黄酮类化合物对托莫西汀体外代谢的影响  

Effect of flavonoids on the metabolism of atomoxetine in vitro

作　　者：梁冰清 朱珺 郑璐璐 LIANG Bing-qing;ZHU Jun;ZHENG Lu-lu(Department of Pharmacy,Tongde Hospital of Zhejiang Province,Hangzhou 310012,China)

机构地区：[1]浙江省立同德医院,杭州310012

出　　处：《药物分析杂志》2020年第7期1226-1235,共10页Chinese Journal of Pharmaceutical Analysis

基　　金：浙江省药学会医院药学专项科研资助项目(2018ZYY10)。

摘　　要：目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)方法测定体外样品中托莫西汀(ATX)及其活性代谢产物4-羟基托莫西汀(4-HAT)的浓度;研究15个黄酮类化合物对ATX在体外代谢的影响,为体内代谢研究提供一定的实验基础。方法:(1)采用Waters ACQUITY UPLC BEH C18色谱柱(2.1mm×50 mm,1.7μm)进行色谱分离,以0.1%甲酸-乙腈为流动相,流速0.4 mL·min^-1采用电喷雾离子源,正离子模式进行检测,用于定量分析的离子对为m/z 256.3→44(ATX)、m/z 272.3→44(4-HAT),建立ATX及其活性代谢产物4-HAT的UPLC-MS/MS检测方法。(2)建立ATX大鼠肝微粒体(RLM)孵育体系。分别在孵育体系内加入二甲基亚砜(DMSO)和15个12.5 mmol·L^-1黄酮类化合物溶液各1.6μL进行孵育,检测4-HAT的生成量并进行分析与比较。结果:ATX质量浓度在1~100 ng·mL^-1范围内与峰面积的线性关系良好(r2=0.9996),定量下限为0.2 ng·mL^-1(S/N=10);4-HAT质量浓度在0.01~2.5 ng·mL^-1范围内与峰面积的线性关系良好(r2=0.9993),定量下限为0.01 ng·mL^-1(S/N=10)。15个黄酮类化合物中,杨梅素对ATX的体外代谢影响最显著,表现为抑制作用;与DMSO组相比,其抑制率为93.44%。结论:该方法可用于测定ATX及其活性代谢产物4-HAT的含量;体外实验表明杨梅素对ATX代谢影响最显著。本实验结果可以为体内代谢研究提供一定的实验基础。Objective:To establish an UPLC-MS/MS method for the determination of the concentration of atomoxetine(ATX)and its active metabolite 4-hydroxyatomoxetine(4-HAT)in samples in vitro.To study the effects of 15 flavonoids on the metabolism of ATX in vitro,and provide a certain experimental basis for the study of metabolism in vivo.Methods:(1)UPLC-MS/MS method was established using Waters ACQUITY UPLC BEH C18 column(2.1 mm×50 mm,1.7μm),0.1%formic acid-acetonitrile as mobile phase,and a flow rate of 0.4 mL·min^-1 for determination the concentration of ATX and its active metabolite 4-HAT.An electrospray onization(ESI+)interface was used for mass spectrometric detection.Quantitation was performed using multiple reaction monitoring of the transitions of m/z 256.3→44,m/z 272.3→44 for ATX and 4-HAT,respectively.(2)To establish a rat liver microsome(RLM)incubation system for ATX incubation.1.6μL dimethyl sulfoxide(DMSO)and 1.6μL 12.5 mmol·L^-1 flavonoid solutions were respectively added to the incubation system for incubation,and the amount of 4-HAT produced was detected,and then analyzed and compared.Results:The linearity of ATX was in the range of 1 to 100 ng·mL^-1(r2=0.9996).The limit of quantitation was 0.2 ng·mL^-1(S/N=10).The linearity of 4-HAT was in the range of 0.01 to 2.5 ng·mL^-1(r2=0.9993).The limit of quantitation was 0.01 ng·mL^-1(S/N=10).Among the 15 flavonoids,myricetin had the most significant effect on the metabolism of ATX in vitro.Compared with the control group,the inhibition rate was 93.44%.Conclusion:This method can be used to determine the concentration of ATX and its active metabolite 4-HAT.In vitro experiments shows that myricetin has the most significant effect on ATX metabolism.These results can provide a certain experimental basis for metabolic studies in vivo.

关 键 词：超高效液相色谱-串联质谱 托莫西汀 4-羟基托莫西汀 选择性去甲肾上腺素再摄取抑制剂 非兴奋型药物 黄酮类 体外代谢 代谢产物 大鼠肝微粒体 孵育体系 

分 类 号：R917[医药卫生—药物分析学]