鱼类鲜度K值的HPLC-UV-MS/MS快速测定方法研究  

Rapid Determination of K Value in Fish by High Performance Liquid Chromatography-Ultraviolet-Tandem Mass Spectrometry

作　　者：张飞 周漪波 邓远强 阮奇珺 付强 郑家概 ZHANG Fei;ZHOU Yibo;DENG Yuanqiang;RUAN Qijun;FU Qiang;ZHENG Jiagai(Guangdong Provincial Engineering Research Center for Efficacy Component Testing and Risk Substance Rapid Screening of Health Food,Guangdong Provincial Key Laboratory of Emergency Test for Dangerous Chemicals,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),广东省化学危害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：《食品工业》2022年第1期300-304,共5页The Food Industry

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);广东省科学院测试分析研究所(中国广州分析测试中心)自筹科研资金项目(202002);河北省重点研发计划项目(21372501D)。

摘　　要：建立了高效液相色谱-紫外-三重四极杆串联质谱快速测定鱼类鲜度K值[三磷酸腺苷(ATP)、二磷酸腺苷(ADP)、腺苷酸(AMP)、肌苷酸(IMP)、肌苷(HxR)和次黄嘌呤(Hx)]的分析方法。研究了不同贮藏温度对K值的影响规律。样品经高氯酸溶液均质-超声提取后,调节pH至6.0~6.4,用水定容。采用C_(18)柱分离,以乙酸铵溶液(20mmol/L,0.1%甲酸)-乙酸铵溶液/甲醇(1∶9,V/V)为流动相进行梯度洗脱,检测波长254 nm,外标法定量,串联质谱法定性确证。ATP和ADP在5.0~100μg/mL范围内线性良好(R2>0.995),其他4种在1.0~100μg/mL范围内线性良好(R2>0.999)。在不同添加水平下(ADP:120,160和200 mg/kg;IMP:1 200,1 600和2 000 mg/kg;其他4种:60,120和180mg/kg),6种化合物的加标回收率为71%~112%。方法的相对标准偏差为8.5%~14.5%,检出限和定量限分别为0.96~18mg/kg和3.2~61 mg/kg。通过对实际样品的测定,结果表明该方法操作简便、分析快速、干扰较小、准确度较高。A novel method was developed for the rapid determination of K value [adenosine triphosphate(ATP),adenosine diphosphate(ADP),adenosine 5’-monophosphate(AMP),inosine 5’-monophosphate(IMP),inosine(HxR) and hypoxanthine(Hx)] to evaluate the freshness of fish by high performance liquid chromatography-ultraviolet-triple quadrupole tandem mass spectrometry(HPLC-UV-MS/MS).The effects of different storage temperatures on K value were studied.After the sample was extracted with perchloric acid solution through homogeneous-ultrasound extraction,the pH was adjusted to 6.0-6.4.The volume was fixed with water.The separation was performed on a C_(18) column using ammonium acetate solution(20 mmol/L,0.1% formic acid)-ammonium acetate solution/methanol(1∶9,V/V) as mobile phase by gradient elution.The detection wavelength was set at 254 nm,and the external standard method was used for quantitation.The qualitative analyses were performed by tandem mass spectrometry.The linear ranges of the method for ATP and ADP were 5.0-100 μg/mL with correlation coefficients larger than 0.995.The method showed a good linearity in the range of 1.0-100 μg/mL for the remaining four compound with correlation coefficients larger than 0.999.The spiked recoveries were in the range of 71%-112% at different levels(ADP:120,160 and 200 mg/kg;IMP:1 200,1 600 and 2 000 mg/kg;the remaining four compound:60,120 and 180 mg/kg).The relative standard deviations were in the range of 8.5%-14.5%.The limits of detection and quantitation were in the ranges of 0.96-18 mg/kg and 3.2-61 mg/kg,respectively.Through the actual sample determination,the results showed that the method was convenience,rapid analysis,less interference and high accuracy.

关 键 词：鱼 鲜度指标 K值 高效液相色谱-紫外-串联质谱法 

分 类 号：O657.63[理学—分析化学] TS254.7[理学—化学]