International Journal of Green Energy

ISSN: 1543-5075 (Print) 1543-5083 (Online) Journal homepage: http://www.tandfonline.com/loi/ljge20

Evaluation of Two-Step Reaction and Enzyme

Catalysis Approaches for Biodiesel Production
from Spent Coffee Grounds

Rachel Burton , Xiaohu Fan & Greg Austic

To cite this article: Rachel Burton , Xiaohu Fan & Greg Austic (2010) Evaluation of Two-Step
Reaction and Enzyme Catalysis Approaches for Biodiesel Production from Spent Coffee Grounds,
International Journal of Green Energy, 7:5, 530-536, DOI: 10.1080/15435075.2010.515444

To link to this article: http://dx.doi.org/10.1080/15435075.2010.515444

Published online: 13 Oct 2010.

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International Journal of Green Energy, 7: 530536, 2010
Copyright Taylor & Francis Group, LLC
ISSN: 1543-5075 print / 1543-5083 online
DOI: 10.1080/15435075.2010.515444

EVALUATION OF TWO-STEP REACTION AND ENZYME

CATALYSIS APPROACHES FOR BIODIESEL PRODUCTION
FROM SPENT COFFEE GROUNDS

Rachel Burton, Xiaohu Fan, and Greg Austic

Piedmont Biofuels Industrial, Pittsboro, North Carolina, USA

Currently, the biodiesel industry is investigating non-food crops with high oil content. The
industry is also evaluating oil sources from deeper into the industrial or commercial waste
streams. Spent coffee grounds could be a feedstock that qualifies for both as a non-food crop
and a waste product. The purpose of this study is to evaluate the feasibility of using coffee oil
extracted from spent coffee grounds as raw material to produce ASTM standard biodiesel.
The two biodiesel production methods that were used are acid-catalyzed esterification fol-
lowed by alkali-catalyzed transesterification and lipase catalyzed transesterification. It was
found that the high level of free fatty acids (FFA) in the coffee oil was reduced from 16.3% to
2.64% by acid-catalyzed esterification with 40/1 molar ratio of methanol to FFA in the pres-
ence of 5% (v/v, based on FFA) hydrochloric acid. However, poor conversion was obtained
for the further alkali-catalyzed transesterification. In comparison, 98.5% conversion was
achieved by using enzymatic catalysis, demonstrating the feasibility of using this approach
to process low quality coffee oil from spent coffee grounds for biodiesel production.

Keywords: Biodiesel; Enzyme catalysis; Esterification; Spent coffee grounds;

Transesterification

INTRODUCTION
At present, biodiesel is considered a real alternative to diesel fuel due to its following
advantages.

1. It can reduce the dependence on crude oil foreign imports and enhance the energy
security.
2. It has favorable energy return on energy invested.
3. It can reduce greenhouse gas emissions in comparison to petroleum diesel.
4. It is biodegradable, nontoxic, and renewable.
5. It can help improve rural economic development since the agricultural surplus and other
waste streams are used as raw material.

Though biodiesel is technically feasible and environmentally beneficial, it should

be noted that not all feedstocks are economically competitive. The high cost of virgin

Address correspondence to Xiaohu Fan, Piedmont Biofuels Industrial, 220 Lorax Lane, P.O.Box 661,
Pittsboro, NC 27312, USA. E-mail: xfan@biofuels.coop

