Evaluation of Ayurvedic Compound Formulation 5- Sama Śarkarã Cûrna

Ashok Kumar Tiwari*, Manoj Kumar Tripathi and Neelesh Dwivedi

Ayurveda Sadan, JRD TATA Foundation for Research in Ayurveda & Yoga Science,

Arogyadham, Deendayal Research Institute, Chitrakoot, Satna – 485780(M.P)

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

ABSTRACT:

Natural products from plant, animal and minerals have been the basis of the treatment of human disease. India is a vast repository of medicinal plants that are used in traditional medical treatments Herbal medicines as the major remedy in traditional system of medicine have been used in medical practices since antiquity. Medicinal plants play an important role in the development of potent therapeutic agents. To keep this view in mind, a polyherbal Sama Śarkarã Cûrna is formulated in house, which is very effective in stomach disorders, asthma, heart diseases and bronchitis. It formulated using eight single drugs viz Zingiber officinale (rhizome), Piper longum (fruits), Piper nigrum (fruits), Mesua ferrea (Stamen), Cinnamomum tamala (leaf), Cinnamomum zeylanicum (stem bark), Elettaria cardamomum (seeds) and Sita. All the ingredients and curna were analyzed in order to assess the authenticity of the drugs based on ayurvedic requirement following a series of physical-chemical, phyto-chemical, chromatography, microbiological parameters. Herbal medicines are inherently different from conventional pharmacological treatments, but presently there is no way to assess their efficacy other than by currently used conventional clinical trial methodologies, in which efficacy is conventionally assessed by clinical, laboratory, or diagnostic outcomes. Systematic study of herbal drug following quality parameters can improve morbidity, mortality, and of course quality of life. There is, however, scope to utilize the flexibility inherent in the modern scientific method for conducting studies on herbal medicines.

KEYWORDS: Ayurvedic formulation, Sama Śarkarã cûrna, quality control parameter.

INTRODUCTION:

Ayurveda, like traditional medical science, is the one of the most ancient medical science of the world. Basic feature of Ayurveda is its holistic approach to treat human beings as a whole and it restore harmony among the human beings, plants and environment. Ayurveda is based on Tridosha principal (Vaya, Pitta, and Kaph) and Panchbhuta therapy (Rasa, Vipaka, Veerya, Prabhava, and Guna). By knowing the standards of a medicine following qualitative and quantitative parameters and method followed in single and formulated drug can improve the efficacy and efficiency of drug as well as confidence of doctor and patient.

Goal of standardization is to assess the authenticity of the drug based on the above principal. Medicinal plants play an important role in the development of potent therapeutic agents. To keep this view in mind, a polyherbal Sama Śarkarã Cûrna is formulated in-house, which is very effective in stomach disorders, asthma, heart diseases and bronchitis(Anonymous, 2000).

It was formulated using eight single drugs viz Suṇṭhi (Zingiber officinale - rhizome), Kana (Piper longum - fruit), Marica (Piper nigrum - fruit), Nãga (Mesua ferrea - stamen), Dala (Cinnamomum tamala - leaf), Tvak (Cinnamomum zeylanicum - stem bark), Elã (Elettaria cardamomum - seed) and Sitã (sugar) in equal quantity (Anonymous, 1996 and Anonymous, 2000). Present study describes the scientific evaluation and standardization of compound drug Sama Śarkarã Cûrna and its single ingredients (Anonymous, 2008). Both are useful in Ayurveda, Siddha and Unani as single or in combination with other drugs. Hence the purpose of standardization of raw drugs and formulation is obviously to ensure the therapeutic efficacy of the drug.

MATERIALS AND METHODS:

Preparation of the Cûrna

All the ingredients were used of Pharmacopoeial quality (Anonymous, 2000). These were washed, dried and ground individually passed through 180 µm separately then weighed separately, mixed in specified ratio and passed through 355 µm to obtain a homogenous blend. It was stored in an airtight container to protect from light and moisture. Five different samples of Sama Sarkarã Cûrna three samples were prepared at research laboratory Ayurveda Sadan, Chtrakoot Batch - A, B and C and  two samples were prepared at Chitrakoot Rsshala Pharmacy, Chitrakoot Batch -D and E were studied. Each sample of Sama Śarkarã Cûrna was formulated using eight ingredients (Anonymous, 2000) i.e. Z. officinale, P. longum, P. nigrum, M. ferrea, C. tamala, C. zeylanicum, E. cardamomum and Sitã in equal proportion.

