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A sensitive, selective, and simple gas chromatography—mass spectrometry method is developed for quantitation of phenazopyridine (PAP) in human plasma using internal standard (diazepam). PAP and IS are extracted from plasma by liquid—liquid extraction and analyzed on a DB-5MS column with mass selective detector. Excellent linearity is found between 5–500 ng/mL ( r  = 0.9992,  n  = 7) for PAP in human plasma. The limit of detection is 0.3 ng/mL. Intra- and Inter-day precisions expressed as the relative standard deviation for the method are 1.37–6.69% and 1.24–6.01%, respectively. Extraction efficiency is more than 90%, and recoveries are in the range of 92.65–96.21%. This method is successfully applied for the pharmacokinetics and bioequivalence of 2 formulations of PAP in 18 healthy male volunteers who received a single 200 mg dose of each formulation.

Determination of Phenazopyridine in Human Plasma by GC—MS and its Pharmacokinetics