Preparation and Evaluation of Classical Ayurvedic Formulation Triphala gritha

Introduction

Our ancient traditional system of Medicine, Ayurveda says “Sarvendriyanam Nayanam Pradhanam”. It means, ‘amongst all the organs of the body, eyes are considered as one of the most important sense organs’. Hence it included a detailed description about prevention of eye diseases, also explained many vision care techniques and procedures. Presently, stressful working environment, sedentary life style, irregular sleeping habits, unhealthy eating habits and extensive use of electronic gadgets, contributes significantly to the development of metabolic disorders like diabetes, hypertension, etc. and these metabolic disorders can adversely affect the eyes and other sense organs.¹

Triphala ghrita is one of the formulations extensively used by Ayurvedic practitioners since ancient times. Charaka mentioned that it is a rasayana for the eyes, and can be used to prevent eye diseases. According to him, Triphala ghrita helps in improvising agni, and thus enhances the absorption of nutrients from the digestive system, which is supported by various studies.^2-4

Triphala ghrita contains Triphala, the three myrobolons (Emblica officinalis, Terminalia chebula and Terminalia belerica) and ghrita (cow ghee) having a variety of classical benefits in Timira (cataract), Visarpa (erysipelas), Netraruja (pain in eyes), Netrasrava (lacrimation), Kandu (itching), Svayathu (oedaema), Visamajvara (intermittent fever), Suklanetraroga (eye disorders related to sclera), Vartmaroga (disorders of eyelids). According to Sharangadhara Samhita, Triphala is beneficial in Netraroga.⁵

Following are the famous yogas containing Triphala as the main ingredient, which are utilized in day-to-day ophthalmic practice:

1. Triphala churna,

2. Sadangakwathaguggulu,

3. Triphaladikwatha,

4. Triphaladianjana,

5. Tripahalavarti,

6. Triphaladirasakriya,

7. Nagarjunavarti,

8. Kokilavarti,

9. Drustipradavarti,

10. Candhanadivarti,

11. Lohadiguggulu,

12. Timiraharalauha,

13. Saptamrutaloha,

14. Triphaladighrita,

15. Mahatriphaladighrita

The aim of the present study was to explore Triphala ghrita formulations appearing in Ayurvedic traditional medicine for establishing the scientific data pertaining to quality assessment.

Materials and Methods

Materials

Procurement of selected formulations and crude drugs

The selected formulations, Triphala ghrita, the crude drugs required for the preparation of laboratory formulation were procured from Soham Ayurved Rasashala, Solapur, Maharashtra.

Reagents and chemicals

Chemicals, solvents and reagents used for the study were of analytical grade.

Methods

In order to develop the methods to assess the quality control parameters of the selected formulations, selected formulations were subjected to determination of physico-chemical constants using reported methods as per WHO guidelines (WHO, 1998) and Indian Pharmacopoeia, 1996.^6,7

The selected formulation Triphala ghrita was prepared as per the methodology described in Ayurvedic texts.

Triphala ghrita

Triphala ghrita mentioned by Sharangdhara is a polyherbal formulation of Ayurveda representing the group of Snehakalpa which is used in the Kriyakalpa procedure called Tarpana, a local eye refreshing and nourishing therapy. This is a significant formulation mentioned in Sharangdhara for the treatment of various eye disorders. Snehakalpana (medicated ghrita) is a unique oleaginous dosage form used for both topical and systemic administration.⁸

Method of preparation of Triphala ghrita

The laboratory formulation of Triphala ghrita was prepared as per the Sharangdhar Samhita.⁵

Ingredients of Triphala ghrita

The ingredients purchased for Triphala ghrita were fruits of Haritaki (Terminalia cheubula), fruits of Bhibitak (Terminalia belerica), fruits of Amalaki (Embelica officinalis) (Table 1).

Preparation of Triphala ghrita laboratory formulation (TGLF)

All the above ingredients were taken into a utensil along with stirrer and kept on a gas stove. The above mixture was heated till ‘Snehasiddhi Lakshana’ appeared in mixture or 320 mL of ghrita remained in utensil, and this was mixed thoroughly to obtain homogeneous blend. Final product was stored in an airtight container.

