Quality Control of the Darvyadi Kvatha Ghana Vati - An Ayurvedic Compound Formulation

Ashok Kumar Tiwari*, Neelesh Dwivedi, Manoj Kumar Tripathi and Sharda Prasad Tripathi

Ayurveda Sadan, Arogyadham, JRD Tata Foundation for Research in Ayurveda and Yoga Sciences, Deendayal Research Institute, Chitrakoot, Satna, Madhya Pradesh, India-485334

*Corresponding Author E-mail: ashokckt77@yahoo.com, gangagargi@gmail.com

ABSTRACT:

The present study deals with the quality control of the Ayurvedic formulation Darvyadi Kvatha Ghana Vati (Tablet) following quality control procedures for the finished product. The results of physico-chemical parameters viz. loss on drying (9.7 %), average weight (5.0 %), friability (0.15 %) and disintegration time (85 min) were found. Microbiological limit test and heavy metals Pb, Cd, As, Hg were also found within the limits set by Ayurvedic Pharmacopeia of India (API). The obtained values can be adapted to lay down new pharmacopoeial standards with batch to batch consistency. The phytochemical constituents found in the raw material used for the preparation of Darvyadi Kvatha Ghana Vati (Tablet) facilitate the desirable therapeutic efficacy of the medicinal formulations a whole in elements and also could help in knowing the underlying mechanism of pharmacological action.

KEYWORDS: Ayurvedic formulation, Darvyadi Kvatha Ghana Vati, quality control.

INTRODUCTION:

In the few decades there has been an exponential growth in the field of herbal medicine. It is getting popularized in developing and developed countries owing to its natural origin and lesser side effect. In olden times, vaidyas used to treat patients on individual basis, and prepare drug according to the requirement of the patient. But the scene has changed now; herbal medicines are being manufactured on a large scale in mechanical units, where manufactures come across many problems such as availability of good quality raw materials, authenticated of raw materials, availability of standards, proper standardization methodology of single drugs and formulation, quality control parameters, etc. All these issues were discussed in the paper (Harish and Patel, 2001).

Standards such as identity, purity and potency of single drugs as well as formulations are important regarding therapeutic efficacy of herbal medicines. If the raw materials to be used in a medicine and stage by stage processes of manufacturer standardized the final product namely, the compound formulations (Rastogi et al., 2008; Rathi et al., 2010; Rajput et al., 2012; Tiwari et al., 2013; Tiwari et al., 2014 and Dwivedi et al., 2015) could be expected to conform to uniform standards. The study was undertaken to develop methods for evaluation of Darvyadi Kvatha Ghana Vati an Ayurvedic compound formulation which is formulated by ten single ingredients (Anonymous, 2000)viz. Dãrvî (Berberis aristata DC.- Stem), Rasãnjana (Berberis aristata DC.-solid extract tem), Vrsa (Adhatoda zeylanica Nees.-Root), Abda (Cyperus rotundus Linn.-Rhizome), Kirãta (Swertia chirata Buch Ham.-whole plant), Bilva (Aegle marmelos Corr. - Fruit pulp), Bhallãtaka (Śuddha) (Semecarpus anacardum Linn. f.-Fruit), Kairava (Nymphaea alba Linn. - Flower) prepared as per AFI(Table 1). The Kvatha Ghana Vati is formulated in house and Chitrakoot Rasshala Pharmacy, Chitrakoot which is very effective in Pradara (excessive vaginal discharge) and its ingrédients are also used to cure several diseases and preparation of ayurvedic compound formulations.

The Darvyadi Kvatha Ghan Vati were analysed following scientific parameters including organoleptic characters, physico-chemical analysis and chromatographic patterns. The work was undertaken is the trust as part of a program of testing and validation of traditional practices of using the Ayurvedic medicine. In this connection, standardization of Darvyadi Kvatha Ghana Vati becomes imperative. This paper dealt with standardization followed according to GMP guideline. Standardization guidelines to be followed for herbals products provided by World Health Organization (Anonymous, 1998), and Ayurvedic pharmacopoeia of India have been considered

MATERIALS AND METHODS:

Collection and Authentication of Raw Materials:

Dãrvî (Berberis aristata DC.- Stem), Rasãnjana (Berberis aristata DC.- solid extract tem) and Bhallãtaka (Śuddha) (Semecarpus anacardum Linn. f.-Fruit) were procured from Karwi, Chitrakoot (U.P.) during 2012. Other plants like Vrsa (Adhatoda zeylanica Nees.-Root, Bilva (Aegle marmelos Corr. - fruit pulp, Kairava (Nymphaea alba Linn. – Flower) and Abda (Cyperus rotundus Linn.-Rhizome, were collected during year 2012 from Chitrakoot forest range. Whereas Kirãta (Swertia chirata Buch Ham.-whole plant was collected from herbal garden, Chitrakoot, Arogyadham in 2012. All the materials were authenticated with the help of taxonomist Dr. RLS Sikarwar at Deendayal Research Institute, Chitrakoot, Satna (M.P.).

Preparation of the Darvyadi Kvatha Ghana Vati (Tablet):

All the ingredients were used of pharmacopoeial quality (Anonymous, 1990).Treated Bhallataka to prepare Suddha-Bhallataka (Anonymous, 2010). Cleaned, washed, dried and grind the Dãrvî (Dãruharidrã), Rasãnjana, Vrsa (Vãsã), Abda (Mustã ), Kirãta (Kirãtatikta), Bilva, Kairava (Kumuda) individually,crush all the ingredients into pieces of 1-3 cm except 4 and 7. Weighed separately and mixed them equal proportions (1:1:1:1:1:1:1:1) to ensure a homogenous mixture, Mix all the ingredients and boil in 16 times of potable water and reduce to 1/8^th of original volume, filter and concentrate further by heating the filtrate to obtain solidified extract mass (Ghana), test the Ghana by TLC, powder the Ghana; bled with approved excipients and preservatives, granulate and compress to tablets of desired weight, these were stored in an airtight containers to protect from light and moisture. Two samples were prepared at research laboratory Ayurveda Sadan, Chitrakoot Batch-A and Batch-B where Batch-C was prepared by Chitrakoot Rasshala Pharmacy, Chitrakoot.

Physico-Chemical Tests:

Organoleptic characters, average weight, friability and physico-chemical analysis of all the samples were carried out. Quantitative analysis for loss on drying at 105⁰C and disintegration time were checked in triplicate according to the prescribed Standard methods in Indian Pharmacopoeia (Lohar and Singh, 2008; Anonymous, 2008 and Anonymous, 1996 ).

High Performance Thin layer chromatography (HPTLC)profile:

Took 0.25 g of dried and powdered Kvatha-Ghan Vati with 10 ml ethanol (10 ml x 4). Filter each of the extracts and combine together. Add 5 g of anhydrous sodium sulphate, keep it for 10 min, filter and concentrate. HPTLC of extracts of all the samples were carried out on silica gel 60 F₂₅₄ precoated plates. The mobile phase used was Toluene: Ethyl acetate (7: 3). The plate was developed and visualized under ultraviolet at 254nm, 366nm and visible light. After spraying with 5% methanolic - sulphuric acid reagent followed by heating at 105⁰C for 5 min (Wagner and Bladt, 2004 and Wall, 2005).

Test for Microbial Limits:

Following tests were carry out as per WHO to determine the microbial load (Anonymous, 2010 and Pelczar et al., 1993) in three batches of Darvyadi Kvatha Ghan Vati (Tablet), a formulated compound drug powder of pharmaceutical substances.

(1) Enumeration of Staphylococcus aureus /gm

(2) Enumeration of Salmonella sp./gm

(3) Enumeration of Pseudomonas aeruginosa/gm

(4) Determination of E.coli

(5) Determination of total bacterial count (TBC)

(6) Determination of yeast and mould.

The microbiological tests were determined using specified agar and enrichment media from Himedia and Privet Limited Mumbai.

Table 1: Ingredients of Darvyadi Kvatha Ghan Vati

S.no	Sanskrit name	Botanical name	Part used	Portion
1.	Dãrvaî(Dãruharidrã)	Berberis aristata DC.	Stem	1 Part
2.	Rasãnjana(Dãruharidrã)	Berberis aristata DC.	Solid Ext.	1 Part
3.	Vrsa (Vãsã)	Adhatoda zeylanica Nees.	Root	1 Part
4.	Abda (Mustã)	Cyperus rotundus Linn.	Rhizome	1 Part
5.	Kirãta (Kirãtatikta)	Swertia chirata Buch Ham.	whole plant	1 Part
6.	Bilva	Aegle marmelos Corr.	Fruit Pulp	1 Part
7.	Bhallãtaka (Śuddha)	Semecarpus anacardum Linn. f.	Fruit	1 Part
8.	Kairava (Kumuda)	Nymphaea alba Linn.	Flower	1 Part
9.	Jala	Potable water	-	128 Parts

