JCBPS; Section B; Nov . 2014 – Jan. 2015, Vol. 5, No. 1 ; 401-412.
E- ISSN: 2249 –1929
Journal of Chemical, Biological and Physical Sciences
An International Peer Review E-3 Journal of Sciences
Available online atwww.jcbsc.org
Section B: Biological Sciences
CODEN ( USA): JCBPAT
Research Article
Evaluation on Edible Oil Quality Parameters As well As
Nutritional Value of Flaxseed (Linseed) Oil in Bangladesh.
Krishna Chowdhury*, Monira Obaid, Sharmin Akter Lisa and Rezaul Karim
*Principal Scientific Officer, Oilseed & Lipid Technology Section, Institute of Food Science &
Technology (IFST), Bangladesh Council of Scientific & Industrial Research (BCSIR).
Received: 10 September 2014; Revised: 25 September 2014; Accepted: 12 November 2014
Abstract: Flaxseed oil, also known as Linseed oil, is incredibly nutritionally rich with
omega-3 fatty acid like alpha linolenic acid. Flaxseed oil was extracted from flaxseed
collected from three locations of Bangladesh- Local Market, Pabna and Bangladesh Rich
Research Institute (BRRI) to investigate the edible oil characteristic parameters and
nutritional value of the extracted oil. The key quality parameters-Acid value, Peroxide
value and Moisture% increased extensively over 12 month period. Acid value increased
48%, 73% and 132% whereas Peroxide value increased 9333%, 1020% and 2833% for
the three samples respectively. There were no such remarkable change in Iodine value,
Saponification value, Ether Insoluble Matter, Refractive Index and Relative Density over
12 month period. Color, Unsaponifiable matter and Heavy metal content (Fe, Cu, Pb)
were found according to the standards of raw linseed oil. 0.059, 0.076 and 0.079 mg/kg
Fe; 0.001, 0.015 and 0.018 mg/kg Cu and 0.059, 0.048 and 0.069 mg/kg Pb were found in
the three samples respectively. Alpha linolenic acid, the core nutritional parameter, was
obtained 46.3%, 52.1% and 49.6% for Market, Pabna and BRRI oil respectively.
However, flaxseed oil from all locations has been found to be nutritionally rich and thus
this study conveys awareness about the edible use of flaxseed oil in Bangladesh.
Keywords: Flaxseed, Alpha linolenic acid, Medicinal supplement, Omega-3 fatty acid,
Oxidative stability, BRRI, Characteristic parameters.
401 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
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INTRODUCTION
Flaxseed oil (Linseed oil), one of the healthy sources of essential fatty acids is obtained from the dried,
ripened seeds of the flax plant (Linum usitatissimum, Linaceae).Flax plant (Linum usitatissimum, L.) is an
annual plant belonging to the genus Linum and the family Linaceae1 . Different varieties of Linum have
been developed for production of fibre and oilseed. Varieties of Linum bred for fibre use are called flax,
whereas the oilseed varieties are called linseed, oilseed flax or just flax 2 . Linseed is an alternative name
used for flax. Crops grown for seed are termed linseed in India and in the United Kingdom and flaxseed
in Canada and the United States, and flax oil or flax seed is used in many European countries3 .
Flaxseed, an edible oilseed/grain and one of the oldest arable crops, was recently acknowledged as a
functional food4 and gained much attention because of its unique nutrient components and potential effect
on the prevention of cardiovascular disease5 . Also it has a strong medicinal value globally for being used
as medicinal supplements for treating rheumatoid arthritis, osteoarthritis, dry eyes, atherosclerosis, high
blood pressure, heart disease, diabetes, and cancer6. In addition to being the richest plant source of alpha
linolenic acid (ALA; 50–62% of flaxseed oil, or ≈22% of whole flaxseed) and lignans (range: 0.2–13.3
mg/g flaxseed), flaxseed is an essential source of dietary fibre (28% by weight), of which 25% is in the
soluble form7 . Alpha-linolenic acid (ALA), a precursor to the essential omega-3 fatty acid converts into
DHA (Docosahexaenoic acid) and EPA (Eicosapentaenoic acid) which are more active omega-3s in the
human body8 . This fatty acid is susceptible to oxidation; it oxidises 20–40 times faster than oleic acid and
2–4 times faster than linoleic acid9 . Unfortunately, a high content of a-linolenic acid induces a poor
oxidative stability of linseed oil 10 .
