Estimation of biphasic isophane insulin 30/70 by using validated RP-HPLC technique

A rapid, specific, reliable, and repeatable RP-HPLC method for estimating biphasic isophane insulin developed. A pH 2.3 sodium sulphate anhydrous buffer is used as mobile phase-A and acetonitrile (76:26) is used as mobile phase-B in the most current verified isocratic RP-HPLC analytical method. The samples were separated using a Zorbax C18 (4.6 x 250 mm), 3.5 m column at a 1.0 ml/min flow rate. The recommended wavelength for ultraviolet detection was discovered to be 214 nm. This technique was evaluated for the investigation of biphasic isophane insulin and its desamido the degradation product in the form of phenol and m-cresol, which are present in small amounts as preservatives in commercial preparations of biphasic isophane insulin. The technique was discovered to be linear over the concentration range of 80-120% with coefficient regression r2 = 0.9995. The mean recovery was determined to be in the range of 99.40% during accuracy tests. A rapid, accurate, linear, low-cost, and reliable RP-HPLC technology is designed and validated in compliance with ICH standards. Since it has been demonstrated to be reliable, this method can now be regularly used to evaluate biphasic isophane insulin.