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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 reversed-phase high performance liquid chromatography (RP-HPLC) method for the determination of Tetrazanbigen in mouse liver tissue. Methods: The liver tissues of mice were weighed accurately and then homogenized by solid phase extraction (SPE). Liquid chromatography was performed on a Lichrospher C18 column (4.6 mm × 250 mm, 5 μm) with methanol-0.1% triethylamine in water (pH 6.5) (90:10 by volume) as the mobile phase at a flow rate of 1.0 ml / min. 260nm, injection volume 20μl. Results: The experimental results showed that there was a good linearity (r = 0.9993) between the range of 2 ~ 20μg / ml and the average recovery of rhodanopig in the liver tissue homogenate with the average recovery of 106.47%. The limit of quantification was 1.0 μg / ml. Conclusion: The RP-HPLC method established in this paper is simple, accurate, selective and reproducible. It is suitable for the detection of hepatic tissue concentrations in zidiazide.