INTRODUCTION:

The contemporary era witnesses an escalating demand for plant-derived commodities among consumers, spanning medicinal, nutraceutical, and cosmetic domains.¹ Consequently, ensuring the quality of these products necessitates the development of robust quality control methodologies that encompass both traditional and modern analytical techniques.² Standardization emerges as a pivotal metric for upholding the quality assurance of herbal drugs. Such standardization protocols must encompass a comprehensive array of evaluations, including organoleptic assessment, pharmacognostic analysis, volatile matter examination, quantitative estimations (such as ash and extractive values), phytochemical profiling, xenobiotic screening,

microbial load assessment, toxicity analysis, and evaluation of biological activity. Among these, the determination of phytochemical profiles holds paramount significance, given its direct influence on the efficacy of herbal drugs. Fingerprint profiling techniques serve as invaluable tools for discerning the phytochemical makeup of a drug, thereby ensuring its quality.

Moreover, the quantification of marker compounds stands as an additional parameter in the assessment of sample quality, further bolstering the reliability of the standardization process.³

Ayurvedic medicine, originating from the ancient wisdom of the Indian subcontinent, stands as a venerable system of healthcare utilized by millions across India, Nepal, Sri Lanka, China, Tibet, and Pakistan. Its increasing adoption in European countries attests to its global recognition and acceptance. Within the realm of alternative medicine, herbal remedies play a pivotal role, and Ayurveda's incorporation of botanical formulations is emblematic of this trend. In this context, the standardization of compound Ayurvedic formulations assumes paramount importance, serving as a crucial step toward establishing authenticity, quality, and efficacy. This imperative is particularly underscored in the present study, which focuses on the standardization of Chaturbhadra Kwatha churna (coarse power).⁴

This polyherbal formulation, comprised of four plant materials, holds promise as a valuable addition to the existing body of knowledge in Ayurveda. Described in Bhaishajya Ratnavali Grahani Roga (Irritable bowel syndrome) context, Chaturbhadra kwatha churna contains Shunti (Zingiber officinale), Amruta (Tinospora cordifolia), Ativisa (Aconitum heterophylum), and Musta (Cyperus rotundus) as its key constituents.

The formulation finds its therapeutic indications in addressing conditions such as fever, diarrhoea, cough, breathing difficulty, and vomiting in children. The standardization process of Chaturbhadra Kwatha churna involved a comprehensive evaluation based on macroscopic and microscopic characteristics, as well as physico-chemical parameters. By scrutinizing these facets, the study seeks to provide a robust foundation for ensuring the consistent quality and effectiveness of this Ayurvedic formulation. As Ayurveda continues to traverse geographical and cultural boundaries, the standardization of its formulations becomes imperative to foster confidence among practitioners and patients alike. The systematic approach undertaken in this study contributes valuable insights to the scientific community, bridging traditional knowledge with contemporary methodologies. This convergence holds the potential to enhance the global integration of Ayurvedic medicine as a viable and standardized healthcare option.

MATERIALS AND METHODS:

Collection of Raw Drugs:

The ingredients of preparation of Chaturbhadra kwatha churna were procured from Bhubaneswar local market. Their identities were confirmed by correlating their morphological and microscopical characters by Research Assistant, Botany of the institute.

Preparation of Chaturbhadra Kvatha Churna:

The preparation of Kwatha Churna was carried out in the pharmaceutical section of the Central Ayurveda Research Institute (CARI) Bhubaneswar. Each ingredient specified in the pharmacopoeia was individually weighed after cleaning, drying, and coarse powdering; following which they were passed through a 710 µm IS Sieve (sieve number 22). The resulting powders of the various drugs were then precisely weighed and thoroughly mixed to achieve the final formulation. To maintain the integrity of the formulation, it was stored in an airtight container, providing protection against both light and moisture. This systematic and standardized preparation process adheres to established pharmaceutical practices, ensuring the quality and stability of Kwatha Churna for subsequent scientific investigations and therapeutic applications.

Table 1: List of Ingredients and Quantity

Ingredients	Botanical name	Part used	Quantity
Shunti	Zingiber officinale	Rhizome	25 %
Amruta	Tinospora cordifolia	Stem	25 %
Ativisa	Aconitum heterophylum	Root tuber	25 %
Musta	Cyperus rotundus	Root tuber	25 %

Organoleptic Evaluation: Organoleptic evaluation refers to the evaluation of the formulation by its appearance, colour, odour, taste etc appearance

Table 2: Organoleptic evaluation of Chaturbhadrika churna

(Power)

Formulation	Observation
Appearance	Powder
Colour	Light brown
Odour	Pungent
Taste	Bitter

Physicochemical Investigations:

In physicochemical investigation, total ash, acid insoluble ash, alcohol and water extractive values, and Ph were estimated as per standard procedures. All the parameters were performed according to the official methods prescribed in Ayurvedic Pharmacopoeia.

