INTRODUCTION:

Herbal medicines are complex chemical mixtures prepared from plants, used for their primary health care needs. Herbal drugs are finished labelled products that contain active ingredients such as aerial or underground parts of plant or other plant material or combination thereof, whether in the crude state or as plant preparations.¹

Herbal medicine is still the mainstay of about 75–80% of the world’s population, mainly in developing countries, for primary health care because of better cultural acceptability, better compatibility with the human body and lesser side effects. It is estimated that approximately one quarter of prescribed drugs contain plant extracts or active ingredients obtained from or modelled on plant substances.²

Herbal drug technology:

Herbal drug technology involves conversion of botanical materials into medicines where standardization and quality control with proper integration of modern scientific techniques and traditional knowledge is employed, and various drug delivery technologies used. The development of these traditional systems of medicines with the perspectives of safety, efficacy and quality will help not only to preserve this traditional heritage but also to rationalize the use of natural products in the healthcare.¹

Advantages of Herbal Medicine³:

· They have better patient tolerance as well as acceptance.

· Improvements in the quality, efficacy and safety of herbal medicines with the development of science and technology.

· Prolong and apparently uneventful use of herbal medicines may offer testimony of their

· Safety, efficacy and cheap in cost.

· They are not harmful and more effective than any synthetic drug.

Traditional Plant based remedies find increasing application as²:

· A direct source of therapeutic agent.

· A raw material for therapeutically active semi synthetics.

· A model for new synthetic compound.

· A new molecule for new synthetic compound.

Globally, the early part of the twentieth century brought an evolution in the pharmaceutical industry. With progress of chemical techniques, crude drugs came to be replaced by pure chemical drugs and developed countries witnessed a decline in popularity of medicinal plant therapy. However, during recent past interest has swung again and there is a resurgence of interest in study and use of medicinal plant. Most of the traditional plant based remedies are back in used and find increasing application.²

Ayurveda is made up of two Sanskrit words: Ayu which means life and Veda which means the knowledge. To know about life is Ayurveda. Ayurveda objective is to help the healthy person to maintain good health and the diseased person to regain good health. The practice of Ayurveda is designed to promote human happiness at physical, mental and spiritual level. By the proper balance of all vital energies in the body, the processes of physical deterioration and disease can be reduced. This is accomplished through proper eating, thinking and living habits as well as the use of herbal remedies to treat illness. According to the ancient Ayurvedic scholar Charaka, "ayu" is comprised of four essential parts, the combination of mind, body, senses and the soul. In U.S., Ayurveda is considered a form of complementary and alternative medicine (CAM).¹

Ayurvedic formulations are of multi component mixtures, containing plant and animal-derived products, minerals and metals. Most of the Ayurvedic therapeutics is polyherbal formulations. This is based on the fact that the therapeutic efficiency of the herbal constituents of plants is enhanced by the synergistic efficacy of other the therapeutic efficiency of the herbal constituents of plants is enhanced by the synergistic efficacy of other plants. Ayurveda is a complex science in which all the components are equally important for the cure of disease and maintaining balance of body, mind, and consciousness.¹

In Ayurveda, Churna is a fine powder of a drug or drugs which is prepared by mixing clean, finely powdered and sieved drugs. The term Churna may be applied to the powder prepared by a single drug or a combination of more drugs.^{4, 5}

Hingwastak Churna is very useful herbal powder mix used in the treatment of Indigestion in Ayurveda practice. Hingwastak Churna mainly contains very safe and easily available herbal spices. Hingu means – Asa foetida, this is the main ingredient. Ashta means 8 indicating that this product contains 8 ingredients. Reference of Hingwastak churna is found in many text books like Bhaishajya Ratnavali, Yoga Ratnakara etc.

Standardization of Ayurvedic formulations is an important step for the establishment of a consistent biological activity, a consistent chemical profile, or simply a quality assurance program for production and manufacturing of herbal drugs. WHO guidelines for the assessment of the safety, efficacy and quality of herbal medicines are a prerequisite for global harmonization ⁶.