530
 BIODIESEL FROM SPENT COFFEE GROUNDS 531

vegetable oil is the most important issue in the economic evaluation of the biodiesel process
(Zhang et al. 2003). Reducing the cost of the feedstock is necessary for the long-term
commercial viability of biodiesel. In order to reduce costs and produce biodiesel that will
be more competitive with petroleum diesel, feedstocks must be less expensive. Non-edible
oils, such as coffee oil from spent coffee grounds, could be used as raw material to reduce
feedstock cost.
It was estimated by the U.S. Department of Agriculture that the worlds coffee pro-
duction is 16.34 billion pounds per year (Pumphrey 2007). Depending on the coffee types,
the coffee source contains about 11%20 wt% oil (Barkenbus and Zimmerman 1927;
Daglia et al. 2004). Previous research shows that an average amount of about 15% oil could
be obtained from the spent coffee grounds (Kondamudi et al. 2008). Therefore, coffee oil
could provide a significant contribution to the biodiesel fuel supply.
However, a direct transesterification process cannot be utilized since high amount
of free fatty acid (FFA) is usually present in the coffee oil extracted from spent coffee
grounds, resulting in poor conversion (Oliveira et al. 2008). Acid-catalyzed transesterifica-
tion is insensitive to FFA in the feedstock, but it requires longer reaction time and higher
temperature. Many researchers recommend using acid catalysis as pretreatment step fol-
lowed by an alkaline-catalyzed step. Wang et al. (2007) adopted this kind of two-step
catalyzed process to prepare biodiesel from waste cooking oil. In the first step of Wangs
method, the FFAs in the waste cooking oil were esterified with methanol catalyzed by
ferric sulfate. In the second step, the triglycerides in the waste cooking oil were trans-
esterified with methanol (methanol/oil molar ratio 6:1) catalyzed by 1.0 wt% potassium
hydroxide at 65 C for an hour. After this two-step catalysis process, the final product with
97.02% conversion of biodiesel was obtained. Another approach to transesterification is
using immobilized lipase as a catalyst. Lipase-catalyzed transesterification involves a two-
step mechanism when looking at a single ester bond. The first step is hydrolysis of the ester
bond and release of the alcohol moiety followed by an esterification with the second sub-
strate. Compared with conventional methods using alkaline or acid catalysts, utilization
of immobilized lipases as catalysts for biodiesel fuel production shows a great potential
because of the following advantages: mild reaction conditions, specificity, reuse, gener-
ating high purity product with little soap and wasterwater produced. Overall, the use of
enzymes for biodiesel production is considered more environmentally friendly (Fukuda
et al. 2001; Akoh et al. 2007).
In this study, both the two-step reaction and enzyme-catalyzed transesterification
were examined, aiming to evaluate the feasibility of using coffee oil from spent coffee
grounds as a raw material for biodiesel synthesis. We also evaluated varying methods
of oil extractionboth cold press oil extraction and hexane extraction were utilized and
compared.

MATERIALS AND METHODS

Materials
Spent coffee grounds were obtained from Chatham Markeplace and Piedmont
Biofuels Community kitchen in Pittsboro, North Carolina, USA. Potassium hydroxide
flake, methanol (analytical reagent grade), and hydrochloric acid (32 wt%) were procured
from Fisher Scientific. A flat-bottom flask was used as a laboratory scale reactor for the
two-step reaction approach, and a hot plate with magnetic stirrer arrangement was used
for heating the mixture in the flask. A shaker bath was used for the enzyme catalysis
532 BURTON, FAN, AND AUSTIC

approach. The 831 KF Coulometer (Metrohm Company, Switzerland) is for moisture anal-
ysis. Soxhlet extractor and Isotemp oven are both from Fisher Scientific. A Type 20 model
Taby Pressen Oilseed Press was used for the cold press oil extraction.

Oil Extraction
Cold press extraction was performed on spent grounds that were dried overnight
at 75 C to remove moisture. This extraction yielded no liquid phase. The only oil from
this extraction procedure was observed on the press plug from the screw press. There was
a slight oily film observed on the plug. This film was insufficient for further analysis.
Thus, the oils of the spent coffee grounds (dried and crushed) were obtained by solvent (n-
hexane) extraction in a soxhlet extractor, with an extraction capacity of 6070 g of spent
coffee grounds (dried and crushed) per batch. About 250 mL of hexane was used for 65 g
of dried and crushed spent coffee grounds. The oils were extracted in batches for 1516 h.
After extraction, the solvent was completely recovered in a rotary evaporator under high
vacuum. The solvents were reused in the next batch of extraction.

Two-Step Biodiesel Production Process

Using coffee oil obtained from the soxhlet extraction process, coffee oil-based
methyl esters were produced. The moisture was measured by direct coulometric Karl
Fischer titration according to ASTM D 6304. The moisture in the filtered coffee oil was
0.39%. The FFA for the coffee oil was 16.3 wt%. Higher amounts of FFA (>1 wt%) in
the feedstock can directly react with the alkaline catalyst to form soaps, and emulsions
thus preventing separation of the biodiesel from the glycerol fraction and decreasing the
yield. Therefore, it is better to select a two-stage catalysis: First acid catalysis followed by
alkaline catalysis (Figure 1). In this study, hydrocholric acid was chosen as acid catalyst
and potassium hydroxide as alkaline catalyst.

Acid-Catalyzed Esterification
About 20 g coffee oil was transferred into the reaction flask and preheated by the
hot plate to the desired reaction temperature before the reaction started. The hydrochloric

HCl
RCOOH + CH 3OH RCOOCH 3 + H 2O

O O
R1 C O CH 2 R1 C O CH 3 CH 2-OH
O KOH O
+ 3CH 3OH + CH-OH
R2 C O CH R2 C O CH 3
O O
CH 2-OH
R3 C O CH 2 R3 C O CH 3

TRIGLYCERIDES M E T H ANO L BIODIESEL G L YC E R O L

Figure 1 Mechanism of biodiesel synthesis by the two-step catalyzed process.

 BIODIESEL FROM SPENT COFFEE GROUNDS 533

acidmethanol solution was heated to the reaction temperature. Then the methanolic-acid
solution was added to the coffee oil in the reaction flask, and at this point, the measurement
of reaction time was started. After the required reaction time, the reaction mixture was
poured into the separation funnel and was allowed to settle for 2 h. The excess methanol,
combined with the hydrochloric acid and impurities, formed a layer on top of the biodiesel
and oil. The lower layer (oil phase) was ready for alkali-catalyzed transesterification.

Alkali-Catalyzed Transesterification
The esterified coffee oil (lower oil phase) was transferred to the reaction flask and
preheated to the desired reaction temperature. The calculated amount of potassium hydrox-
ide and methanol were preheated to the reaction temperature. Finally, the methanolcaustic
solution was added to the esterified coffee oil. The transesterification was carried out on the
hot plate and a constant stirring speed was maintained. After reaction, the reaction mixture
was poured into a separatory funnel and was allowed to settle overnight.

Enzyme Catalysis Biodiesel Production Process

About 20 g coffee oil was transferred into the reaction vessel and preheated in the
water bath to the desired reaction temperature before the reaction started. A 2 g catalyst
(1 g Novozyme 435 and 1 g TL-IM) pre-soaked overnight in the biodiesel was added.
A solvent-free system was used. The reaction was performed in the shaker bath at 35 C
for 72 h.

Analytical Methods
ASTM D 664 was used for FFA determination of coffee oil. The AOCS method
Cc 1795 was used for soap determination in methyl esters derived from coffee oil. The
Karl Fisher coulometric titration method (ASTM D 6304) was utilized to determine the
moisture of coffee oil.
An automatic cool on column injection gas chromatograph (GC; Agilent
Technologies, USA, 5890 Series II) was used for the determination of mono-, di-, and
triglycerides and free glycerol in accordance with ASTM D 6584. A capillary GC column
(Restek, USA, Rtx-biodiesel column (15 m 0.32 mm id 0.10m film)) was used in
the apparatus. The temperature of the flame ionization detector was 380 C. The initial GC
oven temperature was kept at 50 C for 1 min, heated at 15 C/min to 180 C, and then
heated at 7 C/min to 230 C, finally heated at the ramp of 30 C/min to 380 C, at which
it was kept for 10 min. About 100 mg of the biodiesel sample was weighed into a 10 mL
septa vial. Exactly 100 mL of each internal standard and MSTFA silylating agents were
added to the vial. The vial was shaken and allowed to sit for 15 min at room temperature.
Then, 8 mL of n-heptane was added to the vial, which was then briefly shaken. A 1 L
splitless injection volume was used. A calibration curve was generated from four stan-
dards (i.e., triolein (TG), diolein (DG), monoolein (MG), and glycerine) and two internal
standards (i.e., 1,2,4-butanetriol for glycerine and 1,2,3-tricaproylglycerol (tricaprin) for
glycerides). Each standard was injected three times at five different concentrations. The
calibration curves of the standard solutions showed good linearity. (The standard solutions
were a biodiesel package from Supelco Company.)
534 BURTON, FAN, AND AUSTIC

RESULTS AND DISCUSSION

The yield of coffee oil is 9.8% obtained from dry grounds that were processed using
the Soxhlet extraction method. The collected crude oil was filtered under vacuum before
esterification and transesterification.

