ANVESHANA ANALYTICAL STANDARDIZATION OF PANCHSHARA RASA

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1 ANVESHANA Research Article AYURVEDA MEDICAL JOURNAL ISSN: ANALYTICAL STANDARDIZATION OF PANCHSHARA RASA Rajput Pawan 1, Parashar Shalini 2, Sridurga Ch. 3 1 PG Scholar, 3 Asso. Prof., Dept. of Rasashastra and Bhaishajya Kalpana, S.V. Ayurvedic College, Tirupati, India 2 PG Scholar, Dept. of Roga Nidana & Vikriti Vijnana, IPGT & RA, Gujarat Ayurveda University, Jamnagar, Gujarat, India Corresponding Author: dr.pawan0709@gmail.com ABSTRACT Panchshara Rasa was prepared according to Bhaishajya Ratnavali. Panchshara Rasa is the blend of Kharaliya [medicine prepared in mortar and pestle] and Parpatirasayana [preparation which is thin, brittle and has a shape of thin crisp wafer by heating] after preparation its subjected to Organoleptic tests, Physico-chemical tests, Particle size analysis and UV- Spectroscopy for standardization. Physico-chemical analysis of Panchshara Rasa showed moisture value 2.2% w/w, Total Ash value 3.54% w/w. The ph of 1% solution was found to be The particle size analysis found the particle size of Panchshara Rasa was 296.8nm. UV- Spectroscopy showed maximum absorption of Panchshara Rasa at 200nm wave length. Key words: Panchshara Rasa, Standardization, Moisture value, Total Ash value, Particle size INTRODUCTION The aim of an analytical study is to enable prediction about how a change in a system will affect that system's future performance, or prediction as to which plans or strategies for future action on the system will be superior. Analytical study is the key part of any scientific research. It tells us about the correlation between predetermined hypothetical values and actual results obtained. It gives us valuable information about safety, efficacy, stability, and contra-indications etc. of any formulation. Standardization is actually a concept developed basing of analysis. A standardized sequential systematic pattern of any procedure can be established by analyzing the outcomes at every stage. These analysis parameters may vary from simple organoleptic test to advanced Particle size analysis. Organoleptic tests helps in providing basic information about drugs. This generally includes tests that can be done by ones sensory organs and quality of material can be inferred up to some extent. Physico-Chemical tests deal with primary physical and chemical properties of a sample, which can hint about the internal molecular behaviors at different natural conditions. It helps in understanding the stability of a drug when it is stored for long time. Ash tests give idea about organic/inorganic chemical nature of sample. Extractive value tests helps in inferring the presence of various bioactive molecules like lipids and proteins, due to their differential solubility nature in water and alcohol. Particle size analysis of the drug plays major role in its therapeutic action and efficacy.

2 Pharmaceutical process Panchshara Rasa is mentioned in Bhaishjaya Ratnavali.The main ingredients of PanchsharaRasa are Shuddha Parada, Shuddha Gandhaka and Shalmali moola swarasa. Shodhita Parada was taken in khalwa yantra and mardana with Shalmali moola swarasa was done for 21 days. Shodhita Gandhaka was taken in khalwa yantra and triturated with Shalmali moola swarasa for 21 days. After trituration, Mardita Parada and Bhavita Gandhaka were taken in khalwa yantra and mardana was doneto obtain Kajjali. Kajjali was taken in a ghee smeared darvi and melted in mandagni. The molted Kajjali was poured on banana leaf and covered with another banana leaf and compressed by a steel plate to prepare Parpati 1. Parpati churna was subjected to bhavana for 21 days with Shalmali moola swarasa 2 to obtained Panchshara Rasa. After completion of the Pharmaceutical process, Panchshara Rasa was subjected to analysis through organoleptic tests, Physico-chemical tests, Particle size analysis and UV- Spectroscopy. MATERIALS AND METHODS Physico-chemical Test 3 The PanchsharaRasa was subjected to Physico-chemical tests at Department of Animal nutrition, College of Veterinary science, Tirupati. a. Moisture value Materials required: Hot air oven, Digital balance, Porcelain crucibles. check accuracy of weight and is taken in a porcelain crucible. Hot air oven thermostat is adjusted to 105 C and left for certain time to get stabilized at that temperature. Porcelain crucible with sample is kept on oven tray with equidistant from four walls of oven. Sample is dried for one hour. Porcelain crucible is taken out and kept in desiccator to prevent any moisture absorption and let to cool. After self-cooling porcelain crucible with sample is weighed to calculate the loss of weight on drying. The percentage content of moisture value is calculated in percentage (%w/w). b. Total Ash value Materials required: Muffle furnace, Digital balance, Silicon crucibles. check accuracy of weight and is placed in a silicon crucible. This crucible is kept on wire gauze and heated on a gas stove. Heating is continued until fumes get stopped. Then this crucible is kept in muffle furnace equidistant from four walls and furnace temperature is gradually raised up to 450 C. Burning is carried out for 6 hours. After complete incineration and after self-cooling, crucible is taken out and kept in a desiccator. The weight of ash with silica crucible is noted. Then the total ash is calculated in terms of percentage (%w/w). c. Acid insoluble ash value Materials required: 100ml glass conical flask, 100ml glass beaker, Whatman ash less filter paper, 2N HCl, glass pipette, Digital balance, Muffle furnace, Silicon crucibles. check accuracy of weight. Further using 25 ml. of 2N dilute Hydrochloric acid, total ash is washed into a 100 ml beaker. Beaker is boiled for 5 minutes. Contents are filtered through an ash less filter paper, residue is washed twice with hot water. Filter paper is placed in a silica crucible and then incinerated by gradually increasing the heat AAMJ / Vol. 1 / Issue 6 / Nov Dec

