Method Validation of an Ultra-High-Performance Liquid Chromatography (UHPLC) for the Bioequivalence Study of...

Area of Science:

• Analytical Chemistry
• Pharmacokinetics

Background:

• Ensuring bioequivalence of generic drugs, particularly those containing rifampicin, requires validated analytical methods.
• Existing methods may not meet the need for simplicity and speed in quantifying rifampicin in human plasma.

Purpose of the Study:

• To validate an ultra-high-performance liquid chromatography (UHPLC) method for quantifying rifampicin in human plasma.
• To assess the method's selectivity, linearity, sensitivity, accuracy, precision, and carry-over effect according to regulatory guidelines.

Main Methods:

• Plasma samples were prepared using protein precipitation with methanol and ascorbic acid.
• Analysis was performed on a Waters Acquity H-Class UPLC system with a C-18 column, using a gradient elution of water-acetonitrile.
• Method validation followed ICH guidelines for Bioanalytical Method Validation.

Main Results:

• The UHPLC method achieved a 3-minute analysis time with rifampicin eluting at 1.4 minutes.
• The method demonstrated selectivity, linearity (2-25 ug/mL), and acceptable limits of detection (0.64 ug/mL) and quantification (1.94 ug/mL).
• Accuracy and precision were within regulatory limits, and no carry-over effect was observed.

Conclusions:

• A simple, fast, sensitive, reliable, and reproducible UHPLC method for quantifying rifampicin in human plasma was successfully validated.
• This method is suitable for application in bioavailability and bioequivalence studies of rifampicin-containing generic drugs.