A High-Throughput and Green Planar Chromatographic Approach for Furosemide Analysis in Plasma

A high-performance thin-layer chromatography (HPTLC) method was developed and validated to quantify furosemide in human plasma, providing a rapid, cost-effective, and environmentally sustainable alternative to conventional chromatographic techniques. The method employed silica gel ۶۰ F۲۵۴ precoated TLC plates with a mobile phase consisting of isopropyl alcohol, formic acid, ethyl acetate, chloroform, and toluene (۱.۵:۱:۰.۵:۳:۴, v/v/v/v/v), and densitometric detection at ۲۷۱ nm. Paracetamol was used as an internal standard. The method exhibited a strong linear correlation over the ۳۰۰-۲۱۰۰ ng/band concentration range with a correlation coefficient (r²) of ۰.۹۹۴۹. The lower limit of quantification was ۵۰ ng/band, demonstrating high sensitivity. The intra-day and inter-day precision (%CV) were less than ۲%, and the accuracy ranged from ۸۵.۴۹% to ۹۴.۴۶%. Recovery was consistent across all QC levels, with a maximum of ۸۴.۷۰%, and dilution integrity yielded ۱۱۱% recovery. The method proved robust under slight variations in chromatographic conditions and showed acceptable stability under benchtop, freeze-thaw, overnight, and long-term storage conditions. Overall, this HPTLC method is simple, reproducible, and suitable for high-throughput applications, such as pharmacokinetic studies, bioequivalence assessments, and therapeutic drug monitoring of furosemide in human plasma.

Keywords:

Planar Chromatography , Bioanalytical Method Validation , Green Analytical Chemistry , High-Performance Thin-Layer Chromatography (HPTLC) , Analytical Method Development