HPLC-FLD测定大鼠血浆中的10-羟基喜树碱方法学研究  

Determination of 10-hydroxycamptothecin in rat plasma by HPLC-FLD

作　　者：张道旭 刘亮[2] 曲海晶 王洋[3] 海鑫 陈岩[2] 

机构地区：[1]哈药集团技术中心,黑龙江哈尔滨150000 [2]哈尔滨医科大学附属第一医院药学部,黑龙江哈尔滨150000 [3]东北林业大学盐碱地生物资源环境研究中心,黑龙江哈尔滨150000

出　　处：《药学研究》2017年第2期77-80,共4页Journal of Pharmaceutical Research

摘　　要：目的建立一种简单、准确的测定大鼠血浆中10-羟基喜树碱含量的高效液相色谱-荧光检测法(HPLC-FLD)。方法用1%冰醋酸-甲醇溶液处理血浆样品,采用高效液相色谱-荧光检测器,以喜树碱为内标。色谱柱:Phenomenex Luna C18(4.6 mm×250 mm,5μm),流动相:以5%乙腈为流动相A,乙腈为流动相B,流速为1.2m L·min^(-1),荧光检测器激发波长和发射波长分别为Ex=380 nm,Em=515 nm。采用室温梯度洗脱,进样体积为50μL。结果本试验采用两条分段函数,大鼠血浆中10-羟基喜树碱在1.25~20 ng·m L^(-1)及20~320 ng·m L^(-1)范围内线性良好,线回归系数分别为0.999 8和0.999 5。方法准确度为98.9%~103.8%,10-羟基喜树碱高、中、低3个浓度的提取回收率均大于80%。日内精密度RSD<4.2%,日间精密度RSD<4.8%。结论该方法操作简便,灵敏度高,精密度和准确度好,适用于大鼠血浆中10-羟基喜树碱含量的测定。Objective To establish a simple and accurate HPLC-FLD method for the determination of 10-hydroxycamptothecin in rat plasma.Methods The serum sample was performed the precipitation treatment by 1% acetic acid methanol solution,then HPLC-FLD was adopted for detection. With camptothecin as the internal standard,the chromatographic column was the Phenomenex Luna C18（ 4.6 mm×250 mm,5 μm）,the mobile phase was 5% acetonitrile（ A） and acetonitrile（ B）.The excitation wavelength and emission wavelength were at 380 nm,515 nm,respectively and the mobile phase rate was 1.2 mL·min^-1.The injection volume was 50 μL.Results The concentration of 10-hydroxycamptothecin in rat plasma in the range of 1.25 - 20 ng·mL^-1and 20 - 320 ng·mL^-1showed a good linearity.The regression coefficients were 0.999 8and 0.999 5,respectively.The accuracy of the method was 98.9% - 103.8%.The recovery rates of low,middle and high concentrations were more than 80%.The intra-day RSD was less than 4.2% and the inter-day RSD was less than 4.8%. Conclusion The method was simple,sensitive,precise and accurate.It was suitable for the determination of 10-hydroxytryptine in rat plasma.

关 键 词：高效液相色谱-荧光检测法 大鼠 10-羟基喜树碱 血浆 

分 类 号：R965[医药卫生—药理学]