Development of RP-HPLC for analysis of human insulin

A simple and reliable high-performance liquid chromatographic method with diode array detection has been developed and validated for the determination of insulin in human plasma. A good chromatographic separation was achieved on a C18 column with a mobile phase consisting of acetonitrile and 0.2M sodium sulfate (pH 2.4), 25:75 (v/v). Its flow rate was 1.2 mL/min. Calibration curve was linear within the concentration range of 0.15-25 µg/mL. Intra-day and inter-day relative standard deviations for insulin in human plasma were less than 6.3 and 8.5%, respectively. The limits of detection and quantification of insulin were 0.10 and 0.15 µg/mL, respectively. Also, this assay was applied to determine the pharmacokinetic parameters of insulin in eight insulin-dependent diabetes mellitus patients after subcutaneous injection of 25 IU of Actrapid HM.

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Development of RP-HPLC for analysis of human insulin

Cited by 5 publications

References 7 publications

Determination of insulin in innovative formulations by means of LC coupled to fluorescence detection

Determination of insulin in innovative formulations by means of LC coupled to fluorescence detection

Selectivity enhanced cation exchange chromatography for simultaneous determination of peptide variants

Determination of Insulin in Humans with Insulin-Dependent Diabetes Mellitus Patients by HPLC with Diode Array Detection

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