高效液相色谱法测定生物样品中3PO的方法建立与验证（英文）  

Development and validation of an HPLC method for the analysis of 3PO in biological samples

作　　者：Yemarshet K.Gebreyohannes 王旭 Getu Kahsay Weldu Erwin Adams Ann Van Schepdael 

机构地区：[1]荷语天主教鲁汶大学药学院药物分析实验室

出　　处：《药物分析杂志》2013年第11期1873-1876,1886,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立并验证用于大鼠血清、脑脊液、大脑和脊髓中3-(3-吡啶)-1-(4-吡啶)-2-丙烯-1-酮(3PO)含量测定的HPLC-UV法。方法:样品处理使用蛋白沉淀的方法;色谱条件:色谱柱为Chromolith RP-18e(100 mm×4.6 mm),流动相A为乙腈-水(2.5∶97.5,v/v)添加0.1%TFA,流动相B为乙腈添加0.1%TFA,梯度洗脱[0～2.3 min,0%B→100%B(线性梯度);2.3～2.6 min,100%B(等度);2.6～2.7 min,100%B→0%B(线性梯度);2.7～5 min,0%B(等度)],流速2 mL·min-1,检测波长304 nm,进样量10μL。结果:大鼠的血清、脑脊液、大脑和脊髓中内源性物质不干扰3PO的测定,在上述4种基质中,该方法的检测限分别为0.2、0.1、0.1、0.1μg·mL-1;定量限分别为0.8、0.2、0.5、0.5μg·mL-1;线性范围分别为3～150、0.3～150、0.9～150、0.6～150μg·mL-1,相关系数均大于0.999;高、低浓度的回收率均在79.9%～100.3%的范围内;方法重复性(n=5)的RSD均大于0.9%。结论:建立的方法简单,线性范围宽,且选择性、耐用性均佳。To develop and validate a high performance liquid chromatographic method with UV detec- tion for the determination of 3 - （ 3 - pyridinyl） - 1 - （4 - pyridinyl） - 2 - propen - 1 - one （3PO） in rat＇ s serum, eerebrospinal fluid（CSF）, brain and spinal cord. Methods： Protein precipitation was used as sample preparation method. The separation was performed on a Chromolith performance RP - 18e column （100 mm × 4. 6 mm）by gradi- ent elution where mobile phase A consisted of 0. 1% trifluoroacetie acid in aeetonitrile -water（2. 5：97.5,v/v）and B consisted of 0. 1% trifluoroacetic acid in acetonitrile. The gradient program was set as follows ：0 -2. 3 min ,0% 100% of B （ linear gradient） ;2. 3 - 2. 6 min, 100% of B （isoeratie） ;2. 6 - 2. 7 min, 100% ---0% of B （ linear gradi- ent） ;2. 7 -5 min ,0% of B（isoeratic）. Flow rate was set at 2 mL · min-1. UV detection was performed at 304 nm. The injection volume was 10 μL. Results：There was no endogenous interference at the retention time of 3PO in blank brain, spinal cord and CSF. LOD and LOQ of the method in serum, CSF, brain and spinal cord were 0. 2,0. 1, 0. 1,0. 1 μg · mL-1 and O. 8,0. 2,0. 5,0. 5 μg · mL-1 ,respectively. The method was linear in the range from 0. 3 -150μg · mL-1 in CSF,0. 9 -150 μg · mL-l in brain,0. 6 -150 μg ·mL-1 in spinal cord and 3 -150 μg · mL-1 in serum with coefficient of determination values greater than 0. 999. The recovery of high concentration and low concentration were between 79.9% -100. 3%. Conclusion：The method is proved to be simple, robust, selec- tive and linear.

关 键 词：高效液相色谱 生物分析 3PO 3-(3-吡啶)-1-(4-吡啶)-2-丙烯-1-酮 

分 类 号：R917[医药卫生—药物分析学]