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| ORIGINAL ARTICLE |
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| Year : 2022 | Volume
: 10
| Issue : 4 | Page : 221-227 |
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Pharmacognostical and physicochemical profile of Abhayadi Gutika: An polyherbal formulation
Sunny C Patil1, Arjun S Baghel1, Shubhangi B Kamble1, Harisha C Rudrappa2, Vinay J Shukla3, Mehul D Mehta4
1 Department of Basic Principles, Institute of Teaching and Research in Ayurveda (ITRA), Jamnagar, Gujarat, India
2 Pharmacognosy Laboratory, Institute of Teaching and Research in Ayurveda (ITRA), Jamnagar, Gujarat, India
3 Pharmaceutical Chemistry Laboratory, Institute of Teaching and Research in Ayurveda (ITRA), Jamnagar, Gujarat, India
4 QC Department, Pharmacy, Institute of Teaching and Research in Ayurveda (ITRA), Jamnagar, Gujarat, India
| Date of Submission | 20-Aug-2022 |
| Date of Decision | 21-Nov-2022 |
| Date of Acceptance | 01-Dec-2022 |
| Date of Web Publication | 31-Dec-2022 |
Correspondence Address:
Dr. Sunny C Patil
Department of Basic Principles, Institute of Teaching and Research in Ayurveda (ITRA), Jamnagar, Gujarat 361008
India
 Source of Support: None, Conflict of Interest: None
DOI: 10.4103/jism.jism_68_22
Introduction: Quality standardization and maintenance of these standards is the need of the hour in this era which is showing growing demand for traditional medicines. Although Abhayadi Gutika (AG) is an important formulation in Ayurvedic pharmacopeia, it lacks scientific documentation. Aims: The aim was to study organoleptic, microscopic, physicochemical, phytochemical, nutraceutical, and high-performance thin-layer chromatography (HPTLC) profile of AG. Materials and Methods: AG was prepared and subjected to pharmacognostical analysis in which organoleptic and microscopic characteristics were identified. This was followed by analytical study comprising physicochemical, phytochemical, neutraceutical parameters, acid-neutralizing capacity, and HPTLC profile. Results: Pharmacognostical microscopic evaluation revealed the presence of starch grains, acicular crystals, group of fibers, sclereids, stone cells, and epicarp cells. Quantitative physicochemical analysis revealed loss on drying value as 6.37% w/w, ash value as 2.16% w/w, water-soluble extractive value as 44.37% w/w, alcohol-soluble extractive value as 37.91% w/w, and pH value as 6.5. Phytochemical analysis showed the presence of flavonoids, saponins, and tannins in the finished product. In nutraceutical evaluation, functionalities such as total proteins (4.82%), total fats (0.5%), total carbohydrates (70.48%), total sugars (48%), and crude fibers (2.5%) were observed. Acid-neutralizing capacity for AG was found to be 1.3 meq/gm. HPTLC fingerprinting exhibited five peaks (Rf values: 0.18, 0.28, 0.72, 0.81, 0.84) at 254 nm, seven peaks (Rf values: 0.09, 0.22, 0.25, 0.28, 0.53, 0.58, 0.64) at 366 nm and two peaks (Rf values: 0.64, 0.72) at 540 nm on densitogram. Conclusion: Preliminary profiling of AG exhibited striking pharmacognostical as well as analytical characteristics. In nutraceutical analysis, functionalities like proteins, fats, carbohydrates, sugars and crude fibers were observed. Acid-neutralizing capacity which is a measure of efficacy of medicines prescribed for hyperacidity (antacids) was found to be 1.3 meq/gm for AG which supports its therapeutic efficacy in the disease conditions like Vidagdhajirna. Keywords: Abhayadi Gutika, HPTLC, pharmaceutical chemistry, pharmacognosy
How to cite this article:
Patil SC, Baghel AS, Kamble SB, Rudrappa HC, Shukla VJ, Mehta MD. Pharmacognostical and physicochemical profile of Abhayadi Gutika: An polyherbal formulation. J Indian Sys Medicine 2022;10:221-7 |
How to cite this URL:
Patil SC, Baghel AS, Kamble SB, Rudrappa HC, Shukla VJ, Mehta MD. Pharmacognostical and physicochemical profile of Abhayadi Gutika: An polyherbal formulation. J Indian Sys Medicine [serial online] 2022 [cited 2023 Mar 25];10:221-7. Available from: https://www.joinsysmed.com/text.asp?2022/10/4/221/366516 |
| Introduction | |  |
With globalization, many Ayurvedic formulations are being extensively used and promoted worldwide for treatment of various disease conditions. Hence to assure drug quality, standardization of these formulations has become crucial in the current era.[1]Abhayadi Gutika (AG) is one such polyherbal compound formulation containing Haritaki (Terminalia chebula Retz.) and Draksha (Vitis vinifera Linn.) along with Sita (sugar candy) and Madhu (honey). This is the drug of choice indicated for the management of Vidagdhajirna (indigestion induced hyperacidity) in the Ayurvedic text Chakradatta.[2] Although Leha (electuary) form of this formulation is described in classical text, Gutika (chewable tablet) form is generally preferred because of the factors like convenience to carry the drug, feasible timely administration, accurate dose fixation, and palatability. Considering the significant clinical efficacy of this drug, it is essential to maintain the standard quality of this formulation by bringing it in the light of scientific evaluation. Looking into this, it was planned to carry out preliminary scientific evaluation of AG. For this the trial drug was subjected to pharmacognostical, physicochemical and phytochemical evaluations in order to prepare a preliminary profile of this formulation.
