Validation of a new high-performance liquid chromatography assay for nizatidine.

An expedient high-performance liquid chromatography (HPLC) assay for nizatidine measurement in human plasma was developed and validated. After deproteinization of 200 microL of plasma by filtration, nizatidine and 4-amino-antipyrine (internal standard) were separated (capacity ratio 3.0 and 6.63, respectively) on Nova-Pak C18 cartridge at room temperature (RT), and detected spectrophotometrically at 320 nm. The mobile phase, 0.02 mol/L disodium hydrogen phosphate, acetonitrile, methanol, and triethylamine (80:10:10:0.05 vol/vol), was delivered at 1.5 mL/min. Calibration curves were linear (r2 > or = 0.999) in the range 0.02 to 5 microg/mL, detection and quantification limits were 0.01 and 0.02 microg/mL, respectively, intra-run and inter-run coefficients of variation were < or = 3.5% and < or = 4.2%, respectively, and recovery was >90%. Nizatidine was stable for at least 4 hours at RT, 12 weeks at -20 degrees C, and 3 freeze-thaw cycles in plasma; 16 hours at RT and 48 hours at -20 degrees C in deproteinized plasma; and 6 hours at RT and 3 weeks at -20 degrees C in water.