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Item A Preliminary Study on Microbial Quality Standards of an Ayurvedic Compound Preparation "Thalisadee Choorana".(2007) Nageeb, B.M.; Widanapathirana, S.; Amarasinghe, A.P.G.; Kasthuriarchi, K.A.H.Thalisadee choorana is a common Ayurvedic medical preparation widely used by all indigenous medical practitioners in Sri Lanka. It is used for respiratory tract ailments such as cough, common cold, bronchitis, asthmatic conditions and gastro intestinal disorders such as diarrhoea, vomiting, indigestion and loss of appetite. It contains mainly Pipemigram (Gammiris),Piperlongum(Thippili), Abies�wehhiuna (Thalispathra),Cinnamum zylanicum (Kurundupothu), Elettaria repens( Heenensal),Bamboo salt (Unakapuru) and sugar. All plant materials contain a large number of microorganisms.Some are inherent and some are contaminated during the process of harvesting and manufacturing process. Considering these facts, the World Health Assembly in its resolutions WHA-31 :33(1978) 40:33(1987), 42:43(1989) has emphasized the need of ensuring the quality in regard to microbial content of the plant products. Hence this study was carried out to determine the microbial load of this product and the possible sterilization methods of reducing the microbial load. The effect of the method on the drug which reduces the microbial load of the drug also studied. Ten different samples of Thalisadee choorana were subjected to this study. 0.1 gram of the drug sample was dissolved in 10 ml of sterile distilled water. (10�). Using this solution 10-1, 10-2 10-3 dilutions were prepared. Routine sterilization procedures were carried out in all steps.Nutrient agar and Potato dextrose agar were used as general culture media. Pour plate technique and spread plate technique were used to detect the microbial count respectively. 0.1 ml of above dilutions was used on culture plates. Each plate was controlled by using another duplicate culture plate. Plates were sealed and kept under normal room temperature. Colony counts were taken after 24 hours and 72 hours for bacteria and fungi respectively. It was assumed that each colony was formed by a single organism. Same procedure was repeated three times. According to the W.I-I.A standard, aerobic bacteria up to 105 I gram, yeast and moulds up to 103 I gram arc permitted The results ofthe above study indicate that the bacterial count was in between 3x106 to 4xl06 /gram. These results indicate that the limits were exceeding on every sample. The following methods were tried to reduce the microbial load. I 00 grams of the above samples were subjected to (a) Heat treatment in a hot air oven at 80� C for 10 minuets for three consecutive days. (b) Ultra violet radiation at 256 wave length continuously for 24 hours. (c) Steam treatment under atmospheric pressure in a closed container for 10 minuets for three consecutive days. The study of microbial load was thereafter repeated.The plates of the steamed samples were sterile up to 72 hours while the plate of other two methods does not show any reduction in microbial load. The volatile oil content by reflux method using Dead and Stark apparatus and the thin layer chromatographic (T.L.C) patterns of Ethanol extract and Water extracts using Silica gel GF 254 and G06 at the ratio of 1:3 with several solvent systems of both samples (Steamed and un steamed) were studied. The T.L.C. patterns and the volatile oil content of both samples were comparatively same .This preliminary study reveals that the steam treatment method is comparatively an effective method to reduce the microbial load of the above preparation. A detail study of the chemical compounds through other chromatography methods is needed to confirm this.Item A preliminary study of anti-bacterial effect of selected five Ayurvedic compound preparations.(2006) Nageeb, B.M.; Widanapathirana, S.; Amarasinghe, A.P.G.Plant based medicaments have been man?s prime therapeutic weapons to rescue him from disease. Plants are of relevance to pharmacology. Pharmacological properties of medicinal plants may be used as leads in developing modern therapeutic agents. Thalisadee, Thripala,Hinguastaka, Dathree, and Manibadra are common Ayurvedic formulae used in traditional system of medicine in Sri Lanka. Thalisadee and Thripala Choorna are being commonly used in respiratory disorders such as cough, cold bronchitis and fever. The Hinguastaka, Dathree and Manibadra Choorna are being commonly used in gastrointestinal disorders such as diarrhoea dysentery vomiting and indigestion. Most of these conditions may develop due to bacterial infections .The main objective of this study was to evaluate the antibacterial effect of these preparations. Minimum human single dose of these drugs (2.5 gram) was dissolved in sterile distilled water and kept in the shaker at 100 rpm continuously for 04 hours in order to get the maximum soluble liquid extract of these drugs. 0.7 gram of Nutrient broth was dissolved in 50 ml of distilled water and transferred in to five test tubes (10 ml. /tube) and sterilized by autoclaving at 121 C for 20 minuets. These Nutrient broth tubes were inoculated by using inoculating needle with already plated pure test cultures of Pseudomonas aeruginosa, ,Escherichia coli , Salmonella typhi.These tubes were incubated at 37 0 C for 18 to 20 hours. Sterilized Nutrient agar was transferred into ten sterilized Petri dishes at 40 0C and allowed to solidify on a horizontal plane. These plates were sealed and kept in incubator at 37 0C for 24 hours to exclude any contaminations and to reduce the moisture content. A known amount of (0.05 ml) each culture broth containing specific organisms was added to these solidified agar plates and spread evenly using a sterilized glass spreader. On these seeded agar plates sterilized metal cylinders were kept (03 Cylinders/plate) with gentle pressure. These cylinders were filled with 0.1 ml of above liquid extract of drug preparations.De ionized sterilized distilled water 0.1 ml and Chlorampenicol 0.025 mg/0.1 ml were used as controls. These plates were sealed and incubated at 37 0C .This same procedure was repeated for three times for each of the test organism. Chlorampenicol showed 1.0 cm -1.5 cm clear inhibition zones of the bacterial lawns on every test organisms. None of the drug preparations showed any effect on Escherichia coli culture plates. The Ayurvedic compound preparations of Hinguastaka Choorna and Manibadra Choorna extracts showed averagely 0.5cm and 0.3 cm clear inhibition zones of the bacterial lawns on Salmonella typhi plate respectively. Thripala Choorna showed averagely 1.0 cm clear inhibition zone of bacterial lawns on Pseudomonas aeruginosa plates. These zones were clear on every repetition. Theses results were statistically analyzed by using one sample student T-Test. All the means are in between accepted level and P value is <0.05. Comparison to the Chlorampenicol,Higuastka Choorna and Manibadra Choorna are active against Salmonella typhi. Thripala Choorna is active against Pseudomonas aeruginosa. Both, Dathree Choorna and Thalisadee Choorna are not active against any of tested microorganisms. This preliminary study scientifically justifies that the use of the powder preparations of Higuastka Choorna,Manibadra Choorna and Thripala Choorna for infective conditions such as diarrhea, dysentery, vomiting and indigestion.