高效液相色谱-串联质谱法测定油条中羧甲基赖氨酸

孙晓华; 赖克强; 黄轶群; 王婧; 吴轶; 肖竹青

doi:10.13386/j.issn1002-0306.2014.11.057

高效液相色谱-串联质谱法测定油条中羧甲基赖氨酸

1. 上海海洋大学食品学院
2. 上海市食品研究所

基金项目:

国家自然科学基金(61250002,31250006)；上海海洋大学研究生科研基金(B-5008-11-0009102)；

详细信息

作者简介:
孙晓华 (1987-) , 女, 博士研究生, 研究方向:食品质量与安全分析。;

中图分类号: O657.63;TS207.3
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出版历程
- 收稿日期: 2013-09-21

High performance liquid chromatography-tandem mass spectrometry method for the determination of Nε- ( carboxymethyl) lysine in fried bread sticks

College of Food Science and Technology, Shanghai Ocean University
Shanghai Food Research Institute

摘要

摘要: 建立了利用高效液相色谱-串联质谱法（HILIC-HPLC-MS/MS）测定油条中羧甲基赖氨酸（CML）含量的检测方法。样品经硼氢化钠还原、盐酸水解,过混合型阳离子交换小柱萃取净化,样品溶液在HILIC色谱柱中以甲醇-水（均含2mmol/L乙酸铵和0.1%甲酸）为流动相进行梯度洗脱,CML被保留并与样品基质分离。选用电喷雾正离子（ESI+）模式电离,多反应监测模式（MRM）进行串联质谱分析。以内标法定量,D4-CML为内标物,分析时间为12min。本方法添加回收率范围89.5%⁹9.1%,相对标准偏差范围2.1%².6%,仪器检出限为1μg/L,定量限为3μg/L。应用该方法测得15种不同来源的油条中CML含量为69.3±8.2¹82.2±3.1mg/（kg蛋白质）,5.3±0.5¹5.4±0.3mg/（kg样品）。本方法操作简便、快速、灵敏度高、精密度好,为进一步研究食品中CML的产生机理提供了有效的检测方法。
- 羧甲基赖氨酸 /
- 亲水作用色谱 /
- 高效液相色谱-串联质谱 /
- 油条
Abstract: A method for the determination of Nε- ( carboxymethyl) lysine ( CML) content in fried bread sticks was developed using high performance liquid chromatography- tandem mass spectrometry ( HPLC- MS/MS) .Food samples were reduced by sodium borohydride, then hydrolyzed by hydrochloric acid and finally cleaned up by mixed cation exchange ( MCX) solid phase extraction cartridge.CML was remained on an Atlantis HILIC column.Methanol-water ( all containing 2mmoI /L ammonium acetate and 0.1% formic acid) were used as mobile phases.Tandem MS analysis was performed in multiple reactions monitoring ( MRM) mode using the electrospray positive ionization ( ESI +) method.D4- CML was used as internal standard substance, and the analysis time was12min.The average recoveries of CML for this method ranged from 89.5% to 99.1%, with the relative standard deviations ( RSD) were between 2.1% and 2.6%.The instrument detection limit was 1μg /L, and quantitation limit was3μg /L.Using this method, CML contents determined in 15 different sources fried bread sticks were 69.3 ± 8.2¹82.2± 3.1mg / ( kg protein) , and 5.3 ± 0.5¹5.4 ± 0.3mg / ( kg sample) .This method is simple, rapid, highly sensitive and accurate, and it is an effective method for further research on the generation mechanism of CML.

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