反相高效液相色谱法检测带鱼糜中的嘌呤含量  

Determination of Purine Content in Hairtail Surimi by Reversed-phase High Performance Liquid Chromatography

作　　者：吕兵兵[1] 张进杰[1] 储银[1] 程玉[1] 顾伟钢[1] 陈健初[1] 

机构地区：[1]浙江大学生物系统工程与食品科学学院,杭州310058

出　　处：《中国食品学报》2012年第7期192-198,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：浙江省科技计划项目(2010C12012)

摘　　要：目的:建立一种检测水产品中游离嘌呤和水解后总嘌呤含量的反相高效液相色谱法,并将此方法应用到带鱼糜中游离态嘌呤和总嘌呤的含量测定中。方法:以Waters Atlantis dC18色谱柱为固定相,0.02mol/L磷酸二氢钾缓冲溶液(pH3.62)为流动相进行色谱分离;流速:1.0mL/min;柱温:30℃;进样量:10μL;DAD检测波长:254nm。采用水处理样品并在90℃下水解12～14min后取上清液过膜,测定4种游离嘌呤的含量;采用三氟乙酸/甲酸/水(3者体积比5:5:1)处理样品并在90℃下水解12min,水解液浓缩,过膜后测定4种嘌呤总量。结果:在20min内4种嘌呤能得到很好分离,且在质量浓度1～40mg/L范围,4种嘌呤的响应峰面积与其相应浓度相关性良好,r2>0.9998,精密度RSD<2%。结论:该方法简便、快速、准确,可用于检测水产品中游离嘌呤和总嘌呤的含量。objective： A reversed-phase high performance liquid chromatography method for the quantitative determi- nation of free purine and total purine content after hydrolyzing in fishery products was studied, and the contents of free purine and total purines in hairtail surimi were determined. Method： Waters Atlantis dC18 column was used as the sta- tionary phase and 0.02 mol/L KH2PO4 （pH 3.62）as mobile phase. Flow rate： 1.0mL/min; Column temperature： 30 ℃; In- jection volume： 10 μL; DAD detector： 254 nm; the contents of free purine were determined by hydrolyzing with water at 90℃ for 12-14 min; the contents of total purine were determined by hydrolyzing with mixture of trifluoroacetic acid： formid acid： water （5：5：1,V/V/V） at 90℃ for 12 mins; Results： The method had a good resolution in 20 minutes, the peak area of purines and concentration achieved good linear relation ranged from 1 to 40 mg/L, r2〉0.9998, the RSD was 〈2%. Conclusion： The method was simple, rapid and accurate, it can be applied to determining free purine and total purine in fishery products.

关 键 词：反相高效液相色谱 带鱼糜 游离嘌呤 总嘌呤 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程] O657.72[轻工技术与工程—食品科学与工程]