高效液相色谱法测定海水鱼中嘌呤含量

本文建立同时检测海水鱼中腺嘌呤、鸟嘌呤、次黄嘌呤、黄嘌呤的高效液相色谱检测方法。对部分海水鱼可食用部分及内脏中的4种嘌呤含量进行检测。结果表明,当使用Agilent ZORBAX Eclipse XDB-C18(4.6 mm×250 mm,5μm)色谱柱,以水-甲醇-冰乙酸-20%四丁基氢氧化铵(V/V/V/V=879/100/15/6)为流动相,设置流速为0.8 mL/min,柱温30℃,检测波长254 nm,进样量10μL时,4种嘌呤可完全分离且峰型较好。经方法学验证,得出此方法在0.1~400 mg/L范围线性关系良好,相关系数(R)在0.9998~1.0000之间,方法检出限范围0.0465~0.1056 mg/L。高效液相色谱检测方法精密度RSD在0.0200%~0.7000%之间,样品处理重复性腺嘌呤、鸟嘌呤、次黄嘌呤、黄嘌呤依次为1.2421%,0.9711%,0.8836%,1.9727%。加标回收率在94.2888%~102.9188%之间,适用于海水鱼中4种嘌呤的测定。经检测,不同种类海水鱼可食用部分(鱼眼睛、腹部鱼肉、背部鱼肉、鱼皮)嘌呤总含量由高到低依次为海鲈鱼、大菱鲆、金仓鱼、黄花鱼、美国红鱼、踏板鱼、鳕鱼。

Determination of Purines Content of Sea Fish by High Performance Liquid Chromatograp

For identifying the content of purine in different parts of sea fish.In this paper,a high-performance liquid chromatogram(HCLP)method was developed in order to determinate the contents of guanine,hypoxanthine,xanthine and adenine in edible parts and internal organs of seawater fish.Purine bases were separated using Agilent ZORBAX Eclipse XDB-C18(4.6 mm×250 mm,5μm)chromatographic column with water-methanol-ice acetic acid-20%tetrabutyl ammonium hydroxide(V/V/V/V=879/100/15/6)as mobile phase,the flow rate was set at 0.8 mL/min,column temperature was 30℃,and wavelength of UV detection was at 254 nm.The sample volume was 10μL.Four kinds of purines had a good linear relation between the concentration and peak area in the range of 0.1-400 mg/L.The correlation coefficient was 0.9998-1.0000,the limit of detection(LOD)was between 0.0465-0.1056 mg/L,method precision of RSD%was between 0.0200%-0.7000%.The repeatability of sample treatment adenine,guanine,hypoxanthine,xanthine is 1.2421%,0.9711%,0.8836%,1.9727%respectively.The recovery rate was between 94.2888%-102.9188%.So the method was suitable for the determination of four purines in marine fish.The test results showed that the total purine contents of edible part(eye,belly of fish,the back of the fish,fish skin)in sea bass were the highest,which was followed by turbot,golden hamster,yellow croaker,sciaenops ocellatus,Pedal fish and gadus in oeder.