Comparative Standardization Study of Two Marketed Shatavari Churna Formulation

 

Rohan R. Vakhariya*, Swati Talokar, Archana R. Dhole,  Dr. S.K. Mohite, Dr. C.S. Magdum

Rajarambapu College of Pharmacy, Kasegaon. 415404

*Corresponding Author E-mail:

 

ABSTRACT:

Majority of Ayurvedic practitioners uses traditional herbal preparations for their treatment purposes made by them. So it is necessary to improve safety of herbal drugs by developing certain quality control parameters and by following the WHO guidelines for herbal medicines. Our adulteration.  Now days, old methods are  necessary but there are lots of limitations to these methods due to shortage  of many drugs,  unavailability or limited source, adulteration, lack of knowledge of drug identification and adverse effects of drugs etc. Standardization of herbal formulation is essential in order to assess the quality of drugs for therapeutic value. The World Health Organization (WHO) in 1999 has given a detail protocol for the standardization of herbal drugs comprising of a single content, but very little literature is available for the standardization of Shatavari Churna. Two marketed preparations were used for the study. Performed the various parameters including organoleptic characteristics and physicochemical parameters. The set parameters were found to be sufficient to standardize the Shatavari Churna.

 

KEYWORDS: Traditional, Standardization, Shatavari Churna, Physicochemical parameters.

 


 

INTRODUCTION:

India has a rich heritage of traditional medicine constituting with its different components like Ayurveda, Siddha and Unani. Traditional health care has been flourishing in this country for many centuries. Ayurveda and other Indian systems of medicines may be explored with the modern scientific approaches for better leads in the health care. Ayurveda has earth-shattering in audience of worldwide on virtue of its holistic approach of life and its fewer side effects. Ayurvedic herbal and herbo-mineral preparations are used for the treatment of chronic and degenerative diseases without any side-effect1. In the last few decades, there has been an exponential growth in the field of ayurvedic medicine .There are great need of standardization and quality control of ayurvedic formulations.

 

Standardization and quality control depends upon the nature of crude drug and compound drugs, it’s source i.e. factors associated with raw materials which are beyond of human control like seasonal, geographical, age of the plant, time of collection, type of drying etc. due to these natural conditions. The percentage of chemical constituents2,3of the drug does no remain uniform as our expectation. The need of quality control for ayurvedic drug is due to the fact that the preparation of drug according to the ancient method has been reduced due to the commercialization of ayurvedic pharmacy The absence of post-market surveillance and the paucity of test laboratory facilities also make the quality control of ayurvedic medicines exceedingly difficult at this time.4 The individual plant powders of the formulation were subjected to various pharmacognostical parameters. Two formulation, from different manufactures were procured and subjected to various physicochemical analysis.

 

Advantages of Herbal Medicine:

·       They have large amount of use.

·       They have better patient tolerance as well as acceptance.

·       The medicinal plants have renewable source of cheaper medicines.

·       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 and efficacy.

·       They are cheap in cost.

·       They are not harmful.

·       They are more effective than any synthetic drug.

·       Throughout the world herbal medicines have provided many of the most potent medicines to the vast arsenal of drugs available to modern medical science, both in crude form as well as a pure chemical upon which modern medicines are constructed.

 

Herbal Drug Standardization:

Standardization of herbal formulations is essential to assess quality of drugs. It is based on the concentration of their active principles, physical, chemical, Phytochemical, and In-vitro, In-vivo parameters5. The quality assessment of herbal formulations is important to justify their acceptability and safety. One of the major problems faced by the Ayurveda physicians is the unavailability of unique quality control parameters for herbal medicines and their formulations. In India, the department of AYUSH Government of India launched a central scheme to develop standard operating procedures for the manufacturing process to develop pharmacopeia standards for Ayurvedic preparations. The subject of herbal drug standardization is massively wide and deep. There are so many contradictory theories on the subject of herbal medicines and their functions on human physiology and mental function. India needs to explore the medicinally important plants. This can be achieved only if the herbal products are evaluated and analyzed using sophisticated modern techniques of standardization.

