建立同位素稀释-超高效液相色谱-串联质谱法测定烘焙食品中来源于鸡蛋的3种主要致敏蛋白
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(1.首都医科大学公共卫生学院,北京 100069;2.北京市疾病预防控制中心 食物中毒溯源技术北京市重点实验室,北京 100013)

作者简介:

齐开伦 男 硕士生 研究方向为食品安全检测E-mail:qikailun123@sohu.com通信作者:丁晓静 女 教授 研究方向为色谱分析E-mail:dxj_wry@yahoo.com

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基金项目:

北京市自然科学基金(7162088)


Establishment of ultra-high performance liquid chromatography tandem isotope dilution mass spectrometry for the determination of three major allergens from egg in baked food
Author:
Affiliation:

(1.School of Public Health,Capital Medical University,Beijing 100069,China;2.Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning,Beijing Center for Disease Prevention and Control,Beijing 100013,China)

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    摘要:

    目的 建立用于测定烘培食品中来源于鸡蛋的3种主要致敏蛋白溶菌酶、卵转铁蛋白、卵白蛋白的超高效液相色谱-串联质谱法。方法 筛选溶菌酶、卵转铁蛋白、卵白蛋白的特异水解肽段,合成特异肽段及其同位素标记肽段,样品用200 mmol/L Tris-HCl(pH=8.0)匀浆提取2~3次,提取液与胰蛋白酶混匀(胰蛋白酶与蛋白质的质量比为1∶50),37 ℃振摇酶解12 h,采用电喷雾正离子模式下多反应监测(MRM),同位素内标法定量。结果 9条特异水解肽段标准溶液在1~100 nmol/L范围内线性良好,相关系数(r2)均>0.995,烘培食品中3种致敏蛋白的定量限为0.4~10.0 μg/g,回收率在65.0%~108.5%之间,相对标准偏差(RSD)<15.0%。结论 方法灵敏度和特异性高,适用于烘培食品中来源于鸡蛋的3种致敏蛋白的检测。

    Abstract:

    Objective To establish an ultra-high performance liquid chromatography tandem isotope dilution mass spectrometry method for the determination of three major allergens, such as lysozyme, egg transferrin and ovalbumin, from eggs in baked food. Methods The specific peptides of lysozyme, ovotransferrin and ovalbumin were first screened, and then the specific peptides and isotopic markers were synthesized. Samples were homogenized in a buffer solution of 200 mmol/L Tris-HCl (pH=8.0) for 2-3 times. The sample solution was mixed with trypsin in a proportion of 1∶50 (trypsin/protein ratio) and then placed on a shaking bed and hydrolyzed for 12 h at 37 ℃. The enzymatic hydrolysate was detected by the electrospray positive ion (ESI+) with multiple reaction monitoring (MRM) mode and quantified by an isotope dilution method. Results Good linear relationships within the ranges of 1-100 nmol/L with linear correlation coefficients (r2) above 0.995 was achieved for the calibration curves of nine specific peptides standards. The limits of quantitation(LOQ)of the three proteins in baked food were 0.4-10.0 μg/g. The recoveries of target proteins in baked food ranged from 65.0% to 108.5% with relative standard deviations (RSDs) less than 15.0%. Conclusion The method is sensitive, specific and accurate. It is suitable for the detection of egg allergen proteins in baked food.

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齐开伦,杨奕,杨蕴嘉,邵兵,丁晓静.建立同位素稀释-超高效液相色谱-串联质谱法测定烘焙食品中来源于鸡蛋的3种主要致敏蛋白[J].中国食品卫生杂志,2017,29(6):658-665.

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  • 收稿日期:2017-09-07
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  • 在线发布日期: 2018-01-29
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