Development and Validation of A Hyphenated Method for The Determination of Dapagliflozin and Bisoprolol

Abstract

The simultaneous quantification of antidiabetic and cardiovascular drugs is essential for managing comorbid conditions where polypharmacy is prevalent. Dapagliflozin, a sodium-glucose co-transporter 2 inhibitor, and Bisoprolol, a β1-selective adrenergic blocker, were determined in pharmaceutical dosage forms using a unique LC-MS/MS approach that was developed and validated in this study. Method development was guided by physicochemical properties, utilizing a C18 column (250 × 4.6 mm, 5 µm) with a methanol–phosphate buffer (60:40, pH 3.8) mobile phase. Chromatographic separation was achieved within 5 minutes, with retention times of 2.7 and 3.4 minutes, respectively, ensuring high analytical throughput and efficient resolution.

Mass spectrometric finding using electrospray ionization in positive mode with optimized MRM transitions (m/z 408.3 → 135.2 and 326.2 → 116.1) provided high sensitivity and selectivity. The method demonstrated excellent linearity (5-100 ng/mL) with correlation coefficients of 0.998 and 0.999. Low LOD (~1 ng/mL) and LOQ (~3 ng/mL) values confirmed its sensitivity. Validation in accordance with ICH Q2(R2) guidelines showed accuracy (98-102%), precision (%RSD <2%), robustness, and specificity.

The developed method is rapid, reliable, and suitable for routine quality control. Furthermore, its high sensitivity supports bioanalytical applications such as pharmacokinetic profiling and therapeutic drug monitoring. The proposed hyphenated analytical platform represents a significant advancement in pharmaceutical biotechnology, enabling scalable, high-throughput analysis for modern drug development, precision therapeutics, and translational research.