1. INTRODUCTION:

Herbal formulations show the number of problems when quality aspect is considered. This is because of nature of the herbal ingredients and different secondary metabolites present therein. Mainly, variation in the chemical profile of the herbal due to intrinsic and extrinsic factors (growing, harvesting, storage and drying processes).^1-3 The subject of herbal drug standardization is massively wide and deep. There is so much to know and so many seemingly contradictory theories on the subject of herbal medicines and their relationship with human physiology and mental function. For the purpose of research work on standardization of herbal formulations and neutraceuticals, a profound knowledge of the important herbs found in India and widely used in Ayurvedic formulation is of upmost importance. India can emerge as the major country and play the lead role in production of standardized, therapeutically effective ayurvedic formulations. India needs to explore the medicinally important plants.

This can be achieved only if the herbal products are evacuated and analyzed using sophisticated the importance of medicinal plants for public health care in developing nations and has evolved guidelines to support the member states in their efforts to formulate national policies on traditional medicine and to study their potential usefulness including evaluation, safety, and efficacy ⁴.

Standardization is an essential factor for polyherbal formulation in order to assess the quality of drugs based on the concentration of their active principle. It is very important to establish a system of standardization for every plant medicine in the market, since the scope of variation in different batches of medicine is enormous. Plant material when used in bulk quantity may vary in its chemical content and therefore, in its therapeutic effect according to different batches of collection e.g. collection in different season and/or collection from sites with different environmental surrounding or geographical location.⁵ Sitopaladi Churna is mentioned in sarangdhara samhita madhymakhanda adhyay 6,134-135 ½ (an old Ayurvedic classical text). Sitopaladi Churna is made from herbs like Mishri (Candy sugar), Vanshalochana (Silicacius concretion extract of Bambusa bambos Druce), Pippali (Piper longum Linn), Ela (Elleteria cardemomum Maton), Dalchini (Cinnemomum zeylanicum Blume). Sitopaladi churna” is a reputed Ayurvedic polyherbal medicine prescribed for pleurodynia, intercostals neuralgia, cough associated with bronchitis, supportive agent for allergy, viral respiratory infection, and in pharyngeal and chest congestion. Plants containing phenolic compounds have been reported to possess strong antioxidant properties.^6-8 Antioxidants play an important role in protecting cellular damage by reactive oxygen species.⁹ Piper longum (Pippali) is an immuno modulating herb. Anti-allergic activity of the fruit has been studied and is attributed to piperine.^10-11 Ela is having Sukshma Guna which makes this combination having deep penetrating ability and candy sugar makes it one of the best nutritive combinations. It is useful when there is shortness of breath, asthma with wheezing from contact with cold weather, bronchitis and chest congestion due to high kapha. It is helps with sinus congestion, sore throat, high fever and a muzzy head, for seasonal and allergic rhinitis from allergies to dust. Sitopaladi Churna increases appetite, alleviates distaste in tongue. It is useful in burning of palms and sole.¹² Sitopaladi Churna is a totally herbal combination without any side effect. This is the reason is indicated in general debility and weakness in body.

2. MATERIALS AND METHODS:

1) Procurement of raw drugs:

Sitopaladi churna consists of Mishri (Cane sugar), Vanshalochana (Bambusa bambos Druce; Silicacius concretion), Pippali (Piper longum Linn; fruit), Ela (Elleteria cardemomum Maton.; seed), Dalchini (Cinnemomum zeylanicum Blume; Stem bark). All these ingredients were procured from the local market of Patiala, Punjab, India and all the plant materials were authenticated by Department of Pharmacognocy at NIAPR, Patiala.

2) Preparation of Sitopaladi Churna:

Sitopaladi churna was prepared as per Ayurvedic Formulary of India part I, 7:34. All the ingredients were dried below 60ºC, powdered individually in a pulverizer and pass through 85# sieve and stored in air tight containers. Each ingredients were weighed separately required weight, mixed together to obtain a homogeneous blend¹³ details given in Table 1.

Table 1. Sitopaladi churna contains following ingredients.

