INTRODUCTION:

There is increase in acceptability of herbal drugs in present era. Therefore there is need to enlighten users of herbal medicine regarding quality parameters for collection, handling, processing and production of herbal medicine. Also it is necessary to employ such parameters in ensuring the safety .Standardization and quality control depends upon the nature of crude drug and compound drugs.

Following are the problems which may influence the quality of herbal product.¹

1.Herbal drugs are usually mixtures of many constituents.

2.The active principles in most cases are unknown.

3.Selective analytical method ,reference compound may not be available.

4.Plant material are variable.

5.Method of harvesting, drying, storage, transportation and processing may also affect quality of product.

Who guidelines for quality standardized herbal formulations²

WHO Guidelines for standardization of herbal formulation are

1) Quality control of crude drugs material, plant preparations and finished products.

2) Stability assessment and Safety assessment

3) Shelf life, documentation, toxicological studies.

4) Assessment of efficacy proper evaluation.

Standardization and quality control of herbal crude drugs –

Parameters which are paid due attention are

1. Macro and microscopic examination:

To Identify right variety of species and to search adulterants.

2. Foreign organic matter:

This involves removal of matter other than source plant to get the drug in pure form.

3. Ash values:

These criteria are to judge and identify purity of crude drug – Total ash, sulphated ash, water soluble ash and acid insoluble ash etc.

4. Moisture content:

Checking moisture content helps to reduce errors in the estimation of the actual weight of drug material. Low moisture suggests better stability and lowers the degradation of product.

5. Extractive values:

These are indicative weights of the extractable chemical constituents of crude drug under different solvents environment.

6. Crude fiber:

This helps to determine the woody material component.

7. Qualitative chemical evaluation:

It employs different analytical technique to detect and isolate the active constituents. Phytochemical screening techniques involve botanical identification, extraction with suitable solvents, purification, and characterization of the active constituents of pharmaceutical importance.

8. Chromatographic examination:

Include identification of crude drug based on the use of major chemical constituents as markers.

9. Quantitative chemical evaluation:

To estimate the amount of the major classes of constituents.

10. Toxicological studies:

This helps to determine the pesticide residues, potentially toxic elements, safety studies in animals like LD50 and microbial assay to establish the absence or presence of potentially harmful microorganisms.

2. INTRODUCTION OF SAMPLE:

2.1 Sample name :-Gulvel churna

Biological name:- Tinospora cordifolia

Family:- Menispermaceae

Vernacular names:- ³

Table no 1. Vernacular names of Tinospora cordifolia

Hindi	Amrita, giloe, gulancha, gulbel, guloh, gurcha, jiwantika
Marathi	Gulvel
Sanskrit	Amrita, guduchi, jwarari
Telugu	Tippateege
Tamil	Amudom, chindil

2.2 Main constituents of Gulvel ⁴

A. Terpenoids:

Terpenoids which are present in Gulvel are:- Tinosporide, Furanolactone diterpene, Furanolactone clerodane diterpene, furanoid diterpene, Tinosporaside, ecdysterone makisterone and several glucosides isolated as poly acetate, phenylpropene disaccharides cordifolioside A, B and C, cordifoliside D and E, Tinocordioside, cordioside, palmatosides C and F, Sesquiterpene glucoside tinocordifolioside, Sesquiterpene tinocordifolin.

B. Alkaloids:

Following Are The alkaloidal content:- Tinosporine, (S), Magnoflorine, (S), Tembetarine, (S), Berberine, (S, Choline, (S) Palmatine, (S), Jatrorrhizine, (S), 1,2-Substituted pyrrolidine, (S), Alkaloids, viz. jatrorrhizine, palmatine, beberine, tembeterine, choline.

C. Steroids:

sterois which are present in gulvel are Giloinsterol, (S), ß-Sitosterol, (S), 20a- Hydroxy ecdysone, (S).

D. Glycosides:-

neoandrogapholide and andrographiside

E.Flavonoids :-

oroxylin,wogonin,andrographinidine A,B,C,D,E,F

2.3 Uses of Gulvel:

Table no 2. Uses o tinospora cordifolia

i.Anti-cancer/anti-tumour activity	vi. Urinary calculi
ii.Anti Diabetic and Hyperglycaemic activity	vii. Digestive activity
iii.Anti-inflammatory activity:	viii. Hypolipidaemic activity
iv.Antioxidant activity:	ix. Immunobiological activities:
V. Anti-stress activity	x. Anti Ulcer activity:

3.MATERIALS AND METHODS:

The marketed samples of various brands of Gulwel Churna i.e. Baidhyanath and Green Pharmacy were collected from the local market.Other chemicals of analytical grade were used.