3 in a muffle furnace at 450º C for some hours. After complete incineration and self-cooling, crucible is taken out and kept in a desiccator. The weight of ash with silica crucible is noted. Then the total ash is calculated in terms of percentage (%w/w). d. Water soluble ash value Materials required: 100ml glass conical flask, 100ml glass beaker, Whatman ash less filter paper, double distilled water, glass pipette, digital balance, muffle furnace, silicon crucibles. Sample for analysis: 1g of Panchshara Rasa ash. check accuracy of weight. Further using 25 ml. of double distilled water, total ash is washed into a 100 ml beaker. Beaker is boiled for 5 minutes. Filtered through an ash less filter paper, residue is washed twice with hot water. Filter paper is placed in a silica crucible and then, incinerated by gradually increasing the heat in a muffle furnace at 450º C for some hours. After complete incineration, it is kept in a desiccator to cool. The weight of ash with silica crucible is noted. Then the loss of ash in water is calculated and water soluble ash value is quantified in terms of percentage (% w/w). e. ph value Materials: Glass electrode, ph meter, Buffer tablets, Beakers, Panchshara Rasa 1 g Procedure: Instrument is switched on and allowed to warm up. As the instrument is equipped with a manual temperature control, the temperature of the solutions is taken and the control is set to this value. The electrode assembly is inserted into the same beaker, the selector switch of the instrument is set and ph is noted. The Set buffer control is adjusted until the meter reading agrees with the known ph of the buffer solution. The electrode assembly is removed, rinsed in distilled water and placed into a small beaker containing the second buffer solution. Now 1 gram of Panchshara Rasa sample was put in 5 ml. of water and ph is determined for the solution. Particle Size Analysis 4 The Panchshara Rasa was subjected to Particle size analysis at Department of Soil science, Agriculture University, Tirupati. Material: Microtrac Particle size analyzer Sample for analysis: 1g PanchsharaRasa Procedure: The sample was mixed in water and sonicated for 10 minutes. Then it was poured into the sample chamber, where it passes through the laser beam as homogeneous stream of particles. The scattering of light occurs over a wide range of angles upon interacting with the particles in the suspension which are moving by Brownian motion. Based on this scattering pattern of sample, particle size distributions are calculated comparing with appropriate optical model. The scattered light is captured by a detector over the course of the analysis to determine the rate of diffusion (i.e. how fast the particles move within a system due to Brownian Motion) and thus the average Hydrodynamic particle size (referred to as the Z-Average) is calculated on an intensity weighted basis using the Stokes-Einstein equation. In simple terms, small particles move/diffuse more rapidly than larger particles. UV- Spectroscopy 5 The Panchshara Rasa was subjected to UV-Spectroscopy at Department of Soil science, Agriculture University, Tirupati. Material: DU640-UV/Vis Spectrophotometer Sample for analysis: 5g Panchshara Rasa Procedure: 5g, Panchshara Rasa was macerated with 100 ml of solvent in a closed flask for twenty-four hours, shaking frequently during six hours and allowed to stand for eighteen hours. It was filtered, AAMJ / Vol. 1 / Issue 6 / Nov Dec

