摘要
从色谱条件的选择和优化混合酸法提取鸡肉和鸡汤中嘌呤两方面,建立了同时检测鸡肉和鸡汤中腺嘌呤、鸟嘌呤、次黄嘌呤和黄嘌呤的高效液相色谱检测方法。结果表明,使用Waters Atlantis dC 18(4.6 mm×250 mm,5μm)色谱柱,流动相为7×10-3 mol/L的KH 2 PO 4-H 3 PO 4(pH为3.8),流速为1.0 mL/min,柱温为30℃,紫外检测器检测波长为254 nm,进样量为10μL时,4种嘌呤可以完全分开,峰形较好。经方法学验证,4种嘌呤的质量浓度和峰面积在0.5~300 mg/L线性范围内,线性关系良好,相关系数R在0.9998~0.9999之间,检出限在0.024~0.129 mg/L范围内,检测方法精密度RSD为0.4673%~0.8132%。正交实验优化混合酸提取鸡肉和鸡汤中嘌呤实验结果表明,三氟乙酸浓度为95%,在90℃下水解20 min时,鸡肉中总嘌呤的提取效果最好;三氟乙酸浓度为80%,在90℃下水解25 min时,鸡汤中总嘌呤的提取效果最好,样品处理重复性RSD在2.202%~7.293%之间,鸡肉样品平均加标回收率在91.6343%~95.9119%之间,鸡汤样品平均加标回收率在93.3189%~96.6424%之间,适用于检测鸡肉和鸡汤中4种嘌呤含量。
A HPLC method for simultaneous determination of adenine,guanine,hypoxanthine and xanthine in chicken and chicken soup is established by selecting chromatographic conditions and optimizing the mixed acid method to extract purine from chicken and chicken soup.The results show that using Waters Atlantis dC 18(250 mm×4.6 mm,5μm)chromatographic column,the mobile phase is 7×10-3 mol/L KH 2PO 4-H 3PO 4(pH is 3.8),the flow rate is 1.0 mL/min,and the column temperature is 30℃,when the detection wavelength of the UV detector is 254 nm and the injection volume is 10μL,the four purines can be completely separated and the peak shape is better.It is verified by methodology that when the mass concentration and peak area of the four purines are within the linear range of 0.5~300 mg/L,the linear relationship is better.The correlation coefficient R is 0.9998~0.9999,and the detection limit is 0.024~0.129 mg/L,the precision RSD of the detection method is 0.4673%~0.8132%.The orthogonal experiment optimization of mixed acid extraction of purines from chicken and chicken soup shows that the concentration of trifluoroacetic acid is 95%,when being hydrolyzed at 90℃for 20 minutes,the extraction effect of total purines in chicken is the best;the concentration of trifluoroacetic acid is 80%,when being hydrolyzed at 90℃for 25 minutes,the extraction effect of total purines in chicken soup is the best.The sample processing repeatability RSD is 2.202%~7.293%,and the average recovery rates of chicken samples range from 91.6343%to 95.9119%,the average recovery rates of chicken soup samples range from 93.3189%to 96.6424%,which is suitable for detecting the content of four purines in chicken and chicken soup.
作者
方妍
谢定源
丁璐
余永昊
罗鑫
FANG Yan;XIE Ding-yuan;DING Lu;YU Yong-hao;LUO Xin(Key Laboratory of Environment Correlative Dietology,Ministry of Education,College of Food Science and Technology,Huazhong Agricultural University,Wuhan 430070,China)
出处
《中国调味品》
CAS
北大核心
2020年第12期115-123,共9页
China Condiment
基金
国家重点研发计划专项课题(2017YFD0400101)
中央高校基本科研业务费专项基金资助项目(2017JC013)。
关键词
高效液相色谱
嘌呤
提取
鸡肉
鸡汤
high performance liquid chromatography
purine
extraction
chicken
chicken soup
