高效液相色谱-质谱联用测定血浆奥替溴铵浓度

Determination of plasma concentration of otilonium bromide by combined use of liquid chromatography and mass spectrum

目的建立高效液相色谱-质谱联用法测定血浆中奥替溴铵浓度的方法。方法采用Eclipse Plus C18(150mm×2.1mm,3.5μm)色谱柱,柱温30℃,流动相为乙腈-20mmol/L-醋酸铵-乙酸溶液(58:42:0.1,V/V),流速0.35ml/min,电喷雾离子源,正离子检测,检测对象为奥替溴铵和内标氯苄律定的m/z分别为483.3和464.1。结果奥替溴铵和氯苄律定的保留时间分别为5.1min和2.0min,奥替溴铵在0.1-20.0μg/L浓度范围内有良好的线性关系(r=0.9998),定量下限为0.1μg/L,批内和批间精密度的相对标准偏差均小于15%,相对回收率93%-97%%。结论高效液相色谱-质谱联用测定准确、灵敏、简便,可用于人血浆的奥替溴铵浓度检测。

Objective To develop a liquid chromatography-mass spectrometric method for the determination of concentration of otilonium bromide in human plasma. Methods Eclipse Plus C18（150 minx 2.1 mm, 3.5 μm） was employed to determine plasma concentration of otilonium bromide. Column temperature was 30℃. The mobile phase consisted of methyl cyanides-ammonium acetate- acetic acid（58：42..0. 1, v/v） and flow rate was 0.35 ml/min. A tandem mass spectrometer equipped with electrospray ionization source was used as the detector operating in positive ion mode for otilonium bromide and chlorobenzylidine （internal standard, IS）, and the m/z of 483.3 and 464. 1 were used to detect otilonium bromide and chlorobenzylidine. Results The retention time of otilonium bromide and internal standard chlorobenzylidine were approximately 5.1 min and 2. 0 min, respectively. The standard curve of otilonium bromide was linear over the concentration range of 0.1-20. 0 μg/L（r=0. 9998）. The limit of qualification of betamethasone was 0.1 μg/L. Within-day RSD and between-day RSD were less than 15% ,and the relative recovery was all within 93%-97M. ConelusionThe established analytical method is accurate, sensitive, convenient and suitable for the determination of plasma concentration of otilonium bromide.