对氯苯酚提取-液相色谱法分析烟草中的磷酸腺苷  

Measurement of adenosine phosphate in p-Chlorophenol pretreatment flue-cured tobacco by HPLC

作　　者：林小峰[1,2] 吴玉萍[1] 夏振远[1] 孔光辉[1] 郑保忠[2] 

机构地区：[1]云南省烟草农业科学研究院,玉溪653100 [2]云南大学材料科学与工程系,昆明650091

出　　处：《分析试验室》2014年第3期296-299,共4页Chinese Journal of Analysis Laboratory

基　　金：国家烟草专卖局项目(110201002001)资助

摘　　要：建立了新鲜烟叶中三磷酸腺苷（ATP）、二磷酸腺苷（ADP）和单磷酸腺苷（AMP）3种磷酸腺苷同时测定的方法。烟草样品经过对氯苯酚提取、纯化后，采用三乙胺缓冲液作为流动相洗脱，经C18色谱柱分离，在259nm波长对样品进行检测。与高氯酸（PCA）法，以及DNA常用的苯酚提取法进行了比较，结果均表明：低温下以对氯苯酚提取的效果较佳，使用三乙胺缓冲液流动相具有分离效率高、检出灵敏的优点。在线性范围（5～200μg/mL）内，磷酸腺苷浓度与峰面积具有很好的相关系数（r〉0．9999），检出限达到纳克级水平，氯苯酚法对ATP，ADP和AMP的平均回收率分别为96．8％，97．2％和93．3％，RSD均小于5％。A simultaneous analysis method was established to detect adenosine triphosphate （ATP）, adenosine diphosphate （ADP） and adenosine monophosphate （AMP） in fresh tobacco leaves. The tobacco sample was extracted by p-Chlorophenol and separated on a SunFire C18 with triethylamine buffer as mobile phase, then detected by HPLC with detection wavelength at 259 nm. Compared with perchloric acid and phenol-TE extraction methods, the result showed that the optimized Triethylamine buffer as mobile phase has the The peak areas had a linear relationship extraction method was to use p-Chlorophenol at low temperature. advantages of high separation efficiency and detection sensitivity. with concentrations of adenosine phosphate in the range of 5 - 200 μg/mL with good correlation coefficients, （ r 〉 0. 999）. Detection limits reached nanogram level. The spiking recoveries in samples were 96. 78%, 97. 17%, 93.26% respectively. The relative standard deviations （RSD） were less than 5%. This method can quickly and accurately detect adenosine phosphate of tobacco samples, and it provides an important detection way for the influence study of adenosine phosphate to the genetics and breeding, growth and development of tobacco.

关 键 词：高效液相色谱 ATP ADP AMP 对氯苯酚 烟草 

分 类 号：O657.7[理学—分析化学]