A simple and rapid HPLC/UV method for the simultaneous quantification of theophylline and etofylline in human plasma

A simple, sensitive and selective high performance liquid chromatography (HPLC) method with ultraviolet detection (272 nm) was developed and validated for the simultaneous quantification of theophylline and etofylline in human plasma. Following rapid sample preparation, the analytes and internal standard (hydrochlorothiazide) were separated using an isocratic mobile phase on a reverse phase C18 column. The lower limit of quantification was 100 ng/mL for both theophylline and etofylline with a relative standard deviation of less than 6%. A linear dynamic range of 100–10,000 ng/mL for both theophylline and etofylline was established. This HPLC method was validated with between-batch precision of 2.2–6.0 and 1.4–3.7% for theophylline and etofylline, respectively. The between-batch accuracy was 94.3–98.0 and 95.4–98.2%, respectively. Stability of theophylline and etofylline in plasma was excellent, with no evidence of degradation during sample processing (autosampler) and 30 days storage in a freezer. This validated method is simple and rugged enough to be used in pharmacokinetic studies.