INTRODUCTION:

India has a rich heritage of traditional medicine constituting with its different components likeAyurveda, Siddha and Unani. Traditional health care has been flourishing in this country formany centuries. Ayurveda and other Indian systems of medicines may be explored with themodern scientific approaches for better leads in the health care^[1]. In the last few decades, therehas been an exponential growth in the field of Ayurvedic medicine.

There are great need ofstandardization and quality control of ayurvedic formulations. Standardization and qualitycontrol depends upon the nature of crude drug and compound drugs, it’s source i.e. factorsassociated 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 constituents^[2]of 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 theancient method has been reduced due to the commercialization of ayurvedic pharmacy Theabsence of post-market surveillance and the paucity of test laboratory facilities also make thequality control of aurvedic medicines exceedingly difficult at this time. Therefore, an attempt has been made to standardize sitopaladiChurna, an Ayurvedic formulation as prescribed in Ayurvedic Formulary, used as anti tussive, common cold etc.. The individual plant powders ofthe formulation were subjected to various pharmacognostical parameters. ingredients as controls. Ayurvedic science has got its rich heritage in India. People in India believe that natural products are safe compared to synthetic drugs. The development in these traditional systems of medicine leads to maintain proper quality of the product.

These plants are being used for curing many diseases as such in raw condition rather the being prepared as formulation; Standardization is an essential parameter to be done. It is a vital step in formulation since it determines the quality of the product and is essential to develop a protocol on standardization of every product available in the market to avoid variation arising between batch to batch. Plant materials are not like synthetic drugs, they vary in many conditions even in their chemical content depending on the time and season of collection of plant material, the geographical location of the plant being grown etc. the production batches^[3]. Good manufacturing practices and quality control of the ingredients and products can result in ensuring quality assurance of the formulation.

The present study is to standardize a polyherbal formulation available in the market called as Shitopaladi Churna used to treat many ailments of the body. The churna is evaluated for organoleptic properties, physical properties, physiochemical parameters and phytochemical screening to standardise the same. It is recipe of traditional Ayurvedic Pharmacopoeia well known and effective in relieving coughs associated with respiratory disorder. Dispensing and consumption of churna (powder) is inconvenient to the patients. Churna are stick to the tongue and oral cavity due to inherent adhesive nature. Patients are showing less interest to take herbal churna orally because of their astringent, bitter and pungent taste. Churna being in powder form also suffer stability due to their hygroscopicity.

MATERIAL and METHODS:

All the chemicals used in the experiment are analytical gread, like dilute HLC, Benzene, chloroform, acetone,

Plant material:

Sitopaladichurna consist of like Sitopal, Vamsolochana(vamsa) Pippali(Piperlongumfruit), Ela(Elettariacardamomum seed), Tvak(Cinnamomumzeylanicumstem bark) All these ingredients were procured from the local Raw traders of Islampur, Maharashtra, India.

Preparation of Sitopaladi:

In house formulation of sitopaladi churna was prepared as per Ayurvedic Formulary of India Take all the ingredients ^[4]

Table 1: Formulation table

Ingredients	Quantity
Sitopal (cane sugar )	48gm
Vamsolochna (vamsa )	24gm
Pippali ( piper lonugm)	12gm
Ela (elettaria cardamom seed )	6gm
Tavk ( cinnamomumzeylanicum stem bark )	3gm

Procedure:

12gm of Powder separately ingredients and pass through sieve number 80 #. Weigh separately each powdered ingredient and mix together. Pass the churn through sieve number 44 # to prepare a homogeneous blend. Pack it in tightly closed containers to protect from light and moisture.

EVALUATION OF CHURNA:

Orgnoleptic evaluation:

Organoleptic Evaluation refers to evaluation of formulation by

Colour

Odour

Taste

Texture

Physicochemical Investigation^[4]

Determination of total ash:

Total ash determination constitutes detecting the physiological ash (ash derived fromplant(tissue) and non-physiological ash (ash from extraneous matter, especially sand and soiladhering to the surface of the drug). For its detection, 2g of powdered material ofEachformulation and the individual ingredients of the powers were placed separately in a suitabletarred crucible of silica previously ignited and weighed.