Standardization of herbal formulation requires implementation of Good Manufacturing Practices (GMP) (WHO guideline, 1996) In addition, study of various parameters such as pharmacodynamic, pharmacokinetics, dosage, stability, self-life, toxicity evaluation, chemical profiling of the herbal formulations is considered essential. Heavy metals contaminations, Good Agricultural Practices (GAP) in herbal drug standardization are equally important⁶

MATERIAL AND METHODS:

Collection and Processing of Crude Drugs:

Hingwashtak Churna contains the crude drugs Sunthi, Marica, Pippali, Ajamoda, Sveta jirika, Krisna jirika, Hingu-suddha, Saindhav. All these plant crude drugs required for the preparation of the standard formulation of Hingwashtak churna were collected from the local market of Raipur, in the month of March, 2014. All the materials were authenticated and used for formulation. In house formulation of Hingwashtak Churna was based upon the composition given in the book of Ayurvedic Formulary of India, part -1, thoroughly mixed and then sieved through 40 mesh size to get a moderate fine powdered formulation. The prepared churna is stored in airtight container for further processing.

3 laboratory batches were formulated and compared with marketed formulation of (BAIDYANATH).

Organoleptic evaluation of Hingwashtak Churna:

The color, odour and taste of Churna were evaluated manually using sensory organs of our body and results are summarized in Table 1.

Physical Parameters:

5.3:2) Physical Parameter^7-13

Bulk density, Tapped density, Hausner’s ratio, Carr’s index, Angle of repose was determined for evaluating of physical characteristics of the Churna. The results are compiled in Table No.2

a) Bulk density:

It is the ratio of given mass of powder and its bulk volume. It is determined by transferring an accurately weighed amount of powder sample to the graduated cylinder with the aid of a funnel. The initial volume was noted. The ratio of weight of the volume it occupied was calculated.

Bulk density = w/v0 g/ml

Where,

W = mass of the powder,

V0 = untapped volume

b) Tapped density:

It is measured by transferring a known quantity (30g) of powder into a graduated cylinder and tapping it for a specific number of times. The initial volume was noted. The graduated cylinder was tapped continuously for a period of 10-15 min. The density can be determined as the ratio of mass of the powder to the tapped volume.

Tap density= w/vf (g/ml)

Where,

W = mass of the powder,

vf = tapped volume

c) Hausner’s ratio:

It indicates the flow properties of the powder. The ratio of tapped density to the bulk density of the powder is called Hausner ratio.

Hausner’s ratio = Tap density/bulk density

d) Compressibility index:

It is the propensity of the powder to be compressed. Based on the apparent bulk density and tapped density the percentage compressibility of the powder can be determined using the following formula.

Compressibility Index = [(tapped density – bulk density) / tapped density] x 100

e) Angle of repose

The internal angle between the surface of the pile of powder and the horizontal surface is known as the angle of repose. The powder is passed through funnel fixed to a burette at s height of 4 cm. A graph paper is placed below the funnel on the table. The height and the radius of the pile were measured. Angle of repose of the powder was calculated using the formula

Angle of repose= tan^-1(h/r)

Where,

h=height of the pile;

r = radius of the pile

Physiochemical Parameter ^14-22

Ash values, extractive values, loss on drying, pH, swelling index and particle size were determined. The results are compiled in Table no 3.

a) Determination of Ash value- Used to determine quality and purity of a crude drug.

Table No. 1: Organoleptic evaluation

S. No	Organoleptic Parameter	Lab Batch 1	Lab Batch 2	Lab Batch 3	Marketed Formulation
1.	Colour	Yellowish green	Yellowish green	Yellowish green	Yellowish green
2.	Odour	Characteristic and aromatic	Characteristic and aromatic	Characteristic and aromatic	Characteristic and aromatic
3.	Taste	Acrid and pungent	Acrid and pungent	Acrid and pungent	Acrid and pungent

Table No. 2: Physical Parameters

S. No.	Physical Parameter	Lab Batch 1	Lab Batch 2	Lab Batch 3	Marketed Formulation	Result for Lab Batch	Result for Marketed formulation
1.	Bulk density	0.461 gm/ml	0.454 gm/ml	0.468 gm/ml	0.468 gm/ ml	-	-
2.	Tap density	o.6 gm/ml	0.612 gm/ml	0.612 gm/ml	0. 625gm/ml	-	-
3.	Hausner’s ratio	1.30	1.34	1.30	1.33	Passable	Passable
4.	Carr’s index	23.16 %	25.81%	23.52 %	25.12%	Passable	Passable
5.	Angle of repose	50	49.4	49.34	47.46	Poor	Poor

