Docosahexaenoic acid content is significantly higher in ghrita prepared by traditional Ayurvedic method

Abstract

Background:
Ghee (clarified butter) also known as ghrita, has been utilized for thousands of years in Ayurveda. Ghee is mostly prepared by traditional method in Indian households or by direct cream method at industry level. Ayurvedic classics mention that ghrita made from cow milk is superior. However, there is no scientific comparison available on preparation methods and essential fatty acids content of ghrita.

Objective:
To investigate fatty acid composition of ghrita prepared by traditional/Ayurvedic method and commercial method (direct cream method).

Materials and methods:
Fatty Acid Methyl Esters (FAME) extracted from ghrita samples were analysed on Gas Chromatography (GC) Shimadzu B using capillary column BPX70 (0.32 mm*60 m, ID of 0.25 mm). The fatty acids in the samples were identified by comparing peaks with the external standard 68A (Nu-Chek-Prep, Inc.USA). Significant differences between the experimental groups were assessed by analysis of variance.

Results:
Distribution of fatty acids was compared in ghrita samples prepared by traditional method and direct cream method which is commercially used. Saturated fatty acids were predominant in both the groups. Mono unsaturated fatty acids and poly unsaturated fatty acids were in the range of 17-18% and 3-6% respectively. DHA content was significantly higher in ghee prepared by traditional method using curd starter fermentation.

Conclusion:
The findings suggested that ghrita prepared by traditional ayurvedic methods contains higher amount of DHA; Omega-3 long-chain polyunsaturated fatty acids, which is a major component of retinal and brain tissues and remains important in prevention of various diseases.