Physico-chemical parameters:

Organoleptic characters, particle size and physico-chemical analysis of all the samples were carried out (Anonymous, 2008 and Patel et al., 2006). Quantitative analysis for total ash, acid insoluble ash, water soluble ash, extractive values in water soluble and alcohol soluble extractive, loss on drying at 105^°C and pH of filtrate of 10% w/v aqueous solution were checked in triplicate according to the prescribed Quality control methods for medicinal plant material (Anonymous, 1998 and Lohar, 2007).

Microscopic characteristics:

For microscopic analysis (Kokate, 1994) a small quantity represent of the Cûrnas, along with the genuine samples, i.e. Suṇṭhi (Z. officinale, rhizome), Kana (P. longum, fruits), Marica (P. nigrum, fruits), Nãga (M. ferrea, stamen), Dala (C. tamala, leaf), Tvak (C. zeylanicum, stem bark), Elã (E. cardamomum, seeds) and Sitã (sugar) was mixed with water, stained with iodine and mounted in glycerin, were used to examine the starch grains and its type. Another small quantity of samples cleared by heating with chloral hydrate and mounted in glycerin was used to identify diagnostic microscopical characters of the ingredients. Further, small quantity of the Cûrna cleared with dilute KOH (5%) and was mounted in glycerin; was subjected to microscopic examination.

Table 1. Quality test for the finished product, Sama Śarkara Cûrna

Parameter	Sama Śarkara Cûrna					Average value
	Batch-A	Batch-B	Batch-C	Batch- D	Batch -E
Loss on drying at 105°C (%)	4.10	4.20	4.40	4.40	4.40	4.30
Total ash (%)	5.50	5.20	5.20	5.60	5.60	5.52
Acid-insoluble ash (%)	0.70	0.80	0.70	0.60	0.60	0.68
Alcohol-soluble extractive (%)	6.70	6.70	6.60	6.50	6.70	6.64
Water-soluble extractive (%)	62.40	62.70	62.10	62.20	62.50	62.38
pH (10%) aqueous solution	5.60	5.70	5.60	5.80	5.70	5.68

Parameter	Sama Śarkara Cûrna
	Šunthī	Kana (Fr.)	Marica (Fr.)	Naga (Stmn)	Dala (Lf.)	Tvak (st. bk.)	Ela (Sd.)	Sitã
Loss on drying at 105°C (%)	8.68	11.25	6.95	6.96	6.95	8.15	6.35	1.06
Total ash (%)	5.84	6.61	4.28	5.33	6.00	2.90	4.10	-
Acid-insoluble ash (%)	1.50	0.36	0.43	2.56	0.97	1.80	3.61	0.48
Alcohol-soluble extractive (%)	4.14	7.85	7.14	17.63	6.14	3.96	10.37	-
Water-soluble extractive (%)	15.45	19.5	11.45	15.19	15.45	4.73	15.11	-
pH (10%) aqueous solution	-	-	-	-	-	-	-	-

Note - Batch A, B and C - Research laboratory Ayurveda Sadan, Chitrakoot and Batch D and E - Chitrakoot Rasshala Pharmacy, Chitrakoot

Table 2. R_f value of test solution in Sama Śarkara Cûrna at 254 nm (before derivatization)

Rf values	Sama Śarkara Cûrna						Single ingredients
	A	B	C	D	E	1	2	3	4	5	6	7	8
Rf 1 (black)	0.60	0.60	0.60	0.60	0.60	NA	NA	0.60	0.22	NA	NA	NA	NA
Rf 2 (black)	0.64	0.64	0.64	0.64	0.64	NA	NA	0.64	0.26	0.64	NA	NA	NA
Rf 3 (black)	0.68	0.68	0.68	0.68	0.68	0.68	NA	NA	0.64	NA	NA	0.68	NA
Rf 4 (black)	0.70	0.70	0.70	0.70	0.70	NA	NA	0.70	0.73	0.70	NA	0.70	NA
Rf 5 (black)	0.80	0.80	0.80	0.80	0.80	0.80	NA	NA	NA	NA	NA	0.80	NA
Rf 6 (black)	NA	NA	NA	NA	NA	NA	NA	0.90	0.93	NA	NA	NA	NA