Linseed oil is a drying oil, meaning it can polymerise into a solid form. Due to its polymer-forming
properties, linseed oil is used on its own or blended with other oils, resins, and solvents as an impregnator
and varnish in wood finishing, as a pigment binder in oil paints, as a plasticizer and hardener in putty, and
in the manufacture of linoleum11 . In Bangladesh this oil is widely used in furniture manufacture and house
building purposes. Flax plant is cultivated as rabi crop mostly in the districts of Khulna, Jessore,
Dinajpur, Kushtia, Pabna, Dhaka, Mymensingh, Tangail and Faridpur. About 49,000 m tons of seed are
produced annually from about 73,000 ha of land12 . There are some advantages to growing linseed in the
northern latitudes, as the content of linolenic acid and omega-3 fatty acids in the oil are higher in linseed
grown in cool temperatures typical for northern countries compared to southern countries13 . In recent
years, linseed production in Bangladesh has been decreased to 6,313 tons from 9,740 ha area annually14 .
However, Bangladesh has a world share of 0.3% of flaxseed production15 .
The aim of the study was to investigate the edible oil characteristic parameters and nutritional value
especially alpha linolenic acid of the oil extracted from flaxseed collected from three locations (Market,
Pabna and BRRI) of Bangladesh. As flaxseed oil is not being used for edible purpose recurrently in
Bangladesh, the objective of this study was also to bring awareness about the nutritional benefits to the
general mass of Bangladesh.
EXPERIMENTAL
Flaxseeds were collected from three locations i.e. local market, Pabna district of Bangladesh and BRRI
(Bangladesh Rice Research Institute). Oil was extracted from the seeds by traditional ghani. Then the
three types of oil were left for decantation and then separated from impurities and gummy materials. Six
samples from each type were stored in amber colour bottle to monitor the different quality parameters,
402 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
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Chowdhury et al.
fatty acid composition and heavy metal content in zero month, one month, two month, three month, six
month and twelve month during the period from January, 2012 to June, 2013.
The chemical parameters- Acid value (AV), Peroxide value (POV), Iodine value (IV), Saponification
value (SV), Unsaponifiable matter (Nonosap) and the physical parameters- Moisture content, Refractive
Index (RI), Relative density, colour and Ether insoluble matter (EIM) of the stored samples were
measured to assess the edible quality. Fatty acid composition and mineral and heavy metal content – Iron
(Fe), Copper (Cu) and Lead (Pb) of the three samples were also estimated.
Acid value (AV), Peroxide value (POV), Iodine value (IV), Saponification value (SV), Unsaponifiable
matter (Nonosap), moisture content and ether insoluble matter (EIM) were measured following the
standard IUPAC16 method. RI was measured by the ABBE 60 series Refractometer (BELLINGHAM +
STANLEY LIMITED.UK). Density meter DMA35N, manufactured by Anton Paar, Austria was used to
measure the relative density. Colour of the studied samples were measured by Lovibond tintometer,
model F.
Fatty acid composition was analysed by Gas Chromatograph fitted with Flame Ionization Detector (GCFID), Model 14B SHIMADZU, Japan, loaded with software Class GC-10 (version-2.00). The GC was
equipped with Flame ionization detector (FID) and capillary column, with dimension 15m length and
0.25 mm ID. The operating condition was programmed at oven temperature 150˚ (hold time 5 min),
8˚C/min - 190˚C (hold time 0 min), 2˚C/min - 200˚C (hold time 10 min), injection port temperature 250˚C
and detector temperature 250˚C. Nitrogen was used as carrier gas, flow rate 20ml/min and aliquots of 1µl
FAME (formed by esterification of oil samples) was injected and the peaks of fatty acids were recorded
for their respective retention time and areas by the data processor unit of GC.
Iron (Fe), Copper (Cu) and Lead (Pb) was analysed by Flame Atomic Absorption Spectrophotometer
(FAAS), Model AA-6800 SHIMADZU, Japan after digesting the oil samples in fume chamber with
Nitric acid (HNO3 ) and Perchloric acid (HClO4 ) (Assay ≥ 99.7%, Merck, Germany). The FAAS was
equipped with hollow Iron, Copper and Lead cathode lamp respectively and a deuterium lamp for
background correction were used. For Iron (Fe) estimation the spectrometer’s monochromator was
adjusted to 248.3 nm (wavelength), slit width was 0.2 nm and lamp current flow was 12 mA. For Copper
(Cu) estimation wavelength, slit width and lamp current flow were 324.8 nm, 0.5 nm and 6 mA
respectively. The wavelength, slit width and lamp current flow were adjusted to 283.3 nm, 1.0 nm and 10
mA respectively for Lead (Pb) estimation.
