固相萃取RP-HPLC测定小鼠肝脏组织中的德氮吡格

Determination of tetrazanbigen in liver by reversed-phase high performance liquid chromatography

目的:建立小鼠肝脏组织中德氮吡格(Tetrazanbigen)的反相高效液相色谱(RP-HPLC)测定方法。方法:精密称取小鼠肝脏组织,匀浆后固相萃取(Solid phase extraction,SPE)。液相色谱采用Lichrospher C18色谱柱(4.6mm×250mm,5μm),以甲醇-0.1%三乙胺水溶液(pH6.5)(体积比90∶10)为流动相,流速1.0ml/min,检测波长260nm,进样量20μl。结果:实验表明,肝脏组织匀浆液中德氮吡格在2～20μg/ml范围内与峰面积呈良好的线性关系(r=0.9993),平均回收率为106.47%。最低定量限为1.0μg/ml。结论:本文建立的固相萃取RP-HPLC方法简便、准确、选择性和重现性好,适合于德氮吡格的肝脏组织浓度检测。

Objective：To establish a solid phase extraction（SPE）and reversed-phase high-performance liquid chromatographic （RP-HPLC）method for the determination of tetrazanbigen in liver.Methods：Tetrazanbigen in liver samples was determined after a C18 solid phase extraction on a Lichrospher C18 column at 260 nm with an isocratic mobile phase of 90%methanol in 100 ml methanol -deionized water（pH 6.5）,containing 0.1%triethyl amine.The flow rate was set at 1.0 ml/min.Results：The linearity was good over the concentration range of 2～20μg/ml（r=0.9 993）.The average recovery and RSD were 106.47%and 3.0%respectively.The lower limit of quantification was 1.0μg/ml.Conclusion：The method is simple,accurate and can be used to determine the TNBG concentration in liver and study its metabolism.