NA- NA-Major spot not appeared

Figure 1. Powder characteristics of Sama Śarkarã Cûrna

Table 3. R_f value of test solution in Sama Śarkara Cûrna at 366 nm (before derivatization)

Rf values	Sama Śarkara Cûrna					Single ingredients
	A	B	C	D	E	1	2	3	4	5	6	7	8
Rf 1(pink)	0.10	0.10	0.10	0.10	0.10	NA	NA	NA	NA	0.10	NA	0.10	NA
Rf 2(fluorescent )	0.14	0.14	0.14	0.14	0.14	NA	NA	NA	0.14	NA	NA	NA	NA
Rf 3 (brick red)	0.43	0.43	0.43	0.43	0.43	0.43	0.43	NA	NA	0.43	NA	0.43	NA
Rf 4 (white)	0.46	0.46	0.46	0.46	0.46	NA	NA	0.46	NA	0.46	NA	NA	NA
Rf 5(white)	0.55	0.55	0.55	0.55	0.55	NA	NA	0.55	NA	NA	0.55	NA	NA
Rf6(light yellow)	0.62	0.62	0.62	0.62	0.62	NA	NA	0.62	NA	0.62	NA	NA	NA
Rf 7(white)	0.73	0.73	0.73	0.73	0.73	0.73	NA	0.73	0.73	NA	0.73	NA	NA

NA- NA-Major spot not appeared

Table 4. R_f value of test solution in Sama Śarkara Cûrna at 254 nm (after derivatization)

Rf values	Sama Śarkarã Cûrna					Single Ingredients
	003A	003B	003C	003D	003E		Tvak	Elã	Marica,	Nãga	Kana	Śunthî	Dala	Sitã
Rf1 (yellow)	NA	NA	NA	NA	NA		0.10	NA	NA	NA	NA	NA	NA	NA
Rf2 (light yellow)	0.23	0.23	0.23	0.23	0.23		0.23	NA	NA	0.23	0.23	NA	0.23	NA
Rf3 (reddish brown)	0.34	0.34	0.34	0.34	0.34		NA	NA	0.34	NA	0.34	0.34	NA	NA
Rf4 (sky blue)	0.38	0.38	0.38	0.38	0.38		NA	NA	0.38	NA	NA	NA	NA	NA
Rf 5 (brown)	0.42	0.42	0.42	0.42	0.42		NA	NA	0.42	NA	0.42	NA	0.42	NA
Rf 6 (brown)	0.56	0.56	0.56	0.56	0.56		NA	NA	0.56	NA	0.56	0.56	NA	NA
Rf 7 (purple)	0.73	0.73	0.73	0.73	0.73		NA	NA	0.73	NA	NA	NA	NA	NA
Rf 8(black)	0.77	0.77	0.77	0.77	0.77		0.77	NA	NA	NA	0.77	NA	NA	NA
Rf 9(black)	0.93	0.93	0.93	0.93	0.93		NA	NA	NA	0.93	NA	NA	NA	NA

NA- NA-Major spot not appeared

Figure 2. Powder characteristics of Sama Śarkarã Cûrna

Figure 3 . Powder characteristics of Sama Śarkarã Cûrna

Figure 4 . TLC Finger prints in test solution of Sama Śarkara Cûrna at 254 nm (before derivatization)

Test Solutions of Sama Śarkara Cûrna for:

Batch - A, Batch - B, Batch – C, Batch - D, E : Batch - E 1: Tvak, 2: Ela, 3: Marica , 4: Naga, 5: Pippali, 6: Sunthi, 7: Dala(Tejpatra), 8: Sita (Iksu)

Figure 5 . TLC Finger prints in test solution of Sama Śarkarã Cûrna at 366 nm (before derivatization)

Figure 6 . TLC Finger prints in test solution of Sama Śarkarã Cûrna at 254 nm (after derivatization)

High Performance Thin Layer Chromatography (HPTLC) profile:

For HPTLC, 2gm of each sample was extracted with 25 ml of methanol on boiling water bath for 25 min. consecutively of 3 times using fresh portion of 25 ml methanol, filtrate and concentrated. Similarly, methanolic extracts were prepared for all eight ingredients, i.e. Z. officinale rhizome (Anonymous, 2012 and Anonymous, 2010 ), P. longum fruits (Anonymous, 2003), P. nigrum fruits, M. ferrea stamen (Anonymous, 2005), C. tamala leaf (Anonymous, 2005), C. zeylanicum stem bark, E. cardamomum seeds (Anonymous, 2003) and Sitã ingredients for use as reference.