| Materials and Methods | | |
Procurement of Raw Materials
The chief ingredient, Haritaki (Terminalia chebula Retz.) was procured from Pharmacy attached to the Institute where the study was carried out. Other ingredients viz. Draksha (Vitis vinifera Linn.) and Sita (~sugar candy) were purchased from a local market and Madhu (~honey) was obtained from the Gujarat State Forest Development Corporation Limited.
Identification and Authentication of Raw Materials
All the herbal ingredients were authenticated at the Pharmacognosy Laboratory of the institute. The API standards were used for pharmacognostical authentication based on the morphological features, organoleptic characters, and powder microscopy of the ingredients.
Preparation of Abhayadi Gutika[
3]
The formulation composition of AG is summarized at [Table 1]. The whole process of formulation preparation was carried out under controlled conditions at the Department of Rasashastra and Bhaishajya Kalpana of the Institute.
All the four ingredients viz. Haritaki, Draksha, Sita, and Madhu were used in equal proportion [Figure 1]A and B. Haritaki and Sita were pulverized separately into fine powders and sieved through mesh 80#. Draksha was pounded and grinded in the mixer grinder. All the three ingredients were mixed well by grinding them into a mixer grinder [[Figure 1]C and D]. Then, honey was added and mixed in this mixture till formation of homogeneous dough of semisolid consistency [Figure 1]E. Gutika (circular shape) of 1 g each were prepared and dried at room temperature around 30 °C [Figure 1]F–H. | Figure 1: Process of preparation of Abhayadi Gutika. A–B. Ingredients of AG- Haritaki, Draksha, Sita and Madhu; C–D. Mixing and grinding of Haritaki, Draksha, and Sita; E. Addition of Honey; F–H. Preparation and storage of Gutika
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Pharmacognostical Evaluation
AG was subjected to pharmacognostical analysis comprising organoleptic characteristics and microscopic studies. A small quantity of AG was dissolved in distilled water and filtered through Whatman filter paper no. 1. The precipitate was treated with and without stain (Phloroglucinol-HCl) to find out the lignified material along with other cellular components. These findings were compared with the characters of individual components of AG. The microphotographs were taken under Carl Zeiss trinocular microscope attached with camera.[4],[5 [Figure 2] | Figure 2: Pharmacognostical microscopic characteristics of Abhayadi Gutika. A. Stone cells, B. Lignified stone cells, C. Starch grains, D. Sclereids, E. Lignified sclereids, F. Epicarp cells of Haritaki, G. Acicular crystals, H. Epicarp cells, I. Mesocarp cells, J. Rosette crystals, K. Fibers, L. Oil globules, M. Purplish coloring matter of Draksha, N. Pollen grain in honey
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Physicochemical],[ Phytochemical],[ and Nutraceutical Evaluation
AG was analyzed through relevant physicochemical parameters like loss on drying],[ ash value],[ water-soluble extractive value],[ alcohol-soluble extractive value],[ and pH value in the Pharmaceutical Chemistry Laboratory of the Institute.[6],[7],[8] Phytochemical (qualitative) and nutraceutical (qualitative and quantitative) evaluations of the test drug were also carried out.[9] In phytochemical qualitative analysis, presence of saponins, flavonoids, tannins, and alkaloids was assessed by using appropriate qualitative testing methods like Mayer’s test, Wagner’s test, Ninhydrin test, Molish’s test, and Benedict’s test.[10] Acid-neutralizing capacity for AA was assessed by using preliminary antacid test (PAT).[11]
High-Performance Thin-Layer Chromatography