4. STANDARDIZATION PARAMETERS FOR GULVEL CHURNA:

4.1 Study of organoleptic character:

The herbal formulation is studied for organoleptic characters like color, odour and taste using the sensory organs .

4.2 Determination of physico-chemical parameters:

i. Determination of loss on drying:

1.5 g of the sample was weighed and placed in a tarred evaporating dish. It was dried at 105˚C until difference between two successive weighing corresponded to not more than 0.5 mg.

ii. Total ash:⁵

Total ash determination constitutes detecting the physiological ash (ash derived from plant(tissue) and nonphysiological ash (ash from extrageneous matter, especially sand and soil adhering to the surface of the drug). For its detection, 2g of powdered material of each formulation and the individual ingredients of the powers were placed separately in a suitable tare crucible of silica previously ignited and weighed. The powdered drugs were spread into an even layer and weighed accurately The materials were incinerated by gradually increasing the heat, not exceeding 450°C until free from carbon, cooled , weighed.

iii. Acid insoluble ash:

The ash obtained as above was boiled for 5min with 25ml of dilute hydrochloric acid; The insoluble matter was collected on an ash less filter paper, washed with hot water and ignited to constant weight. The percentage of acid-insoluble ash with reference to the air-dried drug was calculated.

iv. Water soluble ash:

This is determined in similar way to acid insoluble ash ,using 25 ml of water instead of dil. HCl.

v. Water soluble extractive:

4g of coarsely powdered air-dried drug was macerated with 100ml of chloroform water in a closed flask for twenty-four hours, shaking frequently during six hours and allowed to stand for eighteen hours. It was then filtered rapidly, taking precautions against loss of solvent. 25ml of the filtrate was evaporated to dryness in a tare flat bottomed shallow dish at 105°C to constant weight and weighed. The percentage of water-soluble extractive was calculated with reference to the air-dried drug and is represented as % value

vi. Alcohol soluble extractive:

5g of coarsely powdered air-dried drug was macerated with 100ml of alcohol in a closed flask for twenty-four hours, shaking frequently during six hours and allowed to stand for eighteen hours. It was then filtered rapidly; taking precautions against loss of solvent. 25ml of the filtrate was evaporated to dryness in a tarred flat-bottomed shallow dish at 105°C to constant weight and weighed. The percentage of alcohol-soluble extractive was calculated with reference to the air dried drug and is represented as% value.

5. Quantitative estimation of selected phyto- constituents

Table no.3 Tests for quantitative estimation of phyto-constituents

Alkaloids	1.Dragendroff test 2.Wagner’s test 3. Mayer’s test
Flanavoids	Shinoda test
Poly phenols	N FeCl3 test
Saponins	Foam test
Steroids	Liebermann’s test
Sugars	Molish test

6 Evaluation of Churna:⁶

i. 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. Theinitial 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

ii. Tap density:

It is measured by transferring a know 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.

iii. 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

vi. 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,

% compressibility=

(tapped density – bulk density) x 100

tapped density]

v. 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

6.1 Determination of pH:

The powder sample of 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

6.2. Chromatographic studies:

Chromatographic studies was done taking chloroform and methanol (9:1) as a solvent system and anisaldehyde–sulphuric acid as spraying reagent⁹.

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

For Cadmium

Table no 4. Tests for evaluating churna for presence of cadmium

Test	Observation
NH4OH added in the sample solution.	White ppt of cadmium hydroxide soluble in excess NH4OH
Potassium ferrocyanide added.	White ppt of cadmium ferrocyanide

For Bismuth

Table no 5. Tests for evaluating churna for presence of bismuth

Test	Observation
H₂S gas added in the sample solution	Dark brown ppt soluble in hot dil. HNO₃ but insoluble in NH₄S
NH₄OH	white ppt insoluble in excess NH₄OH dissolved in dil. HCl.

