高效液相色谱法快速测定青海牛羊肉及内脏组分中尿酸及嘌呤.docVIP

高效液相色谱法快速测定青海牛羊肉及内脏组分中尿酸及嘌呤 　　摘 要：建立青海牛羊肉和内脏组分中尿酸和4 种嘌呤（鸟嘌呤、次黄嘌呤、黄嘌呤和腺嘌呤）的2 种分析方法，一种是采用高效液相色谱（high performance liquid chromatography，HPLC）法同时测定上述5 种组分，并折算出尿酸总含量；另一种是采用生物酶解法将嘌呤全部转化后，采用HPLC法快速测定其降解产物尿酸。2 种方法均采用Agilent ZorBax Eclipse XDB-C18色谱柱分离，浓度为7×10-3 mol/L的KH2PO4-H3PO4（pH=3.83）溶液作为流动相，紫外检测波长为254 nm。结果表明：同时测定法中，被测物质在上述条件下分离良好，在较宽的浓度范围内线性良好，相关系数（r）均在0.998 4以上，平均回收率为92.2%～119.8%，相对标准偏差（relative standard deviation，RSD）为0.78%～10.43%；转化后尿酸快速测定法中，被测物质在1～70 mg/L质量浓度范围内与峰面积间的线性关系良好，r=0.999 4。将2 种分析方法得出的总尿酸含量进行比较，经配对t检验，结果无显著性差异（P=0.7730.05）。2 种方法均可用于牛羊肉和内脏组分中嘌呤类物质的检测。 关键词：高效液相色谱法；尿酸；嘌呤；酶解转化 Abstract： Two analytical methods were developed to determine uric acid and four purines （guanine， hypoxanthine， xanthine and adenine） in meat and viscera of cattle and sheep from Qinghai province. One was based on the simultaneous determination of five individual components by high performance liquid chromatography （HPLC） for subsequent calculation of total uric acid， while the other was based on the enzymatic transformation of all four purines to uric acid for rapid determination of the degradation product by HPLC. In both methods， the chromatographic separation was performed using an Agilent ZorBax Eclipse XDB-C18 column with 7 × 10-3 mol/L KH2PO4-H3PO4 （pH 3.83） as mobile phase， and the analytes were detected at 254 nm. Results shows that the first method achieved good separation and good linearity in a wide concentration range with all correlation coefficients being higher than 0.998 4， and the recoveries were between 92.2% and 119.8% with relative standard deviations （RSD） between 0.78% and 10.43%. Similarly， the second method had a good linearity in the range of 1?C70 mg/L with correlation coefficients of 0.999 4. The values of total uric acid obtained by the two methods did not significantly differ from each other as indicated by the matched pairs t-test （P = 0.773 0.05）. The two methods could be used for the detection of purine compounds in bovine and sheep meat and visc