valorize the food waste.

17 Insects store energy in the form of triglycerides (lipids) and glycogen in their main body

18 cell [7]. Given that the food waste is a feed for insects, a variety of chemical constituents of food

19 waste can be transformed into lipids in the fat body cell of insects by their metabolisms [8, 9].

20 Lipids are important feedstocks for biodiesel production through a well-established industrially

21 used chemical reaction, called transesterification. Soybeans and palms have been broadly used as

22 feedstocks for biodiesel synthesis, and their lipid contents are 18 and 36%, respectively [10]. It is

2
 1 known that insects also have competitive or higher lipid contents (30 – 40%) in reference to the

2 oil-bearing crops [9, 11]. In addition, the high proliferation rate of insects can accelerate the

3 conversion of food waste into lipid. In Table 1, lipid contents of different oil-bearing biomass

4 feedstocks and the estimation of biodiesel productivity are summarized. Thus, the high biodiesel

5 productivity from insects that grew up on food waste is expected, and these can achieve two

6 goals: (a) disposal of food waste and (2) production of energy as a form of biodiesel through an

7 environmentally friendly process.

8 Black soldier fly, referring to Hermetia illucens (H. illucens), is a widespread fly of the

9 family Stratiomyidae and lives in tropical and temperature regions. Black solider fly larvae

10 (BSFL) have three weeks of development time, which is much longer than the development time

11 of house flies (< 5 d). It means that a single larva of black solider fly can consume more food

12 waste and produce larger pupae in reference to house flies [11, 12]. The lipid contents of BSFL

13 and their pupae vary in the range of 11 and 42%, and the average value is 29% (dry basis) [11].

14 Because of the high protein contents of BSFL (≥ 32%, dry basis), the remaining debris of BSFL

15 after lipid extraction can be used as a feed for animals [11]. Prior to entering the pupa stage,

16 BSFL instinctively leave the substrate to move to a high and clean place [13], meaning no need

17 of additional separation step for BSFL from food waste. Such instinctive behavior, also called

18 self-harvesting, can help to remove a labor-intensive collection process.

19 Because of the aforementioned advantages, BSFL have received considerable attentions

20 to be used as a feedstock for biodiesel production. In general, previous studies examined the

21 yields of biodiesel using BSFL or lipids extracted from BSFL depending on the various reaction

22 conditions [11]. They also reported chemical and physical properties of BSFL-derived biodiesel

3
 1 such as density, moisture content, composition of fatty acid methyl esters (FAMEs: biodiesel

2 contents), acid value, viscosity, and phosphorous content. Also, the heating value and cetane

3 number of FAMEs derived from BSFL were competitive with other biodiesels [10, 14-20].

4 However, the previous studies did not fully prove the quality criteria of biodiesel to be

5 used as a transport fuel, and no study has reported all the parameters of quality specification for

6 biodiesel as a liquid transportation fuel. Although lipid contents of BSFL are highly contingent

7 on the feeds of BSFL, the BSFL grown on food waste were also rarely adapted for biodiesel

8 production.

9 In this study, biodiesel was synthesized using lipids extracted from BSFL grown on

10 organic food waste, and all the fuel properties are evaluated to decide the fuel feasibility of the

11 BSFL-derived biodiesel. The relationship between biodiesel yield and reaction conditions (lipid

12 to methanol ratio, catalyst loading, reaction temperature and time) was also examined to improve

13 the reaction efficiency for biodiesel production. After biodiesel production, all the quality

14 requirements of the Korea biodiesel fuel standards (KS M 2965) were evaluated using the

15 biodiesel produced from BSFL lipid. Lastly, the fuel characteristics of BSFL-derived biodiesel

16 were compared with other biodiesel fuels.

17

18 2. Materials and Methods

19 2.1. Lipid Extraction from BSFL

20 BSFL were grown up, feeding food waste for 14 d by Greenteko Co., Ltd. Approximately 25,000

21 larvae were grown per 1 m2, and the weight of the BSFL grown was about 5 kg per m2. The

22 collected BSFL were then dried at 60˚C in a conveyor oven (Tae Yang Tech instrument

4
 1 company, Korea). Lipid was extracted from the dried BSFL with a hexane using a previously

2 reported liquid-liquid extraction method [16]. The extracted lipid was centrifuged at 600 rpm for

3 10 min to separate lipid from the residual solids and decompressed hexane. Protein, lipid,

4 carbohydrate, ash, and moisture contents were analyzed by the analysis methods provided from

5 the Association of Official Analytical Chemists (AOAC 1990). The crude protein and lipid

6 contents were analyzed using nitrogen determination and acid hydrolysis methods [21, 22].

