1. INTRODUCTION:

Medicinal plants have been used in virtually all cultures as a source of medicine, since times immemorial. Herbal Medicine is still the mainstay of health care in several developing countries. The widely used herbal remedies and health care preparations as described in ancient texts such as the Vedas and the Bible are obtained from commonly used traditional herbs and medicinal plants. The medicinal properties of these botanicals are being better understood and are attributable to the phytochemicals that specific plants contain. The efficacy and safety of herbal products therefore rely on the quality and proper identification of the raw material or the original plant source. One major obstacle that might impair the potential use of traditional medicine as medicine of choice is the lack of standardization. Adulterations and substitutions are common in raw material trade of medicinal plants. Unintentional adulterations also exist in herbal raw material trade due to various reasons such as confusion in vernacular names between indigenous systems of medicine and local dialects, lack of knowledge about the authentic plant, non-availability of the authentic plant, similarity in morphology and aroma or careless collection^[1].

Medicinal plants and its therapeutic values are being increasingly recognized in the health care system all over the world. Ayurveda an ancient science postulates that there is no such a substance in the universe which cannot be used for therapeutic usage with the art and skill of formulation, even a poisonous drug can be transformed into a safe and effective drug^[2] and also a simple drug can be transformed into an utmost potent one^[3]. Standardization is an essential factor for polyherbal formulation in order to assess the quality of the drugs based on the concentration of their active principle. The process of evaluating the quality and purity of crude drugs by means of various parameters like morphological, microscopically, physical, chemical and biological observation is called standardization^[4-5]. It is very important to establish a system of standardization for every plant medicine in market, since the scope of variation in different batches of medicine is enormous. Plant materials when used in bulk quantity may vary in its chemical content and therefore, in its therapeutic effect according to different batches of collection e.g. collection of different season or collection from sites with different environmental surroundings or geographical location^[6]. Narasimha Churna is one of the effective classical polyherbal Medicine mentioned in AFI, part-I, 7:18. Narasimha Churna is made up of twelve ingredients like Satavari raja (Asparagus racemosus), Goksura (Tribulus terrestris), Varahi (Dioscorea bulbifera), Guduci (Tinospora cordifolia), Bhallataka - Suddha (Semecarpus anacardium), Citraka (Plumbago zeylanica), Tila (Sesamum indicum), Sunthi (Zingiber officinale), Marica (Piper nigrum), Pippali (Piper longum), Sarkara (Cane sugar), Maksika (Madhu), Vidari kanda raja (Vidari) (Pueraria tuberosa) which are well known vajikarana dravya contain in this formulation. Narasimha churna is a reputed and effective classical ayurvedic polyherbal medicine prescribed for deficiency of semen, senility, wrinkles in the skin, graying of hair, alopecia, excessive flow of urine , anemia, gout, disease of abdomen, fistula-in-ano. Narasimha churna is mainly reputed for the shukra kshaya (Oligospermia). It is a totally polyherbal medicine without any side effects. Vajikarana is the one which provide shukra bala in the person with shukra kshaya, it also indicated for alpa satva persons and sharira kshaya, thus it acts as pushti, urjaskara and also apatyasantanakara. Hence an effort is made to study the clinical evaluation of Narasimha Churna in the management of shukra kshaya (Oligospermia). Narasimha churna have some side effects, people with sensitive stomach and gastritis, special care is needed while administering this medicine to children and diabetic people, since it contains sugar as an ingredient.

2. MATERIALS AND METHODS:

2.1Procurement of raw drugs:

Narasimha churna contains fourteen ingredients like Satavari raja (Asparagus racemosus), Goksura (Tribulus terrestris), Varahi (Dioscorea bulbifera), Guduci (Tinospora cordifolia), Bhallataka – Suddha (Semecarpus anacardium), Citraka (Plumbago zeylanica), Tila (Sesamum indicum), Sunthi (Zingiber officinale), Marica (Piper nigrum), Pippali (Piper longum), Sarkara (Cane sugar), Maksika (Madhu), Vidari kanda raja (Vidari) (Pueraria tuberosa). All these ingredients were collected from local market of Patiala and all the ingredients were identified and authenticated at National Institute of Ayurvedic Pharmaceutical Research (NIAPR), Patiala Punjab (India).

