大鼠肠道菌群对吡嗪酰胺及其活性代谢产物吡嗪酸药动学参数的影响  

Effects of Rat Intestinal Flora on the Pharmacokinetic Parameters of Pyrazinamide and Its Active Metabolite Pyrazinoic Acid

作　　者：刘庆香 武正华 赖雅琳 范国荣[2] 范琦[1] LIU Qingxiang;WU Zhenghua;LAI Yalin;FAN Guorong;FAN Qi(School of Pharmacy,Chongqing Medical University,Chongqing 400016,China;Dept.of Clinical Pharmacy,Shanghai General Hospital Affiliated to Shanghai Jiao Tong University,Shanghai 200080,China)

机构地区：[1]重庆医科大学药学院,重庆400016 [2]上海交通大学附属第一人民医院临床药学科,上海200080

出　　处：《中国药房》2021年第4期412-417,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81973289)。

摘　　要：目的:研究大鼠肠道菌群对吡嗪酰胺及其活性代谢产物吡嗪酸药动学参数的影响。方法:将16只SD大鼠随机分为实验组和对照组,每组8只。实验组大鼠灌胃混合抗生素(硫酸链霉素+硫酸新霉素)构建伪无菌大鼠模型。建模后,两组大鼠灌胃吡嗪酰胺(150 mg/kg),并于给药前和给药后0.167、0.333、0.667、1、1.5、2、3、4、6、9 h时从眼眶静脉丛采血0.1 mL,给药后12、24 h时从眼眶静脉丛采血0.3 mL。以非那西丁为内标,采用液相色谱-串联质谱法测定大鼠血浆中吡嗪酰胺和吡嗪酸的浓度。以Agilent Zorbax SB-Aq为色谱柱,以0.2%甲酸水溶液(含8 mmol/L乙酸铵)-甲醇为流动相进行梯度洗脱,流速为1 mL/min,柱温为30℃,进样量为10μL;以电喷雾电离源为离子源,离子源温度为500℃;碰撞气为氮气,压力为10 psi;质谱传输接口温度为100℃;质谱监测模式为多反应监测,采集模式为正离子模式;用于定量分析的离子对分别为m/z 124.0→79.0(吡嗪酰胺)、m/z 125.1→79.1(吡嗪酸)、m/z 180.0→110.2(内标),去簇电压分别为55、26、85 V,碰撞电压分别为24、23、28 V。使用DAS 2.1.1软件计算药动学参数并比较。结果:吡嗪酰胺和吡嗪酸检测质量浓度的线性范围分别为25~5000 ng/mL(r=0.9976)、100~12500 ng/mL(r=0.9990);定量下限分别为25、100 ng/mL;批内、批间准确度为92.93%~100.50%,批内、批间精密度和基质效应试验的RSD均不高于8.42%(n=6或n=3)。与对照组比较,实验组大鼠体内吡嗪酰胺的tmax显著延长(P<0.01),而其余药动学参数组间比较差异无统计学意义(P>0.05)。结论:大鼠肠道菌群改变可导致单剂量吡嗪酰胺的吸收延迟。OBJECTIVE:To study the effects of rat intestinal flora on the pharmacokinetic parameters of pyrazinamide and its active metabolite pyrazinoic acid.METHODS:Totally 16 SD rats were randomly divided into trial group and control group,with 8 rats in each group.Trial group was given mixed antibiotics(streptomycin sulfate+neomycin sulfate)intragastrically to construct pseudoaseptic rat model.After modeling,both groups were given pyrazinamide intragastrically(150 mg/kg).Before and 0.167,0.333,0.667,1,1.5,2,3,4,6,9 h after administration,0.1 mL blood sample was collected from orbital venous plexus,and 0.3 mL blood sample was collected from orbital venous plexus 12,24 h after administration.Using phenacetin as internal standard,LC-MS/MS method was adopted to determine the plasma concentration of pyrazinamide and pyrazinoic acid.The determination was performed on Agilent ZORBAX SB-Aq column with mobile phase consisted of 0.2%formic acid(containing 8 mmol/L ammonium acetate)-methanol(gradient elution)at the flow rate of 1 mL/min.The column temperature was set at 30℃,and sample size was 10μL.The ion source was ESI and the temperature of ion source was 500℃.The collision gas was nitrogen and the pressure was 10 psi.The temperature of mass transfer interface was 100℃.The mass spectrum monitoring mode was multi reaction monitoring,and the collection mode was positive ion mode.The monitoring transition ion-pairs were m/z 124.0→79.0(pyrazinamide),m/z 125.1→79.1(pyrazinic acid)and m/z 180.0→110.2(internal standard).The de-clustering potential and collision voltage were 55,26 and 85 V,24,23 and 28 V,respectively.The pharmacokinetic parameters were calculated and compared by using DAS 2.1.1 software.RESULTS:The linear ranges of pyrazinamide and pyrazinoic acid were 25-5000 ng/mL(r=0.9976)and 100-12500 ng/mL(r=0.9990).The lower limits of quantification were 25 and 100 ng/mL,respectively.Intra-batch and inter-batch accuracy were 92.93%-100.50%,and RSDs of intra-batch and inter-batch precision and matrix effect tests were

关 键 词：吡嗪酰胺 吡嗪酸 大鼠 肠道菌群 药动学 液相色谱-串联质谱法 

分 类 号：R917[医药卫生—药物分析学] R978.3[医药卫生—药学]