Heavy Metal:

Heavy metal analysis (lead, cadmium, arsenic and mercury) were carried out using Atomic absorption Spectrophotometry (Shimadzu-Model-AA-7000). All samples are digested with concentrated HNO_3: HClO₄ (4:1). Standards solutions are made for different dilution to get linear calibration (Merck). Pb and Cd were performed using graphite oven method, while As (Arsenic) were determined as hydride method and Hg were determined using cold absorption method. (Anonymous, 2010 and Anonymous, 2010a)

RESULTS AND DISCUSSION:

Greyish brown coloured mass (solidified kvãtha) with bitter taste. Physicochemical tests were done and results are given in Table -1.

HPTLC fingerprint profile of the formulations are depicted in (Fig. 1-4) indicates the presence of all the ingredients in proportional quantity in the formulations. This confirms the batch- to- batch consistency of the finished products. Development fingerprint profile would serve as a reference standard of the formulation. The TLC plate was examined under 254nm, 366nm, after derivatization 366nm and visible light. The R_f values and colours of the bands obtained were recorded. It shows major spots at 245 nm R_f 0.36, 0.45, 0.83 all spots were seen as black). It shows major spots at 366 nm R_f 0.09 (red), 0.16, 0.28 (both spots were seen as blue), 0.37(white), 0.52, 0.70, 0.76 (all spots were seen as blue), 0.83 (pink), 0.90(red). And after derivatization the plate shows major spots at 366 nm R_f 0.17, 0. 31, 0.37 (all spots were seen as brown), 0.58 (gray), 0.71(red), 0.77(brown), 0.97(red) and visible light R_f0.51, 0.71, 0.76, 0.97 (all spots were seen as brownish red).

Table 2: Physico-chemical parameters of Darvyadi Kvatha Ghan Vati

Name of Kvatha Ghan vati	Loss on drying at 105⁰ %	Average weight: + %	Friability %	Disintegration time (Minute)
Darvyadi Kvatha Ghan Vati (Batch A)	9.7	5.5	0.15	85
Darvyadi Kvatha Ghan Vati (Batch B)	9.8	5.0	0.15	86
Darvyadi Kvatha Ghan Vati (Batch C)	9.7	5.0	0.16	85
Average value	9.7	5.16	0.15	85

Table 3: Heavy metal of Darvyadi Kvatha Ghan Vati

S.no.	Parameter	Darvyadi Kvatha Ghan Vati			Actual concentration unit	API Limites
S.no.	Parameter	Batch A	Batch B	Batch C	Actual concentration unit	API Limites
1.	Lead (Pb)	0.4505	0.4103	0.4363	ppm	10 ppm
2.	Cadmium (Cd)	ND	ND	ND	ppm	0.3 ppm
3.	Arsenic (As)	0.012	0.012	0.012	ppm	03 ppm
4.	Mercury (Hg)	0.017	0.017	0.018	ppm	01 ppm

Note: ND-Not detected

CONCLUSION:

The present work deals with the physico-chemical, microbial and heavy metals study of the Darvyadi Kvatha Ghan Vati. Observed heavy metals data for Darvyadi Kvatha Ghan Vati (Tablet) presence of Pb, Cd, As, Hg which are within limit set by WHO. HPTLC profile shows 3 spots at 254 nm, 9 spots at 366 nm and after spraying 7 spots at 366 nm, 4 spots at visible light. All the above physicochemical, microbial and heavy metals data can be useful as diagnostic tool for identification and play an important role in quality control for further research.

ACKNOWLEDGEMENT:

Authors are grateful to Sri Abhay Mahajan, Organizing Secretary, Deendayal Research Institute, Chitrakoot for providing the research facilities. Funding authors are also thankful to Department of AYUSH, Ministry of Health and Family Welfare, Government of India, for financial support under the scheme “Centre of Excellence”.

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Received on 03.10.2015 Modified on 27.10.2015

Res. J. Pharmacognosy & Phytochem. 7(4): Oct-Dec. 2015; Page 209-213

DOI: 10.5958/0975-4385.2015.00039.4