HPLC测定血浆中尼美舒利浓度

Determination of nimesulide in plasma by HPLC

目的：建立测定血浆中尼美舒利（Ｎｉｍ）的反相高效液相色谱法。方法：血浆样品用甲醇沉淀蛋白，经酸化后加甲苯提取，氮气挥干，加流动相溶解进样。以ＹＷＧＣ１８柱，甲醇０．０５ｍｏｌ·Ｌ－１磷酸二氢钾（５５∶４５）为流动相，ｐＨ５．０２，甲苯磺丁脲为内标，在波长２３０ｎｍ处检测。结果：在０．２６μｇ·ｍｌ－１～１０．８μｇ·ｍｌ－１浓度范围内，峰面积比与浓度呈良好线性关系，ｒ＝０．９９９５。最低检测限为０．２μｇ·ｍｌ－１，方法重现性好，方法回收率为８６．０％～９２．２％，萃取回收率为７２．２％～８８．４％。结论：用本方法测定８例单剂量ｐｏ１００ｍｇＮｉｍ的血药浓度，结果满意。

OBJECTIVE: To determine the concentration of nimesulide in human plasma. METHOD: After addition of the internal standard (IS; tolbutamide), 0.5 ml of methanol and 0.3 ml of hydrochloric acid (1 mol·L-1) were added to 0.5 ml plasma. The nimesulide was extracted into 2 ml of toluene. The organic layer was evaporated to dryness under a stream of nitrogen, and the residue redissovled in 150 μl of mobile phase for HPLC injection. The drug and IS were chromatographed on C18 column with a mobile phase consisting of 0.05 mol·L-1 potassium dihydrogen phosphate solution (pH 5.02): methanol (55∶45) and detected at 230 nm. Chromatography was carried out at a flowrate of 1.0 ml·min-1 at room temperature (20 ℃). RESULTS: A better linearity was obtained from 0.256 5 μg to 10.78 μg of nimesulide per ml for plasma with a good correlation coefficient (r=0.999 58, n=8). The mininum detectable concentration was 0.2 μg·ml-1 for plasma. CONCLUSION: Drug concentration in plasma was determined by HPLC method following a single oral dose of 100 mg was given to 8 adult male volunteers. The procedure is presently being used in case work.