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Vol.1, No.4, 118-123 (2011)
doi:10.4236/jdm.2011.14016
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opyright © 2011 SciRes. Openly accessible at http://www.scirp.org/journal/JDM/
Journal of Diabetes Mellitus
Anti-diabetic effects of cold and hot extracted virgin
coconut oil
Mahadevappa Siddalingaswamy1*, Arunchand Rayaorth2, Farhath Khanum1
1Biochemistry & Nutrition Discipline, Defense Food Research Laboratory, Mysore, India;
*Corresponding Author: mslswamy@gmail.com
2Department of Medical Biochemistry, Kannur University, Kannur, Kerala, India.
Received 15 July 2011; revised 24 August 2011; accepted 5 September 2011.
ABSTRACT
Virgin coconut oil (VCO) has been shown to po-
ssess insulinotropic effects shown in isolated
perfused mouse islet with hypolipidemic effects.
Hot extracted virgin coconut oil (HEVCO) has
been shown to possess better antioxidant pro-
perties than cold extracted virgin coconut oil
(CEVCO). These properties were exploited to st-
udy the anti-diabetic effects of HEVCO and CE-
VCO in diabetic rats. Four groups 8 rats each,
first group served as non-diabetic control re-
maining groups were made diabetic and force
fed with 2 ml alcoholic extracts of commercial
coconut oil (CCO), CEVCO and HEVCO for 21
days. Blood glucose once in 5 days, body wei-
ght gain, food intake once in a week and water
intake and urine output daily, were monitored.
Animals were sacrificed at the end of 21 days.
The results indicated HEVCO reduced blood gl-
ucose and lipids viz total cholesterol (TC), tri-
glycerides (TG), Low and Very Low Density Li-
poprotein (LDL + VLDL) and thiobarbutyric acid
reactive substances (TBARS) increased the an-
tioxidant status by elevating activities of anti-
oxidant enzymes such as superoxide dismutase
(SOD), catalase, glutathione peroxidase (GSH-
Px), glutathione (GSH) concentration and de-
creased lipid peroxidation in liver than CEVCO.
These beneficial effects may be attributed to in-
creased polyphenolic and other antioxidants
content present in HEVCO.
Keywords: Anti-Diabetic; HEVCO; CEVCO; CCO;
Hypoglycemic; Hypolipidemic; Antioxidant Enzymes
1. INTRODUCTION
Diabetes a metabolic disorder is spreading like an epi-
demic disease and India appears to be the capital. Major
causes of diabetes are consider to be, imbalanced food
habits, change in life style, obesity, lack of physical acti-
vity, uncontrolled oxidative stress, genetic defects etc.
Coconut oil is one of the primary sources of energy, in
tropical countries like costal India, Srilanka, Philippines,
and Indonesia [1]. The oil holds high place of respect in
ayurvedic medicine in India. Virgin coconut oil (VCO) a
preparation of coconut oil without harsh processing such
as refining, hydrogenation, deodorization, bleaching etc
may retain native bioactive compounds present in it [2].
VCO extracted in cold and hot conditions shown to be
rich in polyphenols. Lauric acid, present in coconut oil
has been shown to possess insulino tropic properties in
isolated perfused mouse islet model but not proven in
diabetic animals [3]. It has been shown that hot extracted
virgin coconut oil (HEVCO) possess better antioxidant
potency than cold extracted virgin coconut oil (CEVCO)
[4,5]. These two oils have been shown to inhibit lipo-
protein oxidation with hypolipidemic effects [6]. In dia-
betic patients antioxidants may play a vital role in im-
proving insulin response to the loaded glucose and may
reduce insulin resistance [7]. However information re-
garding the anti-diabetic effects of CEVCO and HEVCO
with their insulino tropic and hypolipidemic properties is
not reported in animal model to the best of our knowl-
edge. Hence experiments were carried out to study the
anti-diabetic effects of CEVCO and HEVCO in strepto-
zotosin induced diabetic rats.
