Phytochemical analysis of chitraka granthikadi kashaya—an ayurvedic polyherbal formulation

PR Dhanya; Neelakanta J Sajjanar; G Gopalakrishna

doi:10.4103/jism.jism_66_22

ORIGINAL ARTICLES

Year : 2023 | Volume : 11 | Issue : 1 | Page : 24-29

Phytochemical analysis of chitraka granthikadi kashaya—an ayurvedic polyherbal formulation

PR Dhanya, Neelakanta J Sajjanar, G Gopalakrishna
Department of PG studies in Kayachikitsa, Sri Sri College of Ayurveda and Research, Bengaluru, Karnataka, India

Date of Submission	30-Jul-2022
Date of Decision	27-Jan-2023
Date of Acceptance	04-Feb-2023
Date of Web Publication	15-Apr-2023

Correspondence Address:
P R Dhanya
Department of PG Studies in Kayachikitsa, Sri Sri College of Ayurveda and Research, Bengaluru 560082, Karnataka
India

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jism.jism_66_22

Abstract

Background: Chromatographic fingerprinting of herbal formulations is a useful tool for explaining the scientific background and to develop a more targeted treatment methodology. Thus, this study has undertaken to report the characteristic parameters of chitraka granthikadi kashaya an ayurvedic polyherbal formulation to validate its quality. Materials and Methods: The root of Shodhita (purified) Chitraka (Plumbago zeylanica L.) and Eranda (Ricinus communis L.), dried rhizome of Shunti (Zingiber officinale Roscoe.), fruit of Pippali (Piper longum L.), and Saindhava lavana (Rock salt) were the ingredients of chitraka granthikadi kashaya. The Kwatha (medicated decoction) was prepared and evaluated for its phytochemical parameters and thin layer chromatography, gas chromatography-mass spectrometry chromatography parameters in accordance with the standard methods. Results: The phytochemical screening, GC–MS study, and TLC of the formulation reported the presence of 13 potential bioactive compounds. The action of these compounds based on the available literature has given a close similarity with the therapeutic action of the formulation. Conclusion: The presence of 13 bioactive compounds was found in this formulation with different peak values. Major compounds present are piperine, B-sitosterol, geranyl isovalarate, hexacosane, etc. Few compounds were very minimal and even no sufficient literature data available for them.

Keywords: Ayurveda poly herbal formulation, Chitraka granthikadi kashaya, GC–MS, Phytochemicals analysis, TLC

How to cite this article:
Dhanya P R, Sajjanar NJ, Gopalakrishna G. Phytochemical analysis of chitraka granthikadi kashaya—an ayurvedic polyherbal formulation. J Indian Sys Medicine 2023;11:24-9

How to cite this URL:
Dhanya P R, Sajjanar NJ, Gopalakrishna G. Phytochemical analysis of chitraka granthikadi kashaya—an ayurvedic polyherbal formulation. J Indian Sys Medicine [serial online] 2023 [cited 2023 Jun 7];11:24-9. Available from: https://www.joinsysmed.com/text.asp?2023/11/1/24/374257

Introduction

Bhaishajya Kalpana the ayurvedic pharmacology deals with the fundamental principles of drug formulation. This covers various aspects like designing the formulation, drug selection, method of preparation, dose fixation, time of consumption, and also the indication of that particular preparation. Following such principles in the field of pharmacology promises the quality of the product and can nullify the effect of compromises in its efficacy.

Analytical study is an inevitable part in the modern pharmaceutical industry, which maintains the quality product. Quality assurance helps to standardize a medicament by authenticating the drugs that are impeccable of adulteration with its top quality. Thus, to standardize and authenticate the classical formulation “chitraka granthikadi kashaya,” this study has planned.

Chitraka granthika kashaya is a classical ayurvedic polyherbal formulation containing Chitraka (Plumbago zeylanica L.), Pippali (Piper longum L.), Eranda (Ricinus communis L.), Shunti (Zingiber officinale Roscoe.), and Saindhava lavana (Rock salt). It is prescribed for the treatment of gastrointestinal condition Table 1].^[1] Presently, the efficacy of this formulation has not been addressed appropriately in previous studies. Thus, it is needed to be updated by considering the lack of research on the same.

Table 1: The latin name and ayurvedic pharmacological action of the drugs in the formulation

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This study aimed to perform a detailed analytical examination of the polyherbal formulation using chromatographic and spectroscopy methods to identify the chemical markers and bioactive compounds present in it.

