Less is more: Validating a single method for comprehensive rh-insulin analysis

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• Current methods for human insulin analysis in the Indian Pharmacopoeia involve separate procedures for potency and related proteins.
• This necessitates multiple analytical runs, increasing time and resource expenditure for quality control.

Purpose of the Study:

• To develop and validate a single, unified method for the simultaneous analysis of human insulin potency and related proteins.
• To establish a more efficient and cost-effective quality control process for insulin formulations.

Main Methods:

• Utilized reverse-phase high-performance liquid chromatography (RP-HPLC) with a C-18 stationary phase.
• Employed a mobile phase of 55% buffer (0.2 M sodium sulfate, pH 2.3) and 45% acetonitrile, with UV detection at 214 nm.
• Validated the method for linearity, accuracy, and limit of detection.

Main Results:

• The method demonstrated excellent linearity over a concentration range of 0.08-4.5 mg/mL (r²=0.999).
• Achieved a limit of detection of 0.094 mg/mL and accuracy ranging from 99% to 102.8%.
• Successfully quantitated both potency and related proteins in a single chromatographic run.

Conclusions:

• A single RP-HPLC method provides a precise and efficient means for analyzing human insulin potency and related proteins simultaneously.
• This unified approach offers significant advantages in terms of time and cost savings for quality analysis in manufacturing and regulatory settings.
• Implementation of this method can enhance the availability of standardized, high-quality insulin preparations for public health.