Validation of a new analytical method for the RP-HPLC quantitative analysis of recombinant human insulin

A rapid, accurate, dependable, and repeatable RP-HPLC method was created for the measurement of
insulin. A sodium sulphate anhydrous buffer with a pH of 2.3 and acetonitrile (76:26) are used as the
mobile phases A and B, respectively, in the most recent validated isocratic RP-HPLC analytical technique
for the determination recombinant human insulin (RHI). The samples were separated using a 150 x 4.6 mm
waters zorbax C18, flow rate of 1.0 ml/min, 3.5 m column. It was found that 214 nm is the wavelength
at which ultraviolet detection occurs. In order to analyse the desamido breakdown product of insulin; this
method was tested with phenol and m-cresol, which are preservatives used in commercial insulin
formulations at low concentrations, and with clear separation between each peak. The technique was
shown to be linear over the 40-60 g/ml concentration range with a regression coefficient of r2
= 0.9994.
During accuracy tests, it was revealed that the mean recovery was around 100.35 per cent. A low-cost,
dependable, exact, linear, and quick RP-HPLC technique was created and verified in accordance with ICH
criteria. As it has been demonstrated to be reliable, this method increasingly frequently used to evaluate
human insulin