Table 1. Narasimha churna contains following ingredients

S. No	Sanskrit name	Scientific name	Parts used	Quantity
1.	Satavari raja (Satavari)	Asparagus racemosus	Rt. Tr.	768g
2.	Goksura	Tribulus terrestris	Fr.	768g
3.	Varahi	Dioscorea bulbifera	Rz.	960g
4.	Guduci	Tinospora cordifolia	St.	1.2kg
5.	Bhallataka – Suddha	Semecarpus anacardium	Fr.	1.536kg
6.	Citraka	Plumbago zeylanica	Rt.	480g
7.	Tila	Sesamum indicum	Sd.	768g
8.	Sunthi	Zingiber officinale	Rz.	128g
9.	Marica	Piper nigrum	Fr.	128g
10.	Pippali	Piper longum	Fr.	128g
11.	Vidari kanda raja (Vidari)	Pueraria tuberosa	Rt. Tr.	768g
12.	Sarkara	Cane sugar	-	3.36kg
13.	Ghrta (Goghrta)	Clarified butter from cow’s milk	-	840g
14.	Maksika (Madhu)	Honey	-	1.680kg

2.2 Preparation of Narasimha churna:

Narasimha churna was prepared as per Ayurvedic formulary of India Part - I, 7:18. All ingredients were dried below 60 ⁰C, powered individually in a pulverizer and pass through 85 # sieve and stored in air tight containers. Each ingredient was weighed separately required weight, mixed together to obtain a homogeneous blend ^[7,
8] details given in Table 1.

Table 2. Organolepatic properties of Narasimha churna.

Appearance	Color	Odor	Taste
Granular powder	Brown	Pleasant	Slightly pungent

3. RESULTS AND DISCUSSION:

3.1 Organoleptic evaluation:

Organoleptic evaluation refers to evaluate that the formation by color, odor, taste and texture. The organoleptic characters of the Narasimha churna were evaluated ^{[8, 9]} and tabulated in Table 2.

3.2 Powder drug analysis of Narasimha churna^[10-14]:

About few mg of Narasimha churna powder warmed with chloral hydrate, washed and mounted in 50 percent glycerin, few mg of Narasimha churna powder washed thoroughly with water and mounted in 50 percent glycerin and few mg of Narasimha churna powder treated with iodine solution and mounted in 50 percent glycerin. Microscopically, covering trichomes (Tribulus terrestris) Fig. 1 (1); vessels (Tinospora cordifolia); Fig.1 (2); oil globules (Semecarpus anacardium) Fig.1 (3); fragments of cork and vessels (Plumbago zeylanica) Fig.1. (4,5); fragment of testa and fatty oil globules (Sesamum indicum) Fig1(6,7); parenchyma with oleo-resin cells and starch grains (Zingiber officinale) Fig 1 (8,9); volatile oil cell and stone cells (Piper nigrum) Fig1 (10,11) and stone cells of Piper longum were observed in different mounts of Narasimha churna.

3.3 Determination of Physical characteristics of powder^[15-19]:

Physical characteristics like bulk density, tap density, Hausner’s ration and Carr’s index were determined for Narasimha churna and results given in Table 3

Table 3. Physical characteristics of samples of Narasimha churna.

S.No	Parameters	Results
1.	Bulk density (gm/ cm³)	0.45
2.	Tap density (gm/ cm³)	0.71
3.	Hausner’s ration	1.57
4.	Carr’s index (%)	36.61

3.4 Evaluation of physicochemical parameters:

Evaluation of physicochemical parameters like pH, total ash, acid insoluble ash and loss on drying at 105^oC, alcohol, and water soluble extractive values were carried out as per the API/WHO guidelines ^[8,19-23] for Narasimha churna results tabulated in Table 4.

Table 4. Physicochemical parameters of Narasimha churna

S. No.	Name of Parameters	Results
1.	pH (10% aqueous solution (v/w)	5.27
2.	Total Ash (% w/w)	8.53
3.	Acid-insoluble ash (% w/w)	1.14
4.	Water-soluble extractive (% w/w)	39.30
5.	Alcohol-soluble extractive (% w/w)	41.69
6.	Loss on drying at 105^oC (% w/w)	5.32

3.4.1 Moisture content / Loss on drying at 105⁰C:

4 g of the sample was taken and heated in an oven at 105°C for 5 hour in a previously weighed 100 ml beaker. It was cooled in desiccators and weighed. The procedure was repeated till constant weight is obtained. The percentage of loss in weight of the sample was calculated.

