INTRODUCTION:

India is rich heritage of traditional medicines. Ayurveda, Siddha, Unani, Homeopathy and Naturopathy are various branches of medicines. According to WHO, about 70% of world population extensively use traditional and alternative medicines for the healthcare¹. The development of these traditional systems of medicines with the perspectives of safety, efficacy and quality will definitely helps not only to preserve the traditional heritage but also to rationalize the use of natural product in healthcare². It is very important to establish a system of standardization for every plant medicine in the market, science the scope for variation in different batches of medicine is enormous.

Plant material when used in bulk quantity may vary in its chemical content and therefore, in its therapeutic effect according to different batches of collection with respect to different geographical and environmental conditions. So as to assess the uniformity in manufacture there is a need for ensuring the quality, efficacy and standard of Ayurvedic formulations³. Quality assurance of herbal product may be ensured by proper quality control of herbal ingredients and by means of good manufacturing practice⁴. As per American Herbal Product Association “standardization refers to the body of information and control necessary to product material of reasonable consistency”⁵.

Shatavari means “who possesses a hundred husbands or acceptable to many”. It is considered as general tonic and female reproductive tonic. In Ayurveda it is known as “Queens of Herb” as it promote love and devotion⁶. Asparagus racemosus is an important medicinal plant of tropical and subtropical India. Its medicinal usage has been reported in the Indian and British Pharmacopoeia and in indigenous systems of medicine. Amongst 22 species reported in India, Asparagus racemosus is one of the most commonly used species in Traditional medicine⁷.

Scientific classification of Shatavari

Kingdom: Plantae

Division: Magnoliophyta

Class: Liliopsida

Order: Asparagales

Family: Asparagaceae, Liliaceae

Genus: Asparagus

Species: racemosus

Botanical name: Asparagus racemosus Willd

Vernacular name:

A. racemosus Willd. is commonly called Shatavari, Satawar or Satmuli in Hindi; Satavari in Sanskrit; Shatamuli in Bengali; Shatavari or Shatmuli in Marathi; Satawari in Gujarati; Toala-gaddalu or Pilli-gaddalu in Telegu; Shimaishadavari or Thanner Vittan Kizhangu or Inli-chedi in Tamil; Chatavali in Malayalam; Majjigegadde or Aheruballi in Kannada; Kairuwa in Kumaon; Narbodh or Satmooli in Madhya Pradesh; and Norkanto or Satawar in Rajasthan⁸.

Racemosus is a well known Ayurvedic rasayana which prevent ageing, increase longevity, impart immunity, improve mental function, vigor and add vitality to the body and it is also used in nervous disorders, dyspepsia, tumors, inflammation, neuropathy, hepatopathy. Reports indicate that the pharmacological activities of A. racemosus root extract include antiulcer, antioxidant, and antidiarrhoeal, antidiabetic and immunomodulatory activities. A study of ancient classical Ayurvedic literature claimed several therapeutic attributes for the root of A. racemosus and has been specially recommended in cases of threatened abortion and as a galactogogue. Root of A. racemosus has been referred as bitter-sweet, emollient, cooling, nervine tonic, constipating, galactogogue, and aphrodisiac, diuretic, rejuvenating, carminative, stomachic, antiseptic and as tonic. Beneficial effects of the root of A. racemosus are suggested in nervous disorders, dyspepsia, diarrhoea, dysentery, tumors, inflammations, hyper dipsia, neuropathy, hepatopathy, cough, bronchitis, hyperacidity and certain infectious diseases^9,10. The root extract of Asparagus racemosus has also been traditionally used in Ayurveda to increase milk secretion during lactation. The aqueous extract of roots increased the weight of mammary glands in post partum and estrogen-primed rats and the uterine weight in the estrogen-primed group¹¹.

MATERIAL AND METHODS:

Plant material:

Raw material i.e. Shatavari roots were collected from Herbal Garden of Priyadarshini J L Chaturvedi College of Pharmacy, 846, New Nandanwan, Nagpur, Maharashtra and was authenticated by Dr. Mrs. Chaturvedi, Professor and Head of Department, Department of Botany, Rashtra Sant Tukdoji Maharaj, Nagpur University, Nagpur. Herbarium is deposited in the Department on 24-03-2013 with number as 9900.