2. MATERIALS AND METHODS
CEVCO and HEVCO were prepared according to the
described method [4]. CEVCO: Matured coconut milk
was prepared by hand pressing the scraped coconut flesh
and squeezed through multi folded mull cloth. The milk
was chilled to 283˚K for 10h to solidify the oil. The
aqueous layer was discarded and the oil was allowed to
stand at 303˚K until it dissolved completely. The mixture
was further centrifuged and the oil layer was separated.
M. Siddalingaswamy et al. / Journal of Diabetes Mellitus 1 (2011) 118-123
Copyright © 2011 SciRes. Openly accessible at http://www.scirp.org/journal/JDM/
119119
HEVCO: Coconut milk emulsion was heated to 373 -
393˚K for 60 minutes until water in the emulsion was
completely evaporated. Commercial coconut oil (CCO)
CEVCO and HEVCO were extracted thrice with metha-
nol (1:4 V: V) and evaporated over a flash evaporator.
Polyphenols [8], DPPH radical scavenging activity [9]
and Fatty acid composition [6] were determined in
HEVCO, CEVCO and CCO.
2.1. Animal Experiment
Animal experiment was carried out as per guidelines
of Institutional Animal Ethics Committee. 32 male Wis-
tar albino rats reared at Defence Food Research Labora-
tory, Mysore, India, with body wt 140 - 150 g were se-
lected based on uniform food intake and weight gain
were maintained with standard laboratory conditions.
The animals were divided into 4 groups of 8 rats, the
first group being non-diabetic control. Remaining 3 gr-
oups were made diabetic by injecting streptozotocin in
biological saline intraperitoneally (55 mg/kg body wt.).
Second, the diabetic control group was force fed with 2
ml extract of CCO. Third and fourth groups were force
fed with 2 ml CEVCO and HEVCO extracts per rat per
day after fasting from 9 AM to11AM. The animals were
fed adlibitum with synthetic diet (Table 1). The experi-
mental duration was for three weeks. Food intake and
body wt gain were monitored weekly while, water intake
and urine output daily. The blood glucose levels were
monitored 5 days once, by obtaining blood samples from
tail vain, after overnight fasting, with standard kit pur-
chased from M/s Krest Biochemical’s, Bombay, India.
At the end, animals were sacrificed, after overnight fast-
ing, by survical dislocation. Blood samples were col-
lected directly from heart, an aliquot was transferred into
heparinized centrifuge tube to obtain plasma (centri-
fuged at 1000 rpm for 5 min) and analyzed for glucose,
urea, creatinine total cholesterol (TC) triglycerides (TG)
and high density lipoprotein (HDL)) with standard kits
marketed by M/s Krest Biochemicals, Bombay, India.
Table 1. Composition of the diet.
Ingredients Quantity (per 100 g)
Casein 20
Coconut oil 9
Mineral mixture USP XIV* 4
Vitamin mix** 2
Shark liver oil 1
α-tocopherol 0.01
Corn starch 63.99
*Purchased from Sisco Research Laboratory, Mumbai, India; **Prepared as
per Indian Standards I.S.7481 (1975).
Low density lipoprotein + very low density lipoprotein
(LDL + VLDL) content was calculated by subtracting
high density lipoprotein (HDL) cholesterol from TC.
Thiobarbutyric acid reactive substances (TBARS) were
assayed in both blood and liver according to Ohkania et
al., [10] and Girroti et al. [11]. Whole blood was ana-
lyzed for haemoglobin, total white and, red blood counts
and plate let counts (Sysmex model KX-21 Trans Asia).
The organs namely liver, kidney and heart were removed
and weighed. Liver and kidney samples were analyzed
for antioxidant enzymes such as Superoxide dismutase
(SOD), [12] catalase [13], and glutathione peroxidase
(GSH-Px) [14] and glutathione (GSH) [15].
2.2. Test of Significance
The data were subjected to ANOVA by “graph pad
prism software” the results mentioned were significant at
p < 0.05.
3. RESULTS
3.1. Total Polyphonic, Radical Scavenging
Active
Polyphenolic and radical scavenging activity were high
in HEVCO than CEVCO and CCO (Table 2, Figure 1).
3.2. Fatty Acid Composition
There was no change in the concentration of fatty acids
in all the three oils namely CCO, CEVCO and HEVCO
(Table 3).
