反相离子对色谱法测定环磷腺苷有关物质

目的建立反相离子对色谱法检测环磷腺苷的有关物质。方法用十八烷基硅烷键合硅胶为填充剂 ;以 0 .0 5mol/L磷酸二氢钾溶液 (含 0 .0 1mol/L四丁基溴化铵 ,pH 4 .3) 乙腈 (85∶15 )为流动相 ;检测波长为 2 5 9nm ;流速为 1.0ml/min。结果腺苷、5′ 腺苷酸、环磷腺苷和降解产物之间的分离度分别为 9.2、11.5、14 .3,腺苷与 5′ 腺苷酸的检测限均为 0 .1ng、定量限均为 0 .3ng。结论此法分离度好、灵敏度高 ,操作简便 ,可有效分离、检测起始原料带入的杂质和降解产物 ,用于环磷腺苷中的有关物质控制 ,优于纸色谱法。

Determination of related substances of cAMP by RP-Ion-Pair chromatography

PurposeTo establish an HPLC method for determination of cAMP′s related substances.MethodsAn optimal HPLC method was set up. The samples were separated on a C 18 column using 0.05 mol/L KH 2PO 4(containing 0.01mol/L TBA,pH 4.3)-acetonitrile(85∶15)as mobile phase and detected at 259 nm. The flow rate was 1.0 ml/min. ResultsAdenosin, AMP, cAMP and unknown substance were well separated. Separation factors were 9.2, 11.5 and 14.3 respectively. The detection limits of Adenosin and AMP were both 0.1 ng, and the determination limits were both 0.3 ng.ConclusionThis method was convenient, reliable and sensitive. It was superior to paper chromatography.