The powdered drugs were spread intoaneven layer and weighed accurately. The materials were incinerated by gradually increasingtheheat, not exceeding 450°C until free from carbon, cooled in a desiccator, weighed and percentage ash was calculated by taking in account the difference of empty weight of crucible andthat of crucible with total ash.

Fig. 1: Total ash

Acid insoluble ash:

The ash obtained as above was boiled for 5min with 25ml of dilute hydrochloric acid thein-soluble matter was collected on an ashless filter paper, washed with hot water and ignitedToconstant weight. The percentage of acid- insoluble ash with reference to the air-dried drugwascalculated.

Fig. 2: Acid insoluble ash

Water soluble ash:

The ash was boiled for 5 minutes with 25 ml of water; collected insoluble matter in an ash lessfilter paper, washed with hot water, and ignited for 15 minutes at a temperature not exceeding450C. Subtract the weight of the insoluble matter from the weight of the ash; the difference inweight represents the water-soluble ash. The percentage of water-soluble ash with referencetothe air-dried drug was calculated.

Loss on drying:

Loss on dying is the loss of mass expressed as percent w/w. About 10g of dug samples of each formulation was accurately weighed in a dried and tare flat weighing bottle and dried at 105 C.

Determination of pH:

The pH of different formulations in 1% w/V and 10% w/V of water soluble portions was determined using standard glass electrode at 240 according to the prescribed standard method in Indian Pharmacopoeia.

Fig. 3: Determination of pH

Phytochemical investigation:

Tests for Carbohydrates:

A reducing sugar is one that can be oxidized. In order to be a reducing sugar, the molecule must contain a free anomeric carbon, since it is the open-chain form of the aldehyde that is able to react (and be oxidized). One test for reducing sugars involves Fehling’s reagent, which contains Cu2+ ions in an aqueous basic solution. If a reducing agent is present, the Cu2+ is reduced to Cu+ and forms a red precipitate of Cu2O. Therefore, if Fehling’s solution is added to a solution containing a reducing sugar, a red precipitate will form. Sometimes the reaction mixture must be heated in order for the precipitate to form the color of the precipitate can vary from red to orange to green (the green color is actually a mixture of an orange and a blue precipitate).

Barfoed’s test is similar to Fehling’s test, except that in Barfoed’s test, different types of sugars react at different rates. Barfoed’s reagent is much milder than Fehling’s reagent. Reducing monosaccharides react quickly with Barfoed’s reagent, but reducing disaccharides react very slowly or not at all. Therefore, it is possible to distinguish between a reducing monosaccharide and a reducing disaccharide using Barfoed’s reagent.

Test for Saponins:

5 ml of extract was shaken vigorously with 5 ml of distilled water in a test tube and warmed. The formation of stable foam was taken as an indication of the presence of saponins.

Test for Alkaloids:

1ml of extract was stirred with 3 ml of 1% HCl on steam bath. 1 ml of mixture was taken separately in two test tubes. Few drops of Dragendorff’s reagent were added in one tube and occurrence of orange red precipitated was taken as positive. Two the second tube Mayer’s reagent was added and appearance of buff colored precipitate was taken as positive test for presence of alkaloids.

Test for flavonoids:

To 1 ml of extract, 1ml of 10% lead acetate solution was added. The formation of a yellow precipitate was taken as a positive test for presence of flavonoids.

Test for Steroids:

Salkowski’s test:

a red color produced in the lower chloroform layer when 2 ml of organic extract was dissolved in 2 ml of chloroform and 2 ml concentrated sulphuric acid was added in it, indicates the presence of steroids.

Physical evaluation^[5]:

Bulk density and Tap density:

The term bulk density refers to a measure used to describe a packing of particles or granules. The equation for determining bulk density (D), Db=M/Vb Where M is the mass of the particles and Vis the total volume of the packing. The volume of the packing can be determined in an apparatus consisting of a graduated cylinder mounted on a mechanical tapping device (Jolting Volumeter) that has a specially cut rotating can. 100gm of weighed formulation powder was taken and carefully added to the cylinder with the aid of a funnel. Typically the initial volume was noted and the sample was then tapped until no further reduction in volume was noted. The initial volume gave the Bulk density value and after tapping the volume reduced, giving the value of tapped density.

