Pathogens Klebsiella pneumonia, Proteus vulgaris, Salmonella typhae, were found to be inhibited by formulation. For the finished product, microbial analysis was done. The results obtained were found to be within acceptable ranges/values and were nearly constant over the tested interval of time of about four months. The results obtained were found to be within acceptable ranges / values and were nearly constant over the tested interval of time of about four months. In Stability study of the finished product, the resin content, eugenol content were checked for four months. In HPTLC procedure, the comparison of finished product with raw material, observed under Visible light and UV light showed distinctive characteristic patterns (Fig. It shows Pieces of pitted vessels, fibers, epithelial cells, cork cells etc. Microscopic analysis of sample showed the presence of identifying diagnostic characters, which are not overlapping. Phytochemical analysis showed the presence of alkaloids, Glycosides, Tannin/ phenols, carbohydrate and fixed oils (Table 3). Results of quantitative analysis for Total ash, Acid insoluble ash, Water soluble extractive, Alcohol soluble extractive, Lipid content, Volatile Oil content, Particle size (80-100 mesh) and Loss on drying at 105 ☌ were calculated (Table 2). Botanical parameters revealed that brownish yellow in color, with pleasant odor, sweet taste. Stability studies: Comparison of the finished product (formulation) stored at room temperature for second, third & fourth month was carried out by conducting tests for the parameters resin content, eugenol content, using HPTLC technique (Gopala et al. 16 Patil, Zafar, Antimicrobial test: Formulation was checked for its antimicrobial activity against Klebsiella pneumonia, Proteus Escherichia coli by Agar diffusion method (Gopala et al. Standard plate count: This method was followed in order to enumerate the total aerobic count in a sample (Gopala et al. Microbial Screening: For the finished product microbial analysis was done. The scanning was performed at 266 nm and 366 nm (Eike, 2006 Sazada et al. The slit dimension setting of length 6.00mm and width 0.45 mm, and scanning rate of 20mm/s was employed for plate. The source of radiation utilized was Mercuric, Deuterium lamp. Densitometric scanning was performed on Camag TLC scanner III in the operated by win CATS planer chromatography version 1.4.3. Subsequent to the development, TLC plate was dried with help of air dryer. 10ml of mobile phase was used for development, which required 10minutes to develop. The length of chromatogram run was 7.5 cm. The optimized chamber saturation time for mobile phase was 15 min at room temperature. Linear ascending development was carried out in 10 cm × 10 cm twin trough glass chamber equilibrated with mobile phase. For stability studies, Toluene: Ethyl Acetate: GAA (7.5 : 3 : 0.2) was used as the mobile phase. Plates were developed using a mobile phase (consisting of Toluene: Ethyl acetate: Methanol (7 : 2 : 1 v/v), for comparing finished product with raw material. The sample, in the form length 8 mm were applied as a band 8mm from bottom, 12 mm from left margin of plate and 10 mm apart from each band, at a constant application rate of 150 nl/s using nitrogen aspirators. The chromatograph was performed by spotting extracted samples on pre-coated silica gel aluminum plate 60F-254 (10 cm × 10 cm with 250 m thickness) using Camag Linomat 5 sample applicator and a 100 l Hamilton syringe. For HPTLC profiling, 1 gm of sample was extracted with 10 ml of methanol in a reflux condenser for 1 hr, filtered and concentrated. Permanent slides were prepared and stained with Safronin (1%) + Glycerin (Selvakumar et al. Microscopic Analysis: The microscopic Character of each ingredient and final product were carried out (Anonymous, 1992). Phyto-chemical Analysis: Preliminary tests were carried out on methanolic extract for the presence / absence of phyto-constituents like alkaloids, carbohydrates, flavanoids, glycosides, saponins, sterols, terpenes and tannins (Sazada et al. Ash value, acid insoluble Ash, water soluble extractive value, ethanol soluble extractive value, fixed oil content, loss on drying were determined (Iyengar, 1995 Trease and Evans Wc., 1989).
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