S. No	Sanskrit name	Scientific name	Parts used	Quantity
1.	Mishri	Candy sugar	-	-
2.	Vanshalochana	Bambusa bambos Dduco.	Silicacius concretion	96 gms
3.	Pippali	Piper longum Linn.	Fruit	48 gms
4.	Ela	Elleteria cardemomum Maton.	Seed	24 gms
5.	Dalchini	Cinnemomum zeylanicum Blume.	Stem bark	12 gms

Table 2. Organolepatic properties of Sitopaladi churna formulations.

Appearance	Color	Odor	Taste
Fine Powder	Dull white	Sharp characteristic	Sweet pungent

3. RESULTS AND DISCUSSION:

1) Organoleptic evaluation:¹⁴

Organoleptic evaluation refers to evaluate that the formulation by color, odor, taste, texture etc. The Organoleptic characters of the Sitopaladi churna were evaluated and tabulated in Table 2.

2) Powder drug analysis of Sitopaladi churna:^15-19

About few mg of Sitopaladi churna powder warmed with chloral hydrate, washed and mounted in 50 percent glycerin; few mg of Sitopaladi churna powder washed thoroughly with water and mounted in 50 percent glycerin and few mg of Sitopaladi churna powder treated with iodine solution and mounted in 50 percent glycerin. Microscopically, thick walled Parenchyma cells containing small acicular raphides are irregularly arranged, long phloem fibers, bunches of Sclereids (Cinnamomum zeylancium Blume) Fig. 1(1-3); numerous epidermal cells, fragments of polygonal Sclerenchyma cells of testa, simple starch grains, prisms of calcium oxalate (Elleteria cardemomum Maton.) Fig. 1(4-7); Dark brown Stone cells, Starch grains (Piper longum Linn) Fig. 1(8-9) and Prismatic crystals (Bambusa bambos Druce) Fig. 1(10); characters were observed in different mounts of Sitopaladi churna.

3) Determination of physical characteristics of powder:

Physical characteristics like bulk density, tap density, angle of repose, Hausner’s ratio and Carr's index were determined for Sitopaladi churna and results given in Table 3. ^20-23

Table 3. Physical characteristics of samples of Sitopaladi churna.

S. No	Parameters	Results
1.	Bulk density ( gm/cm³)	0.56449
2.	Tap density ( gm/cm³)	0.744
3.	Hausner’s ratio	1.3180
4.	Carr’s Index (%)	24.127

a) 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 (Db) is:

Db = M/Vb (1)

Where, M is the mass of the particles and Vb is the total volume of the packing material.

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.

b) Hausner’s ratio:

Hausner, s ratio 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, Where as more cohesive, less flow able powders such as flakes have a Hausner’s ratio greater than 1.6.

The equation for measuring the Hausner’s ratio is:

Hausner’s ratio = Df / Do (2)

Where, Df = Tapped density and Do = Bulk density.

c) Carr’s index:

Carr,s index is another indirect method of measuring the powder flow from bulk density.

The equation for measuring Carr’s index is:

Carr’s index % compressibility = (Df-Do/Df) × 100 (3)

Where, Df = Tapped density and Do = Bulk density.

4) Evaluation of physicochemical parameters:^23-28

Evaluation of physicochemical parameters like total ash, acid insoluble ash and loss on drying at 105 ºC, alcohol, and water soluble extractive values were carried out as per the API/WHO guidelines for Sitopaladi churna results tabulated in Table 4.

Table 4. Physicochemical parameters of Sitopaladi churna.

S. No	Parameters	Results
1.	pH (10% w/v aqueous solution)	6.2
2.	Ash value (% w/w)	23.13
3.	Acid-insoluble ash (% w/w)	16.17
4.	Alcohol soluble extractive (% w/w)	12.23
5.	Water-soluble extractive (% w/w)	39.12
6.	Loss on drying at 105°C (% w/w)	7.19

a) Moisture content/ Loss on drying at 105°C:

4 g of the drug Sitopaladi churna was taken and heated in an oven at 105°C for 5 hour in a previously weighed 100 ml beaker. It was cooled in desiccators and weighed. The procedure was repeated till constant weight is obtained. The percentage of loss in weight of the drug sample was calculated.

