非水毛细管电泳法分离3种单磷酸腺苷及其应用研究

采用非水毛细管电泳方法对2’-AMP、3’-AMP和5’-AMP 3种单磷酸腺苷进行分离研究,考察了电泳溶液pH*值、非水介质、缓冲溶液对分离的影响。以含50%乙腈的Tris-H3BO3体系为缓冲溶液,在pH*10.0、压差进样(50 mbar,5 s)、柱温25℃、25 kV恒压下进行分离,在波长260 nm处负极检测,各组分可达到基线分离。在质量浓度为1～100 mg/L范围内,3种单磷酸腺苷的线性关系良好,平均回收率为88%～106%,RSD小于4%。该方法应用于核苷样品的测定,结果满意。

Separation of Three Single Phosporic Acidic Adenosines by a Nonaqueous Capillary Electrophoresis Method and Its Application

The separation of 2′-AMP,3′-AMP and 5′-AMP were investigated by a nonaqueous capillary electrophoresis(NACE) method.Effects of pH* value,nonaqueous solvent,different category buffer on the separation efficiency were studied.The optimum separation conditions are as follows:running medium:200 mmol·L-1 Tris-H3BO3 containing 50% acetonitrile,pH* value:10.0;applied electric field:25 kV;column temperature:25 ℃;detection wavelength:260 nm,and samples were injected from the cathodal end of the capillary by 50 mbar for 5 s.Under the optimal conditions,there are excellent linearities between peak heights and concentrations of the analytes in the concentration range of 1-100 mg/L.The spiked recoveries ranged from 88% to 106% with RSDs less than 4%.The method was applied in the determination of adenosine samples with satisfactory results.