Advanced Analytical Methods for the Detection and Quantification of Pharmaceuticals in Biological Samples: A Cross-Disciplinary Exploration

Abstract

Pharmacokinetic research, therapeutic medication monitoring, and toxicological evaluations all depend on the identification and measurement of pharmaceuticals in biological samples. This study assesses the effectiveness of high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC–MS), liquid chromatography-tandem mass spectrometry (LC–MS/MS), and advanced analytical techniques in the analysis of plasma, serum, urine, and tissue matrices. One hundred purposively chosen samples from various pharmacological classes were used in a quantitative experimental-analytical design. Descriptive statistics, ANOVA, and multiple regression were used to evaluate and analyze key performance measures, such as accuracy, precision, limits of detection (LOD), and limits of quantification (LOQ). The results showed that LC–MS/MS worked better than GC–MS and HPLC, with the lowest imprecision (mean RSD 3.1%), the highest accuracy (mean recovery 97.8%), and the lowest LOD and LOQ values. While regression analysis showed that sensitivity metrics strongly predicted accuracy (R² = 0.28, p < 0.001), ANOVA demonstrated significant variations in technique accuracy (F(2, 297) = 12.78, p < 0.001). Performance was affected by matrix composition and sample preparation, especially for GC-MS and HPLC. The study highlights the significance of thorough method validation, optimal sample preparation, and sensitivity considerations while concluding that LC–MS/MS is the most dependable platform for pharmaceutical bioanalysis. Future research should look into cross-laboratory reproducibility studies, new analytical technology, and innovative sample preparation techniques.