The collected roots were washed and cut into small pieces and dried for 17 days in shade followed by drying in hot air oven for about 5-6 hrs. Dried roots were powdered and passed through mesh 16.

Formulation Profile:

Formulation 1 (Marketed):

It is in powdered form, purchased from local market in Nagpur. Coded as SCM-I.

Formulation 2 (Marketed):

It is in powdered form, purchased from local market in Nagpur. Coded as SCM-II.

Formulation 3 (Laboratory):

It is in powdered form, prepared in Laboratory of Natural Product, Sudhakarrao Naik, Institute of Pharmacy, Pusad, and Coded as SCL . The raw material (Shatavari root) used for preparation was collected from the Herbal Garden and was authenticated by Department of Botany, Rashtra Sant Tukdoji Maharaj, Nagpur University, Nagpur. The Formulation of Shatavari Churna was prepared as per The Ayurvedic Formulary of India¹². The powders should completely pass through 355 μm I. S. sieve (sieve number 44) and not less than 50 per cent pass through 180 μm I. S. sieve (sieve number 85).

METHODS:

Organoleptic study:

The development of organoleptic character i.e. sensory character provides simplest and quickest means to establish the identity, purity and quality of crude drug in terms of color, odour and taste. The Marketed formulation and In-house formulation were examined for color, odour and taste along with individual ingredient. For determination of color examine the untreated sample under diffuse day light, if required artificial light source with wavelength similar to those of day light. For determination of odour place the small quantity of material in palm of the hand or a beaker of suitable size, slowly and repeatedly inhale the air over the material. If required crush the material between thumb and index finger. For determination of taste if specifically required should be done by placing minimum quantity of material (crushed/powdered) on taste buds of tongue. Interval of 15 minutes between two samples was kept to make available the taste buds fresh every time.¹³

Microscopically study:

Microscopic identity of medicinal plant material is indispensable for the identification of broken of powdered materials. Examinations with microscopy along with other analytical methods supply invaluable supporting evidence. The powder material of SCM-I, SCM-II, SCL and section of root of Shatavari is microscopically analyzed. For powder material, place 2 drops of water, glycerol or ethanol on a glass slide. Moisten the tip of needle with water and dip into the powder. Transfers the quantity of material adheres to the tip of needle into the drop of fluid on the slide and apply the cover slip carefully. Section preparation of dried crude drug i.e. Shatavari root, the drug material is soaked in water or in equal parts of water, ethanol and glycerol for overnight so as to make it soft enough or can be boiled in water for few minutes. Prepare transverse section by cutting with razor blade or microtome at a right angle to the longitudinal axis of the material. Prepare longitudinal section by cutting in parallel with the longitudinal axis, either in radial direction (radial section) or in a tangential direction (tangential section) For detection of various parameters use various micro-chemical reagents such as Iodine, Phloroglucinol, Sudan red, acetic acid, potassium hydroxide, Chinese ink, 1-napthol, sulphuric acid, ferric chloride, etc.¹⁴.

Physical Characteristic:

The development of physical character provides simplest and quickest means to establish the identity, purity and quality of crude drug in terms of Bulk density, Tap density, Angle of repose, Hausner ratio, Carr’s index, etc,. The Marketed formulation and In-house formulation were examined for development of various physical characteristic along with individual ingredient.

Bulk density (ρ_B):

It is defined as the mass of a powder divided by the bulk volume. It was being determined by Fixed funnel method. A sample of 50 cm³ of powder that has been passed through sieve no. 20 is carefully introduced into a 100 ml graduated cylinder. The cylinder is dropped at 2 sec intervals on hard wooden surface three times from a height of 1 inch. The bulk density is obtained by dividing the weight of the sample in gm by the final volume in cm³ of the sample contained in the cylinder¹⁵.

Tap density (ρ_T):

It is defined as the mass of the powder divided by the tapped volume. A powder sample about 5.0g is transferred into the tarred 10 ml cylinder with the help of a funnel. The 250 ml measuring cylinder is placed on the tapping apparatus. The content is tapped and the volume occupied is recorded. The ratio of mass of powder to the tapped volume represents Tapped density¹⁶.