 

Standardization is a system that ensures a predefined amount of quantity, quality and therapeutic effect of ingredients in each dose6. Herbal product cannot be considered scientifically valid if the drug tested has not been authenticated and characterized in order to ensure reproducibility in the manufacturing of the product. Moreover, many dangerous and lethal side effects have recently been reported, including direct toxic effects, allergic reactions, effects from contaminants, and interactions with herbal drugs6. Therapeutic activity of an herbal formulation depends on its phytochemical constituents. The development of authentic analytical methods which can reliably profile the phytochemical composition, including quantitative analyses of marker/ bioactive compounds and other major constituents, is a major challenge to scientists. In view of the above, standardization 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 an herbal drug. The authentication of herbal drugs and identification of adulterants from genuine medicinal herbs are essential for both pharmaceutical companies as well as public health and to ensure reproducible quality of herbal medicine.

 

1 Conventional methods for standardization of herbal formulation:

Standardization of herbal raw drugs include passport data of raw plant drugs, botanical authentification, microscopic and molecular examination, identification of chemical composition by various chromatographic techniques and biological activity of the whole plant5. Macroscopic and microscopic evaluation and chemical profiling of the herbal materials for quality control and standardization have been reported by various workers7,9. Macroscopic identity of medicinal plant materials is based on sensory evaluation parameters like shape, size, colour, texture, odour and taste while microscopy involves comparative microscopic inspection of powdered herbal drug. Further, advances in microscope technology have increased the accuracy and capabilities of microscopy as a mean of herbal crude material identification due to the implication of light and scanning electron microscopes (SEM) in herbal drug standardization. Furthermore, various advanced methods such as chromatographic, spectrophotometric and combination of these methods, electrophoresis, polarography, and the use of molecular biomarkers in fingerprints are currently employed in standardization of herbal drugs 9-11.

 

2 Standardization of herbal formulation:

Standardization of herbal formulation requires implementation of Good Manufacturing Practices (GMP) 7-9. In addition, study of various parameters such as pharmacodynamics, pharmacokinetics, dosage, stability, shelf-life, toxicity evaluation, chemical profiling of the herbal formulations is considered essential. Other factors such as pesticides residue, aflatoxine content, heavy metals contamination, Good Agricultural Practices (GAP) in herbal drug standardization are equally important.

 

Scientific classification of Shatavari-

Kingdom: Plantae

Clade: Angiosperms

Clade: Monocots

Order: Asparagales

Family: Asparagaceae

Subfamily: Asparagoideae

Genus:Asparagus

Species:A. racemes

 

Chemical Constituents of Shatavari-Amino Acid:

Asparagine, Tyrosine and Arginine, plus succinic acid and a methylsulfonium derivative of methionine.

 

Flavonoids- kaempferol, quercitin, rutin

 

Vitamins-A, B1, B2, C, E, Mg, P, Ca, Fe, and folic acid.

Steroidal Saponin.

Resins.

Tannins.

 

Uses of Shatavari:

1.     Shatavari supports reproductive health by toning and nourishing the female reproductive organs

2.     Shatavari treats PMS symptoms by relieving pain and controlling blood loss during menstruation.

3.     Shatavari supports normal production of breast milk for nursing mothers

4.     Shatavari relieves menopausal symptoms such as hot flashes. By producing estrogens it makes up for low estrogen levels in women who are menopausal or have had hysterectomies or oophorectomies

5.     Used in India as a best-known and most often-used herb for women

6.     Shatavari supports normal function of the immune and digestive system

 

PLAN OF WORK:

Comparative standardization of Shatavari Churna formulated by Sharangdhar Pharmaceuticals Pvt.Ltd. and Local Product was planned to carry out development of quality standards for the finished marketed formulation. The method used for the comparative standardization was planned to be carried out as follows:

 

Development of standardization parameters for shatavari Churna:

1 Study of organoleptic characters

·       Colour

·       Odour

·       Taste

 

2 Determination of physicochemical parameters:

·       Moister content

·       Total ash

·       Acid insoluble ash

·       Water soluble ash

·       Water soluble extractive

·       Alcohol soluble extractive

·       Crude fiber contents

 

3 Quantitative estimation of selected phytoconstituents:

Total alkaloids

4 Evaluation of Churna:

·       Powder fineness

·       Bulk density

·       Tap density

·       Angle of repose

·       Compressibility

·       Hausner ratio

 

5 Determination of Ph:

6 Establishing the safety pertaining to Heavy metals and Microbial load:

Experimental Work:

1 Study of Organoleptic Characters:

The herbal formulation is studied for organoleptic characters like color, odour and taste

using the sensory organs of our body.