RESULTS & DISCUSSION
Acid and Peroxide values are used to measure the deterioration in the sensory properties of oil. The acid
value measures free fatty acids, which indicates the extent of hydrolytic rancidity. According to the
standards ISO 15017 and ASTM D23418 and the British Pharmacopoeia19 Raw linseed oil has an acid value
of less than 4.02 . The results of Figure 1 pursue this agreement. Figure 1 summarizes the changes of Acid
value of three location samples (Market, Pabna and BRRI) during storage of 12 months. Acid value of
Market oil, Pabna oil and BRRI oil increased from 1.28 to 1.9 mg/g (as KOH), from 1.06 to 1.84 mg/g (as
KOH) and from 1.24 to 2.88 mg/g (as KOH) respectively after 12 months indicating the progression of
rancidity due to storage.
403 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
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Chowdhury et al.
Change of Acid value with Time
Acid value (as KOH), mg/g
3.5
3
2.5
2
Market
1.5
Pabna
1
BRRI
0.5
0
0
1
2
3
6
12
Time in Month
Figure 1: Change of acid value (degree of rancidity) of three location samples with time
Peroxides are the primary products of oxidation, however, they are relatively short-lived and their
usefulness as oxidation indicators is limited to early stage of rancidity20 .It is known that, with esters of the
more common types of unsaturated fatty acids the initial products of oxidation are mainly
hydroperoxides21, 22. The peroxide value of linseed oil is approximately 210 that goes with the results of
Figure 2 upto 6 months. Peroxide value of Market oil, Pabna oil and BRRI oil increased from 0.12 to
1.28 mEq O2 /kg, from 1.34 to 1.81 mEq O2 /kg and from 0.51 to 1.22 mEq O2 /kg respectively within six
months. The significant change of peroxide values of the samples within 6 months are shown in Figure 2,
but there is a hasty change after 12 months (11.32, 15.01 and 14.96 mEq O2 /kg) that indicates that all oil
samples underwent precarious for human consumption.
Change of Peroxide value with Time
Peroxide value (mEq O2 /kg)
16
14
12
10
8
6
4
2
0
Market
Pabna
BRRI
0
1
2
3
6
12
Time in Month
Figure 2: Change of peroxide value (degree of oxidation) of three location samples with time
404 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
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The iodine value is a function of unsaturation and is used to examine the relative amount of unsaturated
fatty acid contents of lipids20 . The iodine value of linseed oil is 175-177 according to the standards ISO
15017 and ASTM D234-8218 . This is high compared with those of other food oils, such as olive (81), turnip
rape oil (98) or sunflower oil (125) 19, 23 and indicates the highly unsaturated nature of linseed oil 2 . As
shown in Table 1, the iodine value of three location samples varied from 175 to 176; whereas, values
ranging from 170 to 204 have been obtained by different investigators24 . In particular, climate affects the
abundance of the unsaturated fatty acid in the oil; the colder the climate, the higher the iodine value of oil
or the degree of its unsaturation25 .
In this study, there is a slight change in the iodine value after 12 months that the percent change varies
from 0.2 to 0.3 (Table 1). This is for oxidation depresses the iodine value so that old oils usually yield
low values, although, if special precautions are taken to shield them from air and light, they may usually
be kept unchanged for an indefinite period when quite pure24 .
Table 1: Change of Iodine value of three location samples after 12 months
Sample
Iodine Value
Fresh
sample
176.1
Market oil
After 12
months
172.8
Percent Change
1.9
Pabna oil
175.3
174.9
0.2
BRRI oil
176.4
175.9
0.3
175.9±0.57
174.5±1.58
Mean (±SD)
The average fatty acid composition of the samples is shown in Figure 3. The fatty acid composition of
linseed oil makes it interest for food use. Generally plant oils contain less than 25% of ɑ-linolenic acid.
However, linseed oil has an exceptionally high content of ɑ-linolenic acid 26 . In this study, 46.3% to
52.1% ɑ-linolenic acid of three types of sample has been found. Of the other fatty acids, palmitic acid
(5.2-6.4%), stearic acid (3.4-4.2%), oleic acid (27.9-34.8%), linoleic acid (8.5-9.2%) and arachidic acid
(0.5-1.2%) were observed.
C20:0
1.2%
C16:0
5.2%
C18:0
3.4%
C18:1
34.8%
C18:3
46.3%
C20:0
0.9%
C16:0
6.4%
C18:1
27.9%
C18:3
52.1%
C18:2
9.1%
(a)
C18:0
4.2%
C20:0
0.5%
C16:0
6.1%
C18:1
30.8%
C18:3
49.6%
C18:2
8.5%
(b)
C18:2
9.2%
(c)
Figure 3: Fatty acid composition of three location samples-(a) Market oil, (b)Pabna oil
and (c) BRRI oil
405 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
C18:0
3.8%
Evaluati on …
Chowdhury et al.
