高效液相色谱法测定几种热带水果ATP、ADP和AMP含量  

Determination of ATP, ADP and AMP concentrations in several tropical fruits by high performance liquid chromatography

作　　者：莫亿伟[1] 金文龙[1] 刘锴栋[2] 

机构地区：[1]绍兴文理学院生命科学学院,浙江绍兴312000 [2]湛江师范学院生命科学学院,广东湛江524048

出　　处：《果树学报》2013年第6期1077-1082,共6页Journal of Fruit Science

基　　金：海南省自然科学基金(90353);国家自然科学基金(31201586)

摘　　要：【目的】为了更好地了解热带水果能量组分含量和能量供应水平对果实采后保鲜的影响【方法】以番石榴、番荔枝、荔枝、龙眼的果实为试材，通过优化色谱法流动相和检测波长的条件，使用高效液相色谱法测定新鲜和保鲜结束时这4种果实中ATP（三磷酸腺苷）、ADP（Z-磷酸腺苷）和AMP（一磷酸腺苷）的含量。【结果】当色谱条件为安捷伦C18色谱柱（250mmx4．5mm，5μm）；柱温为30℃；进样体积10μL；紫外检测波长259nm；流速为1．0mL·min～，流动相为35mmol·L-1NaHEP04缓冲液时，ATP、ADP和AMP在1750mg·l。内与峰面积呈良好的线性关系，线性相关系数分别为0．9991，0．9988和0．9992，加标回收率分别为97．49％～99．37％，96．59％。99．21％和97．87％。99．26％。日内精密度分别为1．14％、4．09％和3．32％，日间精密度分别为2．90％、2．31％和5．71％。【结论】该方法避免了有机流动相甲醇和乙腈的使用，具有灵敏性高、准确、重复性好等优点，可应用于热带水果能荷组分分析和能荷值的计算。[Objective]The objective of the stady were to explore influences of energy supply and energy components on postharvest preservation in tropical fruits. [Method]A new method was established for de termining contents of ATP, ADP and AMP at harvest and end of storage time in guava, sugar apple, litchi and longan fruits by high performance liquid chromatography. [Result]Standards and extracted compo-nents were eluted by using Extend-C 18 analytical column maintained at constant temperature of 30 ℃ and detected at 259 nm. Mobile phase was 35 retool. L-1 Na2HPO4 and flow rate was 1.0 mL. rain-1, and pH value was adjusted to 6.8 with 1 tool. L-t NaOH. The results showed that calibrated linear curve of ATP, ADP and AMP concentration was within 1-50 mg .L-1, with r=0.9991, 0.9988 and 0.9992, respectively. The detection limit of ATP, ADP and AMP was 0.05, 0.1 and 0.1 rag＂ L-1 （S/N 3）, respectively. The rela- tive and absolute recovery rate of ATP, ADP and AMP was 97.49%-99.37% , 96.59%-99.21% and 97.87%-99.26%, respectively, and RSD of ATP, ADP and AMP was all less than 5.0 %. The within-day RSD of ATP, ADP and AMP were 1.14, 4.09 and 3.32%, respectively. The between-day RSD of ATP, ADP and AMP were 2.90, 2.31 and 5.71%, respectively. [Conclusion]This new method, avoiding using organic solvents methanol and acetonitrile, has properties of accuracy, sensitivity and repeatability, which indicates that it can be applied in determining ATP, ADP and AMP concentrations and calculating energy charge in tropical fruits.

关 键 词：热带水果 三磷酸腺苷 能荷 高效液相色谱 

分 类 号：S667[农业科学—果树学]