Materials and Methods

Collection of Materials

The ingredients were procured and authenticated from a good manufacturing practice certified pharmacy. A fresh decoction was prepared from a sample dose 25 g ingredients, viz., Chitraka (P. zeylanica L.), Eranda (R. communis L.), Pippali (P. longum L.), Shunti (Z. officinale Roscoe.), and S. lavana (Rock salt). The root of Shodita (purified) P. zeylanica L. and R. communis L., dried rhizome of Z. officinale Roscoe., Fruit of P. longum L., and powdered rock salt were taken for the preparation.

Method

This polyherbal fresh decoction was prepared as per the Kwatha vidhi (standard preparation methodology for decoction), where 25 g decoction powder boiled in 400 mL water and reduce to 1/4th and filtered. This filtered part added with 3 g rock salt.^[2]

Phytochemical Evaluation

The various physical parameters like the specific gravity, total solid content, water-soluble extractive value; alcohol-soluble value, and pH were evaluated using the standard pharmacopeia method.

The fresh decoction was evaporated on a water bath for the removal of water; the dried mass was refluxed with methanol for three times and filtered. The methanol extract was subjected to preliminary phytochemical analysis. Ethanol extract was prepared in the same manner, and phytochemical analysis was carried out. Dragendorff’s reagent test for alkaloids, Shinoda test for flavanoids, picric acid test for glycosides, folin ciocalteu phenol reagent test for phenol, foam test for saponins, ferric chloride test for tannins, and Benedict’s test for carbohydrate were conducted.

Thin-Layer Chromatography

The sample fresh decoction of 2.57 g weight dried to get residue and makeup with 5 mL hexane and centrifuged at 300 rpm and carried out thin-layer chromatography with supernatant liquid. Applied 6 µL on TLC plate to a distance of 8 cm using toluene:chloroform:methanol (8:3:1) as mobile phase. After development, allowed the plate to dry in air and examined under ultraviolet light, sprayed the plate with anisaldehyde sulfuric acid followed by heating at 105°C for about 10 min.

GC–MS Analysis

GC–MS analysis of the fresh decoction was done at an analytical study center, using the standard GC–MS model as explained below.

Sample preparation: Methanolic extract of sample decoction of 50 mL was prepared and filtered through a syringe filter (Nylon 13 mm 0.2 µm) into vitals and injected to GC–MS.

Instrument details and GC–MS operating condition: Agilent 7890 instrument was used for GC, with 5975C with triple axis detector.

Column-DB 5MS 30M × 0.250 mm, diameter × 0.25 µm thickness.

GC–MS analysis was performed by splitless injection by injecting 2 mL of sample with split ration 15:1. Helium gas (99.9995%) was used as the carrier gas at a flow rate of 1 mL/min. The analysis was performed in the electron impact (EI) mode with 70 eV of ionization energy. The injector temperature was maintained at 280°C (constant). The column oven temperature program is given in [Table 2].

Table 2: The column oven temperature. The compound was identified after comparing the spectral configuration obtained with that of available mass spectral database (NIST 08 SPECTRAL DATA)

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Observation and Results

Preliminary Photochemical Analysis

The outcome of extracted contents of chitraka granthikadi kashaya in different solvents tested for the presence and absence of various phytochemicals group [Table 3].

Table 3: The list of phytochemicals group present in the fresh decoction sample

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Thin-Layer Chromatography

The major bands are found to be at 254 nm Rf 0.35 (black).

The major bands are found to be at Rf 0.06 (orange), Rf 0.10 (green), Rf 0.19 (green), Rf 0.25 (blue), Rf 0.37 (blue), Rf 0.42 (blue), Rf 0.52 (green), Rf 0.58 (violet), and Rf 0.70 (red) 366 nm.

The major bands are found to be at Rf 0.06 (red), Rf 0.16 (orange), Rf 0.21 (yellow), Rf 0.35 (blue), Rf 0.47 (yellow), and Rf 0.71 (black) 366 nm after derivitization.

The major bands are found to be at Rf 0.07 (purple), Rf 0.18 (purple), Rf 0.37 (green), Rf 0.47 (purple), and Rf 0.71 (green) white light after derivitization. The major bands found were given in [Figure 1] and [Figure 2].

Figure 1: Thin layer chromatography results description

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Figure 2: Thin layer chromatography plates of the freshly prepared decoction sample

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GC–MS Analysis

The chromatogram of GC–MS spectra analysis showing the peaks of the number of compounds from GC fraction of the methanol extracts of the fresh decoction is represented in [Table 4].

