高效液相色谱法同时测定肉类食品中的嘌呤和尿酸

目的建立同时测定肉类食品中4种嘌呤(腺嘌呤、鸟嘌呤、次黄嘌呤、黄嘌呤)和尿酸的高效液相色谱分析方法,并对常见肉类食品中的含量进行测定。方法样品用10%(V/V)高氯酸在沸水浴中水解60min,调节pH至4,离心,上清液过膜,以7×10-3mol/L KH2PO4-H3PO4(pH 4.0)为流动相,选用Agilent ZORBAXEclipse XDB-C18(4.6mm×250mm,5μm)色谱柱进行色谱分离,流速1.0ml/min,柱温25℃,检测波长254nm。结果各组分在相应检测浓度范围内与其响应值呈良好的线性关系(r>0.9999);方法回收率在90.0%～107.5%,相对标准偏差在1.7%～13.3%之间。几种肉类食品中除含有4种嘌呤外,同时还含有尿酸(133.7～86.2μg/g),尿酸约占嘌呤和尿酸总量的7%。鸡肉中嘌呤总含量(1759.3±64.6)μg/g]高于兔肉、羊肉、猪肉和牛肉(1440～1000μg/g)。结论该方法用于肉类食品嘌呤检测简便快速,准确可靠。

Measurement of purines and uric acid simultaneous in meat with high performance liquid chromatographys

Objective To determinate adenine,guanine,hypoxanthine,xanthine and uric acid simultaneously in meat,a reversed-phase high performance liquid chromatography(HPLC) was developed.Methods The meat were hydrolyzed with perchloric acid 10%(v/v)in boiling water for 60min.After the hydrolysate was adjusted to pH 4,centrifuged,and filtrated with a 0.45μm membrane,the supernatants were separated on an Agilent ZORBAX Eclipse XDB-C18 column(250mm× 4.6mm i.d.,5μm)at 25℃ with a mobile phase of 7×10-3mol/L KH2PO4-H3PO4(pH 4.0),a flow rate of 1.0 ml/min,and UV detection at 254 nm.Results Each component in the corresponding concentration range showed a good linear relation with its peak area,correlation coefficient r>0.9999,recovery was 90.0%~107.5%,RSD was 1.7%~13.3%.In addition to containing four kinds of purines,there was quite amount of uric acid(about 133.7-86.2μg/g) in the mentioned meat.The ratio of uric acid to total purine and uric acid was about 7%.The content of total purine in chicken was(1759.3±64.6)μg/g higher than in rabbit,mutton,pork and beef(1440-1000μg/g).Conclusion The validated method is simple,rapid,accurate and reliable to the determination of purines and uric acid in meat.