It can be noted that the experimental values do not vary significantly from other literature data. Flax
varieties grown in Western Canada, average from 495 samples analysed, contained 5% palmitic acid
(16:0), 3% stearic acid (18:0), 17% oleic acid (18:1), 15% linoleic acid (18:2), and 59% linolenic acid
(18:3)27 . Although similar varieties were grown in North Dakota, the 11 cultivars assessed showed the
following fatty acid composition: 5–6% of 16:0, 3–6% of 18:0, 19–29% of 18:1, 14–18% of 18:2, and
45–52% of 18:3 28 .
Saponification value is a measure of the average molecular weight (chain length of the fatty acids)2 . The
results of saponification value of the experimental samples are given in Table 2 which shows that the
initial saponification values were from 191.4-192.2 mg/g (as KOH), where they changed 0.3%-1.7% after
12 months.The saponification value of linseed oil (188-195 according to the standard ISO 150) is similar
to that of many other food oils19, 23
Table 2: Change of Saponification value of three location samples after 12 months
Sample
Saponification Value, as KOH (mg/g)
After 12
months
194.71
Percent Change
Market oil
Fresh
sample
191.45
Pabna oil
192.0
192.56
0.3
BRRI oil
192.23
193.62
0.7
191.9±0.4
193.6±1.07
Mean (±SD)
1.7
Saponification number is inversely proportional to the average molecular weight or chain length of the
fatty acids present in fat or oil29 . So, the decrease in saponification value shown in Table 2 concludes that
the chain length of fatty acids decreased during oxidation30 .
Unsaponifiables are components of an oily (oil, fat, wax) mixture that fail to form soaps when blended
with sodium hydroxide. The unsaponifiables of the raw linseed oils were found 0.39%, 0.42% and 0.4%
in Market, Pabna and BRRI oil respectively. However, literature says that linseed oil usually contains
about 0.8 percent of unsaponifiable matter24 and according to Baião and Lara31 it is maximum 1.5%.
The change of moisture content in the studied samples with time has been expressed by the Figure 4,
where it increased in every case. Water is an unusual component of oils and fats, as the two are nonmiscible and the presence of water can be compatible only at very low proportions32 .Water is a catalyst of
almost all chemical degradation reactions and the presence of high moisture content enhances oxidative
degradation and thus PV32 .
The moisture content lies between 0.13% to 0.22% for Market oil, 0.17% to 0.26% for Pabna oil and
0.14% to 0.22% for BRRI oil and it increased with time as moisture penetrates into oil from surrounding
during storage. Maximum accepted values are between 0.5 and 1.0%, since moisture interferes directly
with the energy content33 . Moreover, in the presence of moisture content above the critical limit of 0.2%,
FFA are also formed by a chemical process called autocatalytic hydrolysis where the FFA moieties
initially present act as catalysts and highly enhance subsequent formation of other FFA34-38 .
406 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
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Ether insoluble matter (EIM) or impurities is another characteristic parameter showing the purity of oil.
Impurity is calculated as the percentage of the insoluble fraction of the lipid in petroleum ether at
temperatures between 40-60o C. Impurity contents should be lower than 1% 31 . In this study, EIM varied
from 0.48% to 0.6% for Market oil, 0.56% to 0.6% for Pabna oil and 0.62% to 0.68% for BRRI oil which
is displayed in Figure 5. Figure 5 shows that there is no such significant change in EIM% with time of
the three location samples.
Moisture% change with Time
Moisture (%)
0.3
0.25
0.2
Market oil
Pabna oil
0.15
BRRI oil
0.1
0
1
2
3
6
12
Time in Months
Figure 4: Moisture content change with time
EIM(%)
EIM% change with Time
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
Market oil
Pabna oil
BRRI oil
0
1
2
3
6
12
Time in Months
Figure 5: Ether insoluble matter (EIM)change with time
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The colour of technical linseed oils is normally yellowish, a greenish colour is considered undesirable in
food use. According to the standards ISO 150 17 and ASTM D234-8218 raw linseed oil should be clear and
transparent, with no sediment at 650 C2 . Figure 6 shows the colour of crude linseed oil measured by
Lovibond Tintometer. According to Hosseinian et al.39 , chlorophyll content in linseed is responsible for
dark colour. Microbiological quality also affects the colour of oil, according to e.g. Mondal and Nandi40
fungi of the genus Aspergillus affect the colour changes of oil plants.
