![RJPS Journal Cover Page](../../../../files/rjps/cover/cover5.png)
RJPS Vol No: 14 Issue No: 3 eISSN: pISSN:2249-2208
Dear Authors,
We invite you to watch this comprehensive video guide on the process of submitting your article online. This video will provide you with step-by-step instructions to ensure a smooth and successful submission.
Thank you for your attention and cooperation.
1Dr. J J Magdum Pharmacy College, Kolhapur, Maharashtra, India
2Dr. Krishna Murthy, Professor & Head, Department of Pharmacognosy, Al Ameen College of Pharmacy, Bengaluru, Karnataka, India.
3Department of Pharmacognosy, Al Ameen College of Pharmacy, Bengaluru, Karnataka, India
*Corresponding Author:
Dr. Krishna Murthy, Professor & Head, Department of Pharmacognosy, Al Ameen College of Pharmacy, Bengaluru, Karnataka, India., Email: drkm289@gmail.com![CC BY NC 4.0 ICON](https://rjps.journalgrid.com/images/cc-by-nc.png)
Abstract
Objective: The objective of this study was to prepare and evaluate Triphala gritha.
Methods: The selected formulation Triphala ghrita was prepared as per the methodology described in Ayurvedic texts. The prepared and marketed formulations were analyzed for their physico-chemical constants using established methods in accordance with WHO guidelines, the Indian Pharmacopoeia, and standard protocols from the Ayurvedic Pharmacopoeia of India, Herbal Pharmacopoeia, and other relevant sources.
Results: A profile consisting of distinguishing parameters was developed to provide means of correct identification. Some important organoleptic features of Triphala ghrita laboratory and market formulations like color, odour, taste, appearance, touch, etc. were documented. Physico-chemical constants were determined and documented. The formulations 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. High performance thin-layer chromatography (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.
Conclusion: The studies carried out on Triphala ghrita can serve as a valuable tool and provide suitable standards for its identification. 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 this formulation in a scientifically accepted manner.
Keywords
Downloads
-
1FullTextPDF
Article
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
Supporting File
References
- Rani S, Siddapur AL. Vision care through Ayurveda. International Ayurvedic Medical Journal 2018:6(1):150-155.
- Mishra V, Agrawal M, Onasanwo SA, et al. Antisecretory and cyto-protective effects of chebulinic acid isolated from the fruits of Terminalia chebula on gastric ulcers. Phytomedicine 2013;20(6): 506-11.
- Baliga MS, Meera S, Mathai B, et al. Scientific validation of the ethnomedicinal properties of the Ayurvedic drug Triphala: a review. Chin J Integr Med 2012;18:946-954.
- Gautam MK, Goel S, Ghatule RR, et al. Curative effect of Terminalia chebula extract on acetic acid induced experimental colitis: role of antioxidants, free radicals and acute inflammatory marker. Inflammopharmacology 2013;21:377-83.
- Pandit Parashurama Shastri. Sharangadhara Samhita. Uttarakhanda 13/2. Varanasi: Chauk-hambha Orientalia Publications; 2013. p. 379.
- AYUSH, Government of India. Ayurvedic Pharmacopoeia of India, Part-I, Vol II. New Delhi: Govt. of India, Ministry of Health and Family Welfare; 1999. p. 85-86, 116-117, 207.
- World Health Organization. Quality control methods for herbal materials. Geneva: World Health Organization; 2011. p. 51-83.
- Pandit Parashurama Shastri. Sharangadhara Samhita. Uttarakhanda 13/52-55. Varanasi: Chauk-hambha Orientalia Publications; 2013. p. 387.
- Zhang LZ, Zhao WH, Guo YJ, et al. Studies on chemical constituents in fruits of Tibetan medicine Phyllanthus emblica. Zhongguo Zhong Yao Za Zhi 2003;28(10):940-3.
- Bhattacharya SK, Bhattacharya D, Sairam K, et al. Effect of bioactive tannoid principles of Emblica officinalis on ischemia-reperfusion induced oxidative stress in rat heart. Phytomedicine 2002;9(2):171-4.
- Yi-Fei W, Ya-Fenga W, Xiao-Yana W, et al. Phyllaemblicin B inhibits Coxsackie virus B3 induced apoptosis and myocarditis. Antiviral Res 2009;84(2):150-58.
- Habib-ur-Rehman, Yasin KA, Choudhary MA, et al. Studies on the chemical constituents of Phyllanthus emblica. Nat Prod Res 2007;21(9):775-81.
- Jain SK, Khurdiya DS. Vitamin C enrichment of fruit juice based ready-to-serve beverages through blending of Indian gooseberry (Emblica officinalis Gaertn.) juice. Plant Foods Hum Nutr 2004;59: 63-6.
- Scartezzini P, Antognoni F, Raggi MA, et al. Vitamin C content and antioxidant activity of the fruit and of the Ayurvedic preparation of Emblica officinalis Gaertn. J Ethnopharmacol 2006;104(1-2):113-8.
- Kilaje SV, Shirsat MK. Simultaneous estimation of ascorbic acid and gallic acid in Triphala ghrita formulation by HPTLC. Nat Volatiles Essent Oils 2021;8(6):3240-9.