RP HPLC Method for the determination of Tamsulosin in bulk and Pharmaceutical Formulations.

A simple, sensitive, precise and specific reverse phase high performance liquid chromatographic method was developed and validated for the determination of Tamsulosin in bulk and tablet dosage forms. It was found that the excipient in the tablet dosage forms does not interfere in the quantification of active drug by proposed method. The HPLC separation was carried out by reverse phase chromatography on Shimadzu HPLC, 10-At detector with hypersil ODS C18 Column 250 X 4.6 mm (particle size of 5μ) and constant flow pump. Rheodyne injector with 20 μl loop with a mobile phase composed in the ratio acetonitrile: (0.05M) KH2PO4 buffer (45:55) at flow rate 1.8 ml /min. The detection was monitored at 240nm. The calibration curve for Tamsulosin was linear from 10-50g/ml and internal standard (Bromhexine) 10g/ml were prepared by suitable dilutions of the stock solution with appropriate mobile phase. The interday and intraday precision was found to be within limits. The proposed method has adequate sensitivity, reproducibility and specificity for the determination of Tamsulosin in bulk and its tablet dosage forms. LOD and LOQ for Tamsulosin were found to be 0.495 and 0.461.Accuracy (recoveries: 98.5-98.55%) and reproducibility were found to satisfactory.

Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies

Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape tissue. We describe a protocol for isolating intact and high quality RNA from various grape tissues as evident by high A260/A280 absorbance ratio (1.8 to 1.9) and electrophoretic profile on denaturing formaldehyde agarose gel. On an average, 205 µg RNA per g of fresh tissue were obtained using this modified protocol. RNA quality was further assessed through RT-PCR, differential display RT-PCR and subtractive hybridization, and found to be suitable for molecular studies.