Angle of repose:

Angle of Repose has been used as indirect methods of quantifying powder flowability because of its relationship with inter particle cohesion. As a general guide, powders with angle of repose greater than 50 degree have unsatisfactory flow properties, whereas minimal angle close to 25degrees correspond to very good flow properties. The fixed funnel and the free standing cone method employs a funnel that is secured with its tip at a given height, which was taken 2.5 cm(H), above the graph paper that is place on flat horizontal surface. Powder or granulation wascarefully poured through the funnel until the apex of the conical pile just touched the tip ofthe funnel. Tan= H/ R or = arc tan H/R Where is the angle of repose, R beingthe radius of the conical pile.

Table 2: Angle of repose

Sr. No	Angle of Repose	Type of Flow
1	< 20	Excellent
2	20 – 30	Good
3	30 – 34	Passable
4	> 34	Very Poor

Fig. 4: Angle of repose

Hausner ratio:

It is related to interparticle friction and as such can be used to predict the powder flow properties.Powders with low interparticle friction such as coarse spheres have a ratio of approximately 1.2,whereas more cohesive, less flowable powders such as flakes have a Hausner ratio greater than1.6. The equation for measuring the Hausner ratio is: Df / Do, where Df = Tapped density and Do = Bulk density.

Carr’s index (or) % compressibility:

It indicates powder flow properties. It is expressed in percentage and is give

Dt – D

I = ------------ x 100

Where,

Dt is the tapped density of the powder and

Db is the bulk density of the powder.

Table 3: Carr’s index (or) % compressibility

% Compressibility	Flow ability
5 – 12	Excellent
12 – 16	Good
18 – 21	Fair Passable
23 – 35	Poor
33 – 38	Very Poor
< 40	Very Very Poor

Analytical Evaluation ^[6]:

Thin Layer Chromatography:

Chromatography is the separation of two or more compounds or ions by the distributionbetween two phases, one which is moving and the other which is stationary. These two phasescan be solid-liquid, liquid-liquid or gas-liquid. Although there are many different variations of chromatography, the principles are essentially the same. If you took Cham 15 at University Park,you may remember using paper chromatography to separate inks and food dyes. The cellulose paper was the stationary or solid phase and the 1-propanol/water mixture was the mobile or liquidphase. This chapter explores a very similar micro scale technique for separating organic molecules, thin-layer chromatography.

Procedure:

1. Wash the glass slide and dry it hot air oven.

2. Prepare a slurry of silica gel in beaker with water

3. Apply a slurry on glass slide by pouring method

4. Dry the glass slide in hot air oven at 110 c

5. prepare a solution of given amino acid in a suitable solvent

6. Spot of amino acid is applied on that slide by using a capillary tube

7. That slide for 1 min in air and keep in solvent chamber and allow it to run

8. Remove the glass slide from the solvent chamber and dry in air

9. Keep the slide in iodine chamber for visualization of spot

10. Calculate and report the RF value of amino acid.

Formula:

Rf Value = Distance traveled by solute

Distance traveled by solvent

Table 4: Microscopical Evaluation

Sr. No	Reagent	Observation	Characteristics
1	Phoroglusinol +conc.HCL (1:1)	pink	Lignified sclernchymatous cells
2	Dil .iodine solution	blue	Starch present in perisperm
3	Sulphuric acid (60 %w/w)	Calcium sulphate crystals	Calcium oxalate
4	Alcoholic picric acid	yellow	Aleurone grains
5	Sudan red III	Red	Oil globules

RESULTS AND DISCUSSION:

Organoleptic investigation:

The organoleptic parameters were performed and the results were shown in Table no1.

Table 5: Organoleptic investigation

Different formulation	Appearance	Colour	Taste	odor
Formulation A	powder	Whitish brown	Sweetish sweet	Fragrant
Formulation B	powder	Whitish brown	Sweetish sweet	Fragrant
Formulation C	powder	Whitish brown	Sweetish sweet	Fragrant
Formulation D	powder	Whitish brown	Sweetish sweet	Fragrant

Physicochemical evaluation:

Physicochemical evaluation were performed and the results were shown in Table 2.