Deterioration time of the plant material depends upon the amount of water present in plant material. If the water content is high, the plant can be easily deteriorated due to fungal attack. The loss on drying at 105°C of Narasimha churna was found to be 5.32.

3.4.2 Determination of Total ash value:

2 g of the sample was taken accurately in a previously ignited and tarred Silica dish. The material was spread evenly and ignited in a muffle furnace by gradually increasing the temperature to 600^oC until it is white, indicating the absence of carbon. The crucible was cooled in desiccators and allowed to stand for 30 minutes and weighed.

Total ash value of plant material indicated the amount of minerals and earthy materials attached to the plant material. Analytical results showed total ash value of Narasimha churna was 8.53.

3.4.3 Determination of Acid insoluble ash value

To the dish containing the total ash, 25 ml of 20 % Hydrochloric acid was added covered with a watch glass and boiled gently for 5 minutes. The watch was rinsed with a hot water and added to the crucible. The residue was washed with the hot water till the washings were neutral to the litmus. The insoluble material was collected and again placed in a same crucible and again ignited for 6 hr. to constant weight. The residue was cooled a desiccators for 30 minutes and weighed.

Percentage of acid insoluble as was calculated. The amount of acid-insoluble siliceous matter present in the Narasimha churna was 1.14.

3.4.4 Determination of Water soluble extractive value

4 g of the sample was taken in a glass stoppered flask.100 ml of distilled water was added. The flasks were shaken occasionally for 6 hours and then allowed to stand for 18 hours. The extract was filtered and 25 ml of the filtrate was pipette out in a pre-weighed 100 ml beaker and evaporated to dryness on a water bath. It was kept in a hot air oven for 5 hr at l05°C, cooled in desiccators for 30 minutes and weighed. The procedure was repeated till constant weight.

The water-soluble extractive value indicated the presence of sugar, acids and inorganic compounds. The water soluble extractive value in the Narasimha churna sample was found to be 39.30.

3.4.5 Determination of Alcohol soluble extractive Value

Same procedure as for the water soluble extractive value was followed. Instead of water, rectified spirit was taken as a solvent.

The alcohol soluble extractive values indicated the presence of polar constituents like phenols, alkaloids, steroids, glycosides, flavonoids and secondary metabolites present in the plant sample. The alcohol soluble extractive value was found to be 41.69 in the Narasimha churna.

3.4.6 Determination of pH Value

10% aqueous solution of sample was prepared and used for determining the pH value by pH meter. The pH value of Narasimha churna was found to be 5.27 (acidic).

3.4.7 Thin Layer Chromatography Methodology:

4g of the sample was soaked in 40 ml of rectified spirit (90%) with occasional shaking for 18 hrs, boiled for 10 minutes, filter, and concentrate the extract to 10 ml and carry out the thin layer chromatography. Apply 10 μl (micro liters) of the extract on (Aluminum plates precoated with Silica gel 60 F ₂₅₄ of thickness) TLC plate and develop the plate to a distance of 8 cm using toluene: ethyl acetate: formic (5: 1.5: 0.5) as mobile phase. After development, allow the plate to dry in air and examine under ultraviolet light. It shows 3 major spots at R_f0.56, 0.76, 0.92 (pale green) under 254 nm; and 0.23 (brown), 0.60, 0.74 (blue), 0.91(pale blue) under 366 nm. The plates were dried and then dipped in Vanillin- Sulphuric acid and heated at 105°C till the spots appeared. It shows 3 major spots at R_f0.34 (brown), 0.65 (blue), 0.82 (pale pink) in visible light after derivatisation.

4 ACKNOWLEDGEMENTS:

The authors are very grateful to Dr. Ramesh Babu Devalla, Director General, Dr. M. M. Padhi, Assistant Director (Technical) of CCRAS, New Delhi and Dr. M. M. Rao, Director, ACRI, New Delhi for providing encouragement and facilities for carrying out this work.

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Received on 06.10.2012 Modified on 15.10.2012

Asian J. Research Chem. 5(11): Nov., 2012; Page 1368-1371