Full text

Journal of Ayurveda & Integrative Medicine 1
Address for Correspondence:
Dr. Kalpana S. Joshi, Department of Biotechnology, Sinhgad College
of Engineering, Pune, Maharashtra, India. E-mail: joshikalpana@
gmail.com
Received: 26-Sep-2013
Revised: 14-Nov-2013
Accepted: 25-Nov-2013
Docosahexaenoic acid content is signifi cantly
higher in ghrita prepared by traditional
Ayurvedic method
Kalpana S. Joshi
Department of Biotechnology, Sinhgad College of Engineering, Pune, Maharashtra, India
INTRODUCTION
Ghee or Ghrita is obtained by clarifi cation of milk fats by
heating and has longer shelf life at room temperature as
compared with butter. Fatty acid profi le of ghee is complex
and shown to contain high levels of saturated fats and
cholesterol. Composition varies from source of ghee such
as goat milk, sheep milk, cow milk, and buffalo milk.
[1,2]
Ghee is mostly prepared by traditional method in Indian
households or by direct cream method at industry level.
[3]
The quality of ghee depends on the type and quality of milk
and method of preparation.
[4,5]
Ghee is fairly shelf-stable
largely because of its low moisture content and possible
anti-oxidative properties. Shelf life of ghee is also dependent
on the method of preparation. The storing quality of desi
ghee is better than that of direct cream or creamery butter
ghee because of the presence of phospholipids.
[6]
The physical and chemical characteristics of the butter
and ghee from cow, goat, and sheep milk are reported.
[1,7]
Ghee contains 99-99.5% fats and less than 1% moisture,
unsaponifiable matter and traces of charred casein,
carotene, and fat-soluble vitamins.
[8]
Fatty acid profi le
determines many of the physical, organoleptic, and
nutritional characteristics of milk and dairy products.
[9]
Milk fat contains over 400 individual fatty acids and their
isomers. Cow milk contains large amounts of saturated
fatty acids (SFA), particularly C14:0 and C16:0, and small
amounts of mono unsaturated fatty acids (MUFA), poly
unsaturated fatty acids (PUFA) and omega-3 fatty acids
with benefi cial effects on human health.
[10]
Background: Ghee (clarifi ed butter) also known as ghrita, has been utilized for thousands of years in Ayurveda. Ghee is
mostly prepared by traditional method in Indian households or by direct cream method at industry level. Ayurvedic classics
mention that ghrita made from cow milk is superior. However, there is no scientifi c comparison available on preparation
methods and essential fatty acids content of ghrita. Objective: To investigate fatty acid composition of ghrita prepared by
traditional/Ayurvedic method and commercial method (direct cream method). Materials and Methods: Fatty Acid Methyl
Esters (FAME) extracted from ghrita samples were analysed on Gas Chromatography (GC) Shimadzu B using capillary
column BPX70 (0.32 mm*60 m, ID of 0.25 mm). The fatty acids in the samples were identifi ed by comparing peaks with
the external standard 68A (Nu-Chek-Prep, Inc.USA). Signifi cant differences between the experimental groups were assessed
by analysis of variance. Results: Distribution of fatty acids was compared in ghrita samples prepared by traditional method
and direct cream method which is commercially used. Saturated fatty acids were predominant in both the groups. Mono
unsaturated fatty acids and poly unsaturated fatty acids were in the range of 17-18% and 3-6% respectively. DHA content
was signifi cantly higher in ghee prepared by traditional method using curd starter fermentation. Conclusion: The fi ndings
suggested that ghrita prepared by traditional ayurvedic methods contains higher amount of DHA; Omega-3 long-chain
polyunsaturated fatty acids, which is a major component of retinal and brain tissues and remains important in prevention
of various diseases.
Key words: Ayurveda, Docosahexaenoic acid, ghrita, Omega-3 fatty acids, traditional method
ABSTRACT
ORIGINAL RESEARCH ARTICLE
EXPERIMENTAL
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Joshi: DHA in ayurvedic ghrita
2 Journal of Ayurveda & Integrative Medicine
Many attempts have been made to alter the fatty acid
composition of milk fat from dairy cows to improve
the long chain poly unsaturated fatty acids (LCPUFA)
content.
[11,12]
The biosynthesis of LCPUFAs such as
docosahexenoic acid (DHA) and arachidonic acid (AA)
takes place by desaturation and elongation of precursors
such as α-linolenic acid (ALA, 18:3n-3) and linoleic
acid (LA, 18:2n-6), respectively. Precursors ALA and LA
supplied with forage undergo bioconversion in rumen and
mammary glands forming DHA and AA.
[9]
Major sources
of DHA include fi sh such as mackerel, salmon, tuna, fi sh
oil, and deep-sea algae.
[13]
Ghee also known as “Ghrita” is extensively used in
Ayurveda practice. Ghee is strongly recommended in daily
diet and Ayurvedic therapeutics such as “Panchakarma” or
“Anupana” (Vehicle) for various medicines. Traditionally
ghee is made from cow milk or any other milk after
churning curdled whole milk, separating the butter
after fermentation, and clarifying it by heating it in pan
on low fl ame. Alternatively, it is prepared by clarifying
cream collected from raw whole milk at industrial scale.
Ayurvedic classics describe eight kinds of ghee from eight
different animal milk; among them ghee made from cow
milk is said to be the superior.
[14]
The present study was
carried out to investigate fatty acid composition of “ghrita”
prepared by traditional household/Ayurvedic method
[15]
and commercial method (direct cream method).
[8]
MATERIALS AND METHODS
Preparation of “Ghrita” by traditional ayurvedic method
Cow milk was boiled and cooled at room temperature.
About 10% curd was added as starter culture, mixed, and
incubated for 8-10 hrs at room temperature. Curd formed
was manually churned until butter fl oated on top of the
buttermilk. Butter was washed 3-4 times using water to
remove residual butter milk and then heated in stainless
steel container till it is clarifi ed. Clear liquid “ghrita” was
then decanted in a glass container.
Preparation of “Ghee” by direct cream method
