RP-HPLC Bio-Analytical Method for Precise Quantification and Validation of Bupropion Hydrochloride in Human Plasma

Abstract

Importance:

Bupropion is a widely prescribed antidepressant and smoking cessation aid. Monitoring its plasma concentration is crucial for assessing therapeutic efficacy and safety. Current methods for quantifying Bupropion in plasma are either complex or not widely accessible. This study presents the development and validation of a robust, sensitive, and efficient analytical method for quantifying Bupropion in plasma.

Research Gap:

While several methods exist, there is a need for a simple, reliable, and sensitive method that provides accurate results with minimal sample preparation, specifically suitable for routine clinical analysis.

Objective:

The primary objective of this study was to develop a High-Performance Liquid Chromatography (HPLC) method for the quantification of Bupropion HCl in plasma, ensuring high specificity, sensitivity, and reproducibility.

Methodology:

The method involves plasma extraction followed by analysis using HPLC with UV detection. Calibration curves were constructed and stability studies were conducted under various conditions. The method was validated according to US-FDA guidelines, assessing parameters such as precision, accuracy, linearity, LOD, LOQ, and matrix effects.

Key Findings:

The method demonstrated excellent linearity (R² = 0.999) over the range of 50–750 ng/mL. The limits of detection and quantification were 8 ng/mL and 25 ng/mL, respectively. The precision and accuracy of the method were validated, with intra- and inter-day variability <2%. Stability studies showed that Bupropion was stable under various conditions, with freeze-thaw recovery rates between 92%–97%, room temperature stability up to 6 hours (93%–96%), and long-term stability at -20°C (94.2%–96.8%).

Results:

Chromatograms for blank and spiked plasma confirmed the specificity and sensitivity of the developed method. The calibration curve exhibited excellent linearity with an equation of y = 0.023x + 0.15 and a high correlation coefficient (R² = 0.999). Intra-day and inter-day precision studies yielded % RSD values well within the acceptable limits, ensuring the method’s robustness. Recovery studies showed satisfactory mean recoveries (99.45%) for Bupropion at various concentrations, confirming the method's accuracy.

Implications and Future Potential:

This validated HPLC method offers a reliable approach for routine quantification of Bupropion in clinical settings. The method's simplicity, sensitivity, and reproducibility make it ideal for therapeutic drug monitoring and pharmacokinetic studies, with potential applications in other clinical and research environments. Future work will focus on expanding the method's application to other related compounds and improving the automation of the process.