HPLC-DAD测定血浆拉莫三嗪、奥卡西平及10-羟基卡马西平浓度效果观察
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摘要
目的观察采用二极管阵列检测器的高效液相色谱法(HPLC-DAD)测定人血浆中拉莫三嗪(LTG)、奥卡西平(OXC)及活性代谢物10-羟基卡马西平(MHD)浓度的效果。方法以氯唑沙宗为内标,甲醇为蛋白沉淀剂,Agilent Zorbax Eclipse Plus C18(4.6×100mm,3.6μm)为分析柱,流动相为乙腈-10 mmol/L KH2PO4溶液(V/V,24:76,pH 5.5),流速为0.8ml/min,进样量为10μl,检测波长220nm,参比波长360 nm。结果 LTG、OXC和MHD在1.0～40.0μg/ml内线性关系良好,LTG线性方程为y=0.171x+0.036(r=0.99974),MHD的线性方程为y=0.091x+0.018(r=0.99969),OXC的线性方程为y=0.130x+0.036(r=0.99969)。LTG、OXC和MHD在低、中和高浓度质控(2.0、10.0和30.0μg/ml)的方法回收率在93.4%～100.95%,日内和日间精密度均<6%。结论采用HPLC-DAD方法测定人血浆中LTG、OXC及MHD浓度的专属性强、灵敏度高,适用于监测LTG、OXC及MHD的血药浓度。
Objective To develop a new HPLC-based method for the simultaneous determination of lamotrigine(LTG),Oxcarbazepine(OXC) and 10-monohydroxycarbazepine(MHD) in human plasma. Methods LTG, MHD and OXC were extracted from human plasma samples by methanol. The analytes were separated on the Agilent Zorbax Eclipse Plus C18column(4.6×100mm 3.6um) with a mobile phase consisting of acetonitrile-10 m M KH_2PO_4(24:76V/V pH 5.5) and the flow rate of 0.8 ml/min. The injection volume was 10 μl and detective wavelength was set 220 nm. The chlorzoxazone was used as the internal standard(IS). Results The method was validated over the concentration range of 1.0-40.0 μg/ml for LTG, MHD and OXC, the regression equation was(y=0.171x+0.036, r=0.99974) for LTG,(y=0.091x+0.018, r=0.99969) for MHD and(y=0.130x+0.036, r=0.99969) for OXC. The extraction recoveries with low, medium and high quality controls(2.0,10.0 and30.0μg/ml) ranged from 93.4% to100.95%. The Inter-and intra-day precisions were<6%. Conclusion The specificity and accuracy HPLC-DAD method have been validated for detection of LTG, MHD and OXC in human plasma, indicating that the method can be applied for monitoring of LTG, MHD and OXC clinically.
Objective To develop a new HPLC-based method for the simultaneous determination of lamotrigine(LTG),Oxcarbazepine(OXC) and 10-monohydroxycarbazepine(MHD) in human plasma. Methods LTG, MHD and OXC were extracted from human plasma samples by methanol. The analytes were separated on the Agilent Zorbax Eclipse Plus C18column(4.6×100mm 3.6um) with a mobile phase consisting of acetonitrile-10 m M KH_2PO_4(24:76V/V pH 5.5) and the flow rate of 0.8 ml/min. The injection volume was 10 μl and detective wavelength was set 220 nm. The chlorzoxazone was used as the internal standard(IS). Results The method was validated over the concentration range of 1.0-40.0 μg/ml for LTG, MHD and OXC, the regression equation was(y=0.171x+0.036, r=0.99974) for LTG,(y=0.091x+0.018, r=0.99969) for MHD and(y=0.130x+0.036, r=0.99969) for OXC. The extraction recoveries with low, medium and high quality controls(2.0,10.0 and30.0μg/ml) ranged from 93.4% to100.95%. The Inter-and intra-day precisions were<6%. Conclusion The specificity and accuracy HPLC-DAD method have been validated for detection of LTG, MHD and OXC in human plasma, indicating that the method can be applied for monitoring of LTG, MHD and OXC clinically.
引文
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[3]Hiemke C,Baumann P,Clement HW,et al.Consensus Guidelines for Therapeutic Drug Monitoring in Neuropsychopharmacology:update 2017[J].Pharmacopsychiatry,2018,51(1-02):9-62.DOI:10.1055/s-0043-116492.
[4]中华医学会儿科学分会临床药理学组.儿童治疗性药物监测专家共识[J].中华儿科杂志,2015,53(9):650-659.DOI:10.3760/cma.j.issn.0578-1310.2015.09.005
[5]Dupouey J,Doudka N,Belo S,et al.Simultaneous determination of fourantiepileptic drugs in human plasma samples using an ultra-high-performance liquid chromatography tandem mass spectrometry method and its application in therapeutic drug monitoring[J].Biomed Chromatogr,2016,30(12):2053-2060.DOI:10.1002/bmc.3789.
[6]阙万才,丘宏强,程昱,等.固相萃取-高效液相色谱法测定人血浆中奥卡西平及其代谢物10-羟基卡马西平的浓度[J].中国临床药理学杂志,2015,31(22):2254-2256.DOI:10.13699/j.cnki.1001-6821.2015.22.025.
[7]沈芊,王彦改,曾红,等.RP-HPLC法同时测定人血清中拉莫三嗪、苯巴比妥、奥卡西平及其活性代谢物单羟基卡马西平的浓度[J].中国药房,2016,27(11):1493-1496.DOI:10.6039/j.issn.1001-0408.2016.11.17.
[8]国家药典委员会.中华人民共和国药典第四部(2015年版)[M].北京:中国医药科技出版社,2015:341-344.DOI:10.11669/cpj.2015.20.013.
[9]何大为,王丽,王寅初.反相高效液相色谱法同时测定拉莫三嗪和卡马西平血药浓度[J].药物分析杂志,2006,26(2):212-214.DOI:10.16155/j.0254-1793.2006.02.036.