液相色谱-串联质谱法测定SD大鼠全血中新型亲环蛋白D抑制剂RN-0001浓度  

Determination of a New Cyclophilin D Inhibitor,RN-0001,in SD Rat Whole Blood by LC-MS/MS

作　　者：赵磊 孙丽华 俞巧玲 邱云良 ZHAO Lei;SUN Lihua;YU Qiaoling;QIU Yunliang(China State Institute of Pharmaceutical Industry,Shanghai InnoStar Bio-tech Co.,Ltd.,Shanghai 201203;Farsight Medical technology(Shanghai)Co.,Ltd.,Shanghai 201206)

机构地区：[1]中国医药工业研究总院,上海益诺思生物技术股份有限公司,上海201203 [2]睿诺医疗科技(上海)有限公司,上海201206

出　　处：《分析科学学报》2023年第4期377-382,共6页Journal of Analytical Science

基　　金：国家“十三五”重大新药创制科技重大专项(2019ZX09732001-020)。

摘　　要：建立了液相色谱-串联质谱法测定新型亲环蛋白D抑制剂RN-0001在SD大鼠全血中的浓度。样品前处理采用简单的蛋白沉淀法,色谱分离在Gemini C18110,色谱柱(50 mm×2 mm,5μm)上进行,以含10 mmol/L乙酸铵和0.1%甲酸的水溶液作为流动相A,含0.1%甲酸的甲醇溶液作为流动相B,在0.8 mL/min流速下梯度洗脱。待测物RN-0001和内标环孢菌素A的检测采用正离子电喷雾多反应监测模式,其检测离子对分别为m/z 645.4/156.2和m/z 601.8/156.1。RN-0001在30.0 ng/mL~30.0μg/mL浓度范围内线性关系良好(r 2>0.9961),定量下限为30.0 ng/mL,批内和批间的精密度小于5.2%,批内和批间准确度在-3.6%~6.7%之间。RN-0001基质样品在室温放置18 h,-80℃冰箱放置40天以及经过5次冻融循环后均稳定。SD大鼠尾静脉注射3 mg/kg的RN-0001注射液进行药代动力学研究,RN-0001的半衰期t 1/2为3.53 h,峰浓度C 0为9370 ng/mL,血药浓度-时间曲线下面积AUC 0-24 h为5300 h·ng/mL。结果表明,所建方法适用于RN-0001在SD大鼠体内的药代动力学研究。A simple and fast LC-MS/MS method was developed and validated for determination of a novel cyclophilin D inhibitor,RN-0001,in SD rat whole blood.Protein precipitation as a fast and simple sample treatment was applied to this method.A Gemini C 18110,LC column(50×2 mm,5μm)was used in chromatographic separation with mobile phase A(10 mmol/L NH 4 OH and 0.1%formic acid in water)and mobile phase B(0.1%formic acid in MeOH).Following a sample injection(2μL),a gradient elution mode delivered mobile phases to the column at a flow rate of 0.8 mL/min.Multiple reaction monitoring and electron spray ionization technique in positive mode were applied to detection of RN-0001 and cyclosporine A(CsA)used as internal standard on a mass spectrometer with triple quadrupole.For RN-0001 and CsA,quantification was performed using the m/z 645.4/156.2 and m/z 601.8/156.1 transitions,respectively.Calibration curve covered a range of 30.0 ng/mL[WT5,7]-[WT]30.0μg/mL using a linear regression mode weighted by 1/x 2.Low limit of quantitation was 30.0 ng/mL.The precision of inter-and intra-batches was less than 5.2%and the accuracy of inter-and intra-batches was from-3.6%to 6.7%.The matrix samples of RN-0001 stored at room temperature for 18 h,-80℃for 40 days and following five freeze-thaw cycles were stable.With injection of 3 mg/kg RN-0001 to tail vein of SD rats,the major pharmacokinetic parameters of RN-0001 were discovered with t 1/2 of 3.53 h,C 0 of 9370 ng/mL and AUC 0-24 h of 5300 h·ng/mL.The method is simple and rapid,which can fit the needs of pharmacokinetic studies of RN-0001 in SD rats.

关 键 词：亲环蛋白D抑制剂 急性肾损伤 液相色谱-串联质谱 方法学验证 

分 类 号：O657.63[理学—分析化学] R917[理学—化学]