Angle of repose (θ):

A glass funnel is held in place with a clamp on ring support over a glass plate. The glass plate is placed on a micro-lab jack. Approximately 100g of powder is transferred in to the funnel which has been passed through number 10 size mesh, keeping the orifice of funnel blocked by the thumb. The lab-jack is so adjusted so that the gap of about 6-7 mm is maintained between top of powder pile and bottom of funnel stem. When the powder is emptied from the funnel, the angle of heap to the horizontal plane is measured with a protector. Measure the height of the pile (h) and the radius of the base (r) with the ruler. The angle of repose is thus estimated by the formula.

θ = tan^-1( h / r )

Hausner ratio:

The Hausner ratio is the ratio of Tap density divided by Bulk density, it is named after the engineer Henry H. Hausner, A Hausner ratio greater than 1.25 is considered to be an indication of poor flowability. It is calculated by the formula¹⁷.

H = ρT / ρB

Carr’s index:

The Carr index represents the flowability and compressibility of powder. It is named after the pharmacologist Charles Jelleff Carr (1910–2005). It measures the relative significance of interparticle interactions. A Carr index greater than 25 is considered to be an indication of poor flowability, and below 15, of good flowability. It is calculated by the formula¹⁷.

Physico- chemical Study:

The activity of any herb is dependent on the class of phytoconstituents or specific phytoconstituents being present in it. In majority, of the herbals which are in use the knowledge about these is fairly known. Therefore, it is necessary to devise a method of standardization based upon the presence of these chemicals. Physico-chemical constants like ash value, water soluble extracts, alcoholic extracts, loss on drying and pH values were determined of all the formulations and dried crude drug as per method described in The Ayurvedic Pharmacopoeia of India¹⁸_.

Determination of Ash Value:

The total ash method is designed to measure the total amount of material remaining after ignition, including physiological and non-physiological ash. Acid insoluble ash is a parameter obtained after boiling the ash with dilute hydrochloric acid¹⁹.

Determination of Extractives:

The percent extractive values were determined in water and alcohol. The extractive values were determined for all the Formulations SCM-I, SCM-II, SCL and dried crude drug¹⁹.

Determination of Foreign matter:

500 g of the drug sample was examined by spread it out in a thin layer. The foreign matter was detected by inspection with the unaided eye or by the use of a lens (6x). Separated and weighed the foreign matters and calculate the percentage of foreign matter present²⁰.

Determination of Moisture content:

LOD of the powdered drug was carried out to find out the percentage of moisture present in the drug since moisture facilitates the enzyme hydrolysis or growth of microbes lead to deterioration. 10 g of sample (without preliminary drying) were placed after accurately weighing it in a tarred evaporating dish. After placing the above said amount of the sample in the tarred evaporating dish dry at 105^oC and continue the drying and weighing at 10 minutes interval until difference between two successive weightings corresponds to not more than 0.25 per cent. Constant weight is reached when two consecutive weightings after drying for 30 minutes and cooling for 30 minutes in a desiccators, show not more than 0.01 g difference²¹. Finally moisture content was measured directly in percentage.

Determination of pH:

The pH of different formulations in 1% w/v and 10% w/v of water soluble portions were determined using pH paper (Range 3.5–6) and (6.5–14) with standard glass electrode²².

Determination of Crude fiber:

2 gm of drug was taken in a beaker and 50ml of 10% nitric acid was added. It was heated to boil with stirring (30 sec.).This was strained through fine cloth on a buchner funnel. The residue was washed with boiling water and transferred to a beaker. 50ml of 2.5% v/v sodium hydroxide solution was added. It was strained and washed with hot water. The residue was transferred in a clean and dried crucible. The residue was weighed and the crude fiber content was determined²³.

Qualitative photochemical studies:

The detection of presence of various phytoconstituents in the formulations and in dried crude drug is performed on the various extracts of all the formulation along with crude drug. The qualitative analysis of alkaloid, carbohydrate, tannins, Saponins, phenols, phytosterol, oils, fat, gum, mucilage, etc., was done²⁴.