8 2.2. Biodiesel Synthesis and Purification

9 5 g of extracted BSFL lipid was used to synthesize biodiesel through (trans)esterification.

10 Different experimental conditions (lipid to methanol ratio, catalyst loading, reaction temperature,

11 and reaction time) were used for biodiesel production. Lipid to methanol ratio was in the range

12 of 1:4 to 1:10, and the amount of acid catalyst (H2SO4) loading was changed from 2 to 6%,

13 relative to lipid. The reaction temperature was between 50 and 80 ˚C, and the reaction times were

14 2, 4, 6, and 8 h. The resulting products were cooled down to 25 ˚C, and methanol was separated

15 through distillation process. Glycerol in the lower layer of biodiesel product solutions was

16 removed through layer separation and decantation. The crude biodiesel was washed with distilled

17 water, and then its weight was measured gravimetrically. Total FAME contents were analyzed

18 with chromatography methods to measure biodiesel yield according to EN 14103. For the

19 purification of BSFL-derived biodiesel, a decompression distillation was conducted. The quality

20 parameters specified in Korea biodiesel fuel standards (KS M 2965) were then measured. The

21 overall experimental procedures are drawn in Fig. 1. Experiments were performed in triplicate,

22 and the errors of all the results were less than 2%.

5
 1

2 2.3. Calculations

3 The yield of biodiesel synthesized from BSFL was calculated as the amount of FAME produced

4 against the amount of lipids used as reactant. The yield of the synthesized biodiesel was

5 measured using the crude biodiesel obtained after synthesis was completed. The FAME content

6 was analyzed using EN14103 and calculated using Eq. (1), (2), and (3).

𝑊𝐴
𝐶𝑟𝑢𝑑𝑒 𝑏𝑖𝑜𝑑𝑖𝑒𝑠𝑒𝑙 (%) = × 100 (1)
𝑊𝐵

∑ 𝐴 − 𝐴𝐸𝐼 𝑊𝐸𝐼
𝐹𝐴𝑀𝐸 𝑐𝑜𝑛𝑡𝑒𝑛𝑡 (%) = × × 100 (2)
𝐴𝐸𝐼 𝑊

𝐶𝑟𝑢𝑑𝑒 𝑏𝑖𝑜𝑑𝑖𝑒𝑠𝑒𝑙 × 𝐹𝐴𝑀𝐸 𝑐𝑜𝑛𝑡𝑒𝑛𝑡

𝐵𝑖𝑜𝑑𝑖𝑒𝑠𝑒𝑙 𝑦𝑖𝑒𝑙𝑑 (%) = (3)
100

8 Where WB is the weight of lipid before reaction (mg),

9 WA is the weight of crude biodiesel after reaction (mg),

10 ΣA is the sum of FAME area from C6:0–C24:1,

11 AEI is the peak area of methyl nonadecanoate,

12 WEI is the weight of methyl nonadecanoate as internal standard (mg), and

13 W is the weight of crude biodiesel (mg).

14

15 2.4. Analysis of Biodiesel Characteristics

16 The FAME content in biodiesel was determined following the EN 14103, using gas

17 chromatography (GC) (Agilent, Palo Alto, USA) equipped with a flame ionization detector (FID)

18 and a chromatography data management system. The fused silica capillary column (SP-2380
6
 1 model, Supelco, Inc., USA) used in the GC analyzer was 30 m in length and 0.25 mm in inside

2 diameter, with a film thickness of 0.2 μm. Methyl nonadecanoate was used as an internal

3 standard solution. The cold filter plugging point (CFPP) measurement was carried out following

4 KS M 2411 on the model FPP5Gs (ISL, Verson, Carpiquet, France). About 45 mL of sample was

5 cooled under the specified conditions. At 1 ˚C of interval, a 200 mm H2O vacuum was applied to

6 draw up the sample into a pipette through a filter. As the sample was cooled, the amount of

