136 Journal of Food, Agriculture & Environment, Vol.

7 (2), April 2009

Detection and determination of furfural in crude palm oil
Abdul Azis Ariffin
1
, Rosnah Mat Soom
2
, Huey-Chern Boo
1
, Chia-Chun Loi
1
, Yuen-Hwa Chai
1
and S. M. AbdulKarim
1*
1
Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
2
Malaysian Palm Oil Board, Bandar Baru Bangi, 43000 Kajang Selangor, Malaysia.
*e-mail: ak@food.upm.edu.my, hcboo@putra.upm.edu.my
Received 10 December 2008, accepted 6 April 2009.
Abstract
In the palm oil mill, fresh fruit bunch (FFB) undergoes various thermal and mechanical treatments to produce the crude palm oil (CPO). FFB consists
of many fruits attached to the spikelets that are spirally arranged on the main bunch stalk. Each fruit is made up of a nut enveloped by the fleshy
mesocarp, which is reinforced by strands of fibers running from the base towards the fruit tip. A ripe fruit mesocarp contains oil-rich cellulosic cells.
These cells are bound together by hemicellulose. Whilst cellulose is very stable, the hemicellulose is easily hydrolyzed. This hydrolysis occurs during
sterilization of the FFB when it is exposed to temperatures of 140-145C and pressure of 40-45 pound per square inch (psi) or 275.8-310.3 kPa for
1-1 hours. This condition aims at and ensures the detachment of fruits from the bunch. The in-depth chemical changes that occur in the FFB during
sterilization are not fully understood and continuously being investigated. Xyloses form one of the products of hydrolysis, and furfural is another
product that results from the dehydration of pentose formed also upon the hydrolysis of hemicellulose. Presence of furfural was tested in six
extracted samples, namely CPO, mill-pressed crude, condensate oil, sludge oil, sterilized FFB oil and unsterilized FFB oil, using aniline acetate
colorimetric method, thin-layer chromatography (TLC) and UV-visible spectrophotometry. The color formation was compared to that of standard
furfural. Furfural was detected in CPO, crude, condensate oil, sludge oil and sterilized FFB oil, while it was undetected in the unsterilized FFB. The
amount of furfural was quantified in CPO, condensate oil and sludge oil using high-performance liquid chromatography (HPLC).
Key words: Crude palm oil, hemicellulose, xylose, furfural.
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Introduction
Palm oil processing involves many physical and mechanical
activities circumventing in the intention to the extraction of crude
palm oil (CPO) due to the nature of its fruits. Upon harvesting of
fresh fruit bunch (FFB) in the plantation, FFB is sent to the mill for
processing. In the conventional mill, fresh fruit bunches are first
placed in cages prior to high-pressure steam cooking in the
sterilizer. The steam at 40-45 psi and 130-135C is pumped into the
sterilizer for 1 to 1 hours.
In the milling process, sterilization is the only process that allows
maximum chemical reactions in the FFB
1
and hence is a very
critical and important process to ensure successful deliberations
of series of events. Sterilization plays an important role in
deactivating oil-splitting enzyme (lipase), which is responsible
for the formation of free fatty acids in palm oil. The quality of palm
oil is dependent on its free fatty acid content where low free fatty
acid represents better quality. Sterilization is also important to kill
microorganisms and insects that may be present in the fruit
consignment. Small damages created by insects and enzyme
released by microorganisms may help increase the free fatty acid
content. Sterilization process affects eventual fruit kernel
detachment from the nut shell.
During the sterilization process, cellulosic materials, especially
hemicellulose, undergo various chemical breakdowns resulting in
detaching and freeing of the fruits from the spikelets of the fruit
bunch stalk. Hemicellulose is a long carbohydrate polymer
consisting of 5-carbon pentose and 6-carbon hexose units.
Hemicellulose is responsible for holding the cells in the fruit
together and attaches the fruits to the spikelets demarked by the
abscission layer. Hemicellulose is less stable compared to alpha-
cellulose and is prone to chemical breakdown during sterilization
where the hemicellulose is first hydrolyzed to pentoses and then
dehydrated with the release of 3 water molecules to furfural.
Furfural, also known as 2-furaldehyde, 2-furanaldehyde, fural,
furfuraldehyde, pyromucic aldehyde or 2-furancarboxyaldehyde
2, 3
is a heterocyclic aldehyde with the chemical formula C
5
H
4
O
2
.
Furfural is a colorless liquid with a sweet, pungent almond-like
aroma and easily oxidizes by air to a dark red/brown color
compound. It is soluble in most organic solvents, except saturated
aliphatic hydrocarbons
3
, and it has low solubility in water. Furfural
is an industrial organic chemical derived from a variety of
agricultural by-products containing pentosans
4
, and it is not a
natural product in the plant system. In theory, furfural can be
derived from any material containing pentosans
2, 4
in condition
that the raw materials undergo a hydrolysis to pentoses and the
pentoses are subsequently dehydrated to furfural
3
. Pentosans
like araban and xylan, which are found in cereal straws and seed
husks, also serve as raw materials for commercial production of
furfural
5
. Some of the common raw materials used for furfural
production are corncobs, oat hulls, sugar cane bagasse and rice
hulls
3, 4, 6
.
Furfural has many essential uses in the industry and one of the
most widely used solvents
7, 8
. Besides being used in fungicides,
Journal of Food, Agriculture & Environment, Vol.7 (2), April 2009
137
germicides, herbicides and insecticides, furfural acts as an
intermediate in the manufacture of many polymers. Furfural acts
as an accelerator in the vulcanization of rubber polymer, as
plasticizer in the plastic industry and as binder in foundry brick
and runway road. Furfural is often used as a selective solvent in
petrochemical industry to produce lubricating oils
3, 4, 7
and as a
reactive wetting agent for the resin binder system in the production
of abrasive wheels and refractory components
3
. Furfural is one of
the important additives in the formulation of rocket fuel. Besides
