氯甲酸异丁酯衍生结合气相色谱-质谱法测定葡萄酒和黄酒中7种生物胺的含量  

Determination of 7 Biogenic Amines in Wine and Huangjiu by Gas Chromatography-Mass Spectrometry Coupled with Isobutyl Chloroformate Derivatization

作　　者：周江平 木玉仙 和春映 翟江舸 赵永信[2] 吴平谷[2] ZHOU Jiangping;MU Yuxian;HE Chunying;ZHAI Jiangge;ZHAO Yongxin;WU Pinggu(Gucheng District Center for Disease Control and Prevention,Lijiang 674100,China;Zhejiang Province Center for Disease Control and Prevention,Hangzhou 310051,China)

机构地区：[1]古城区疾病预防控制中心,丽江674100 [2]浙江省疾病预防控制中心,杭州310051

出　　处：《理化检验(化学分册)》2025年第3期338-344,共7页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

摘　　要：目前国家标准中测定食品中生物胺的方法操作繁琐,检测时间长,且我国尚未制定酒中生物胺的标准检测方法,基于此进行了题示研究。取1 mL葡萄酒或黄酒样品,加入100μL 50 mg·L^(−1)1,7-二氨基庚烷(1,7-DAP)内标溶液、300μL饱和碳酸氢钠溶液和0.01 g交联聚乙烯吡咯烷酮(PVPP),涡旋净化5 min,离心1 min,取0.5 mL上清液,加入衍生试剂氯甲酸异丁酯70μL,室温涡旋反应5 min。加入100μL乙醇、150μL二氯甲烷,涡旋提取5 min,离心1 min,取下层二氯甲烷提取液,经无水硫酸钠脱水后,采用气相色谱-质谱法(GC-MS)测定其中2-苯乙胺(2-PEA)、腐胺(PUT)、尸胺(CAD)、组胺(HIS)、色胺(TRY)、酪胺(TYR)、章鱼胺(OA)等7种生物胺的含量。7种生物胺的衍生物在DB-5MS毛细管色谱柱上按照柱升温程序分离,采用电子轰击离子源以选择离子监测模式检测,内标法定量。结果表明,7种生物胺标准曲线的线性范围均为0.2~10.0 mg·L^(−1)。对实际样品进行0.5,2.0,5.0 mg·L^(−1)等3个浓度水平的回收试验,回收率为83.3%~116%,测定值的相对标准偏差(n=6)小于10%,并确定测定下限均为0.2 mg·kg^(−1),方法完全可以满足澳大利亚等国家生物胺(HIS)限量规定要求(2~10 mg·L^(−1))。At present,the method for the determination of biogenic amines in food in the national standard has cumbersome operation and long detection time,and China has not yet developed a standard detection method for biogenic amines in wine.Based on this,the title research was carried out.100μL of 50 mg·L^(−1)1,7-diaminoheptane(1,7-DAP)internal standard solution,300μL of saturated sodium bicarbonate solution and 0.01 g of cross-linked polyvinylpyrrolidone(PVPP)were added into 1 mL of wine or huangjiu samples,and the mixture was vortex purified for 5 min and centrifuged for 1 min.0.5 mL of supernatant was taken,70μL of isobutyl chloroformate as the derivatizing agent was added,and the mixture was vortex reacted for 5 min at room temperature.Then 100μL of ethanol,and 150μL of dichloromethane were added,and vortex extraction was performed for 5 min.After centrifugation for 1 min,the lower dichloromethane extract was taken and dehydrated by anhydrous sodium sulfate.7 biogenic amines including 2-phenylethylamine(2-PEA),putrescine(PUT),cadaverine(CAD),histamine(HIS),tryptamine(TRY),tyramine(TYR)and octopamine(OA)were determined by gas chromatography-mass spectrometry(GCMS).The derivatives of 7 biogenic amines were separated on DB-5MS capillary chromatographic column according to the column temperature program,detected by electron impact ion source in selected ion monitoring mode,and quantified by internal standard method.As shown by the results,the linear range of the standard curves of 7 biogenic amines was 0.2-10.0 mg·L^(−1).Test for recovery at 3 concentration levels of 0.5,2.0,5.0 mg·L^(−1)was carried out on the actual samples,and the recoveries were in the range of 83.3%-116%.RSDs(n=6)of the determined values were less than 10%,and the lower limit of determination was determined to be 0.2 mg·kg−1.This method could fully meet the limit requirements of biogenic amines(HIS)in Australia and other countries(2-10 mg·L^(−1)).

关 键 词：生物胺 气相色谱-质谱法(GC-MS) 组胺 葡萄酒 黄酒 

分 类 号：O657.63[理学—分析化学]