7 crystals increased. The temperature at which either the sample ceased to flow through the filter

8 within 60 s or failed to return into the test jar was recorded as the CFPP. The cloud point (CP)

9 was measured by a Mini Pour/Cloud Point Tester MPC-602 (Tanaka, Japan) covering a

10 temperature range from – 60 to 51˚C. This tester was in accordance with ASTM D2500 and

11 ASTM D637.

12 The oxidation stability measurements were carried out following European standard EN

13 14112 with the model 743 Rancimat (Metrohm, Herisau, Switzerland). About 3 g of samples

14 were analyzed under constant air flow of 10 L/h at 110˚C heating block temperature. The

15 temperature correction (∆T) was set to 1.5˚C. The KS M ISO 6245 method using a Brinkman

16 Metrohm 809 Titrando instrument (Riverview, FL) was utilized to measure acid value (AV). The

17 kinematic viscosity was determined according to KS M 2014 by measuring the time for a volume

18 of liquid to flow under gravity through a calibrated capillary viscometer at 40˚C. The test method

19 was done using Automated Kinematic Viscometer CAV-2100F (Cannon instrument company,

20 USA).

21

22 3. Results and Discussion

7
 1 3.1. Characteristics of BSFL Lipid

2 FAMEs, biodiesel components, were produced from the conversion of lipids, in particular,

3 transesterification of triglycerides and esterification of free fatty acids. Because the

4 (trans)esterification reactions are susceptible to the presence of organic and inorganic impurities

5 that can be presented in biomass feedstocks [23], lipid content was extracted from BSFL prior its

6 conversion to biodiesel. The lipid was extracted using liquid/liquid extraction method. In Table 2,

7 major chemical constituents of lipid extracted from BSFL and left-over cake (pressed cake) are

8 shown. Lipid extracted had 85.49% of purity having 9.91% protein, 2.65% carbohydrate, 1.37%

9 ash, and 0.58% of water. The major compounds of left-over BSFL cake after lipid/lipid

10 extraction were protein (64.20%), followed by carbohydrate (19.67%), lipid (8.24%), ash

11 (6.76%), and water (1.13%). From the lipid extraction process, high lipid content, a feedstock for

12 biodiesel production, was obtained. Also, other nutrients (protein and carbohydrate) abundant

13 cake, which can be used as a feed for animals, were prepared.

14 To further understand the presence of other contents in the lipid extracted, iodine and

15 acid values were examined (Table 3). Note that the iodine value is associated with unsaturated

16 fatty acids contained in the lipid. The iodine value of BSFL (73.4 g Iodine 100 g-1 lipid) was

17 lower than that of soybean oil (117 – 143 g Iodine 100 g-1 lipid) and waste cooking oil (108.4 g

18 Iodine 100 g-1 lipid), but higher than that of palm oil (48 – 58 g Iodine 100 g-1 lipid) [10, 24, 25] .

19 This demonstrates that lipid extracted from BSFL has a lower fraction of unsaturated fatty acids

20 than soy and waste cooking oils. The unsaturated fatty acids can suffer from chemical bond

21 scissions, resulting in undesired side products such as volatile fatty acids and alkanes. The side

22 products increase the impurity in the final product, otherwise requiring additional separation

8
 1 processes. Therefore, the low quantity of unsaturated fatty acids in lipid extracted from BSFL is

2 beneficial in terms of its fuel quality.

3 Acid value (AV) is a parameter that indicates the amount of free fatty acid and other acids (e.g.,

4 residual catalyst) contained in the samples. AV of lipid extracted from BSFL (1.65 mg KOH g-1

5 lipid) was placed in the middle of the AV ranges of other feedstocks for biodiesel production as

6 shown in Table 3. Considering relatively lower iodine value and moderate AV of BSFL, it is

7 inferred that BSFL can be considered a competitive lipid source for biodiesel production.

9 3.2. Biodiesel Synthesis and Purification

10 For biodiesel production from the BSFL-derived lipid, (trans) esterification was performed with

11 a broadly used acid catalyst (i.e., H2SO4). The acid catalyst was chosen because alkaline catalysts

12 can lead to formations of soap and alcohol through the saponification reaction of free fatty acids

13 (acid value > 1). To obtain high yield of biodiesel production, different reaction conditions (lipid

14 to methanol ratio, catalyst loading, reaction temperature and time) were used for

15 (trans)esterification of BSFL-derived lipid. The results are depicted in Fig. 2.

16 The relationship between biodiesel yield and lipid to methanol ratio (1:4, 1:6, 1:8, and

17 1:10) was monitored at 70˚C for 6 h with 5% (0.25 g) of H2SO4 catalyst. The highest yield

18 (86.51%) of biodiesel was shown at 1:8 ratio (Fig. 2(a)). When the methanol ratio was lower, the

19 biodiesel yields significantly decreased, and the higher methanol loading did not improve

20 biodiesel yield. To further expedite (trans)esterification reactions for higher yields of biodiesel,

21 more catalyst loading, higher temperature, and longer reaction time were used. However, no

22 remarkable improvement of biodiesel yield was shown. In contrast, the significant decrease of

9
 1 biodiesel yield was shown when the lower methanol or catalyst loadings, reaction temperature,

2 and shorter period of reaction time were used.

3 Table 4 shows the chemical compositions of FAMEs in biodiesel. BSFL-derived

4 biodiesel was composed of 12 types of FAMEs, and major constituents of FAMEs were methyl

5 laureate (C12:0, 23.6%), methyl oleate (C18:1, 23.2%), methyl palmitate (C16:0, 20.3%), and methyl

6 linoleate (C18:2, 15.9%). 55.9% of biodiesel were saturated FAMEs, while monosaturated and

7 polyunsaturated FAMEs were 26.0% and 18.1%, respectively. BSFL-derived biodiesel had

8 relatively higher portion of saturated FAMES, comparing to other biodiesels. These results have

9 good agreement with iodine values as described in Table 3. Other biodiesels derived from edible

10 oils were composed of C16 and C18 FAMEs (Table 4), while BSFL-derived biodiesel showed

11 high quantity of C12 FAME (methyl laureate).

12

13 3.3. Characteristics of Biodiesel Fuel

14 To evaluate the feasibility of BSFL-derived biodiesel as a transportation fuel, its properties were

15 further examined. As discussed earlier, the AV of biodiesel is an important parameter to evaluate

16 whether biodiesel will be degraded and cause damages to the internal combustion engines. Also,

17 the measurement of sulfur content in biodiesel should be investigated because sulfur can be

18 oxidized into sulfuric oxides during combustion process of engines. Sulfuric oxides are harmful

19 chemicals and highly regulated by governments. AV (1.04 mg KOH g-1 fuel) and sulfur content

20 (146 mg sulfur kg-1 fuel) of the BSFL-derived biodiesel were higher than Korea (KS M 2965)

21 and EU (EN 14214) standards: AV (< 0.50 mg KOH g-1 fuel) and sulfur (10.0 mg sulfur kg-1

10
 1 fuel). Thus, it is necessary to have further purification processes to upgrade BSFL-derived

2 biodiesel.

3 In other previous reports of biodiesel production from insects, AV and sulfur content of

4 produced biodiesel also did not meet the standards. According to Li et al. [16] the AV (1.1 mg

5 KOH g-1) of biodiesel produced from BSFL was higher than EU standard. Nguyen et al. [18]

6 produced biodiesel from BSFL, and they found that the sulfur content in the biodiesel was 1,200

7 mg kg-1, which was much higher than the quality standards of various countries. The high AV

8 and sulfur contents can be attributed to the presence of unreacted/degraded free fatty acids,

9 sulfuric acid catalyst, or/and intrinsic sulfur content in BSFL lipid. Therefore, the separation of

10 them from biodiesel should be performed to meet the fuel standards in Korea and EU.

11 The lipid extracted from BSFL had a high sulfur content (430 mg kg-1 lipid), and it

12 decreased to 146 mg kg-1 after evaporation and washing steps. In addition, the AV of the lipid

13 dropped from 1.65 to 1.04 mg KOH g-1 after both processes. Thus, it was hypothesized that

14 remaining free fatty acids and intrinsic sulfur in lipid were presented in the crude biodiesel.

15 To obtain high purity biodiesel, decompression distillation was further conducted to

16 obtain evaporated FAMEs fractionally at lower boiling points. Through the fractional distillation

17 approximately 90% of FAMEs (purified biodiesel) was collected from the crude biodiesel. The

18 sulfur content in the purified biodiesel was reduced to 5.6 mg kg-1 biodiesel, and the AV was

19 0.16 mg KOH g-1 biodiesel, thus satisfying both Korea and EU biodiesel standards.

20 When the acid and sulfur contents were derived from sulfuric acid, decompression

21 distillation could not work because the boiling point of sulfuric acid is placed between that of C14

22 and C 16 methyl esters. Thus, the sulfur content was likely derived from protein contained in

11
 1 BSFL-derived lipid (Table 2). Note that some types of protein such as methionine and cysteine

2 contain sulfur containing amino acids.

3 A variety of chemical and physical properties of purified BSFL-derived biodiesel are

4 compared with Korea and EU fuel standards, and those of other biodiesels in Table 5. All the

5 properties met the quality criteria of both Korea and EU biodiesel fuel standards except oxidation

6 stability. This proved that the BSFL-derived biodiesel can be considered a promising biodiesel

7 with some stabilization process. Oxidation stability of biodiesel is an indicator to decide the

8 long-term storage stability of biodiesel and its fuel efficiency. Saturated FAMEs are likely to be

9 more reactive upon exposure to oxygen in the air [26]. The oxidized products can cause various

10 problems in diesel engines such as corrosion of metals, fuel filter plugging, injector fouling, and

11 others [27]. As discussed in Table 4, the fraction of saturated FAMEs in BSFL-derived biodiesel

12 was higher than that of other biodiesels. The high quantity of saturated FAMEs is indeed

13 desirable to have higher tolerance against oxidation reactions with air. However, the oxidation

14 stability of BSFL-derived biodiesel was 0.8 h. Although the fuel itself did not meet the quality

15 standards of Korea and EU, the oxidation stability was significantly improved to 7.7 h in the

16 addition of a practically used antioxidant, 200 mg of tert-butylhydroquinone in 1 kg of biodiesel.

17 Nevertheless, the further investigation to determine the factors, leading to low oxidation stability

18 of BSFL-derived biodiesel, would be important future study. The oxidation stability can be also

19 susceptible to heat, and other impurities presented in the fuel.

20 One of the major concerns when using biodiesel in internal combustion engine is its low

21 temperature operation ability. To measure the cold flow properties, cloud point (CP), pour point

22 (PP), and cold filter plugging point (CFPP) are examined. The CP is the temperature at which a

12
 1 liquid component within biodiesel precipitates on cooling test, and PP refers to the lowest

2 temperature that a fuel can migrate when the container is tipped. CFPP for biodiesel is the

3 highest temperature when a given amount of it fails to pass through a filter. Among them, Korea

4 biodiesel standard has a quality criterion of CFPP (0˚C), and the BSFL-derived biodiesel met the

5 CFPP criterion. Comparing to other cold flow properties of BSFL biodiesel with other biodiesels,

6 it had the second best cold flow properties in reference to five representative biodiesels (Table 6).

8 4. Conclusions

9 Biodiesel was synthesized from black soldier fly larvae (BSFL) grown on food waste. Various

10 fuel properties of the biodiesel were examined. The maximum biodiesel yield was 86.51% at 70

11 ˚C for 6 hr when lipid to methanol ratio was 1:8 with 5% H2SO4 catalyst. High acid value and

12 sulfur contents in the biodiesel were significantly reduced through decompression distillation.

13 The properties of the biodiesel met the Korea standard except oxidation stability, but the

14 oxidation stability was improved in the addition of an antioxidant. These proved the feasibility of

15 BSFL biodiesel to be used as a transportation fuel.

16

17 Acknowledgments

18 This work was supported by the Korea Institute of Planning and Evaluation for Technology in

19 Food, Agriculture, Forestry and Fisheries (IPET) through the Agri-Bioindustry Technology

20 Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs

21 (MAFRA)(318018033HD040).

22

13
 1 Author Contributions

2 J.-Y.P. (Research scientist) conceptualized, conducted formal analysis, and wrote original

3 manuscript. S.J. (Postdoctoral researcher) conducted formal analysis and data interpretations,

4 visualized, and wrote original manuscript. Y.-G.N. (Research scientist) conducted experiments,

5 reviewed and edited manuscript. C.-H.J. (Senior research scientist) conducted experiments,

6 reviewed and edited manuscript. H.-Y.C. (Researcher) conducted data interpretations, reviewed

7 and edited manuscript. E.-Y.Y. (Professor) conducted data interpretations, reviewed and edited

8 manuscript. S.-h.L. (Company president) conducted data interpretations, reviewed and edited

9 manuscript. E.E.K. (Professor) conceptualized, conducted formal analysis, reviewed and edited

10 manuscript. J.-K.K. (Principal research scientist) conceptualized, conducted formal analysis,

11 reviewed and edited manuscript.

12

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19

17
 BSFL

Dry & Extraction

Lipid extracted from BSF

L
MeOH with H2S
O
(Trans)esterification

Evaporation

MeOH

Separation

Glycer
ol
Washing

Water

Crude biodiesel

Evaporation

Upgraded biodiesel
1
2 Fig. 1. Experimental procedures for biodiesel synthesis and purification.

18
 100 100
(a) (b)
80 80
Biodiesel yield(%)

Biodiesel yield(%)
60 60

40 40

20 20

0 0
20 30 40 50 0.15
0.10 0.20 0.25 0.30
Methanol(g) Catalyst loading(g)
100 100
(c) (d)
80 80
Biodiesel yield(%)

Biodiesel yield(%)
60 60

40 40

20 20

0 0
50 60 70 80 0 2 4 6 8 10
o
Reaction time(h)
1 Temperature( C)

2 Fig. 2. Biodiesel synthesis from transesterification of BSFL under various reaction conditions.
3 Reaction conditions for transesterification of BSFL were 5 g of lipid, 40 g methanol, 0.25 g
4 catalyst loading, 70˚C and 6 h reaction time. Other parameters were changed to study effects of
5 (a) methanol ratio (20 to 50 g), (b) catalyst loading (0.10 to 0.30 g), (c) reaction temperature (50
6 to 80˚C), and (d) reaction time (2 to 8 h).
7

8 Table 1. Lipid Content in Oil-bearing Biomass Feedstocks and Estimation of Their Biodiesel
9 Productivity [10, 11]
Oil (lipid) contents
Oil yield Biodiesel productivity
Feedstocks (% oil by wt. in dry
(L oil ha-1 year-1) (kg biodiesel ha-1 year-1)
biomass)
Soybean 18 636 562
Rapeseed 41 974 862
Palm 36 5,366 4,747
Jatropha 28 741 656
Microalgae 50 97,800 86,515
BSFL 30 – 40 162,000 145,800a
a
10 Based on 40% lipid in BSFL

19
 1

2 Table 2. Chemical Constituents of Lipid Extracted from BSFL and Left-over Cake

Chemical constituents Lipid extracted (%) Left-over cake (%)

Water 0.58 1.13
Carbohydrate 2.65 19.67
Protein 9.91 64.20
Fat (lipid) 85.49 8.24
Ash 1.37 6.76
3

4 Table 3. Iodine Value and Acid Value of Lipid Extracted from BSFL and Other Bio-oils
Lipid or oil Iodine value Acid value
Refs.
extracted (g Iodine 100 g-1 lipid) (mg KOH g-1 lipid)
BSFL 73.4 1.65 this study
Soybean 128-143 0.1-0.2
Palm 48-58 6.9-50.8
Waste cooking oil 108.4 0.15 [10, 24, 25]
Rapeseed 94-120 0.2
Jatropha 82-98 15.6-43
5

10

11

12

20
1 Table 4. The FAME Compositions in BSFL Biodiesel and Biodiesel Derived from Other
2 Biomass Feedstocks