TLC of extracts of all the samples and the reference ingredients was carried out on silica gel 60 F₂₅₄ precoated plates (0.2 mm thickness; from Merck India Limited Mumbai). An Applicator from Camag Linomat -5 (Camag Switzerland: 140443) was used for band application and photo documentation unit (Camag Reprostar-3:140604) was used for documentation of chromatographic fingerprints. The mobile phase used was Toluene: Ethyl acetate: Formic acid (7: 2.5: 0.5). The plate was developed over a distance of 9 cm in a saturated development chamber (Twin trough chamber (10×10 cm with SS lid, and visualized under visible light, 254 nm and 366nm. After spraying with 5% methanolic sulphuric acid followed by heating at 110⁰C for 5-10 min (Anonymous, 1998 and Lohar, 2007).

RESULTS AND DISCUSSION:

Sama Śarkarã Cûrna samples from 2 different manufactures, Batch-A, Batch-B, Batch-C, Batch-D, Batch-E, were subjected to analysis as above. All samples were dusty cream colored powder with a characteristic odour, sweetish bitter taste. The powder completely passes through 355 µm and not less than 50 percent through 180 µm. Results of loss on drying at 105 ⁰C 4.30%, total ash content 5.32%, acid insoluble ash 0.68%, alcohol soluble extractive 6.64%, water soluble extractive 62.68%, and pH 5.68 were calculated and are given in Table 1.

Microscopic examinations were also carried out to see presence of the different ingredients in all 5 samples of Sama Śarkarã Cûrna (Figures1-3). Groups of cork cells, fragment of fibres, pitted, septate fibres with reticulate and spiral thickening, oleo-resin canal, parenchymatous cell filled with starch grains (Sunthi); fragments of parenchyma, oval to elongated stone cells, oil globules, starch grains (Kana); perisperm cells filled with starch grains, slightly elongated stone cells, beaker-shaped stone, aleurone grains (Marica); spongy parenchyma, palisade tissue, trichomes, fibres (Dala); perenchymatus cells, elongated cells, pollen grains (Naga); cork cells, acicular crystals of calcium-oxalate, stone cells, parenchymatous cell, sclereids (Tvak); perisperm cells filled with starch grains, schlerenchymatous cells, prismatic crystals of calcium oxalate, oil globules, parenchymatous cells filled with oil globules and aleurone grains (Ela).

TLC of methanolic extracts of all the samples and there reference ingredients were carried out on silica gel plates (Figures 4-5). Several bands were observed, some of which were specific to the individual ingredients. These bands were chosen to identify the presence or absent of the individual ingredients in each of the five samples. It was observed that all these identifying bands were present in all of Cûrna samples thereby indicating the presence of all the eight ingredients in each of them. Tables 2-4 enlist identifying bands of the individual ingredients along with their R_f values.

CONCLUSION:

From the present studies, it can be calculated that the characteristic microscopical characteristics and the distinguishing band in the HPTLC profiles are very important for monitoring the quality of the Cûrna formulation as well as for establishing whether all the required ingredients are present in them. Also, standardization and development for reliable quality protocols for Ayurvedic formulations are important for keeping a check on batch to batch variations. Hence, the physiochemical parameters, quantitative analysis, HPTLC fingerprinting profiles and the microscopic characteristics together may be used for quality evaluation and the standardization of compound formulations and maintaining their quality, purity and efficacy.

ACKNOWLEDGEMENT:

Authors are thankful to Dr. Bharat Pathak, General Secretary Deendayal Research Institute, Chitrakoot for providing the research facilities. Authors are also thankful to Department of AYUSH, Ministry of Health and Family Welfare, and Government of India for providing the financial support.

REFERENCES:

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Received on 16.11.2013 Modified on 10.12.2013

Accepted on 15.12.2013 ©A&V Publications All right reserved

Res. J. Pharmacognosy & Phytochem. 6(1): Jan.-Mar. 2014; Page 09-15