widely used in the non-food industries, furfural is also used as a
flavoring agent in the food and beverage industry and is detected
in cocoa, coffee, tea, beer, wine, milk products and bread
9
. Furfural
also has been identified in the essential oil of citronella, lime,
lavender, sassafras and camphor
10
. The presence of furfural in
the processed food and beverages is the result of thermal treatment
during processing that causes decomposition of carbohydrates
to furfural
3
. The aim of this study was to detect and reaffirm the
presence of furfural in CPO using various analytical methods such
as well-known aniline acetate colorimetric method, thin layer
chromatography (TLC) and UV-visible spectrophotometry. High
Performance Liquid Chromatography (HPLC) was employed to
quantify furfural content.
Materials and Methods
Materials: A total of 6 types of oil obtained from fruits of palm,
Elaeis guineensis, were used in this investigation. These were
crude palm oil (CPO), mill press crude, condensate oil, sludge oil,
sterilized fruit bunch oil and unsterilized fruit bunch oil.
CPO, the intended product of palm oil mill, was extracted from
palm fruit mesocarp at the mill. Mill waste discharges, including
condensate oil from the sterilizer and sludge oil, the final mill waste
discharge, were obtained from the mill. Unsterilized FFB oil was
extracted from ripe, fresh fruits whilst sterilized FFB oil was
extracted from fruits that were sterilized but without going through
the complete milling process.
Reagents and standard: Aniline acetate solution was prepared
using aniline, glacial acetic acid and 95% ethanol in 1:1:8 (v/v/v)
ratios. Standard furfural was purchased from Merck (Darmstadt
Germany).
Extraction of furfural: Twenty g of oil sample was placed into a
250 ml round bottom flask and 100 ml of distilled water was added.
A separation funnel containing 100 ml of distilled water was
attached to the top of the flask, and water was allowed to drip at
the rate of 1.5 to 2 ml/min. The water level in the flask was maintained
at the 100 ml throughout the extraction. The mixture was boiled
and stirred on a hotplate magnetic stirrer. A condenser was used
to connect the flask to another distillate-receiving flask. The
extraction was stopped once 100 ml of distillate was collected.
Aniline acetate colorimetric method and UV-visible
spectrophotometry: From the distillate obtained during the
extraction process, 0.1 ml was added to 5 ml aniline acetate solution.
The color changes were observed and recorded at 200-700 nm
using a Shimadzu UV-1650 UV-VIS Spectrophotometer (Shimadzu,
Kyoto, Japan) and using ethanol as a blank.
Thin Layer Chromatography (TLC) analysis: Ten microlitres of
each extracted distillate were pipetted and spotted onto a glass
TLC plate pre-packed with a layer thickness of 250 m silica gel 60
(Merck, Darmstadt, Germany). Chloroform was used as the mobile
phase. Once the separation was completed, aniline acetate solution
was sprayed on the plate. The color change was observed. The
retention factor (R
f
) value of the furfural-aniline acetate complex
was 0.68.
High-Performance Liquid Chromatography (HPLC) analysis:
A Hypersil ODS C
18
5 m column (250 x 4.6 mm. i.d.) (Thermo
Fisher Scientific, Waltham, USA) was used. Separations were
carried out isocratically at room temperature using a mixture of
acetonitrile-water (5:95, v/v) at a flow rate of 1 ml/min as the
mobile phase. Detection using UV detector at 284 nm was carried
out
11
. The injection volume was 20 l.
Furfurals were quantified by referring to a calibration curve in
the range of 0.2-10 ppm assay of standard furfural. The value from
the calibration curve was to multiply by 5 to obtain the furfural
content.
Statistical analysis: Means and standard deviations were
calculated with SPSS statistical software (Version 14.0, SPSS Inc.
Chicago, IL, USA). SPSS was used to perform one-way-analysis
of variance (ANOVA) with a Tukeys posteriori test to the average
furfural content to detect statistically significant differences
(p<0.05) between distillates of CPO, sludge oil and condensate
oil.
Results and Discussion
The furfural extracted from different oil samples was allowed to
react with aniline acetate. The presence of furfural was indicated
by the color changes from yellow to pink and finally to red when
more furfural was present
12
. From the study, furfural was detected
in CPO, crude, condensate oil, sludge oil and sterilized FFB oil,
which corresponds to the reaction between standard furfural and
aniline acetate, giving a pink color solution, while the unsterilized
FFB oil remained yellow as is the color of aniline acetate.
Furfural and aniline acetate have a very specific reaction, and
this is not influenced by interference from other aldehydes such
as hydroxymethylfurfural, although these two compounds are very
similar in their formation process. The amount of furfural in the oil
samples is minute and requires a very sensitive detection method.
Hence, the reaction between furfural and aniline was utilized to
detect furfural at low concentrations
12
. The results of the
spectrophotometer absorbance at 518 nm (Fig. 1) that is specific
for furfural-aniline acetate reaffirm that a peak was observed in
the range of 515 to 520 nm for all samples containing furfural. The
variable content of furfural in the CPO of the mills may be
associated with the intensity of the mill sterilization process.
From TLC analysis, the samples containing furfural showed
separation on the plate close to the standard furfural. The
completed chromatographic separation sprayed with aniline
acetate solution also corresponded with the standard furfural-
aniline acetate retention factor (R
f
) value. It was clearly shown the
separation of furfural compound in all distillates except that from
unsterilized FFB oil by showing a pink color spot.
Acid and hemicelluloses are the two crucial compounds in the
formation of furfural during thermal treatment employed in the
food processing industry, in this case, the sterilization process of
138 Journal of Food, Agriculture & Environment, Vol.7 (2), April 2009
0.500
0.450
0.400
0.350
0.300
0.250
0.200
0.150
0.100
0.050
0.000
2
0
0
.
0