 

2 Physico-chemical analysis12:

Determination of loss and drying:

10 g of the sample (without preliminary drying) was weighed and placed in a tarred evaporating dish. It was dried at 105˚ C for 5 hours, and at 1 hour interval until difference two successive weighing corresponded to not more than 0.25%.

 

Determination of Total ash:

About 2 to 3 g of sample was accurately weighed in a tarred silica dish at a temperature not

exceeding 450 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.

 

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 and ignited to constant weight. The percentage of acid in soluble ash was calculated with reference to the acid dried drug.

 

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 a temperature not exceeding 450 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.

 

Determination of Water-soluble extractive

5 g of test sample was weighed and macerated with 100 ml of chloroform 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.

 

Determination of Alcohol-soluble extractive

Procedure for water soluble extractive was followed for the determination of alcohol soluble

extractive but 90% ethanol was used instead of chloroform water.

 

3 Qualitative Phytochemical Screening13,14,15 Detection of alkaloids

50 mg of solvent free extract was hydrolyzed with dil.HCL and filtered. The filtrates were tested

carefully with various alkaloid test reagents as follows

 

1. Dragendroff’s test

To few ml of filtrates, 1 to 2 ml of dragendroff’s reagent was added. A prominent yellow precipitate indicates the test is positive.

 

2. Wagner’s test

To few ml of filtrates, few drops of wagner’s reagent were added by the side of the test tube. A reddish-brown precipitate confirms the test as positive.

 

3. Mayer’s test

To few ml of filtrates, few drops of mayer’s reagent were added by the side of the test tube. White or creamy precipitates if obtained indicate the presence of alkaloids.

 

4. Hager’s test

To few ml of filtrates, few drops of hager’s reagent were added. A prominent yellow precipitate indicates the test is positive.

 

4 Determination of physical characteristics16:

Bulk density:

It is the ratio of given mass of powder and it’s 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

 

Tapped density:

It is measured by transferring a known quantity (25g) 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.

 

Tapped volume= w/vf g/ml

Where,

W = mass of the powder

Vf = tapped volume.

 

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=[(v0-vf)/v0] x 100,

Or

% compressibility=[(tapped density – bulk density)]/ tapped density] x 100

 

Hausner 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 ratio= Tapped density/bulk density

 

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 was 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

 

5 Determination of PH range

The powder sample of Shatavari Churna was weighed to about 5g and immersed in 100 ml of water in a beaker. The beaker was closed with aluminum foil and left behind for 24 hour s 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.

 

6 Heavy metals test17

For Cadmium

Experiment

Observation

Result

NH4OH added in the sample solution.

White ppt of cadmium hydroxide soluble in excess NH4OH

P/0 cadmium

Potassium ferrocyanide added

White ppt of cadmium ferrocyanide

P/0 cadmium

 

For Bismuth

Experiment

Observation

Result

H2S gas added in the sample solution

Dark brown ppt soluble in hot dil. HNO3 but insoluble in NH4S

p/o bismuth

NH4OH

white ppt insoluble in excess NH4OH dissolved in dil. Hcl.

p/o bismuth

 

For Lead

Experiment

Observation

Result

Dil. HCL added in sample solution

White ppt of CaCl2 soluble in boiled water and conc. HCL

p/o lead

KI is added in sample solution

Yellow ppt soluble in boiling water

p/o lead

 

RESULTS AND DISCUSSIONS:

1. Determination of organoleptic characters

Organoleptic property

Shatavari Churna of Sharangdhar

Shatavari Churna Of Local Market

Appearance

Powder

Powder

Color

Whitish-yellow

Whitish-yellow

Odor

None

None

Taste

Slightly Bitter

Slightly Bitter

 

2. Physicochemical Standards

Parameters

Shatavari Churna of Sharangdhar

Shatavari Churna of Local Market

Loss on Drying

7.3%

9.2%

Total Ash value

3.7 %

3.8%

Acid insoluble ash

0.23 %

0.37%

Water soluble ash

2.2%

2.6%

Water soluble extractive

14.73%

16.23%

Alcohol soluble extractive

6.76%

5.35%

 


 

3. Quantitative estimation of selected phyto-constituents in both the formulations

Sr.

no.