Table 4: The total percentage report of the compounds with its peak height

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In this analysis, presence of a total 13 different bioactive compounds was identified as the major potent compounds of the fresh decoction, namely, piperine; β-sitosterol; dodecanoic acid, 1, 2, 3-propanetriyl ester; octacosane; hexacosane; heptacosane; 2-propenoic acid, 3-phenyl-, methyl ester; 3-decanone, 1-(4-hydroxy-3-methoxyphenyl); naphthalene, decahydro-1,1-dimethyl-; β-guaiene; nerolidyl acetate; benzene, 1, 2, 3-trimethoxy-5-(2-propenyl)-; geranyl isovalerate (GIV) [Figure 3].

Figure 3: Chromatogram representing the presence of bioactive compounds present in the fresh decoction sample

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Discussion

The phytochemical screening of chitraka granthikadi kashaya was performed to identify the dominant chemical constituent present in the medicine. Qualitative analysis revealed the presence of carbohydrates, tanins, phenols, and flavanoids. The chromatography technique adopted for the phytochemical analysis includes both TLC and GC–MS. On TLC evaluation, the various bands formed at different Rf were recorded. On GC–MS evaluation, a total of 13 bioactive compounds were found in the fresh decoction.

Piperine—Digestive stimulant action of piperine was well established by various research works. A study revealed that the influence of piperine on intestinal motility and food transit time, where it delay the gastrointestinal motility.^[3] Another study revealed that the gastrointestinal food transit time shortened by prolonged dietary piperine in rats.^[4] Effect of piperine on gastric mucosa has been established by various research work, where the use of Black pepper caused increases in gastric parietal and pepsin secretion and increased gastric cell exfoliation in humans.^[3] Another study is suggestive of inhibition action of piperine on diarrhea produced by castor oil, MgSO₄, and arachidonic acid in mice. Piperine reduced castor oil induced intestinal fluid accumulation in mouse intestine.^[5] Other effects includes thermogenic action of piperine via adrenal catecholamine secretion in rats.^[3]

B-sitosterol—It is a bioactive phytosterols present in plant cell membranes naturally with chemical structure similar to the mammalian cell-derived cholesterol. Lipid-rich plant foods were the major source of it. β-Sitosterol (SIT) is one of the major bioactive compound found in majority of plants. In many of the in vitro and in vivo studies, SIT showed to possess a varied biological actions such as anxiolytic and sedative effects, analgesic, immunomodulatory, antimicrobial, anticancer, anti-inflammatory, lipid-lowering effect, hepatoprotective, protective effect against NAFLD and respiratory diseases, wound healing effect, antioxidant, and antidiabetic activities. It also appeared to have a protective effect on the digestive system diseases.^[6]

GIV is routinely used as a food flavoring agent. Some study proposes GIV as a potential lead or supplementary molecule in treating and preventing colorectal cancer.^[7]

Dodecanoic acid, 1, 2, 3-propanetriyl ester trilaurin is a triglyceride obtained by formal acylation of the three hydroxyl groups of glycerol by lauric (dodecanoic) acid. It is a triglyceride and a dodecanoate ester.^[8]

Octacosane—Octacosane is a straight-chain alkane containing 28 carbon atoms. It act as a plant metabolite. Octacosane is a natural product found in Hypericum olympicum, Euonymus latifolius, and other organisms.^[8]

Hexacosane—Hexacosane is a straight-chain alkane comprising of 26 carbon atoms.^[9] It has a role as a volatile oil component and a plant metabolite. Another study suggested antibacterial activity of it.^[10]

β-Guaiene—These are terpenes with three consecutive isoprene units. Based on a literature review, a small amount of articles have been published on beta-guanine.^[11]

Nerolidyl acetate—A sesquiterpenoid, naturally found in Artemisia annua.^[12] As per some study it reduces colon adenoma in rats.^[13]

Probable Mode of Action

The efficacy of the formulation was attributed to the presence of these potent bioactive compounds. Piperine is the major component in the kashaya and has the stimulatory effect, effect on motility and transit time. This might be helped to ease the gastric activity and also improves the absorption of nutrients due to its bioavailability. Moreover, the formulation has a protective action over gastrointestinal system and also gives lubrication to the gastric system. Thus it can be considered as effective for gastric condition and is justifiable to its indication.

Conclusion

The analytical study of chitraka granthikadi kashaya showed the presence of potent phytochemicals compounds, this substantiate the medicinal value of the freshly prepared medicated polyherbal decoction as per ayurveda. Since this is a preliminary report, further investigation on the isolation and characterization of the molecules for their medicinal value is required.

Acknowledgements

We thank care Keralam Pvt. Ltd. for performing the analytical study, Farmers pharmacy Pvt. Ltd. for providing the raw materials. We also thank Dr. Ravishankar B for the constant support and guidance throughout the study.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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