Development of a Reverse-Phase High-Performance Liquid Chromatography Method for Simultaneous Quantification of...

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• Simultaneous quantification of tofacitinib citrate (TFC) and berberine chloride (BBC) is crucial for pharmaceutical quality control and pharmacokinetic studies.
• Existing analytical methods may lack the sensitivity or specificity required for complex biological matrices.

Purpose of the Study:

• To develop and validate a novel reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of TFC and BBC.
• To ensure the method's suitability for analyzing TFC and BBC in both pharmaceutical injection formulations and blood plasma.

Main Methods:

• An isocratic RP-HPLC method was employed using a mobile phase of phosphate buffer saline (pH 5.5) and acetonitrile (70:30) at a flow rate of 1 mL/min.
• Detection was performed at 306 nm, with linearity assessed over a 5-30 μg/mL concentration range.
• System suitability, recovery, sensitivity, and robustness were evaluated according to standard analytical guidelines.

Main Results:

• The method demonstrated excellent linearity (R² > 0.999) and high sensitivity with low detection and quantification limits.
• Recovery studies yielded high mean recovery rates (99.4-100.7% for TFC, 99.9-100.4% for BBC).
• System suitability parameters and robustness testing confirmed the method's reliability and stability.

Conclusions:

• The developed RP-HPLC method is accurate, precise, sensitive, and robust for the simultaneous quantification of TFC and BBC.
• This validated method is suitable for routine quality control of pharmaceutical formulations and for pharmacokinetic investigations in biological samples.
• The method offers a valuable tool for therapeutic drug monitoring and research involving TFC and BBC.