Table No. 3: Physicochemical Parameters

S. No.	Physiochemical parameter	Lab Batch 1	Lab Batch 2	Lab Batch 3	Marketed Formulation
1.	Ash value % Total ash	18.5%	18 %	19%	19.5%
	Acid insoluble	0.5%	0.5%	1%	0.6%
	Water soluble	99%	99.05%	99.15%	99.5%
2.	Extractive value % Alcohol soluble	13.6%	12.8%	15.2%	14.4%
2.	Water soluble	28.8%	27.2%	36.8%	35.2%
3.	Loss on drying	6.4%	7.2%	8.8%	9.6%
4.	pH	7.0	7.0	7.5	7.0
5.	Particle size (80 sieve)	60% Mode. Fine	56% Mode. Fine	64% Mode. Fine	100% (Fine)
6.	Swelling factor	1 ml	1 ml	1 ml	0

Table No.4: Powder microscopy

	Powdered microscopic structures of marketed formulation
	Powdered microscopic structures of marketed formulation
	Powdered microscopic structures of marketed formulation
	Powdered microscopic structures of marketed formulation
	Powdered microscopic structure of laboratory formulation(batch 1)
	Powdered microscopic structure of laboratory formulation (Batch 2)
	Powdered microscopic structure of laboratory formulation ( Batch-3)

Table No.5: Chemical Test

S.No.	Phytoconstituent	Lab Batch 1	LAB Batch 2	Lab Batch 3	Marketed Formulation
1.	Alkaloids	--	--	--	--
2.	Saponins	+	+	+	+
3.	Cardiac glycoside	--	--	--	--
4.	Anthraquinone glycosides	--	--	--	--
5.	Carbohydrates	+	+	+	+
6.	Phenolic compound	+	+	+	+
7.	Tannins	--	--	--	--

Table No.6: Thin Layer Chromatography

S. NO.

Chemical constituent

Solvent system

Detecting agent

Rf of Lab batch 1

Rf of Lab batch 2

Rf of Lab batch 3

Rf of marketed formulation

6- gingerol

n-Hexane: Ether(40:60)

Vanillin sulfuric acid

0.43

0.42

0.45

0.41

Piperine

Benzene : Ethyl acetate : Diethyl ether(2:1:10)

Vanillin sulfuric acid

0.43

0.47

0.48

0.4

Determination of Total ash:

About 2 g of sample was accurately weighed in a tarred silica dish and kept in a muffle-furnace at a temperature not exceeding 600 C until it was free from carbon. Then it was cooled and weighed. The percentage of total ash was calculated with reference to the air dried drug.

Total Ash value % = [Wt. of total ash / air dried drug] ×100

Determination of Acid insoluble ash:

The total ash obtained was boiled for 5 minutes with 25 ml of dilute hydrochloric acid , the insoluble matter obtained was collected on an ash less filter paper, washed with hot water until the filtrate is neutral and ignited to constant weight. The percentage of acid insoluble ash was calculated with reference to the air dried drug.

Acid insoluble ash value % = [Wt. of acid insoluble ash / air dried drug] ×100

Water-soluble Ash:

The ash obtained in the determination of total ash was boiled for 5 minutes with 25 ml of water. The insoluble matter was collected on an ash less filter paper and washed with hot water. The insoluble ash was transferred into a tarred silica crucible and ignited for 15 minutes at temperature not exceeding 600 C. The weight of the insoluble matter was subtracted from the weight of the total ash. The difference in weight was considered as the water- soluble ash was calculated with reference to the air dried drug.

Water soluble ash value % = [Wt. of water soluble ash / air dried drug] ×100

b) Determination of extractive value:

Used for estimation of specific constituents, soluble in that particular solvent used for extraction.

Determination of Water-soluble extractive:

5 g of test sample was weighed and macerated with 100 ml of water in a closed flask for twenty-four hours, shaking frequently during six hours and allowing standing for eighteen hours. It was filtered rapidly, taking precautions against the loss of solvent.25 ml of the filtrate was taken and evaporated to dryness in a tarred flat bottomed shallow dish at 1050 C, to constant weight and weighed the percentage of water soluble extractive was calculated with reference to the air dried sample.

Water soluble extractive value% = [water soluble residue /air dried drug] ×100

Determination of Alcohol-soluble extractive:

5 g of test sample was weighed and macerated with 100 ml of alcohol of specified strength in a closed flask for twenty-four hours, shaking frequently during six hours and allowing standing for eighteen hours. It was filtered rapidly, taking precautions against the loss of solvent.25 ml of the filtrate was taken and evaporated to dryness in a tarred flat bottomed shallow dish at 1050°C, to constant weight and weighed the percentage of water soluble extractive was calculated with reference to the air dried sample.

