Analytical Standards of Vrukshamla (Garcinia indica Choisy) Beeja Taila - Kokum Butter

Mohammed, Joshi, Bairy, and Koppala Narayana: Analytical Standards of Vrukshamla (Garcinia indica Choisy) Beeja Taila - Kokum Butter

Authors

Garcinia indica Choisy (Figure 1), syn. G. purpurea Roxb. And Brindonia indica Dupetit-Th. (Family Clusiaceae) is the most commonly accepted source of Vrukshamla, one of the commonest discussed drugs in the classics of Ayurveda. The species was firstly described by Dr. Garcia in 1574, hence the genus name is Garcinia and as it’s endemic to India, the species name is indica. The word purpurea is originated from a latin word Purpura, means purple colour; as the colour of its ripe fruit.[1-3] It’s a small, slender, fruit-bearing tree, having very strong and sturdy growth; leaves are approximately 10 cm × 5 cm in dimensions; having red-tinge, when young, which turns in dark green on maturation; oblong, ovate or elliptic-lanceolate, rarely obtuse; glabrous, membranous, mucronate; with rounded, acute or abruptly acuminate apex and narrowed it base. Petiole is up to 0.7 cm long. Flowers are polygamous. Male flowers are 3 to 8 in number, axillary and/or terminal fascicles, solitary or in a spreading fascicles; with 4-sepals, decussate, outer ones smaller than inner sepals; whereas female flowers are sessile/sub-sessile, solitary or 2 to 3 together, terminal, shortly peduncled.[4,5] Flowering season is November to February. Fruits, commonly known as kokum fruits, are globose and resemble a small apple in size, not furrowed/grooved.[4]

Figure 1

Garcinia indica Choicy

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It’s green, when unripe and turns dark red/purple on maturation, which usually happens by April/May in each year. Fruit-pulp is dark in colour and little-sweet in taste. Each fruit contains 5 to 8 big, flat seeds embedded in juicy pulp.[1] The seed coat is fibrous from outside; each seed is dicotyledonous, reniform-shaped and has a dimension of about 1.875 cm × 1 cm, which contains 23% to 26% (c. 44% on the weight of kernels) oil.[6] Extracted seed-oil, known as kokum butter, remains solid at room temperature.[1] It’s sold in market in form of oval-shaped moulds/lumps or cubical cakes. Its colour is grayish/yellowish white and granular, greasy consistency.[6] There are two methods of extraction of kokum butter from the seeds, viz. the traditional boiling method and the churning method.[7] Kokum butter is a specific remedy in dysentery and mucous diarrhea, administered in doses of 1 tola in a quarter seer of milk thrice a day until complete recovery; it’s useful in Phthisis pulmonalis and some scorbutic (skin) diseases. It has been recommended as a substitute for cod-liver oil, and is eaten by poor people as a substitute for ghee. Externally, oil has a healing property and might be usefully employed as an application to ulcerations, fissures of lips, hands, chapped skin, etc.[8]

According to classical Ayurvedic texts, its unripe fruit is Amla (sour), whereas ripe fruit is Katu (hot and spicy), Kashaya (Pungent)/Amla (Sour), Alpa madhura (Little sweet).[3] It’s Guru, Ruksha and Sangrahee.[3] It’s Veerya is Ushna and Vipaka is Amla. Its bark and oil is Vranaropaka (accelerates the wound healing) and Stambhanakara.[10] Its fruit rind possesses weight reduction properties due to 3-hydroxy citric acid.[9] It’s also semen coagulant and CNS depressant.[9]

In this study the kokum butter was prepared by traditional boiling method and analyzed for standard oil parameters.

Plant Material

Seeds of Vrukshamla (Garcinia indica Choisy) were collected from Udupi district and surrounding forest area of South Kanara.

Preparation of VBT

Traditional boiling-method was followed in this study; for which the collected seeds were dried completely under sunlight. Thereafter, the kernels were obtained by removing the seed-coat manually. Kernels were further subjected to trituration with sufficient quantity of water, for the preparation of paste. That was mixed with 3 litres of water (per kg of seed-kernel) in a vessel and subjected to heat on mild flame for very long time. After some time, a layer of oil started floating on the water, which was skimmed off and taken into another vessel. This second vessel was also simultaneously subjected to the heat on the other stove, until the remnant of water evaporates and the oil remains alone. This oil was allowed to cool down at room temperature and solid Kokum butter was obtained. Heating the first vessel was continued until no or negligible quantity of oil floated. Obtained quantity of Kokum butter was 40 gm per kg of seed-kernel.

