反相高效液相色谱法测定人血浆中腺苷浓度

Determination of adenosine in human plasma by RP-HPLC

目的：建立人血浆中腺苷浓度的RP-HPLC测定方法，考察腺苷在人血浆中的稳定性。方法：色谱柱为Agilent Zorbax SB-C18（4．6mm×250mm，5μm），以磷酸缓7中液（含50mmol·L^-1 NaH2PO4，50mmol·L^-1 Na，HPO4，0．05mol·L^-1四丁基溴化铵）：乙腈=95：5为流动相，检测波长为260nm。结果：腺苷在0．7—40μg·mL^-1范围内线性关系良好；回归方程为A=30．302C＋2、8423（r=0．9998）；最低定量限为0．7μg·mL^-1；平均回收率为94％。腺苷在人血浆中静置45min，降解率为40％；静置2h，降解率为90％。结论：本法专属性高、样品处理过程简单，可用于测定人血浆中腺苷的浓度；除了红细胞可以摄取灭活腺苷外，人血浆中存在的多种酶类对腺苷也有较强的代谢能力，腺苷在人血浆中具有一定的不稳定性，测定时样品必须在采集后立即处理。

Objective：To determine the concentration of adenosine by RP-HPLC and to study the stability of adenosine in human plasma. Methods： An Agilent Zorbax SB-C18 column （4.6 mm×250mm,5μm） was used with the mobile phase consists of phoshpate buffer solution （50mmol· L^-1 Nail2 PO4：50 mmol·L^- 1 Na2 HPO4 ： 0.05 mol· L^- 1 tetrabutyl ammonium bromide） - acetonitrile （95： 5, v： v） at the detection wavelength of 260 nm. Results： The calibration curve showed a good linearity with the range of 0.7 - 40μg·mL^-1 （ r = 0. 999 8）. The average recovery was 94%. The limit of quantification of adenosine was 0.7 μg·mL^-1. Forty percent adenosine was metabolized in human plasma within 45 min and ninety percent within 2 h. Conclusion ： The assay in this study is sensitive, simple and rapid. It could be used to determine the concentration of adenosine in human plasma. As adenosine was not stable in human plasma, plasma sample should be measured as soon as it was collected.