Alcohol soluble extractive value= [alcohol soluble residue /air dried drug] ×100

c) Determination of loss on drying:

2.5 g of the sample (without preliminary drying) was weighed and placed in a tarred evaporating successive weighing corresponded to not more than 0.25%.

Loss on drying % = [Initial wt.-final wt. / initial wt.] ×100

d) Determination of pH:

The powder sample of Hingwashtak Churna was weighed to about 5g and immersed in 100 ml of water in a beaker. The beaker was closed with aluminium foil and left behind for 24 hours in room temperature. Later the supernatant solution was decanted into another beaker and the pH of the formulation was determined using a calibrated pH meter.

e) Determination of particle size:

25 g of sample was placed in a sieve of suitable nominal mesh aperture (sieve no.80). Sieve was shaken for not less than 30 minutes in horizontal direction and vertically by tapping on a hard surface. Weights of amount remaining on sieve were taken and determine the particle size.

Percent of sample passing through each sieve = (Wt. of sample taken - wt. of sample remaining on the sieve)/wt. of sample taken] × 100

f) Powder microscopy^{21, 22}

The Churna was taken and was examined microscopically to identify the various features like phloem fibres, parenchyma, starch grains, endosperms, calcium oxalate crystals, oil glands etc by using various reagents.Results are compiled in Table 4.

Phytochemical screening of constituents:

Chemical test-^{18, 23}

Laboratory and market formulations of Hingwashtak Churna were subjected to the following tests separately for the presence of various phytoconstituent like alkaloids, saponins, carbohydrates, terpenoids, anthraquinone glycosides, tannins and phenolic compounds.Results are summerized in Table 5.

Thin Layer Chromatography (TLC)^{18, 23}

Thin layer chromatography is particularly valuable for the qualitative determination. TLC is a technique in which a solute undergoes distribution between two phase, a stationary phase acting through adsorption and a mobile phase in the form of liquid. The adsorbent is relatively thin uniform layer of dry finely powdered material, applied to a glass plate are the most commonally used. Separation may also be achieved on the basis of partition or a combination of partition and adsorption depending on the particular types of support its preparation and its use with different solvent system.

TLC Study:

TLC is an important tool by which the quality control and fingerprint of herbs can be maintained. It also helps to identify the presence or absence of the individual herbs in combination herbal formulations. Hence, the TLC profiling of Hingwashtak Churna was carried out to develop unique TLC spots in the formulation as an identifier of its every ingredient. Results of the same are summerized in Table 6.

Estimation of 6-gingerol in HINGWASHTAK CHURNA by TLC procedure:²³

Sample preparation-The powdered sample extracted with chloroform. The extract was concentrated and dissolved in chloroform.

Solvent system - n-Hexane: Ether (40: 60)

Detecting agent -Vanillin sulfuric acid

Rf value - 0.41

Estimation of Piperine in HINGWASHTAK CHURNA by TLC procedure:²⁴

Sample preparation-The powdered sample extracted with methanol. The extract was concentrated and dissolved in methanol.

Solvent system - Benzene: Ethyl acetate: Diethyl ether (2:1:10)

Detecting agent -Vanillin sulphuric acid

Rf value - 0.40

Formula, Rf value = Distance travelled by solute / Distance travelled by solvent

CONCLUSION:

In the present work, classical Ayurvedic formulation Hingwashtak Churna was characterized by Pharmacognostic, physicochemical, pharmaceutical, microbiological, toxicological and chromatographic parameters as per the present standards of Ayurvedic Pharmacopoeia of India (API). Standardization guidelines provided by World Health Organization (WHO), European Agency for Evaluation of Medicinal Products (EMEA) and the United States Pharmacopoeias (USP) were also considered and the results obtained was found to be within the standard for crude drugs and if the crude drug evaluation is observe under the limit so the formulation can be prescribed as standard [22-24]. This study may be utilised for rapid authentication fingerprints for the formulation, achieving batch-to-batch consistency and preparing a concise quality monograph for Hingwashtak Churna. The result of present study will also serve as reference monograph for the quality control of Hingwashtak churna which will also assist the regulatory authorities, scientific organizations and manufacturer in developing standards in the preparation of drug formulation.

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Received on 28.03.2018 Modified on 19.04.2018

Research J. Pharm. and Tech 2018; 11(4):1319-1326.

DOI: 10.5958/0974-360X.2018.00246.9