4 taking for UV spectroscopical study. The Spetra was taken at nm from the peak obtained the λmax value was calculated. OBSERVATIONS AND RESULTS UV-Spectroscopy Fig.2: Showing UV-Spectrum of Panchshara Rasa Organoleptic tests Table 1: Organoleptic tests of Panchshara Rasa Sl. No. Parameter Observation 1. Colour Black 2. Taste Tasteless 3. Odour Shalmali moola 4. Touch Smooth 5. Appearance Lusterless Physico-chemical tests Table 2: Physico-chemical tests of Panchshara Rasa Sl. No. Parameter Observation 1. Moisture value 2.2% w/w 2. Total Ash value 3.54% w/w 3. Acid insoluble ash value 0.379% w/w 4. Water soluble ash value 3.71% w/w 5. ph 8.07 Particle Size Analysis Table 3: Particle size of Panchshara Rasa Measurement type Particle size Sample name Panchshara Rasa Scattering angle 90 Temperature of the holder 25 C T% before means Viscosity of the dispersion medium mpa-s Form of distribution Standard Representation of result Scattering light intensity Count rate 453 kcps Table 4: Particle size calculation of Panchshara Rasa Peak No. S.P. Area ratio nm Cumulant operations Mean S.D. Mode 29.7 nm nm Z- Average : nm PI : Fig. 1: Graph of particle size of Panchshara Rasa UV Spectrum of Panchshara Rasa showed maximum absorption at 200 nm. DISCUSSION Organoleptic tests: Organoleptic properties are the aspects of a substance as experienced by the senses including taste, sight, smell and touch. It gives idea about physical condition of the sample. It plays a major role when medicine is ingested orally in tablet form as the patients experience the same sensations while taking it. Black colour of the final product is due to Meta Cinnabar. Medicine was soft to touch as it was in the form of fine Shalmali moola like odour is due to bhavana of Parpati churna with Shalmali moola swarasa. Panchshara Rasa was tasteless and lustureless. Physico-chemical tests Investigating Physico-Chemical systems makes possible to determine the nature of interactions between the components of sample through a study of relationship between sample physical properties and composition. Moisture value of the final product was less i.e. 2.2% w/w which indicates the preparation is almost free from the moisture, suggesting its long shelf life. The excess moisture content can lead to growth of microbes, fungi etc which can deteriorate the drug. Total Ash value of Panchshara Rasa was 3.54% w/w. Panchshara Rasa contains AAMJ / Vol. 1 / Issue 6 / Nov Dec

5 mainly Mercury and Sulphur. These elements evaporated and very little amount of ash found. Acid insoluble Ash value is less (0.379% w/w) as the Panchshara Rasa sample is highly soluble in HCl. Hence, little quantity was remained on filter paper. Water-soluble ash value (3.71 % w/w) was less due to presence of less quantity of water-soluble salts. ph value of Panchshara Rasa is 8.07 suggests that it is alkaline in nature. Particle size analysis The size of the particles in the drug plays major role in its therapeutic action and efficacy. The mean particle size of the particles in Panchshara Rasa was nm. This nano size of the drug proves the fact that the processes like Sodhana and Bhavana reduces particle size. The nano size of drug is indicative of its quick absorption and faster dispersion into body resulting in better therapeutic efficacy. UV- Spectroscopy UVS refers to absorption spectroscopy or reflectance spectroscopy in the ultraviolet-visible spectral region. Different molecules absorb radiation of different wavelengths. An absorption spectrum will show a number of absorption bands corresponding to structural groups with the molecule. UV- Spectrum of Panchshara Rasa showed maximum absorption at 200 nm. CONCLUSION ACKNOWLEDGMENT Authors would like to acknowledge Prof. Dr. V. Ramana, Department of Animal Nutrition, Sri Venkateswara Veterinary University, Tirupati and Dr. T.N.V.K.V.Prasad, Senior scientist, Department of Soil science, Agriculture University, Tirupati for their cordial support and cooperation during the analytical study. REFERENCES 1. Sen Govinda Das, Bhaishajya Ratnavali. Edited by Lalchandraji Vaidya. Vajikaranadhikara, verse th ed. Delhi: Motilal Banarasi Das Publication; Sharma Vd. Pandit Hariprapana, Rasa Yoga Sagar. Vol.2, Pakaradirasa Varanasi: Krishnadas Academy; Reprint Government of India, The Ayurvedic Pharmacopoeia of India Part 1, vol.6, 1 st edition Ministry of Health and Family welfare Department of Ayurveda, Yoga & Naturopathy, Unani, Siddha and Homoeopathy, New Delhi; p , Available from: nalysis. [Last modified on 2015 Apr 26]. 5. Available from: ible_spectroscopy. [Last modified on 2015 Sep 1]. Moisture value of Panchshara Rasa was 2.2% w/w which indicates that the preparation was free from moisture and suggesting its long shelf life. Particle size analysis report showed the mean particle size of Panchshara Rasa was nm which indicates that the drug is easily absorbed in the body. UV-S of Panchshara Rasa showed maximum absorption at 200 nm. How to cite this article: Rajput Pawan et. al: Analytical Standardization of Panchshara Rasa. AAMJ 2015; 1: Source of Support: Nil. Conflict of Interest: None declared AAMJ / Vol. 1 / Issue 6 / Nov Dec