Acid-Catalyzed Esterification and Alkali-Catalyzed Transesterification

The molar ratio of alcohol to oil, catalyst concentration, reaction temperature, and
time are the main factors affecting acid-catalyzed esterfication. Among these, the molar
ratio of alcohol to oil has a great effect on reducing FFA of vegetable oil. The molar ratios
between 4.5:1 and 18:1 were reported in the literature (Veljkovic et al. 2006). In this study,
a 40:1 molar ratio of methanol to FFA was preferred and 5% (based on FFA) hydrochloric
acid was added. The FFA in the coffee oil was decreased from 16.3% to 2.64%. About
84% reduction was achieved after 2 h acid-catalyzed esterification at 55 C.
After esterfication, the transesterification of the esterified coffee oil was carried out at
55 C with a 6/1 molar ratio of methanol to oil in the presence of 1% (wt/wt, based on oil)
potassium hydroxide, plus the amount needed to neutralize the FFA (Marchetti et al. 2007;
Meher et al. 2006a, 2006b). Unlike the results reported by other researchers (Oliveira et al.
2008), after completion of the reaction, two layers were separated after 1 h. This is due to
the fact that relatively low FFA was obtained after esterification and less soap was formed
during the transesterification process. However, the conversion was lower than 90%. This
may be attributed to the presence of unsaponifiable matter in the coffee oil, which could
affect the interactions between the reactants. It is also possible that soap formation yielded
a larger loss to the glycerin phase.

Lipase-Catalyzed Transesterification Process

TL-IM (Thermomyces lanuginose) is preferable to transesterification, while
Novozyme 435 is preferable to esterification based on manufacturers recommendations.
Since coffee oil showed high amount of FFA, 50% Novozyme 435 and 50% TL-IM were
selected. A stepwise addition of methanol was utilized due to the fact that one-time addition
in large quantities of methanol can inactivate the catalyst. A 3/1 molar ratio of methanol
to the coffee oil was used in this study. The first-step methanolysis was conducted at 35 C
in a mixture of 1/3 molar equivalent of methanol and coffee oil. The second and third 1/3
molar equivalent of methanol was added after 24 and 48 h of the reaction, respectively.
After total 72 h reaction, the conversion reached 98.5%. No soap was detected for the final
product.
Figure 2 showed the corresponding gas chromatography. It can be seen from Figure
2 that free glycerol, mono-, di-, and triglyceride peaks were all identified. Bound glycerol
(monoglycerides + diglycerides + triglycerides) is the indicator of the completeness of
the biodiesel production process. Lower bound glycerol indicates a more complete trans-
esterification process. Bound glycerol content of 0.16% was obtained in this study and
was within the ASTM D 6751 standard requirement, demonstrating the feasibility of using
enzymes to produce biodiesel from spent coffee grounds.
Hence, it was found that enzymatic production was more favorable than the two-
step reaction approach for biodiesel production from spent coffee grounds. Though the
 BIODIESEL FROM SPENT COFFEE GROUNDS 535

Figure 2 Gas chromatography of biodiesel from coffee oil by the enzyme-catalyzed approach.

enzyme catalyst is more expensive than the commonly used sodium hydroxide and potas-
sium hydroxide, it can be reused for several cycles and no further purification process is
needed, which could reduce the overall production cost.

CONCLUSION
In summary, the mechanical cold press oil extraction of both wet and dry spent
grounds was unsuccessful. In the evaluation of the spent coffee grounds, it was noted that
the grounds are at least 56% water content by weight. Therefore, the only means of oil
extraction from spent coffee grounds feasible was by hexane-soxhlet extraction. The oil
content after hexane extraction was a 10% by dry weight of the grounds and 5% by wet
weight. The coffee oil contained a FFA content of 16% by weight. Due to its high FFA
content in this coffee oil obtained, acid pretreatment followed by alkali-catalyzed transes-
terification and enzyme catalysis approach are the optimal methods to process coffee oil.
The enzymatic approach was the most effective with a higher yield at 98.5% conversion
and simpler sampling and processing paradigm. For future consideration, a larger volume
of coffee oil might be obtained for production. Therefore, further analysis of the coffee
oil-based methyl esters may be evaluated against the ASTM D 6751 standard specification.

ACKNOWLEDGMENTS
The authors wish to acknowledge Piedmont Biofuels Industrial for financing this research study.

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