Chemical constituents

Tests

Shatavari Churna of Sharangdhar

Shatavari Churna of Local Market

1

Alkaloids

Dragendroff test

 - ve

- ve

2

Flavanoids

Shinoda test

+ ve

+ ve

3

Steroids

Liberman test

+ ve

+ ve

4

Polyphenol

Neutral FeCl3 test

- ve

- ve

5

Saponin Glycoside

Foaming test

+ ve

+ ve

6

Sugar

Molish test

+ve

+ve

 

4. Determination of Physical Characteristics of Churna

Sr. No

Parameter

Shatavari Churna of Sharangdhar

Shatavari Churna of Local Market

1

Bulk density

0.45 g/mL

0.47 g/mL

2

Tap density

0.52 g/mL

0.55 g/mL

3

Hausner’s ratio

1.16

1.17

4

Compressibility Index

13.46%

14.54%

5

Angle of repose

32.5o

34.2o

 

5. Determination of PH

Sr. No

Parameter

Shatavari Churna Of Sharangdhar

Shatavari Churna Of Local Market

1

PH

6.8

5.3

 

6. Heavy metal tests for both the formulations

Sr.

no

Test

observation

Shatavari Churna of Sharangdhar

Shatavari Churna of Local Market

1

Lead

White ppt of CaCl2 is absent. When  dil.HCL is added

Lead absent

Lead absent

Yellow ppt was absent when KI is added

2

Cadmium

White ppt was Absent when NH4OH  and potassium ferrocyanide is added

Cadmium absent

Cadmium absent

3

Bismuth

Dark brown ppt was  absent. When H2S gas is added

Bismuth absent

Bismuth absent

White ppt is Absent when NH4OH is added

 


 

CONCLUSION:

From the present investigation various standardization parameters such as physicochemical standards like total ash, acid insoluble ash, water and alcohol soluble extractive values, loss on drying, phyto-chemical analysis, flow properties and safety evaluation were carried out. The comparative study of two marketed preparation did not show much deviation in their results therefore it can be concluded that they contain character of ideal Churna. The study shows that the contents of formulation presented are within the permissible limits.

 

The advancement of analytical techniques will serve as a rapid and specific tool in the herbal research, thereby, allowing the manufacturers to set quality standards and specifications so as to seek marketing approval from regulatory authorities for therapeutic efficacy, safety and shelf- life of herbal drugs.

 

REFERENCES:

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2.     P. K. Mukherjee, A. Wahile, Journal of Ethnopharmacology, 2006, 103, 25–36.

3.     L. V. Asokar, K. K. Kakkar, O. J. Chakra, Glossary of Indian medicinal plants with active principles, Publication and Information Directorate, New Delhi, 1992.

4.     Anonymous , The Ayurvedic Formulary of India, Govt. of India, Ministry of Health and Family Welfare, New Delhi, 1976.

5.     Arun Rasheed, A Review on standardization of herbal formulations, Inter. J. of Phytotherapy / Vol. 2 / Issue 2 / 2012 / 74-88.

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12.  Anonymous, “Indian Pharmacopoeia” 1996. Govt. Of India, Ministry of Health, Controller of Publication, Delhi, India.

13.  Dr. Khandelwal K.R, Practical Pharmacognosy, Techniques and Experiments. Nirali Prakashan, Twelfth edition, Pages 149-155.

14.  J.B.Harborne, Phyotochemical methods- A Guide to Modern Techniques of Plant Analysis. Third edition. Pages 3-31.

15.  Evans 1997. Trease and Evans Pharmacognosy. 16th edition. Harcourt Brace and Company Asia Pvt. Ltd. Singapore. PP.343.

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Received on 29.02.2016          Accepted on 20.03.2016        

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Ana. 6(1): January- March, 2016; Page 1-6

DOI: 10.5958/2231-5675.2016.00001.6