A new high performance thin-layer chromatography (HPTLC) method for determination of theophylline in plasma

A new high-performance thin-layer chromatography (HPTLC) method for the determination of theophylline in plasma has been developed. Plasma samples were prepared by the internal reference method. Denaturation was carried out through cold centrifugation. Separation of the drug from the complex matrix was accomplished within a short 50 mm migration distance and 25 min migration time on silica gel 60 F 254 layers using toluene-isopropanol-acetic acid (80:20:5) as the mobile phase. The analyte was quantified by absorbance/reflectance densitometry using peak-area ratio analysis. Validation parameters were investigated considering the special features of the method.