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超高效液相色谱-串联质谱法测定鸡肉中肌苷及其关联物

2021-11-12陈超王真任南刘梦瑶纪晓萌李莹莹郭文萍

肉类研究 2021年10期

陈超 王真 任南 刘梦瑶 纪晓萌 李莹莹 郭文萍

摘 要:建立一种利用超高效液相色谱-串联质谱法检测鸡肉中5-腺苷酸(5-adenosine monophosphate,AMP)、5-肌苷酸(5-inosine monophosphate,IMP)、肌苷(inosine,I)、次黄嘌呤(hypoxanthine,Hx)的方法,并进行方法学考察。结果表明:优化后的方法为样品中加入体积分数3%高氯酸-10%甲醇水溶液后添加匀浆均质子涡旋,离心,调节提取液pH值为5.0后用PEIME HLB柱净化,采用SB-Aq色谱柱(3.0 mm×100 mm,1.8 ?m),流动相为体积分数0.1%甲酸水溶液-乙腈,梯度洗脱,流速0.3 mL/min,正离子多反应监测模式测定,外标法定量;经方法学考察,4 种核苷类成分(AMP、IMP、I、Hx)线性关系良好,相关系数均大于0.996,加标回收率为79.63%~91.08%,相对标准偏差为1.0%~4.5%(n=6)。

关键词:超高效液相色谱-质谱联用技术;鸡肉;正离子多反应监测;5-腺苷酸;5-肌苷酸;肌苷;次黄嘌呤

Determination of Inosine and Its Related Compounds in Chicken by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

CHEN Chao, WANG Zhen, REN Nan, LIU Mengyao, JI Xiaomeng, LI Yingying*, GUO Wenping

(China Meat Research Center, Beijing 100068, China)

Abstract: This study aimed to set up an analytical method to detect 5-adenosine monophosphate (AMP), 5-inosine monophosphate (IMP), inosine (I), and hypoxanthine (Hx) in chicken by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and to assess its figures of merit. The results showed that the experimental conditions of this method were optimized as follows: samples were homogenized using a vortex mixer by addition of 3% perchloric acid-10% methanol in water followed by centrifugation, and the supernatant extract was purified with a PEIME HLB cartridge after being adjusted to pH 5.0. The analytes were separated on an SB-Aq column (3.0 mm × 100 mm, 1.8 ?m) using a mobile phase consisting of 0.1% formic acid in water and acetonitrile by gradient elution at a flow rate of 0.3 mL/min, detected in the positive ion multiple reaction monitoring (MRM) mode, and quantified by the external standard method. The calibration curves for the four nucleoside components had good linearity with correlation coefficients of greater than 0.996. The spiked recoveries ranged between 79.63% and 91.08%, with relative standard deviations (RSDs) of 1.0%–4.5% (n = 6).

Keywords: ultra performance liquid chromatography-mass spectrometry; chicken; positive ion multiple reaction monitoring mode; 5-adenosine monophosphate; 5-inosine monophosphate;  inosine; hypoxanthine

DOI:10.7506/rlyj1001-8123-20210709-185

中圖分类号:TS201.2                                        文献标志码:A 文章编号:1001-8123(2021)10-0041-07

引文格式:

陈超, 王真, 任南, 等. 超高效液相色谱-串联质谱法测定鸡肉中肌苷及其关联物[J]. 肉类研究, 2021, 35(10): 41-47. DOI:10.7506/rlyj1001-8123-20210709-185.    http://www.rlyj.net.cn

CHEN Chao, WANG Zhen, REN Nan, et al. Determination of inosine and its related compounds in chicken by ultra performance liquid chromatography-tandem mass spectrometry[J]. Meat Research, 2021, 35(10): 41-47. DOI:10.7506/rlyj1001-8123-20210709-185.    http://www.rlyj.net.cn

禽肉是我国主要食用肉类之一,广受消费者欢迎[1]。

禽肉营养丰富,味道鲜美,富含蛋白质、脂肪、风味物质等,是人们膳食中不可或缺的重要组成部分[2]。鸡肉在炖煮过程中可释放小肽、游离氨基酸、无机盐、核酸代谢物等物质,赋予鸡汤一定的风味,核酸代谢物是炖肉中与鲜味相关的主要化合物,其中5-肌苷酸(5-inosine monophosphate,IMP)对鲜味贡献大[3-4]。核苷酸属于一类有机化合物,由嘌呤碱基或嘧啶碱基、核糖或脱氧核糖以及磷酸3 种物质组成,五碳糖通过β-糖苷键连接,与有机碱合成核苷,核苷与磷酸合成核苷酸,核苷和核苷酸是胞内化合物,参与各种调节和代谢功能[5]。……

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