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:
1.
Rohan R. Vakhariya , Swati S Talokar
, Archana R Dhole, C.S Magdum. Herbomineral Formulations - A Review 2015. IJSRST ;
Volume 1 : Issue 3:161-169.
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.
6.
www.phytotherapyjournal.com
7.
Indian Herbal
Pharmacopoeia, Indian Drug Manufacturers’ Association, Mumbai, 2002.
8.
British Herbal
Pharmacopoeia, British Herbal Medicine Association, 1996.
9.
Quality Control
Methods for Medicinal Plant Materials, WHO, Geneva, 1996.
10. Bhutani KK. Herbal medicines enigma and a challenge for
science and guidelines for new initiatives. J Nat Prod 2003;19(1):
3-8.
11. Svicekova M, Havranek E, Novak V.
Determination of heavy metals in samples of herbal drugs using differential
pulse polarography. J Pharm
Biol 1993; 42(2):68-70.
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.
16.
“Indian
Pharmacopoeia”. 3rd edition, 1985. Pharmaceutical Chemistry, Inorganic- G.R Chatwal, PP.419-422.
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