Determination of Total Ash:

Incinerate about 2 to 3 g accurately weighed, of the ground drug in a tared platinum orsilica dish at a temperature not exceeding 600^℃until free from carbon, cool in a desiccator for 30 minutes and weigh. If a carbon free ash cannot be obtained in this way, exhaust the charred mass with hot water, collect the residue on an ash less filter paper, collect the residue on an filter paper, incinerate the residue and filter paper, add the filtrate, evaporate to dryness, and ignite at a temperature not exceeding 600^℃. Calculate the percentage of ash with reference to the air-dried drug. Formula for Ash value: = 100/a x(c-b) Where, a is initial weight of sample b is weight of empty crucible c is Weight of crucible along with ash ⁵

Determination of Acid-Insoluble Ash:

To the crucible containing total ash, add 25 ml of dilute hydrochloric acid. Collect the insoluble matter on an ash less filter paper (Whatman 41) and wash with hot water until the filtrate is neutral. Transfer the filter paper containing the insoluble matter to the original crucible, dry on a hot-plate and ignite to constant weight. Allow the residue to cool in a suitable desiccator for 30 minutes and weigh without delay. Calculate the content of acid-insoluble ash with reference to the air-dried drug.⁵

Determination of Alcohol Soluble Extractive:

Macerate 5 g of the air dried drug, coarsely powdered, with 100 ml of alcohol the specified strength in a closed flask for twenty-four hours, shaking frequently during six hours and allowing to stand for eighteen hours. Filter rapidly, taking precautions against loss of solvent, evaporate 25 ml of the filtrate to dryness in a tared flat bottomed shallow dish, and dry at 1050, to constant weight and weigh. Calculate the percentage of alcohol-soluble extractive with reference to the air-dried drug.

Determination of Water Soluble Extractive:

Proceed as directed for the determination of alcohol-soluble extractive, using Chloroform-water (22.5 ml chloroform in purified water to produce 1000 ml) instead of ethanol.⁶

Determination of pH Values:

The pH value of an aqueous liquid may be defined as the common logarithum of the reciprocal of the hydrogen ion concentration expressed in g per litre. Although this definition provides a useful practical means for the quantitative indication of the acidity or alkalinity of a solution, it is less satisfactory from a strictly theoretical point of view. No definition of pH as a measurable quantity can have a simple meaning, which is also fundamental and exact. The pH value of a liquid can be determined potentiometrically by means of the glass electrode, a reference electrode and a pH meter either of the digital or analogue type.⁷

RESULTS AND DISCUSSION:

Microscopic Evaluation:

The powdered formulation has shown the presence of starch grains, fibers, crystal fibers, on microscopically evaluation.

Table 3: Microscopic Evaluation of Chaturbhadra churna (Power)

Sl. No.	Name of ingredient	Microscopic finding
1	Shunti	Parenchymal cells containing starch grains, fragmented separated fibers, fragmented vessels bearing annular thickening.
2	Amruta	Cork cells, crystals of calcium oxalate, crystal fibers, bordered pitted vessels.
3	Atibisa	Cortical parenchyma with starch grains, xylem vessels with reticulate thickening
4	Mustha	Vessel element with spiral thickening tissue fragments with thin walled cells.

Physico-Chemical Parameters:

Ash value of a drug provides an idea of the earthy matter or the inorganic components and other impurities present along with the drug. Extractive values are useful for the determination of exhausted or adulterated drugs. The water soluble extractive was high in the formulation. The results of physico-chemical constants of the drug powder are presented in Table below.

Table 4: Physico-chemical parameters of Chaturbhadra churna (Power)

Parameter	Value %w/w
Total ash value	5.704%w/w
Acid insoluble ash	1.8873%w/w
Alcohol soluble extractive	43.3679%w/w
Water soluble extractive	48.9880%w/w
Ph(1%Aq.)	5.76

Therapeutic uses: Agnimandya (Loss of appetite), Grahani ^5-10(Chronic dysentry)

CONCLUSION:

The comprehensive investigation conducted on Chaturbhadra Kvatha Churna through pharmacognostical and physico-chemical evaluations establishes its potential as a reliable reference standard for quality control and assurance purposes. The findings not only contribute valuable insights into the formulation's characteristics but also pave the way for future studies by serving as a crucial source of information. For testing more specifically about the quality of the formulations we can further test it through various sophisticated instruments like HPLC, Mass Spectroscopy, IR spectroscopy, which will give us a brief idea about the presence and quantity of all the chemical constituents present in the formulation. The sample shows satisfactory results, but the efficacy of the products can only be judged by doing the pharmacological studies.

ACKNOWLEDGEMENT:

The authors are thankful to the Director General, Deputy Director General of CCRAS and Director of CARI, Bhubaneswar for the support and guidance provided for carrying out the study.

CONFLICT OF INTEREST:

The authors declare that they have no competing interests.

REFERENCES:

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4. The Ayurvedic Formulary of India, Part-I, 4:6, 1^st Edi-1978, AYUSH, Govt of India, p-46.

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6. The Ayurvedic Pharmacopoeia of India, Part-II, Vol-IV, 1^st Edi-2017, AYUSH, Govt. of India, p-118

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Received on 05.01.2024 Modified on 09.05.2024

Res. J. Pharmacology and Pharmacodynamics.2024;16(3):153-155.

DOI: 10.52711/2321-5836.2024.00026

Physico-Chemical Analysis of Ayurvedic Formulation -Chaturbhadra Kwatha Churna