Standardization of Triphala ghrita laboratory (TGLF) and marketed formulation (TGMF)

The laboratory and marketed formulations of Triphala ghrita were standardized as per WHO guidelines under the various headings using standard protocol obtained from Ayurvedic Pharamacopoeia of India, Herbal Pharmacopoeia.⁶

Determination of Phenolic Content (AP part-I, 2001; API part-I, 1999; AFI part-I, 2003)

The total phenolic content of the formulations was determined by the Folin-Ciocalteau method with some modifications. Five grams of TGLF and TGMF were mixed in 50 mL water and then filtered with Whatman No.1 paper. 0.5 mL of the filtrate was added to 2.5 mL of 0.2 N Folin Ciocalteau reagent and placed for five minutes. Two mL of 75 g/L of Na₂CO₃ was then added and the total volume was made up to 25 mL using distilled water. The above solution was then kept for incubation at room temperature for two hours. Absorbance was measured at 760 nm using 1 cm cuvette in a Perkin Elmer UV-VIS lambda 25 spectrophotometer. A standard calibration curve was generated using tannic acid concentrations ranging from 0 to 800 mg/L. The total phenolic content was expressed as mg of tannic acid equivalents (TAE) per gram of extract.

Considering wide therapeutic applications of Triphala ghrita and presence of the marker constituents, to ensure identity and quality of Triphala ghrita, a simple, sensitive, specific and reproducible High performance thin-layer chromatography (HPTLC) method was developed for the quantification of markers in the formulation.

Results

Standardization of finished product

The prepared sample was standardized using various parameters, including organoleptic, physicochemical, qualitative, and quantitative analyses. The data were also compared with commercially available product. The organoleptic characteristics of both prepared and the commercially available product of Triphala ghrita were analyzed on the basis of observational criteria (appearance, color, taste, and odor). The physicochemical parameters such as loss on drying, refractive index, viscosity, iodine value, saponification value, acid value and peroxide value were determined as per the guidelines mentioned in Ayurvedic Pharmacopeia of India (Table 2).⁶ The phytochemical screening was performed using different qualitative assay methods described previously. Quantitative estimation of total polyphenolic content was performed using spectroscopic method and was expressed in equivalent of Gallic acid (Table 3). Assessment of heavy metal contamination, microbial contaminants, pesticides, and aflatoxins are mentioned in Tables 4, 5 and 6.

Discussion

Some important organoleptic features of Triphala ghrita laboratory and market formulations like color, odour, taste, appearance, touch, etc were documented.

Determination of physico-chemical constants indicated a loss on drying of 0.15% and 0.18%, refractive index of 1.4536 and 1.4534, Iodine value of 36.7 and 35.2, saponification value of 201.9 and 210.3, acid value of 01.34 and 02.13, peroxide value (meq/kg) of 00.23 and 00.25, Phenolic content (9 mg/100g) of 08.04 and 05.27, respectively for laboratory and marketed formulations. The Triphala ghrita laboratory formulation and marketed formulation were tested for the presence of heavy metals, microbial content, pesticide residues and aflatoxin which were also found within the limits of WHO specifications.

Preliminary phytochemical studies of Triphala ghrita revealed the presence of terpenoids and phenolics as chief constituents.

Amla is one of the most extensively studied plants. Reports suggest that it contains tannins, alkaloids and phenols.⁹ Fruits have 28% of the total tannins distributed in the whole plant. The fruit contains two hydrolysable tannins, Emblicanin A and B, both of which possess antioxidant properties. Upon hydrolysis, Emblicanin A yields gallic acid, ellagic acid, and glucose, while Emblicanin B produces ellagic acid and glucose.¹⁰ The fruit also contains Phyllemblin.¹¹ Activity directed fractionation revealed the presence of several phytochemicals like gallic acid, corilagin, furosin and geraniin.¹² Flavonoids like quercetin, alkaloids like phyllantine and phyllantidine were found. Its fruit juice contains the highest concentration of vitamin C (478.56 mg/100 mL). Vitamin C levels are more than those in oranges, tangerines and lemons.^13,14

The phytochemical studies showed presence of phenolics and terpenoidal moieties in the Triphala ghrita while performing co-TLC studies with some of similar compounds  available. The Rf of one of the components corresponded with that of Ascorbic acid and Gallic acid, thus revealing their presence. Therefore, simple, sensitive, specific and reproducible HPTLC method was developed and reported for the quantification of ascorbic acid and gallic acid as marker constituents to ensure identity and quality of Triphala ghrita.¹⁵

Conclusion

Thus, from the present study, it can be concluded that the pharmacognostical studies carried out on Triphala ghrita can serve as a valuable tool and provide suitable standards for the identification of Ayurvedic formulations. Phytochemical screening and the quantitative HPTLC assay method developed for quantification of ascorbic acid and gallic acid in Triphala ghrita can serve for the routine quality control of commercial samples and help in justifying the traditional claims endowed upon the formulations in a scientifically acceptable manner.

Conflict of Interest

Nil