Table 6: Physicochemical Evaluation

Test	Formulation A	Formulation B	Formulation C	Formulation D
Total ash content	0.3- 0.5	0.2- 0.4	0.4- 0.6	0.3-0.5
Acid insoluble ash value	21.0-0.05	15.9- 0.06	20.6-0.32	23.5-0.40
Water soluble ash	26.3-0.59	19.25-0.35	21.3-0.29	18.4-0.560
Loss on drying	2.141-0.0023	1.633-0.0225	3.171.0.0025	2.142-0.00210

Phytochemical Evaluation:

Phytochemical Evaluation were performed and the results were shown in Table 3.

Table 7: Phytochemical Investigation

Test	Formulation A	Formulation B	Formulation C	Formulation D
Test for Reducing sugar	-	-	-	-
Test for Hexose sugar	+	+	+	+
Test for Proteins	+	-	+	+
Test for alkaloids	+	+	+	+
Test for Steroids	+	+	-	+
Test for saponins	+	+	+	+
Test for Flavonoids	+	-	+	-

Physical evaluation of Churna:

Physical evaluation were performed and the results were shown in Table no.4

Table 8: Physical Evaluation

Parameter	Formulation A	FormulationB	Formulation C	Formulation D
Angle of repose	38.41 ± 0.31	23.65 ± 0.95	33.02 ± 0.23	36.12 ± 0.25
Tap Density	0.83 ± 0.04	0.78 ± 0.01	0.75 ± 0.03	0.64 ± 0.04
Bulk Density	0.59 ± 0.03	0.54 ± 0.04	0.58 ± 0.04	0.51 ± 0.02
Carrs index	28.91 ± 0.04	30.76 ± 0.06	22.66 ± 0.05	20.31 ± 0.06
Hausner ratio	1.40 ± 0.04	1.44 ± 0.03	1.29 ± 0.05	1.12 ± 0.03

MATERIAL and METHODS:

Colour

Acid insoluble ash:

Fig. 2: Acid insoluble ash

Water soluble ash:

Loss on drying:

Determination of pH:

Fig. 3: Determination of pH

Phytochemical investigation:

Tests for Carbohydrates:

Hausner ratio:

Carr’s index (or) % compressibility:

It indicates powder flow properties. It is expressed in percentage and is give

Table 3: Carr’s index (or) % compressibility

Analytical Evaluation ^[6]:

Thin Layer Chromatography:

Fig. 5: Thin Layer Chromatography

Microscopical Evaluation:

Table 4: Microscopical Evaluation

RESULTS AND DISCUSSION:

Organoleptic investigation:

Physicochemical evaluation:

Phytochemical Evaluation:

Phytochemical Evaluation were performed and the results were shown in Table 3.

Physical evaluation of Churna:

Physical evaluation were performed and the results were shown in Table no.4

Analytical evaluation:

Microscopical Evaluation:

CONCLUSION:

REFERENCES:

Formulation	RF value
Formulation A	0.4
Formulation B	0.5
Formulation C	0.6
Formulation D	0.3

MATERIAL and METHODS:

Colour

Acid insoluble ash:

Fig. 2: Acid insoluble ash

Water soluble ash:

Loss on drying:

Determination of pH:

Fig. 3: Determination of pH

Phytochemical investigation:

Tests for Carbohydrates:

Hausner ratio:

Carr’s index (or) % compressibility:

It indicates powder flow properties. It is expressed in percentage and is give

Table 3: Carr’s index (or) % compressibility

Analytical Evaluation [6]:

Thin Layer Chromatography:

Fig. 5: Thin Layer Chromatography

Microscopical Evaluation:

Table 4: Microscopical Evaluation

RESULTS AND DISCUSSION:

Organoleptic investigation:

Physicochemical evaluation:

Phytochemical Evaluation:

Phytochemical Evaluation were performed and the results were shown in Table 3.

Physical evaluation of Churna:

Physical evaluation were performed and the results were shown in Table no.4

Analytical evaluation:

Microscopical Evaluation:

CONCLUSION:

REFERENCES:

Analytical Evaluation ^[6]: