Assessing the suitability and efficiency of high performance liquid chromatography instrument - A new aproach

Abstract

A unique, cheap, precise and a simple analytical instrument qualification (AIQ) method was developed for a high performance liquid chromatography (HPLC) by using hypoxanthine, a universal nucleobase. This method could be used to determine the suitability of any HPLC that will be used for biomedical, environmental and food sample analysis. The volumetric flow of the mobile phase delivered by the pumps was consistent for given flow rate, with percent relative standard deviation (RSD %) of ± 0.05 % for pump A and B. The hypoxanthine calibration curve for the combined HPLC components in 1% acetonitrile and 0.05 M ammonium phosphate buffer at pH 6, as mobile phase was linear over a concentration range of 4 μM to 20 μM. A correlation coefficient of 0.95 and a sensitivity of 1824.2 AU/μM was obtained. A log-log analysis of the hypoxanthine calibration curve was linear. Column efficiency determined from toluene and biphenyl in 90: 10 % (v/v) methanol/water mixtures at a flow rate of 1 mL/min. was comparable to the results obtained by the manufacturer. In conclusion, it was determined that the HPLC instrument operated within the manufacturer specified and user approved parameters.