纸色谱-分光光度法测定生物转化体系中脱氧腺苷含量

目的建立纸色谱-分光光度法测定生物转化液中脱氧腺苷(dAR)含量。方法用不同极性的展开剂进行纸色谱从混合物中分离dAR,分光光度法测定分离的dAR浓度。结果测试了4种展开剂对dAR的分离效果,发现用展开剂正丁醇-异丙醇-氨水-水(3∶3∶2∶2)一次色谱可将dAR与其它成分分离。将分离并重新溶解的dAR在258 nm处测定其吸光度值,根据标准曲线即可计算出转化反应液中的产物浓度。结论该法测定结果与高效液相色谱法测定结果一致,是一种简易有效的快速测定生物转化体系中dAR含量的方法。

Determination of deoxyadenosine in biotransformation system by paper chromatography-spectrophotometry

Purpose To establish a paper chromatography-spectrophotometry method for the determination of deoxyadenosine content quickly and easily in biotransrormation system.MethodsThe deoxyadenosine product was separated from mixture by means of the paper chromatography method with different polarity chromatographic solutions,and the concentration of separated deoxyadenosine was measured with spectrophotometer.ResultsThe separation effect of 4 kinds chromatographic solutions were tested.The separation of deoxyadenosine could be achieved effectively,when solution n-butanol-isopropyl alcohol-ammonia solution-water(3∶3∶2∶2) was used as solvent to do single-dimensional paper chromatography.The separated deoxyadenosine was re-dissolved,and its absorption value was measured at 258 nm.The concentration of deoxyadenosine in biotransformation liquid could be calculated according to the absorption value and standard curve.ConclusionThe determining results by of paper chromatography-spectrophotometry method was the same as that by HPLC method.This method was simple and effective with a good reproducibility for the determination of deoxyadenosine product in biotransrormation system.