High-performance thin-layer chromatography (HPTLC) of AG was carried out with the aim to obtain unique quantitative results for its identification in terms of reference values.[12] 2.5 g sample of drug was taken in a beaker and 50 mL of Hexane was added to it. It was refluxed for 30 min. After cooling, the solution was filtered with Whatman’s filter paper. 50 mL chloroform was added to the filtrate and refluxed for 30 min. The solution was allowed to cool and chloroform was discarded. 50 mL of methanol was then added and refluxed for 30 min. After cooling, the solution was filtered and methanol was evaporated up to10 mL. The obtained methanol was filtered with a 0.45-micron membrane filter. The solution thus obtained was used as test solution for HPTLC fingerprinting. HPTLC fingerprinting was carried out with CAMAG Linomat 5-applicator and stationary phase used was MERCK-HPTLC Silica gel 60 F254 on aluminum sheets. Sample application volume was 2.0 µL and application (Y axis) start position was set at 10 mm while development end position was set at 80 mm from plate base. Chamber saturation time was set at 30 min. Mobile phase used for the study was Toluene: Ethyl acetate: Glacial acetic acid (7:2:1v/v). HPTLC plate was preheated at 100 ± 5 °C for 3 min. Visualization was made at 254 nm, at 366 nm, and at 540 nm after derivatization.
| Results and Discussion | | |
Pharmacognostical Analysis
Detailed pharmacognostical evaluation was carried out in order to obtain information about biochemical and physical properties of the test drug AG. Results of organoleptic characteristics of AG are depicted at [Table 2]. Powder microscopy of the finished product revealed striking characters of all its individual components. The study revealed features such as stone cells [Figure 2A], lignified stone cells [Figure 2B], starch grains [Figure 2C], sclereids [Figure 2D], lignified sclereids [Figure 2E], and epicarp cells [Figure 2F] of Haritaki. Acicular crystals [Figure 2G], epicarp cells [Figure 2H], mesocarp cells [Figure 2I], rosette crystals [Figure 2J], fibers [Figure 2K], oil globules [Figure 2L], and purplish coloring matter [Figure 2M] of Draksha were also observed along with pollen grain in honey [Figure 2N]. The vital characteristics revealed in the microscopic study of all the ingredients were matching with their mentioned standards in API.[13],[14],[15],[16] This confirms the presence of all ingredients in the finished product.
Physicochemical Analysis
AG was subjected to pharmaceutical analysis using relevant physicochemical parameters. The analysis was carried out with the aim to gain information about the qualitative and quantitative composition of the test drug. The observations obtained are presented in [Table 3]. Ash value of the test drug was found to be 2.16% w/w. Ash value determination is one of the salient parameters for standardization of herbal drugs. High ash value is indicative of contamination, substitution, and adulteration in preparing the drug or drug formulation provided there are no mineral ingredients in it. Ash value was found low indicating no contamination in test drug. All the physicochemical parameters of AG like loss on drying, ash value, water-soluble and alcohol-soluble extractive values, and pH can be used as standard, as this formulation has not been mentioned in Ayurvedic Pharmacopoeia of India.
Phytochemical Analysis
In phytochemical qualitative analysis, the presence of saponins, flavonoids, tannins, and alkaloids was assessed using appropriate qualitative testing methods. The results are summarized in [Table 4]. The test drug was found to contain saponins, flavonoids, and tannins while the presence of alkaloids could not be established in it. Qualitative data regarding these phytoconstituents in AG will certainly help in determining bioactive agents present in the drug as well as further standardization and quality control of this drug.