For Lead

Table no 6. Tests for evaluating churna for presence of Lead

Test	Observation
Dil HCl added in sample solution.	White ppt of CaCl₂ soluble in boiled water and conc. HCl
KI is added in sample solution.	Yellow ppt soluble in boiling water.

7. RESULTS AND DISCUSSIONS

7.1 Determination of organoleptic characters

Table no.7 Organoleptic characters of both formulations

Organoleptic property	Formulation A	Formulation B
Appearance	Powder	Powder
Color	Whitish-yellow	Whitish-yellow
Odor	None	None
Taste	Bitter	Bitter

7.2 Physico Chemical Standards

Table no.8 Physico chemical characteristics of both formulation

Parameters	Formulation A	Formulation B
Loss on Drying	6.7%		6.2%
Total Ash value	3.8 %		3.68%
Acid insoluble ash	0.19 %		0.28%
Water soluble extractive	13.78%		12.75%
Alcohol soluble extractive	4.56%		5.5%
Foreign matter	0.5%		0.8%

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

Table no 9.Quantitative estimation of phytoconstituents

Sr.no	Chemical constituents	Tests	Observation
1	Alkaloids	Dragendroff test	+ ve
2	Flavanoids	Shinoda test	+ ve
3	Steroids	Liberman test	+ ve
4	Polyphenol	Neutral FeCl₃ test	+ ve
5	Saponin	Foaming test	+ ve
6	Sugar	Molish test	+ve

7.4 Evaluation of Churna

Table no.10 Powder characteristics of both formulations

Sr.no	Parameter	Formulation A	Formulation B
1	Bulk density	0.416	0.58
2	Tap density	0.714	0.82
3	Hausner’s ratio	1.717	1.413
4	Compressibility Index	13.13	11.26
5	Angle of repose	29⁰	31⁰

7.5 Determination of pH

pH of water extract of Formulation A was found to be 6.8

pH of water extract of Formulation B was found to be 5

7.6 Heavy metal tests for both the formulations

Table no.11 Results of heavy metal tests of both formulation

Sr. no	Test	observation	Results
1	Lead	White ppt of CaCl₂ is absent. when dil. HCl is added	Lead absent
		Yellow ppt is absent when KI is added
2	Cadmium	White ppt is Absent when NH₄OH and potassium ferrocyanide is added	Cadmium absent
3	Bismuth	Dark brown ppt is absent.when H₂S gas is added	Bismuth absent
		White ppt is Absent when NH₄OH is added

7.7 Chromatographic studies

On TLC identity test the drug showed major molecules having Rf value 0.24 (yellow)

8. CONCLUSION:

In 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, TLC profile 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, to seek marketing approval from regulatory authorities for therapeutic efficacy, safety and shelf- life of herbal drugs.

9. REFERENCES:

1. Kunle, Oluyemisi Folashade, Egharevba, Henry Omoregie. Standardization of herbal medicines - A review. International Journal of Biodiversity and Conservation Vol. 4(3), , March 2012 pp. 101-112

2. Arun Rasheed, Sravya Reddy B, A review on standardization of herbal formulation. International Journal of Phytotherapy 2 ( 2) 2012 p 75

3. S. Selvarajan, V. Gayathri Devi Pharmacognostical studies of Tinospora cordifolia (miers.) Hk. F and th. (stem) International Journal of Pharmacy and Technology 6 (3) jan-2015. 7065-7066

4. Kirti Sinha, N P Mishra Tinospora cordifolia (Guduchi), A Reservoir Plant For Therapeutic Applications: A Review Indian Journal of Traditional Knowledge Vol. 3(3), July 2004, Pp. 267-270

5. Sangram Keshari Panda Standardization of Sitopaladi Churna: A Poly-Herbal Formulation Scholars Research Library Der Pharmacia Lettre, 2012, 4 (1):207-209

6. Pallab Dasgupta and Amartya De. Comparative Standardization Study of Two Marketed Ashwagandha Churna Formulation International Journal of Research in Pharmaceutical and Biomedical Sciences Vol. 3 (2) Apr – Jun2012 p 736-737

Received on 04.01.2017 Modified on 28.01.2017

Res. J. Pharm. Dosage Form. & Tech. 9(1): Jan.-Mar. 2017; Page 24-28.

DOI: 10.5958/0975-4377.2017.00005.2