Deterioration time of the plant material depends upon the amount of water present in plant material. If the water content is high, the plant can be easily deteriorated due to fungal attack. The loss on drying at 105°C of Sitopaladi churna was found to be 7.19 %.

b) Determination of ash values:

i) Total ash value:

2 g of the Sitopaladi churna was taken accurately in a previously ignited and tarred Silica dish. The material was spread evenly and ignited in a muffle furnace by gradually increasing the temperature to 600^oC until it is white, indicating the absence of carbon. The crucible was cooled in a desiccators and allowed to stand for 30 minutes and weighed.

Total ash value of plant material indicated the amount of minerals and earthy materials attached to the plant material. Analytical results showed total ash value of Sitopaladi churna was 23.13%.

ii) Acid insoluble ash value:

To the dish containing the total ash, 25 ml of 20 % Hydrochloric acid was added covered with a watch glass and boiled gently for 5 minutes. The watch glass was rinsed with a hot water and added to the crucible. The residue was washed with the hot water till the washings were neutral to the litmus paper. The insoluble material was collected and again placed in a same crucible and again ignited for 6 hr. to constant weight. The residue was cooled a desiccators for 30 minutes and weighed.

Percentage of acid insoluble as was calculated. The amount of acid-insoluble siliceous matter present in the Sitopaladi churna was 16.17%.

c) Determination of solvent Extractive values:

i) Water soluble extractive value:

4 g of the Sitopaladi churna was taken in a glass stoppered flask.100 ml of distilled water was added. The flasks were shaken occasionally for 6 hours and then allowed to stand for 18 hours. The extract was filtered and 25 ml of the filtrate was pipette out in a pre-weighed 100 ml beaker and evaporated to dryness on a water bath. It was kept in a hot air oven for 5 hr at l05°C, cooled in desiccators for 30 minutes and weighed. The procedure was repeated till constant weight.

The water-soluble extractive value indicated the presence of sugar, acids and inorganic compounds. The water soluble extractive value in the Sitopaladi churna sample was found to be 39.12%.

ii) Alcohol soluble extractive Value:

Same procedure as for the water soluble extractive value was followed. Instead of water, rectified spirit was taken as a solvent.

The alcohol soluble extractive values indicated the presence of polar constituents like phenols, alkaloids, steroids, glycosides, flavanoid and secondary metabolites present in the plant sample. The alcohol soluble extractive value was found to be 12.23% in the Sitopaladi churna.

d) Determination of pH Value:

10% w/v aqueous solution of sample was prepared and used for determining the pH value by pH meter. The pH value of Sitopaladi churna was found to be 6.2.

5) Thin layer chromatography:

Extract 10 g of churna sample with 40 ml ethanol under reflex on a water bath 30 min, filter, and concentrate the extract to 10 ml and carry out the thin layer chromatography. Apply 10 µl (micro liters) of the extract on (Aluminum plates precoated with Silica gel 60 F ₂₅₄ of thickness) TLC plate and develop the plate to a distance of 8 cm using toluene: ethyl acetate (5: 2) as mobile phase. After development, allow the plate to dry in air and examine under ultraviolet light. It shows 4 spots at R_f. 0.29, 0.43, 0.47, 0.57 (pale green) under 254 nm; and 0.32, 0.47, 0.60, 0.73 (dark blue) under 366 nm. Spray the plate with vanilin sulphuric acid reagent followed by heating at 105°C till spots apeared. It shows 9 major spots at R_f 0.14 (pale yellow), 0.26, 0.32, 0.35, 0.60, (pale blue) 0.79 (pale yellow), 0.85, 0.93, 0.95 (blue) in visible light.

4. CONCLUSION:

Sitopaladi churna and its ingredients are identified and authenticated through Organoleptic, physical characteristics, Physicochemical studies. Phamacognostic characters established for the raw materials could be employed as quality control standards for evaluating its identity and can be used for routine analysis. Purity and potency of the materials and formulations following the procedure given could be performed in quality control laboratory of pharmacy.

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Received on 19.09.2011 Modified on 10.10.2011

Asian J. Research Chem. 4(12): Dec., 2011; Page 1867-1871