Nutraceutical Evaluation
The test drug was screened for qualitative and quantitative analysis of some nutraceutical parameters such as total proteins, total sugars, total fat, total crude fibers, total carbohydrates, and acid-neutralizing capacity (ANC). The results of nutraceutical analysis are presented in [Table 5] which illustrated the nutraceutical functionalities present in the drug. Presence of carbohydrates, fats, proteins, sugars, and crude fibers was established in the formulation among which total carbohydrates were found in maximum proportion, that is, 70.48% which included a total sugar content of 48%. | Table 5: Proximate nutraceutical composition and acid-neutralizing capacity of Abhayadi Gutika
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Acid-Neutralizing Capacity
In the modern medical science, the efficacy of medicines prescribed for hyperacidity (antacids) is assessed on various parameters such as PATs, ANC test, and buffering pH profile test. ANC is a measure for the overall buffering capacity against acidification. Neutralizing capacity of the antacids is expressed as meq (milliequivalents) per minimum dose of tablets, tea-spoons, or capsules. In a study, it was found that ANC for various brands of antacids was ranging from 0.4 to 21.7 meq/gm.[17],[18] In the present study, ANC for AG was found to be 1.3 meq/gm. [Table 5]
High-Performance Thin-Layer Chromatography
In HPTLC fingerprinting, when the drug was analyzed under densitometer at 254 nm, the chromatogram showed five peaks at Rf values 0.18, 0.28, 0.72, 0.81, and 0.84. It showed seven peaks at 366 nm with Rf values 0.09, 0.22, 0.25, 0.28, 0.53, 0.58, and 0.64 while densitometer at 540 nm displayed two peaks of Rf values 0.64 and 0.72. [Figure 3][Figure 4][Figure 5] This data of HPTLC fingerprinting of AG is very crucial as HPTLC analysis is one of the most potent techniques for quality control of herbal medicines. This will serve as a standard for identification, authentication, and quality control of this formulation of AG. | Figure 3: High-performance thin-layer chromatography of methanolic extract of Abhayadi Gutika at 254 nm
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 | Figure 4: High-performance thin-layer chromatography of methanolic extract of Abhayadi Gutika at 366 nm
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 | Figure 5: High-performance thin-layer chromatography of methanolic extract of Abhayadi Gutika at 540 nm
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Standardization of herbal formulations is essential in the present era of modernization. Ayurveda is a pioneer pathy among the traditional systems of medicine practiced worldwide. Numerous single and compound formulations have been described in ancient Ayurveda texts. Research on scientific profiles of these formulations is the need of the time for their better acceptance on global platform.
In the present study AG was prepared following pharmaceutical procedure mentioned in classical Ayurveda text Sharangadhara Samhita. Microscopical evaluation of the final product is very important in the initial identification of ingredients as well as in the detection of adulterations in the product. Identification of original ingredients is the first step to maintain the quality of the final product. In the present study all the ingredients of AG were authenticated with the help of characters mentioned in the API. Powder microscopy of AG showed striking characters of all individual drugs of AG. It showed the presence of starch grains, acicular crystals, group of fibers, sclereids, stone cells, and epicarp cells. This confirms the ingredients present in the finished product and there is no major change in the microscopic structure of the raw drugs during the pharmaceutical processes of preparation of the drug.
In physico-phytochemical analysis crucial data related to analytical and phytological parameters of AG was obtained which will be useful in future studies carried on the drug. Quantitative physicochemical analysis showed loss on drying value as 6.37% w/w, ash value as 2.16% w/w, water-soluble extractive value as 44.37% w/w, alcohol-soluble extractive value as 37.91% w/w, and pH value as 6.5. Among these, very low ash value found in AG indicates negligible contamination in the test drug as high ash value is indicative of contamination, substitution, adulteration in preparing the drug or drug formulation provided there are no mineral ingredients in it.
Phytochemical analysis revealed the presence of flavonoids, saponins, and tannins in the finished product. Nutraceutical profiling along with estimation of ANC and HPTLC study of AG were also carried out which provided the exact information that will be beneficial in future studies on AG. In nutraceutical evaluation, functionalities such as total proteins (4.82%), total fats (0.5%), total carbohydrates (70.48%), total sugars (48%), and crude fibers (2.5%) were observed. ANC for AG was found to be 1.3 meq/gm. HPTLC fingerprinting exhibited five peaks (Rf values: 0.18, 0.28, 0.72, 0.81, 0.84) at 254 nm, seven peaks (Rf values: 0.09, 0.22, 0.25, 0.28, 0.53, 0.58, 0.64) at 366 nm, and two peaks (Rf values: 0.64, 0.72) at 540 nm on densitogram.
Documentation, substantiation, and maintenance of quality standards of herbal formulations are the crucial factors in this era of globalization of Ayurveda. The present study may prove to be a significant preliminary step towards establishing pharmacognostical and physicochemical standards of AG, an important formulation in the Ayurvedic pharmacopoeia.
| Conclusion | | |
Preliminary profiling of AG exhibited striking pharmacognostical as well as analytical characteristics. In nutraceutical analysis, functionalities such as proteins, fats, carbohydrates, sugars and crude fibers were observed. ANC which is a measure of efficacy of medicines prescribed for hyperacidity (antacids) was found to be 1.3 meq/gm for AG which supports its therapeutic efficacy in the disease conditions like Vidagdhajirna. As no published data is available on pharmacognostical, analytical and nutraceutical evaluation of AG, this preliminary profile can be used as reference in future studies.
Acknowledgements
Authors express their sincere gratitude to Prof. A. B. Thakar, Hon’ble Director, ITRA; Prof. H. A. Vyas, Dean, ITRA; and Prof. B. J. Patgiri, HOD, Dept. of RSBK, ITRA for providing the facilities, kind support, and encouragement during the work. Authors are also thankful to Dr. Ushnesh Bhat, Pharmaceutical Chemistry Laboratory, ITRA for valuable technical inputs during this work.
Financial Support and Sponsorship
Institute of Teaching and Research in Ayurved (ITRA) provided the financial and administrative support for the conduction of research project.
Conflicts of Interest
There are no conflicts of interest.
| References | | |
| 1. | WHO. Guidelines on safety monitoring of herbal medicines in pharmacovigilance systems, Part I, Chapter 3.1. Geneva: World Health Organization; 2004. p. 13-4.  |
| 2. | Sharma PV, editor. Chakradatta of Chakrapanidatta, Chapter 6, Verse 74. Reprint ed. Jaipur: Swami Jayaramdas Ramprakash Trust; 1993. p. 210. |
| 3. | Vidyasagar P, editor. Sharangadhara Samhita of Sharangadhara, Madhyama Khanda. Chapter 8, Verse 1–2. Reprint ed. Varanasi: Chaukhambha Surbharati Prakashan; 2018. p. 210-11. |
| 4. | Khandelwal KR Examination of powdered drugs. In: Khandelwal KR, editor. Practical Pharmacognosy Techniques and Experiments. 19th ed. Pune; Nirali Prakashan; 2008. p. 162-6. |
| 5. | Anonymous. The Ayurvedic Pharmacopoeia of India. Part II, Vol. I, 1st ed. New Delhi: Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India; 2001. p. 71. |
| 6. | Anonymous. The Ayurvedic Pharmacopoeia of India. Part II, Vol. I, 1st ed. New Delhi: Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India; 2001. p. 83. |
| 7. | WHO. Quality Control Methods for Medicinal Plant Materials. Geneva: World Health Organisation; 1998. p. 32. |
| 8. | CCRAS, Parameters for Qualitative Assessment of Ayurveda and Siddha Drugs, Part A. New Delhi: CCRAS; 2005. p. 31. |
| 9. | CCRAS. Basics of Analytical and Phytochemical Analysis of Medicinal Plants, Analytical Methods for ASU Single/Compound Drugs Sponsored by WHO, CSMDRI for Ayurveda and Siddha. Chennai: CCRAS, Govt. of India; 2008. p. 11. |
| 10. | Sahira Banu K, Cathrine L General Techniques involved in phytochemical analysis. Int J Ad Res in Chem Sciences 2015;2:25-32. |
| 11. | Yafout M, Elhorr H, Otmani ISE, Khayati Y Evaluation of the acid-neutralizing capacity and other properties of antacids marketed in Morocco. Med Pharm Rep 2022;95:80-7. |
| 12. | Stahl E Thin Layer Chromatography a Laboratory Hand Book. Berlin: Springer-Verlag; 1969. p. 125-41. |
| 13. | Anonymous. The Ayurvedic Pharmacopoeia of India. Part I, Vol. I. 1st ed. New Delhi: Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India; 2001. p. 80. |
| 14. | Anonymous. The Ayurvedic Pharmacopoeia of India. Part I, Vol. III. 1st ed. New Delhi: Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India; 2001. p. 75. |
| 15. | Anonymous. The Ayurvedic Pharmacopoeia of India. Part I, Vol. VI, 1st ed. New Delhi: Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India; 2001. p. 215. |
| 16. | Anonymous. The Ayurvedic Pharmacopoeia of India. Part I, Vol. VI, 1st ed. New Delhi: Department of AYUSH, Ministry of Health and Family Welfare, Govt. of India; 2001. p. 218. |
| 17. | Lin MS, Sun P, Yu HY Evaluation of buffering capacity and acid neutralizing-ph time profile of antacids. J Formos Med Assoc 1998;97:704-10. |
| 18. | Hankin L, Hanna JG Neutralizing capacity of antacid products. In Bulletin 815, The Connecticut Agricultural Experiment Station. New Haven: Connecticut Department of Consumer Protection; 1983. p. 11-13. |
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]
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