Table 2. Polyphenolic concentration.
TOTAL POLYPHENOL (µg/g)
SAMPLE Concentration
CCO# 64 ± 5
CEVCO## 75 ± 7
HEVCO### 242 ± 13
Figure 1. Free radical scavenging activity of coconut oils by
DPPH assay.
M. Siddalingaswamy et al. / Journal of Diabetes Mellitus 1 (2011) 118-123
Copyright © 2011 SciRes. Openly accessible at http://www.scirp.org/journal/JDM/
120
3.3. Effect on Food Intake Body Weight
Water Intake Urine Output and Liver,
Heart and Kidney Weights
Increased food intake with body weight loss was ob-
served in all the diabetic groups. Lowest weight loss was
observed in HEVCO. Higher weight loss was observed
in CCO group with no change in water intake urine out-
put and liver, heart and kidney weights (Table 4).
3.4. Effect on Blood Glucose
Gradual decrease in blood glucose was observed in all
the 3 groups of animals. CCO extract was able to decr-
ease the blood glucose from 381mg to 318 mg during 3
weeks (4%, 11% and 16% at the end of 1st, 2nd and 3rd
week respectively). Significant decrease in CCO extract
fed was observed at the end of 2nd week and remained
unchanged up to 3rd week. In CEVCO group significant
decrease from 1st and 2nd week was observed but with no
change at the end of 3rd week. (The percent decreased
was 21%, 33% and 39% at the end of 1st, 2nd and 3rd
week respectively). HEVCO group decreased the blood
glucose 21% (p < 0.05) 33% and 47% at the end of 1st,
2nd and 3rd week respectively Complete inhibition of
lipid peroxidation was observed in HEVCO group but
was marginal decrease (not significant) was observed in
CE-VCO group and CCO (Tables 5 and 7).
Table 3. Effect of different extracts of coconut oils on Food
intake, body weight, water intake and urine output.
Groups Food intake
(g/day)
Weight
gain/loss
(g/21 days)
Wat e r
intake
(ml/day)
Urine output
(ml/day)
Control Non
diabetic 13.4 ± 1.4a 22.5 ± 1.5 38.5 ± 6.5a 42.5 ± 1.5a
Control diabetic 22.6 ± 1.7b –28.5 ± 4.5a 104.5 ± 5.5b 112 ± 3b
CEVCO# 21.7 ± 1.5b –19.5 ± 3.2b 87 ± 2.5b 109.5 ± 6.3b
HEVCO## 21.1 ± 1.13b –11.5 ± 0.2c 89 ± 5.2b 95.6 ± 4.6b
Values are mean of ± SD of 8 rats; Values bearing different superscripts in
the same column are significantly different (p < 0.05).
Table 4. Effect of different extracts of different virgin coconut
oils on organ weight.
Organ weight (g)
Groups
Liver Kidney Heart
Control non diabetic 3.44 ± 0.52 0.80 ± 0.10 0.35 ± 0.02
Control diabetic 4.45 ± 0.79 0.98 ± 0.12 0.36 ± 0.04
CEVCO# 3.82 ± 0.5 0.77 ± 0.08 0.36 ± 0.06
HEVCO## 3.73 ± 0.43 0.73 ± 0.09 0.38 ± 0.01
Values are mean of ± SD of 8 rats.
3.5. Effect on Blood Lipids
Diabetes increased TC and TG levels (70% and 25%)
with respect to non diabetic control. The increased lipid
levels were brought down to non diabetic control with
HEVCO feeding. CEVCO and CCO had no significant
effect. HDL levels remained unchanged. Significant in-
crease in the concentration of urea was observed in dia-
betic group which was brought down to normal with
HEVCO extract feeding (Table 7).
3.6. Effect of Antioxidant Enzymes,
Glutathione Concentration and Lipid
Peroxidation in Liver
Activities of all the enzymes decreased significantly
in diabetic control animals with an increase in lipid per-
oxidation. The decrease in activities of catalase, SOD,
GSH-Px was 54%, 69% and 54% respectively. The decr-
ease in the activities of these enzymes with CEVCO
were 33%, 55% and 18% and 9%, 41% and 8% with res-
pect to HEVCO group HEVCO fed animals maintained
catalase and GSH-Px activity to that of non-diabetic con-
trol. The glutathione concentration was also reduced from
12.64 to 8.36 mmol (33%) in diabetic control but in-
creased in HEVCO group than CEVCO and CCO groups.
Lipid peroxidation was effectively inhibited by both the
Table 5. Effect of different extracts of different virgin coconut
oils on plasma glucose.
BLOOD GLUCOSE mg/dl
Groups
5th day 10th day 15th day 21st day
Control Non
diabetic 84 ± 9 82 ± 4 75 ± 08 79 ± 4
Control
diabetic 381 ± 24a 367 ± 19a 339 ± 18b
(11%)
328 ± 12b
(16%)
CEVCO## 444 ± 32a348 ± 28b
(21%)
295 ± 2c
(33%)
273 ± 1c
(38%)
HEVCO### 485 ± 22a386 ± 20b
(20%)
325 ± 18c
(41%)
256 ± 09d
(48%)
Values are mean of ± SD of 8 rats; Values bearing different superscripts in
the same row are significantly different (p < 0.05).
Table 6. Fatty acid composition of CCO, CEVCO and HEVCO
(%).
Fatty acid CCO CEVCO HEVCO
Caprilic 5.8 6.3 6.2
Capric 6.3 7.3 7.1
Lauric 47.8 48.6 47.9
Myristic 17.2 15.9 16.7
Palmitic 8.2 7.7 7.9
Stearic 3.1 2.9 2.8
Linoleic 1.94 1.79 2.1
Linolenic ND ND ND
M. Siddalingaswamy et al. / Journal of Diabetes Mellitus 1 (2011) 118-123
Copyright © 2011 SciRes. http://www.scirp.org/journal/JDM/Openly accessible at
121121
extract (p < 0.05). The increase in lipid peroxidation
(151%) was brought down to 64% and 27% with HEVCO
and CEVCO (Table 8).
3.7. Effect on AO Enzymes in Kidney
The activities of catalase, SOD and GSH-Px decreased
by 61%, 54% and 65% respectively in diabetic control.
The decrease was only 23%, 43% and 17% in CEVCO
and 17%, 45% and 3% in HEVCO group. Glutathione
concentration was decrease by 38% in diabetic control
group. Non diabetic control group and HEVCO group
had same GSH concentration. Increased lipid peroxida-
tion (69%) was brought down efficiently by HEVCO to
32 % (Table 9). All haematological parameters remained
unchanged in all the groups (Table 10).
4. DISCUSSION
Diabetes, a silent killer, with many metabolic disorders
is spreading like an epidemic disease. Anti-diabetic drugs
for a longer period may have undesirable effects. In ad-
dition affordability for a common man on long run could
be an economical burden and may affect the economy of
both developed and underdeveloped countries. In this
context natural anti-diabetic foods with least ill effects
and low cost are most desirable.
It has been reported that lauric acid in coconut oil has
insulino tropic properties but reported in isolated islet of
mouse [3]. Kapila et al. [4] have demonstrated HEVCO
fed animals had better antioxidant status than CEVCO
due to high polyphenolic (PP) content. Medium chain
fatty acids and TG with these fatty acids metabolizes fast
and may assist in preventing obesity and stimulate
weight loss in diabetic obese patients [16].
The fatty acid present may not be packaged into lipo-
protein and do not circulate in the blood string which is
true in vegetable fat. Antioxidant may enhance the sensi-
tivity to insulin or otherwise may also reduce insulin re-
sistance and injury to pancreatic beta cells by scavenging
reactive oxygen species (ROS) in diabetic patients.
These observations made us to exploit the advantages of
CEVCO and HEVCO for their anti-diabetic effects.
It is clear from the present study that HEVCO had bet-
ter PP content and radical scavenging activity than CE-
VCO (Figure 1, Table 2), agrees very well reported lit-
erature [4]. Increased PP content in HEVCO could be
due to increased release of bounded PP with heat proc-
essing. The fatty acid composition remained unchanged
in all the three oils (Table 6). In addition lauric acid with
insulinotropic properties may have synergistic effects in
combination with PP in reducing blood sugar [7]. Better
hypoglycemic effect was observed in HEVCO (47%) than
CEVCO (39%) (p < 0.05) with same lauric acid content
suggests the vital role of PP in protecting the pancreatic
beta cells from apoptosis which is possible in diabetic
condition due to enhanced formation of ROS. Better hy-
poglycemic effect of CEVCO than CCO could be due to
available bioactive molecules retained by manual proc-
essing [17] (Table 5). The availability and concentration
of these molecules may be more in HEVCO and CE-
VCO than CCO. However, CCO has also reduced blood
glucose significantly could be due to lauric acid. Increa-
sed food intake, water intake and urine output are well-
established diabetic symptoms, were not altered signifi-
cantly. May be by increasing the quantity of extracts these
complication could be prevented. However body weight
loss was effectively prevented by HEVCO than CEVCO
and was due to better hypoglycemic effect (Table 3 ).
Table 7. Effect of different extracts of different virgin coconut oils on serum lipids and urea.
Groups TC (mg/dl) TC (mg/dl) HDL (mg/dl) LDL + VLDL
(mg/dl) TBARS (µ mol/dl) Urea (mg/dl)
Control
non diabetic 68.9 ± 3.8a 131.2 ± 6.9a 33.8 ± 2.8 a 30.3 ± 2.9a 58.72 ± 2.8a 16.8 ± 2.5a
Control diabetic 118.6 ± 5.4b 164.6 ± 8.2b 49.6 ± 3.4b 69.8 ± 5.2b 94.62 ± 8.2b 24.8 ± 2.5b
CEVCO# 99.9 ± 8.9b 152.7 ± 4.8b 48.9 ± 4.1b 52.5 ± 4.1c 78.2 ± 6.6c 25.7 ± 3.5b
HEVCO## 76.5 ± 2.4a 136 ± 5.7a 40.5 ± 2.5b 32.3 ± 2.9a 60.1 ± 3.9a 15.6 ± 5.2a
Values are mean of ± SD of 8 rats; Values bearing different superscripts in the same column are significantly different (p < 0.05).
Table 8. Effect of different extracts of different virgin coconut oils on liver antioxidants enzymes and lipid peroxidation.
Group Catalase* × 103 SOD** × 102 GSH-Px** GSH µ mol/g TBARS × 10–8 mol/g
Control non diabetic 0.93 ± 0.08a 0.937 ± 0.06a 0.89 ± 0.02a 12.64 ± 7.6a 0.741 ± 0.03a
Control diabetic 0.4 ± 0.03b 0.285 ± 0.03b 0.41 ± 0.02b 8.36 ± 0.38b 1.86 ± 0.05b
CEVCO# 0.626 ± 0.04c 0.419 ± 0.02c 0.73 ± 0.05c 9.48 ± 0.69c 1.215 ± 0.06c
HEVCO## 0.85 ± 0.02a 0.545 ± 0.02c 0.82 ± 0.03a 10.39 ± 0.78a 1.15 ± 0.6c
Values are mean of ±SD of 8 rats; Values bearing different superscripts in the same column are significantly different (p < 0.05); *ΔA of 0.1/min/mg protein;
**Units/min/mg protein.
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122
Table 9. Effect of different extracts of different virgin coconut oils on kidney antioxidants enzymes and lipid peroxidation.
Group Catalase* × 103 SOD** × 102 GSH-Px
** TBARS ×10–8 mol/g µ mol/g GSH
Control Non diabetic 0.47 ± 0.003a 0.97 ± 0.04a 0.52 ± 0.02a 0.34 ± 0.07a 3.92 ± 0.44a
Control diabetic 0.18 ± 0.004b 0.44 ± 0.06 b 0.18 ± 0.002b 1.105 ± 0.1b 2.42 ± 0.52b
CEVCO# 0.36
± 0.005c 0.53 ± 0.01b 0.40 ± 0.03c 1.082 ± 0.03b 2.91 ± 0.27c
HEVCO## 0.39 ± 0.003c 0.54 ± 0.02b 0.54 ± 0.03a 0.752 ± 0.01c 3.77 ± 0.47a
Values are mean of ±SD of 8 rats; Values bearing different superscripts in the same column are significantly different (p < 0.05); *ΔA of 0.1/min/mg protein;
**Units/min/mg protein.
Table 10. Effect of different extracts of different virgin coco-
nut oils on hematological parameters.
HEMATOLOGY
Groups
Hemoglobin WBC count RBC count Platelet
count
Control Non
diabetic 17.5 ± 1.6 6500 ± 385 9.56 ± 2.8 8.1 ± 1.2
Control diabetic 16.7 ± 1.5 7200 ± 250 9.25 ± 1.2 6.9 ± 1.5
CEVCO# 16 ± 0.8 8700 ± 550 9.1 ± 1.5 7.1 ± 1.1
HEVCO## 16.2 ± 0.9 7500 ± 550 9.43 ± 2.1 7.6 ± 1.5
Values are mean of ± SD of 8 rats.
Hyperlipidemia and elevated oxidative stress due to dia-
betes may lead to cardiac complications in diabetic pa-
tients. It has been reported that VCO reduces blood lip-
ids even under normal condition [5]. VCO rich in me-
dium chain triglycerides metabolizes quickly thus accu-
mulation of fat is inhibited [16,18]. In the present study
TC, TG and LDL+VLDL level reached the control in
HEVCO group whereas CEVCO reduced only LDL +
VLDL levels. It has been shown antioxidants reduce TG
[19] correlates well with the observations made in the
present study since HEVCO had better PP content than
CEVCO. It may also be possible that other vital antioxi-
dant vitamins present in these oils due to no saponifica-
tion may have role in reducing lipids [17]. Antioxidants
present in HEVCO, may also have a role in regulating
cholesterol synthesis by regulating HMG CoA reductase
activity. [20]. CEVCO with lesser PP content has re-
duced marginally but not significantly both TC and TG.
Lipid peroxidation in blood was also efficiently brought
down by HEVCO and CEVCO. Decreased blood glu-
cose, lipid peroxidation with hypolipidemic effects is de-
sirable in diabetic patients. Urea and creatinine levels as-
sess the kidney function. Creatinine levels did not show
any change (data not presented) but urea levels in
HEVCO group was similar to that of control non diabe-
tic animals suggesting less disturbed protein catabolism
(Table 7).
SOD, catalase and GSH-Px constitute mutually sup-
portive and defensive agents against reactive oxygen sp-
ecies (ROS). Decrease in the activity of these enzymes
in liver may indicate increased production of free radi-
cals. Activity of these enzymes particularly catalase and
glutathione peroxidase reached the non-diabetic level with
HEVCO and increased significantly with CEVCO. He-
patic lipid peroxidation was also effectively inhibited by
both the extracts indicating better anti-diabetic effects of
HEVCO and CEVCO (Table 8).
Decreased activities of antioxidant enzymes in kidney
were reversed with both the extracts. The activities of GSH-
Px the animal fed with HEVCO reached the control val-
ues suggesting better antioxidant properties (Table 9).
A study conducted by Nevin et al. [5], has shown the
VCO, compared with coconut and peanut oil in both in
vitro and in vivo enhanced the activities of antioxidant
enzymes and reduced lipid peroxidation in normal rats.
In this study similar observations were made in diabetic
animals may be attributed to quantity and bioavailability
of antioxidant sin HEVCO.
Thus the study has clearly demonstrated the anti-dia-
betic effects of VCO and HEVCO for the first time. The
overall effect of HEVCO was better than CEVCO in en-
hancing the antioxidant status, reducing the blood glu-
cose and lipid levels. The better health benefits of HE-
VCO may be attributed to its higher PP content and also
possible increased bioavailability of nutrients.
5. CONFLICT OF INTEREST
The authors have declared that there is no conflict of
interest.
6. ACKNOWLEDGEMENTS
The authors are thankful to Dr. A. S. Bawa, Director, Defence Food
Research Laboratory for his constant encouragement in carrying out
this research. We would like to thank Mrs. S. Sukanya, for typing the
manuscript.
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