Colour in Lovibond scale
Colour of Crude Linseed Oil
25
20
15
Yellow
10
Red
5
0
Market
Pabna
BRRI
Figure 6:Colour of Crude Market, Pabna and BRRI Linseed Oil by Lovibond Tintometer
Table 3 represents the density and refractive index of the samples where according to Vegetable Oils
Grading and Marking Rules 41 , the specific gravity of raw linseed oil should be 0.923 to 0.928 at 300 C and
the same rule says that the refractive index of raw linseed oil should be 1.4720 to 1.4750 at 400 C. The
density of raw linseed oil varies according to its origin24 .
Table 3 shows that after storage for 12 months there was very slight decrease in the value of density.
Under favorable conditions, pure raw oil may be kept for an indefinite time without appreciable change. If
the oil is not pure, the density frequently alters slightly in consequence of the settling out of the impurities
24
. Refractive index was also measured shown in Table 3. The refractive index is a valuable indication of
the purity of linseed oil 24 .
Table 3: Relative density and Refractive Index of three location samples
Sample
Relative Density at 300 C
Refractive Index at 400 C
Fresh sample
After 12 months
Fresh sample
After 12 months
Market oil
0.9229
0.9228
1.4727
1.4728
Pabna oil
0.9231
0.9229
1.4729
1.4730
BRRI oil
0.9234
0.9231
1.4729
1.4731
408 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
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The concentration of metals- iron (Fe), copper (Cu) and lead (Pb) of crude linseed oil have been
determined so that the harmful effect of these metals would occur if they are overdosed. Trace levels of
metal ions (Cu, Fe, Mn, Co, Cr, Pb, Cd, Ni, and Zn) are known to have adverse effect on the oxidative
stability of edible oils. Transition metals such as copper and iron catalyze the decomposition of
hydroperoxides and lead to more rapid formation of undesirable substances42 .
Figure 7 shows the concentration of Fe, Cu and Pb of crude linseed oil at a glance where Fe is between
0.059 to 0.079 mg/kg, Cu- 0.001 to 0.018 mg/kg and Pb- 0.048to 0.069 mg/kg. The results agree with the
Codex Standard for Named Vegetable Oils where the maximum limits for Fe, Cu and Pb are 1.5, 0.1 and
0.1 mg/kg respectively43 . The presence of heavy metals in edible oils is due to both endogenous factors,
connected with the plant metabolism, and hexogenous factors due to contamination during the agronomic
techniques of production and the collection of olives and seeds during the oil extraction and treatment
processes, as well as systems and materials of packaging and storage 44, 45 .
Metal concentration (mg/kg)
Metal concentration of
Crude Linseed Oil
Fe
Cu
Pb
0.079
0.076
0.059 0.059
0.069
0.048
0.015
0.018
Pabna
BRRI
0.001
Market
Figure 7: Metal concentration of Crude Market, Pabna and BRRI Linseed Oil
CONCLUSION
Linseed is one of the most important nutritionally rich oilseed crops for its high linolenic acid content. In
Bangladesh, most of the edible oils such as Soybean oil, Sunflower oil, Canola oil, Safflower oil are
imported from different foreign countries which are economic burden for this country. But if the public
can consume their self-produced edible oil like linseed oil, the public as well as the whole country would
be economically benefited. Throughout this study, the quality parameters and nutritional value of crude
linseed oil have been observed for twelve months. If it can be possible to refine the crude linseed oil at a
certain level where the quality parameters would be within the permeable limit for edible purpose, it
would be very useful to the health and financial benefit of the general mass. On the other hand, linseed oil
can also be used for nutritional supplement for its universal medicinal value. Therefore, further study can
be done for refining the linseed oil so that linseed oil can be consumed for edible purpose.
409 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.
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ACKNOWLEDGEMENT
This paper is an outcome of R&D project work “Investigation on Linseed oil (Flaxseed oil) quality and
stability” of Institute of Food Science & Technology (IFST), Bangladesh Council of Scientific &
Industrial Research (BCSIR), Dhaka. The instruments used in this work have been supported by an ADP
project funded by the Ministry of Science & Technology, Government of Bangladesh. We are grateful to
the BCSIR authorities for the financial support to conduct the study.
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* Corresponding author:Krishna Chowdhury
*Principal Scientific Officer, Oilseed & Lipid Technology Section, Institute of Food Science
& Technology (IFST) Bangladesh Council of Scientific & Industrial Research (BCSIR),
412 J. Chem. Bio. Phy. Sci. Sec. B, Nov. 2014 – Jan. 2015; Vol.5, No.1; 401-412.