RESULT AND DISCUSSION:

Organoleptic study:

All the formulations of Shatavari churna were evaluated as per WHO guidelines. The sensory and organoleptic study reveals that the formulations were grayish white in color, having characteristic odour and sweet in taste. (Table No. 1)

Table No. 1: Organoleptic Study

Sr. No.	Name	Color	Odour	Taste
1	SCM-I	Grayish White	Characteristic	Sweet
2	SCM-II	Grayish White	Characteristic	Sweet
3	SCL	Grayish White	Characteristic	Sweet
4	CRUDE DRUG	Creamish white	characteristic	sweet

Microscopically study:

The microscopical studies of all the formulation along with dried crude drug were done as per standards given by WHO in Quality Control Methods for Medicinal Plants Materials. The study revealed the authenticity of the crude drug. The transverse section of Shatavari roots shows the outermost layer is the 6-8 layered cortex having outer cells collenchymatous and inner cells parenchymatous, some of which may be pitted, wavy-walled, overlapping and may have mucilage content. Endodermis is 1-2 layered, lignified, pitted and continuous. Pericycle is 1-2 layered and consists of thin walled parenchymatous cells. Vascular bundles are radial, a characteristic of monocot root. Pith consists of pitted, lignified or non-lignified cells having intercellular spaces. The microscopic features of all the powders are very similar and show the presence of pericyclic fibers, prismatic and acicular calcium oxalate crystals (isolated or in bundles); pitted, annular and reticulate-thickened xylem vessels. (Figure 1and 2 illustrates the details)

Figure 1: Transverse Section of Asparagus racemosus wild. (A) Epidermis, (B) Cortex, (C) Endodermis, (D) Pericycle, (E) Xylem vessels, (F) Phloem, (G) Pith

CONCLUSION:

As per the Ayurvedic Pharmacopoeia of India all the formulations i.e. SCM-I, SCM-II, SCL and Crude drug were standardized. The identity, purity and quality of the formulations were done and all the formulations were

found of good quality and purity. All the physico-chemical values were as per the standards given in The Ayurvedic Pharmacopoeia of India.

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Received on 24.08.2015 Modified on 16.09.2015

Research J. Pharm. and Tech. 8(11): Nov., 2015; Page 1495-1501

DOI: 10.5958/0974-360X.2015.00267.X

Sr. No.	Parameter		SCM-I	SCM-II	SCL	CRUDE DRUG
1	Total Ash [Mean (n=3) ± SD]		4±0.54	4.5±0.47	4.7±0.34	3.9±0.56
2	Acid insoluble Ash[Mean (n=3) ± SD]		0.5±0.12	0.6±0.09	0.5±0.11	0.4±0.15
3	Water soluble extractive [Mean (n=3) ± SD]		33.5±1.57	34.0±1.53	35±1.14	34.5±1.37
4	Alcohol soluble extractive [Mean (n=3) ± SD]		6.5±1.2	7.0±1.1	6.9±0.9	7.0±1.0
5	Foreign matter [Mean (n=3) ± SD]		NIL	NIL	NIL	1.21±.08
6	Moisture content [Mean (n=3) ± SD]		10±0.25	9.5±0.32	8.0±0.18	8.2±0.35
7	pH	1% w/v [Mean (n=3) ± SD]	5.1±0.01	5.1±0.12	5.3±0.11	5.3±0.15
7	pH	10% w/v [Mean (n=3) ± SD]	5.3±0.02	5.3±0.10	5.5±0.12	5.5±0.14
8	Crude fiber [Mean (n=3) ± SD]		17.93±0.95	18.23±1.05	19.45±0.95	19.25±0.99

Sr. No.	Phytochemical	SCM-I	SCM-II	SCL	CRUDE DRUG
1	Alkaloid	-	-	-	-
2	Glycoside	+	+	+	+
3	Carbohydrate	+	+	+	+
4	Gum and Mucilage	+	+	+	+
5	Tannin	+	+	+	+
6	Saponin	+	+	+	+
7	Phytosterol	+	+	+	+
8	Fat	-	-	-	-