Cream is separated from whole cow milk and heated in
stainless steel pan. Clear molten “ghrita” is decanted and
stored in glass container.
Analysis of fatty acids
Modifi ed Bligh and Dyer method was used for fatty acid
analysis.
[16]
Briefl y, 0.1 ml of molten “ghrita” was taken
into glass test tube, followed by addition of 10 ml of 0.6
N methanolic HCl. Samples were incubated at 80°C for
2 hrs. Samples were extracted with hexane three times.
Hexane extracts were mixed together and concentrated
under vacuum evaporation. Analysis of fatty acid methyl
esters (FAME) was done on GC Shimadzu B using capillary
column BPX70 (0.32 mm*60 m, ID of 0.25 mm). The fatty
acids in the samples were identifi ed by comparing peaks with
the external standard 68A (Nu-Chek-Prep, Inc. USA).
[17]
Statistical analysis
Data was analyzed using the Statistical Package for the
Social Sciences (SPSS software version: 11, Chicago, IL).
Signifi cant differences between the experimental groups
were assessed by analysis of variance. All data are expressed
as mean ± standard error of mean (SEM); P value less than
0.05 was considered to be signifi cant.
RESULTS
Distribution of fatty acids was compared with ghrita
samples prepared by traditional method and direct cream
method, which is commercially used. Saturated fatty acids
were predominant fatty acids in both the groups. Mono
unsaturated fatty acids (MUFA) and poly unsaturated
fatty acids (PUFA) were in the range of 17-18% and
3-6%, respectively. No signifi cant difference was observed
between the two acids [Table 1].
Distribution of selected n-6 and n-3 fatty acids was
analyzed in ghrita samples prepared by both the methods.
It was observed that there was no signifi cant difference
in precursor fatty acids such as LA and ALA contents in
ghrita samples. DHA content was signifi cantly higher in
ghee prepared by traditional method using curd starter
fermentation [Table 2].
DISCUSSION
DHA contributes to reduced risk of diseases like heart
attack, cancer, insulin resistance, arthritis,
[18]
and ADHD.
[19]
Ghee is generally used in Ayurveda. According to Ayurveda
classics ghrita promotes longevity and protects body
from various diseases.
[20]
It increases digestive fi re agni
and improves absorption and assimilation. It nourishes
dhatus, improves memory, and has lubricating activity in
joints. Most of these activities are known to be imparted
Table 1: Distribution of fatty acids in Ghrita
prepared by traditional method and direct
cream method
Fatty acid %
distribution
Ghrita prepared by
traditional method
Ghrita prepared by
direct cream method
SFA 72.4 73.2
MUFA 18.6 17.5
n-6 5.38 6.3
n-3 3.7 3.1
n-3: n-6 ratio 0.69 0.5
SFA=Saturated fatty acids, MUFA=Mono unsaturated fatty acids
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Joshi: DHA in ayurvedic ghrita
Journal of Ayurveda & Integrative Medicine 3
by anti-oxidants and essential fatty acids such as DHA.
We observed consistent and reproducible increase in
DHA content in ghee prepared by traditional method in
controlled laboratory conditions, however, this needs to
be confi rmed at fi eld level. Whether traditional method
of ghee preparation protects unsaturated fatty acids from
trans fat formation is unknown. Limitation of this study is
that the trans fats and anti-oxidant levels of ghee samples
prepared by both the methods were not measured.
We observed increase in DHA content of ghrita prepared by
traditional fermentation method, which may be attributed
to rich microbial fl ora used in starter curd culture. Lactic
acid bacillus cultures along with added lactose and fructose
are shown to promote formation of conjugated linoleic
acid.
[21]
Further analyses of microbial community and fatty
acid profi le are required to establish presence of desaturase
enzyme producing bacteria responsible for conversion
of precursor fatty acid ALA to DHA. DHA along with
fat-soluble vitamins, anti-oxidants, and conjugated linoleic
acid (CLA) could be responsible for health benefi ts of ghrita.
There are other considerations for the traditional
processing of ghee. For example, mango seed kernels have
been used to increase the shelf life of ghee. It has now
been found that phenolics and phospholipids isolated from
mango seed kernel, when added jointly to buffalo ghee,
prevented peroxidation of fats and helped in extending
the shelf life of ghee.
[22]
Also, Ayurveda mentions about
enhanced therapeutic qualities of very old cow ghee. It
will be interesting to study the composition of very old
cow ghee as well as physicochemical and organoleptic
characteristics at various time intervals. Use of traditional
ayurvedic methods to prepare ghrita needs to be studied
for its possible role in promoting health.
CONCLUSION
The fi ndings suggested that ghrita prepared by traditional
ayurvedic method contains higher amount of DHA with
benefi cial effects on human health.
ACKNOWLEDGEMENT
The author would like to thank Dr. Bhushan Patwardhan for his
encouragement to undertake this work. Thanks are also due to
Ms. Shalini Jaiswal, M. Sc student for her technical help.
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Table 2: Distribution of selected Omega-6
and Omega-3 fatty acids in Ghrita prepared
by traditional method and direct cream
method (n=3)
Fatty acid %
distribution
Ghrita prepared by
traditional method
Ghrita prepared by
direct cream method
LA 5.1±0.544 6.2±1.29
AA 0.157±0.061 0.132±0.037
ALA 3.66±0.88 3.0±0.7
AA 0.218±0.096 0.169±0.95
DHA 0.083*±0.003 0.062±0.002
Values are Mean±SEM, * Value signifi cant with P≤0.05, LA=Linoleic acid,
AA=Arachidonic acid, ALA=α-linolenic acid, DHA=Docosahexaenoic acid
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Joshi: DHA in ayurvedic ghrita
4 Journal of Ayurveda & Integrative Medicine
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How to cite this article: Joshi KS. Docosahexaenoic acid content
is signifi cantly higher in ghrita prepared by traditional Ayurvedic
method. J Ayurveda Integr Med 0;0:0.
Source of Support: Nil, Confl ict of Interest: None declared.
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