High performance liquid chromatography also called high pressure liquid chromatography (HPLC) is one of such sophisticated techniques by means of which we can do quantitative as well as qualitative measurement of different types of samples at a very low concentration even in the order of pictogram and nanogram level. The technology is an important one particularly in the field of Pharmaceutical Technology. Chromatography technique effects the separation of two or more component in a mixture. Our goal is to develop a unique method so that specific compound as mentioned can be identified in a single run from BSA protein qualitatively as well as quantitative from a BSA dilute solution. A Rheodyne model 7125 six port injection valve fitted with a 20 μl sample loop and a Novapak C18 column (150×39 mm, waters, USA) packed with 5 micrometer particles were used. The column was fitted with a Guard column (5 cm×4.6 mm) packed with the same packing material as in Novapak column. Single protein BSA can be easily be enriched or separated with a optimum chromatographic condition, at a flow rate of 0.8 ml/min, run time 8 min, injection volume 20 μl, mobile phase 0.9% NaCl +10 mM Tris buffer, pH 7.4. As a result BSA produced a peak at 1.406 min with the area 66 944 and the peak height was 6291. Comparing with the standard it can easily conclude that the peak was only for BSA sample.
Keywords: HPLC, BSA standard, BSA dilute solution.
Declaration of interest The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.