Analytical Study

Following physic-chemical parameters of VBT were studied, as per the testing protocol of ASU medicines. Specific gravity, Rancidity test (Kreis Test), Refractive index, Determination of Acid value, Determination of Saponification value, Determination of unsaponifiable matter, Iodine value, Ester value, Colour, Odour, Viscosity, Determination of peroxide value and Chromatographic analysis (HPTLC).[11]

The specific gravity of VBT was found to be 0.9478. It was not oxidized when rancidity rest (Kreis Test) was performed. Its refractive index was found to be 1.46944. Its determined Acid value and Saponification value were 75.69 and 165.49 respectively; whereas, the unsaponifiable matter was found to be 0.59%. Its calculated Iodine value and Ester value were 24.40 and 89.80 respectively. Colour was Yellowish/off-white and the odour was characteristic. The viscosity test could not be performed due to the very thick consistency of the sample, as it could not pass through the tube of viscometer. The calculated peroxide value of VBT was found to be 0.85 (Table 1).

TLC photo documentation revealed presence of many phyto-constituents with different Rf values and HPTLC densitometric scan of the plates showed numerous bands under 254 nm, 366 nm and 620 nm (after derivatisation). On Photo documentation, there were 3 spots under 254 nm, 4 spots under 366 nm and 12 spots under 620 nm post-derivatisation with vanillin sulphuric acid spray reagent. Densitometric scan at 254 nm revealed 4 peaks corresponding to 4 different compounds in the chloroform extract, compounds with Rf 0.02 (24.43%), 0.42 (4.29%), 0.76 (33.63%) and 0.93 (37.64%) were 4 peaks. The densitometric scan at 366 nm showed 2 peaks; the first with Rf 0.02 (77.08%) and the second with Rf 0.76 (22.92%). Then, at 620 nm following derivatisation, the densitometric scan showed 10 peaks corresponding to 10 different compounds, compounds with Rf 0.28 (28.50%), 0.13 (24.40%), 0.44 (10.45%), 0.94 (6.61%) and 0.03 (5.89%) were the major peaks (Table 2 and Figure 2).

Table 1.

Standard oil parameter results of Vrukshamla beeja taila (Kokum butter)

ParameterResult
Specific Gravity0.9478
RancidityNot oxidized
Refractive Index1.46944
Acid value75.69
Saponification value165.49
Unsaponifiable matter0.59 %
Iodine value24.40
Ester value89.80
ColourOff-white/Yellowish white
OdourCharacteristic odour
Peroxide value0.85
Figure 2.

HPTLC of chloroform extract of Vrukshamla beeja taila (Kokum butter)

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Solvent system: Toluene: Ethyl acetate (8:0.5)

Table 2.

Rf values of Chloroform extract of Vrukshamla beeja taila (Kokum butter)

At 254 nmAt 366 nmAfter derivatisation
--0.07 (Dark purple)
--0.10 (Light purple)
--0.15 (Light purple)
--0.18 (Light purple)
--0.25 (Dark purple)
-0.30 (Fluorescence-aqua blue)-
--0.34 (Light purple)
0.36 (Light green)--
--0.39 (Dark purple)
-0.50 (Fluorescence blue)-
--0.52 (Light purple)
0.65 (Dark green)0.65 (Fluorescence-Dark blue)0.65 (Light purple)
--0.68 (Light purple)
--0.72 (Light purple)
0.82 (Dark green)-0.82 (Light purple)
-0.91 (Fluorescence-Dark blue)-

This documented analytical study of VBT can be used to judge the adulteration and purity of it, as it was performed by standard oil parameters. HPTLC fingerprinting will help to supplement information in regard to its identification and standardization.

ACKNOWLEDGEMENTS

The authors are thankful to SDM Centre for Research in Ayurveda and Allied Sciences, Udupi for providing essential facility and support for the study.

Notes

[1] Conflicts of interest SOURCE OF SUPPORT

Nil

[2] Conflicts of interest CONFLICT OF INTEREST

None declared

Biography

Contributors Dr Sreekant contributed to design, literature study and data acquisition and analysis. Dr Faisal and Dr. TS Bairy contributed to the conceptualization of the topic, manuscript review and analysis. Dr. Sunil guided in analysis and manuscript preparation.

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