摘要
旨在建立一种利用液质联用(liquid chromatography mass spectrometry,LC-MS)技术实现对猪肠道食糜中多种生物胺进行同步检测的方法。试验首先优化了丹磺酰氯(dansyl chloride,DNS)对生物胺的衍生条件,并进一步确定了肠道食糜样品中生物胺的最佳净化方法。结果显示,衍生条件在衍生剂浓度为8 mg·mL^-1,衍生温度为40℃的条件下,衍生90 min为最佳。生物胺净化方法确定为先用乙酸乙酯进行提取,衍生结束后再用乙酸乙酯进行萃取的组合方式提取生物胺的效果最好;11种生物胺在5~1000μg·L^-1的质量浓度范围内定量结果呈良好的线性关系(R^2>0.99),检出限和定量限分别为0.01~0.23μg·L^-1和0.03~0.76μg·L^-1;各生物胺的日内相对标准偏差在0.73%~7.92%,日间相对标准偏差在0.07%~8.67%。最后采用本方法对仔猪回肠、盲肠、结肠和直肠食糜样品中的生物胺进行检测,初步确定了11种生物胺在不同肠段食糜中的含量分布。通过对仔猪回肠、盲肠、结肠和直肠中的食糜样品进行检测,发现食糜中的生物胺主要为亚精胺、腐胺、尸胺、精胺和组胺,而酪胺、1,7-二氨基庚烷、5-羟色胺和胍基丁胺4种生物胺含量较少,色胺和苯乙胺未检出。综上所述,本试验建立的方法具有结果准确、灵敏度高、重现性好等优点,可用于仔猪不同肠道段食糜中多种生物胺的同步检测。
This experiment aimed to establish a method for simultaneous detection of multiple biogenic amines in pig intestinal chyme using liquid chromatography mass spectrometry(LC-MS).The experiment firstly optimized the conditions for derivatization of biogenic amines by dansyl chloride(DNS),and further determined the best method for purifying biogenic amines in intestinal chyme samples.The results showed that the derivatization conditions were optimal when the derivatization agent concentration was 8 mg·mL^-1 and the derivatization temperature was 40℃for 90 min.The biogenic amine purification method was determined to be the best combination of extraction with ethyl acetate first and extraction with ethyl acetate after derivatization.The quantification results of 11 biogenic amines in the mass concentration range of 5-1000μg·L^-1 showed a good linear relationship(R^2>0.99).The detection limit and limit of quantification were 0.01-0.23μg·L^-1 and 0.03-0.76μg·L^-1,respectively.The intra-day relative standard deviation of each bioamine was 0.73%-7.92%,and the inter-day relative standard deviation was 0.07%-8.67%.Finally,this method was used to detect biogenic amines in chyme samples from piglets’ileum,cecum,colon and rectum.The content distribution of 11 biogenic amines in different intestinal chyme was preliminarily determined.Detection of chyme samples in piglets’ileum,cecum,colon and rectum found that the biogenic amines in the chyme were mainly spermidine,putrescine,cadaverine,spermine and histamine.Tyramine,1,7-diaminoheptane,serotonin,and agmatine were detected in very small amounts,and no tryptamine and phenethylamine were detected.In summary,the method established in this test has accurate results,high sensitivity,and good reproducibility,and can be used to detect multiple biogenic amines in chyme of different intestinal sections of piglets simultaneously.
作者
任梦雅
冯潇慧
解竞静
文晓宾
孟庆石
刘丽莉
REN Mengya;FENG Xiaohui;XIE Jingjing;WEN Xiaobin;MENG Qingshi;LIU Lili(College of Life Science,Hunan University of Science and Technology,Xiangtan 411201,China;State Key Laboratory of Animal Nutrition,Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization,Xiangtan 411201,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2020年第6期1320-1331,共12页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31672428)
中央级公益性科研院所基本科研业务费专项(Y2020PT19)
湖南省自然科学基金(2016jj3059)。
关键词
肠道食糜
生物胺
检测方法
液质联用
intestinal chyme
biogenic amine
detection method
LC-MS