HPLC-荧光法测定人血浆中伊立替康及其活性代谢产物的浓度
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篇名: HPLC-荧光法测定人血浆中伊立替康及其活性代谢产物的浓度
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摘要: 目的:建立同时测定人血浆中伊立替康(CPT-11)及其活性代谢产物7-乙基-10-羟基喜树碱(SN-38)浓度的方法。方法:血浆样品经乙腈沉淀蛋白及盐酸酸化后,以喜树碱为内标,采用高效液相色谱-荧光法测定。色谱柱为Waters Luna C18,流动相为0.05 mol/L磷酸二氢钠溶液-乙腈(70 ∶ 30,V/V,用磷酸调节pH至4.0),流速为1 mL/min,激发波长为380 nm,发射波长为480 nm(CPT-11)、535 nm(SN-38),柱温为25 ℃,进样量为20 μL。结果:CPT-11和SN-38血药浓度分别在200~1 000、5~45 ng/mL范围内线性关系良好(r分别为0.999 4、0.999 2,n=5),定量下限分别为200、5 ng/mL;日内、日间RSD为1.68%~5.57%;CPT-11和SN-38的相对回收率分别为90.12%~106.93%(RSD<8%,n=5)、92.07%~102.56%(RSD<6%,n=5),提取回收率分别为72.23%~86.56%(RSD<6%,n=5)、71.98%~83.44%(RSD<7%,n=5)。采用该方法测得5例结肠癌患者体内CPT-11和SN-38的血药浓度分别为431.13~617.19、13.97~31.89 ng/mL(静脉滴注结束后1 h),398.14~584.43、11.61~29.94 ng/mL(静脉滴注结束后2 h)。结论:该方法样品处理简单、快速,且灵敏度高、重复性好,适用于临床常规监测CPT-11及其代谢物SN-38的血药浓度及药动学研究。
ABSTRACT: OBJECTIVE: To develop a method for simultaneous determination of irinotecan (CPT-11) and its active metabolite 7-ethyl-10-hydroxycamptothecin (SN-38) in human plasma. METHODS: After precipitated by acetonitrile and acidified with hydrochloric acid, using camptothecin as internal standard, the plasma sample was determined by HPLC-FLD. The determination was performed on Waters Luna C18 column with mobile phase consisted of 0.05 mol/L sodium dihydrogen phosphate-acetonitrile (70 ∶ 30,V/V, adjusted pH to 4.0 by phosphoric acid) at flow rate of 1 mL/min. The excitation wavelength was set at 380 nm; the emission wavelengths of CPT-11 and SN-38 were set at 480 nm and 535 nm, respectively. The column temperature was 25 ℃ and the sample size was 20 μL. RESULTS: The linear ranges were 200-1 000 ng/mL for CPT-11 (r=0.999 4,n=5) and 5-45 ng/mL for SN-38 (r=0.999 2,n=5). RSDs of inter-day and intra-day were 1.68%-5.57%. The relative recoveries of CPT-11 and SN-38 were 90.12%-106.93%(RSD<8%,n=5) and 92.07%-102.56%(RSD<6%,n=5); the extraction recoveries of CPT-11 and SN-38 were 72.23%-86.56%(RSD<6%,n=5) and 71.98%-83.44%(RSD<7%,n=5),respectively. The plasma concentrations of CPT-11 and SN-38 in 5 patients with colon cancer were 431.13-617.19, 13.97-31.89 ng/mL(1 h after intravenous dripping) and 398.14-584.43, 11.61-29.94 ng/mL(2 h after intravenous dripping). CONCLUSIONS: The method is simple, rapid, sensitive, reproducible and suitable for the determination of plasma concentration and pharmacokinetic study of CPT-11 and its metabolite SN-38.
期刊: 2017年第28卷第29期
作者: 张晓霈,刘晓明,徐今宁,王爱萍,姜爱雯
AUTHORS: ZHANG Xiaopei,LIU Xiaoming,XU Jinning,WANG Aiping,JIANG Aiwen
关键字: 高效液相色谱-荧光法;伊立替康;7-乙基-10-羟基喜树碱;血药浓度
KEYWORDS: HPLC-FLD; Irinotecan; 7-ethyl-10-hydroxycamptothecin; Plasma concentration
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