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酶辅助提取-液相色谱-质谱法测定鸡肉中的砷形态。

Enzyme-assisted extraction and liquid chromatography mass spectrometry for the determination of arsenic species in chicken meat.

机构信息

Department of Laboratory Medicine and Pathology, Faculty of Medicine and Dentistry, 10-102 Clinical Sciences Building, University of Alberta, Edmonton, Alberta T6G 2G3, Canada.

Department of Laboratory Medicine and Pathology, Faculty of Medicine and Dentistry, 10-102 Clinical Sciences Building, University of Alberta, Edmonton, Alberta T6G 2G3, Canada.

出版信息

Anal Chim Acta. 2015 Aug 12;888:1-9. doi: 10.1016/j.aca.2015.05.001. Epub 2015 May 7.

Abstract

Chicken is the most consumed meat in North America. Concentrations of arsenic in chicken range from μg kg(-1) to mg kg(-1). However, little is known about the speciation of arsenic in chicken meat. The objective of this research was to develop a method enabling determination of arsenic species in chicken breast muscle. We report here enzyme-enhanced extraction of arsenic species from chicken meat, separation using anion exchange chromatography (HPLC), and simultaneous detection with both inductively coupled plasma mass spectrometry (ICPMS) and electrospray ionization tandem mass spectrometry (ESIMS). We compared the extraction of arsenic species using several proteolytic enzymes: bromelain, papain, pepsin, proteinase K, and trypsin. With the use of papain-assisted extraction, 10 arsenic species were extracted and detected, as compared to 8 detectable arsenic species in the water/methanol extract. The overall extraction efficiency was also improved using a combination of ultrasonication and papain digestion, as compared to the conventional water/methanol extraction. Detection limits were in the range of 1.0-1.8 μg arsenic per kg chicken breast meat (dry weight) for seven arsenic species: arsenobetaine (AsB), inorganic arsenite (As(III)), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), inorganic arsenate (As(V)), 3-nitro-4-hydroxyphenylarsonic acid (Roxarsone), and N-acetyl-4-hydroxy-m-arsanilic acid (NAHAA). Analysis of breast meat samples from six chickens receiving feed containing Roxarsone showed the presence of (mean±standard deviation μg kg(-1)) AsB (107±4), As(III) (113±7), As(V) (7±2), MMA (51±5), DMA (64±6), Roxarsone (18±1), and four unidentified arsenic species (approximate concentration 1-10 μg kg(-1)).

摘要

鸡肉是北美人消费最多的肉类。鸡肉中的砷含量范围为μg kg(-1) 到 mg kg(-1)。然而,人们对鸡肉中砷的形态知之甚少。本研究的目的是开发一种能够测定鸡肉中砷形态的方法。我们在此报告了一种从鸡肉中提取砷形态的酶增强提取方法,使用阴离子交换色谱(HPLC)进行分离,并使用电感耦合等离子体质谱(ICPMS)和电喷雾串联质谱(ESIMS)同时进行检测。我们比较了几种蛋白水解酶(菠萝蛋白酶、木瓜蛋白酶、胃蛋白酶、蛋白酶 K 和胰蛋白酶)对砷形态的提取效果。使用木瓜蛋白酶辅助提取,可以提取和检测到 10 种砷形态,而水/甲醇提取物中可检测到 8 种砷形态。与传统的水/甲醇提取相比,结合超声处理和木瓜蛋白酶消化,整体提取效率也得到了提高。七种砷形态的检测限为每公斤鸡胸脯肉(干重)1.0-1.8μg 砷,分别为:砷甜菜碱(AsB)、无机亚砷酸盐(As(III))、二甲基砷酸(DMA)、一甲基砷酸(MMA)、无机砷酸盐(As(V))、3-硝基-4-羟基苯胂酸(Roxarsone)和 N-乙酰-4-羟基-m-苯胂酸(NAHAA)。对六只摄入含 Roxarsone 饲料的鸡的胸脯肉样本进行分析,结果表明存在(平均值±标准偏差μg kg(-1)) AsB(107±4)、As(III)(113±7)、As(V)(7±2)、MMA(51±5)、DMA(64±6)、Roxarsone(18±1)和四种未鉴定的砷形态(近似浓度为 1-10μg kg(-1))。

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