5
0
0
.
0

7
0
0
.
0

6
0
0
.
0

4
0
0
.
0

3
0
0
.
0

A
b
s
o
r
b
a
n
c
e
Wavelength nm
Figure 1. The chart shows the absorbance spectrum of standard furfural-
aniline acetate [1] complex. [2], [3] and [4] are absorbance at 518 nm of
CPO from palm oil mill A, B and C, respectively.
Figure 2. Chromatogram of furfural in CPO distillate.

Furfural

V
2.0E+03
1.2E+04
4.0E+03
6.0E+03
8.0E+03
1.0E+04
0.0E+00
2.00
[min]
14.00 12.00 10.00 8.00 6.00 4.00
Furfural
CPO Dengkil Condensate oil Sludge oil
14.00
0.00
2.00
4.00
6.00
8.00
10.00
12.00
F
u
r
f
u
r
a
l

c
o
n
c
e
n
t
r
a
t
i
o
n

(
p
p
m
)
Sample
11.67 0.13a
2.78 0.06c
3.78 0.04b
X
X
X
3.78 0.04
b
11.67 0.13
a
2.78 0.06
c
Figure 3. Average furfural content of different oil samples from the
mill. Results are means standard error (n = 3). Bars that have no
common letters are significantly different (p<0.001).
FFB at the palm oil mill. The readily available free fatty acids (FFA)
in the fruit act as catalysts in contributing a proton, which will
then react with hemicellulose to produce furfural at a higher
pressure and temperature. During the sterilization process,
hemicellulose will first hydrolyze to xylose and then dehydrate to
furfural. Weak acids like FFA show little reaction with xylose under
normal conditions
10
but when heated in the sterilizer, the reaction
is amplified.
In the case of preparation of unsterilized FFB oil, cold hexane
was used to extract the oil from the fruits and thus avoiding the
formation of furfural during the extraction process. Therefore, no
furfural was detected in the oil.
The HPLC chromatogram of furfural of CPO, shown in Fig. 2,
corresponds to 11.67 ppm (Fig. 3). Similar HPLC chromatograms
of furfural of condensate and sludge oils were observed to be
similar to the HPLC pattern of furfural of CPO. From the one-way
ANOVA, furfural content in CPO, sludge oil and condensate oil
was shown to be significantly different (p<0.01). With the Tukeys
posteriori test, it was shown that furfural content in all the oils
was significantly different from one another. Furfural content in
CPO exhibits highest followed by condensate oil, and sludge oil
exhibits the lowest furfural content amongst the oils. The reason
for this behavior has not been fully understood.
Conclusions
Furfural was detected in CPO, crude, condensate oil, sludge oil
and sterilized FFB oil. The reaction of furfural with aniline acetate
resulted in the formation of pink furfural-aniline acetate complex.
The function of furfural-aniline acetate complex was further
confirmed by the absorbance at 518 nm. Sterilization process of
FFB resulted in the formation of furfural, which is eventually
trapped in the final CPO of the palm oil mill. The quantity of furfural
varied in different palm oil mills and it is also varied in different oil
samples from the same mill.
Acknowledgements
This study was supported by Research University Grant Scheme
(RUGS), Universiti Putra Malaysia (91013).
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