BSFL (%) Soybean (%) Palm (%) Waste cooking Rapeseed (%) Jatropha (%)
FAMEs
this study [10] [10] oil (%) [25] [28] [28]
C8:0 - - 0.8 - - -
C10:0 0.7 - 0.5 - - -
C12:0 23.6 0.1 0.3 - - -
C14:0 6 0.1 1.1 0.1 0.1 0.1
C14:1 0.1 - - - - -
C16:0 20.3 11.6 42.5 8.2 5.6 13.7
C16:1 2.7 0.2 0.2 5.2 0.3 0.9
C17:0 0.5 - - - - -
C18:0 4.6 3.9 4.2 2.1 1.9 6.8
C18:1 23.2 23.7 41.3 59.7 55.1 44.3
C18:2 15.9 53.8 9.5 19.3 26.2 33.5
C18:3 2.2 5.9 0.3 6.8 6.8 0.2
C19:0 - - - - - -
C20:0 0.2 0.3 - - 0.6 0.2
C20:1 - 0.3 - - 1.0 0.1
C22:0 - 0.3 0.1 - 0.3 -
C22:1 - 0.1 - - - -
Saturated
fatty 55.9 16.3 49.5 10.4 8.5 20.8
FAMEs
Monounsa
turated 26.0 24.0 41.5 64.9 55.4 45.2
FAMEs
Polyunsatu
rated 18.1 60.0 9.8 26.1 34.0 33.8
FAMEs
3

21
1 Table 5. Fuel Quality Criteria of Korea and EU Standards, and Fuel Properties of BSFL-derived Biodiesel and Other Biodiesels [10,
2 24, 25]
Other biodiesel references
BSFL-
KS M EN derived Soybean Palm oil Waste Rapeseed Jatropha
BSFL fed
BSFL fed BSFL fed
Property oil restaurant
2965 14214 Biodiesel [10, 24, cooking oil oil oil manure wheat bran
(this study) [10, 24, 25] [10, 24, 25] [10, 24, 25] [10, 24, 25]
waste
[19] [18]
25] [19]
Ester content, wt%, min 96.5 96.5 97.5 - - - - - - - -
Sulfur, mg/kg, max 10 10.0 6 2 2 - 4 5 - - 0.12
Density @ 15oC, kg/m3 860-900 860-900 875 882 873 877 879 876 860 885 875
Viscosity @ 40oC, 4.9 5.8 5.2
1.9-5.0 3.50-5.00 4.0 4.26 4.61 5.23 4.50 4.75
mm2/s
Flash Point, oC, min 120 101 138 159 163 157 169 152 128 123 121
Water, wt%, max 0.05 0.05 < 0.01 < 0.00005 - - 0.0003 - 0.02 0.03 0.03
Acid value, mg KOH/g, 0.6 1.1 < 0.8
0.50 0.50 0.16 0.266 0.24 0.21 - 0.4
max
CFPP, oC, min 0 - 0 -4 9 - -12 0 - - -
Methanol, wt.%, max 0.2 0.20 0.12 - - - - - - -
Oxidation stability @ - - -
6 8 0.1 (7.7)a 2.1 10.3 - 7.6 2.3
110oC, h, min
Copper corrosion @ - - -
1 1 1 1 1 - - 1
50oC, 3 h, max
Carbon residue, wt%, - - -
0.1 - < 0.1 - - - - -
max
Ash, wt%, max 0.01 - < 0.01 - - - - 0.012 - - -
Sediment, mg/kg, max 24 24 <1 - - - - - - - -
Total glycerol, wt%, - - -
0.24 0.25 0.02 0.149 0.01 - - 0.1
max
Monoglyceride wt%, 0.80 0.70 < 0.01 0.473 - - - 0.291 - - -
22
max
Diglyceride wt%, max 0.20 0.20 < 0.01 0.088 - - - 0.104 - - -
Triglyceride wt%, max 0.20 0.20 < 0.01 0.019 - - - 0.022 - - -
Glycerol wt%, max 0.02 0.02 0.02 0.012 0.01 - - 0.006 - - -
Metals, Na+K 5 5.0 <1 - - - - - - - -
mg/kg, - - -
Ca+Mg 5 5.0 <1 - - - - -
max
Phosphorous, mg/kg, - - -
10 4.0 <1 < 0.1 < 0.001 - - < 0.1
max
a
1 7.7 h after adding antioxidant (TBHQ, 200ppm)
2

3 Table 6. Cold Flow Properties of BSFL Derived Biodiesel in Comparison with Other Biodiesels
Cold filter
Biodiesel feedstocks Cold point (˚C) Pour point (˚C) plugging point Ref.
(˚C)
BSFL 2 -1 0 this study

Soybean 0 -4 -4

Palm 14 13 9

Waste cooking oil 3 0 - [10]

Rapeseed -3 -10 -12

jatropha 5 0 -
4

23