高效液相色谱法快速测定鱼类鲜度指标K值及方法学优化  

Rapid determination of K value of fish freshness index by high performance liquid chromatography and optimization of methodology

作　　者：周漪波 胡雪玉 梁洁怡 潘云山 张飞 ZHOU Yibo;HU Xueyu;LIANG Jieyi;PAN Yunshan;ZHANG Fei(Guangdong Provincial Key Laboratory of Chemical Measurement and Emergency Test Technology,Guangdong Provincial Engineering Research Center for Efficacy Component Testing and Risk Substance Rapid Screening of Health Food,Institute of Analysis,Guangdong Academy of Sciences(China National Analytical Center,Guangzhou),Guangzhou 510070)

机构地区：[1]广东省科学院测试分析研究所(中国广州分析测试中心),广东省化学测量与应急检测技术重点实验室,广东省保健食品功效成分检测与风险物质快速筛查工程技术研究中心,广州510070

出　　处：《中国食品添加剂》2022年第8期189-194,共6页China Food Additives

基　　金：广东省科学院建设国内一流研究机构行动专项资金项目(2020GDASYL-20200102007);河北省重点研发计划项目(21372501D)。

摘　　要：对行业标准SC/T3048-2014测定鱼类鲜度K值(三磷酸腺苷,二磷酸腺苷,腺苷酸,肌苷酸,肌苷和次黄嘌呤)的分析方法进行优化。优化后的条件:以高氯酸溶液为提取剂,样品经高速均质结合超声波辅助提取后,用氢氧化钠溶液调节pH至6.0~6.4,一级水定容。采用C_(4)柱(4.6mm×250mm,5μm)分离,以磷酸二氢钾溶液(0.048mol/L,pH=5.7)~甲醇/磷酸二氢钾溶液(9/1,v/v)为流动相进行梯度洗脱,检测波长254nm,流速1.0mL/min,柱温30℃,外标法定量。优化结果表明:6种化合物的分析时间由行业标准方法的38min减少至11min。该方法的加标回收率(77.2%~112%),相对标准偏差(4.69%~7.81%),检出限(0.648~2.15mg/kg)和定量限(2.16~7.18mg/kg)均优于行业标准方法。结果表明该方法操作简便,分析快速,干扰较小,灵敏度高和重现性好,具有一定的实用价值。A method for determination of K value(adenosine triphosphate,adenosine diphosphate,adenosine 5′-monophosphate,inosine 5′-monophosphate,inosine and hypoxanthine)of fish freshness in the industry standard SC/T 3048-2014 was optimized.The optimized conditions were as follows:Perchloric acid solution was the extractant.The samples were treated with high-speed homogenization and ultrasonic assisted extraction.The pH was adjusted to 6.0~6.4 with sodium hydroxide solution then diluted to scale with gradeⅠwater.The separation was performed on a C_(4) column(4.6mm×250mm,5μm)using potassium dihydrogen phosphate solution(0.048mol/L,pH=5.7)~methanol/potassium dihydrogen phosphate solution(9/1,v/v)as mobile phase by gradient elution.The detection was carried out at a wavelength of 254 nm and the flow rate was 1.0 mL/min.The column temperature was 30℃and the quantification was performed by external standard method.The optimized results showed that the analysis time of 6 compounds was decreased from 38 min of the industry standard method to 11 min.These parameters including spiked recoveries(77.2%~112%),relative standard deviations(4.69%~7.81%),limits of detection(0.648~2.15mg/kg)and quantitation(2.16~7.18mg/kg),were better than the ones under industry standard method.The method is found to be convenient,rapid,with less interference and high accuracy,and has certain potential for practical applications.

关 键 词：K值 鱼 高效液相色谱法 方法学优化 

分 类 号：TS207.3[轻工技术与工程—食品科学]