UPLC-MS/MS同时测定大鼠血浆中美沙酮、文拉法辛及其代谢物浓度与药动学研究
x

请在关注微信后,向客服人员索取文件

篇名: UPLC-MS/MS同时测定大鼠血浆中美沙酮、文拉法辛及其代谢物浓度与药动学研究
TITLE:
摘要: 目的:建立测定大鼠血浆中美沙酮和文拉法辛及其代谢产物浓度的方法并用于其药动学研究。方法:大鼠血浆样品经乙腈沉淀蛋白后,以地西泮为内标,采用超高效液相色谱-串联质谱法测定。色谱柱为Waters Acquity UPLC BEH C18,流动相为乙腈-0.1%甲酸(梯度洗脱),流速为0.4 mL/min,柱温为40 ℃,进样量为2 μL。采用电喷雾电离源,以多反应监测方式进行正离子扫描,用于定量分析的离子对分别为m/z 310.4→265.4(美沙酮)、m/z 278.2→234.1[2-亚乙基-1,5-二甲基-3,3-二苯基吡咯烷(EDDP)]、m/z 278.1→57.8(文拉法辛)、m/z 263.9→57.9(O-去甲文拉法辛)、m/z 285.1→193.1(内标)。取8只SD大鼠分别灌胃盐酸美沙酮6 mg/kg、盐酸文拉法辛10 mg/kg(每个药物各4只大鼠),分别于给药前及给药后0.083、0.167、0.25、0.5、0.75、1、1.5、2、3、4、6、8、10、12、24 h经尾静脉取血0.3 mL进样测定,采用DAS 3.0软件计算药动学参数。结果:美沙酮、EDDP、文拉法辛和O-去甲文拉法辛的质量浓度线性范围分别为0.5~250 ng/mL(r=0.999 7)、0.5~250 ng/mL(r=0.999 2)、0.4~200 ng/mL(r=0.999 9)、0.4~200 ng/mL(r=0.999 9),定量下限分别为0.5、0.5、0.4、0.4 ng/mL;日内、日间RSD均小于9.0%(n=6);方法回收率分别为94.20%~102.87%、90.93%~102.94%、92.95%~101.61%、90.33%~101.97%(RSD均≤5.5%,n=6);提取回收率分别为85.90%~94.45%、85.97%~91.66%、87.97%~93.58%、88.53~94.54%(RSD均≤5.9%,n=6);基质效应分别为95.96%~97.78%、92.33%~97.40%、95.28%~97.71%、95.33%~95.74%(RSD均≤4.9%,n=3);t1/2分别为(1.42±1.02)、(2.59±0.76)、(0.63±0.08)、(1.29±0.14) h;cmax分别为(52.21±5.42)、(25.68±3.45)、(52.64±2.29)、(47.63±13.09) μg/L;MRT0-24 h分别为(3.55±0.21)、(3.98±0.41)、(1.44±0.21)、(2.01±0.17) h;AUC0-24 h分别为(201.95±51.14)、(86.09±15.95)、(75.38±23.95)、(82.90±23.44) μg·h/L。结论:建立的方法专属性强、分离完全、快速灵敏,可用于同时测定美沙酮及文拉法辛及其代谢物的血药浓度及其在大鼠体内的药动学研究。
ABSTRACT: OBJECTIVE: To establish a method for simultaneous determination of methadone, venlafaxine and their metabolites in rat plasma, and to use it for the study of pharmacokinetic in rats. METHODS: UPLC-MS/MS method was adopted to determine plasma after precipitated with acetonitrile using diazepam as internal standard. The determination was performed on Waters Acquity UPLC BEH C18 column with mobile phase consisted of acetonitrile-0.1% formic acid (gradient elution) at the flow rate of 0.4    mL/min. The column temperature was set at 40 ℃, and sample size was 2 μL. ESI was used for positive ion scanning by multiple reaction monitoring (MRM) mode. The ion pairs for quantitative analysis were m/z 310.4→265.4(methadone), m/z 278.2→234.1(EDDP), m/z 278.1→57.8(venlafaxine), m/z 263.9→57.9(O-desvenlafaxine), m/z 285.1→193.1(internal standard). Eight SD rats were given methadone hydrochloride 6 mg/kg and venlafaxine hydrochloride 10 mg/kg intragastrically (4 rats for each drug); blood samples (0.3 mL) were collected from the tail vein before and 0.083, 0.167, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, and 24 h after medication. Pharmacokinetic parameters were calculated by using DAS 3.0 software. RESULTS: The linear ranges of methadone, EDDP, venlafaxine and O-desvenlafaxine were 0.5-250 ng/mL (r=0.999 7), 0.5-250 ng/mL (r=0.999 2), 0.4-200 ng/mL (r=0.999 9), 0.4-200 ng/mL (r=0.999 9), respectively. The limits of quantitation were 0.5, 0.5, 0.4, 0.4 ng/mL. RSDs of intra-day and inter-day were all lower than 9.0% (n=6). The method recoveries were 94.20%-102.87%, 90.93%- 102.94%, 92.95%-101.61%, 90.33%-101.97%(RSD≤5.5%,n=6). The extraction recoveries were 85.90%-94.45%, 85.97%- 91.66%, 87.97%-93.58%, 88.53%-94.54%(RSD≤5.9%,n=6). The matrix effects were 95.96%-97.78%, 92.33%-97.40%, 95.28%-97.71%, 95.33%-95.74%(RSD≤4.9%,n=3). The pharmacokinetic parameters included that t1/2 were (1.42±1.02), (2.59±0.76),(0.63±0.08),(1.29±0.14) h;cmax were (52.21±5.42), (25.68±3.45),(45.68±2.29),(47.63±13.09) μg/L; MRT0-24 h were (3.55±0.21), (3.98±0.41),(1.44±0.21),(2.01±0.17) h;AUC0-24 h were (201.95±51.14), (86.092±15.95), (75.38±23.95),(82.90±23.44)μg·h/L. CONCLUSIONS: The method is specific, absolute separated, rapid and sensitive, and can be used for the simultaneous determination of methadone, venlafaxine and their metabolites in plasma and pharmacokinetics research.
期刊: 2018年第29卷第11期
作者: 李好,陈连国,尤玮玮
AUTHORS: LI Hao,CHEN Lianguo,YOU Weiwei
关键字: 超高效液相色谱-串联质谱;大鼠;美沙酮;文拉法辛;代谢产物;血药浓度;药动学
KEYWORDS: UPLC-MS/MS; Rat; Methadone; Venlafaxine; Metabolite; Plasma concentration;Pharmacokinetics
阅读数: 243 次
本月下载数: 5 次

* 注:未经本站明确许可,任何网站不得非法盗链资源下载连接及抄袭本站原创内容资源!在此感谢您的支持与合作!