QuEChERS技术在食品真菌毒素检测中的研究进展

真菌毒素是丝状真菌产生的有毒代谢产物,对人类和动物的健康构成严重威胁。因此,建立食品中多种真菌毒素的高效提取和净化方法,有利于降低真菌毒素的暴露风险,保障食品安全。QuEChERS方法具有简单、快速、回收率高、试剂用量少、安全等优点,可运用于真菌毒素的检测。该文概述了该方法在食品中真菌毒素分析检测方面的应用,根据QuEChERS技术的特点,探讨应用QuEChERS技术对真菌毒素进行预处理过程中,提取、盐析和净化步骤所应用的方法及对真菌毒素提取效率的影响,总结QuEChERS技术在食品真菌毒素检测中的应用,并对该技术现阶段应用中亟待解决的问题提出了展望和建议。     

QuEChERS联合UPLC-Orbitrap/MS同时测定检测坚果和干制水果中60种真菌毒素

目的 本文针对混合坚果和干制水果类休闲食品，建立同时测定该类食品中60种真菌毒素的方法。方法 选择富含油脂和糖分的花生和葡萄干两种典型食品基质，样品经QuEChERS（900 mg MgSO₄、150 mg PSA、150 mg C₁₈）净化，采用超高效液相色谱-串联静电场轨道阱高分辨质谱的一级全扫描结合二级数据依赖扫描模式进行分析。以保留时间和母离子精确质量数定性，母离子色谱峰面积定量，同时以二级特征碎片离子进行确证。结果 提取溶液pH值会显著影响真菌毒素的解离状态，进而影响其提取回收率。通过QuEChERS净化能有效除去样品中糖类和脂类等组分的干扰，提高定性定量的准确性。采用空白基质匹配标准曲线进行校正可以减小基质效应以达到定量分析要求，真菌毒素在各自的质量浓度范围内线性关系良好（决定系数R²>0.95），回收率60%~120%，相对标准偏差小于20%，另外有7种真菌毒素线性决定系数小于0.95，进行定性分析。结论 该方法灵敏度高，结果准确、可靠，适用于坚果和干制水果60种真菌毒素的定性定量分析。     

QuEChERS技术在食品安全中的应用进展

该文对样品前处理QuEChERS技术在食品安全检测中的应用进展进行综述.QuEChERS是以固相萃取技术和基质固相分散技术为基础的前处理技术,对该技术的改良、发展等方面进行详细介绍,综述近10年QuEChERS在食品安全领域中农药残留、兽药残留、真菌毒素、非法添加物等领域的应用.QuEChERS样品前处理技术具有快速、...     

基于磁性ZIF-67纳米材料的QuEChERS-超高效液相色谱-串联质谱技术同时检测小麦中17种真菌毒素

目的 建立一种QuEChERS（Quick，easy，inexpensive，effective，rugged，and safe）-超高效液相色谱-串联质谱（Ultrahigh-performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS）同时测定小麦中17种真菌毒素的方法。方法 样品经乙腈-水-柠檬酸（80：19：1，V:V:V）溶液提取，采用自制的Fe3O4@ZIF-67磁性纳米材料作为QuEChERS 吸附剂进行净化，实验对提取条件、缓冲盐的选择、材料用量等因素进行了优化，并对建立的方法进行了方法学评价。结果 实验结果表明，在0.1~1000 ng/mL的范围内，17种真菌毒素线性良好（r2≥0.998），定量限在0.1~10 μg/kg之间，平均回收率在80.2%~103.0%之间，相对标准偏差（RSD）为2.1 %~8.4 %。结论 该方法简便、准确，可满足小麦中真菌毒素的检测需要。     

QuEChERS-超高效液相色谱-串联质谱法在食品真菌毒素检测中应用的研究进展

真菌毒素是一类由产毒丝状真菌产生的有毒次生代谢产物,能诱发人畜各种生理损害,是世界各地食品和农产品的主要污染物之一。建立高效的真菌毒素分析方法将有效降低真菌毒素的暴露风险,复杂基质中多种真菌毒素的联合提取净化和检测是建立这类分析技术的前提保证。Qu ECh ERS(quick,easy,cheap,effective,rugged and safe)方法具有简单、快速、回收率高、试剂用量少、安全等优点,适用于大批量样品的检测。超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)可同时用于定性和定量分析,具有检出限低、灵敏度高等优点。本文根据Qu ECh ERS前处理技术的特点,结合样品真菌毒素的理化性质和污染特点,对Qu ECh ERS-UPLC-MS/MS技术在真菌毒素分析中的应用做了简要综述。     

QuEChERS净化-超高效液相色谱-串联质谱法测定婴幼儿谷类辅助食品中12种真菌毒素

目的建立QuEChERS净化-超高效液相色谱-串联质谱法测定婴幼儿谷类辅助食品中12种真菌毒素的方法。方法样品用乙腈-水-乙酸(84∶15∶1,V/V)提取,提取液经QuEChERS方法净化,采用Synergi 4μ Fusion-RP C18色谱柱(50 mm×2.0 mm,4μm)进行分离,用超高效液相色谱-串联质谱仪进行检测,基质外标法定量。结果12种真菌毒素在各自的线性范围内线性关系良好,相关系数(r)均不低于0.999,回收率在80.5%~106.4%之间,相对标准偏差在5.3%~9.5%之间。结论该方法快速简便,可用于婴幼儿谷类辅助食品中真菌毒素多残留成分的检测。     

梧州六堡茶中真菌多样性分析及真菌毒素残留量的测定

为研究六堡茶中真菌的多样性及真菌毒素残留量情况，验证其饮用安全性，该试验选取10批次不同年份、不同企业生产的梧州六堡茶样品进行研究，先提取样品的总DNA，再利用高通量测序技术进行ITS测序分析，然后采用QuEChERS（quick、easy、cheap、effective、rugged、safe）-超高效液相质谱联用方法测定样品中 16 种常见真菌毒素等残留量。研究结果表明，梧州六堡茶中真菌的优势菌为曲霉属（Aspergillus）、青霉属（Penicillium），10个样品全部含有曲霉属，其平均相对含量为50.6%～99.9%，10个样品曲霉属平均相对含量86.4%；试验样品中真菌毒素残留量全部低于方法检出限。     

磁性固相萃取技术在食品真菌毒素分析中的应用

真菌毒素常见于食品和各种粮食作物中，具有致癌性、诱变性、致畸性等危害，严重威胁人类和动物的健康。由于真菌毒素在食品基质中存在种类多、浓度低、极性范围广等特点，因此建立快速高效的食品样品前处理方法对真菌毒素的痕量分析十分重要。磁性固相萃取（magnetic solid-phase extraction，MSPE）是一种基于磁相互作用的样品前处理技术，凭借简单快速、绿色安全、高效经济等特点，已广泛应用于食品样品真菌毒素分析的前处理中。本文介绍了MSPE的萃取过程和磁性纳米材料的制备和修饰，综述了以无机材料（氧化硅基和碳基）、有机材料（印迹型、有机高分子、表面活性剂和有机小分子）和其他材料（离子液体和单克隆抗体）为代表的MSPE吸附剂在食品样品真菌毒素检测中的应用，并对MSPE技术的发展趋势进行了展望。     

白酒酒醅中真菌毒素的检测

该实验建立了白酒生产原料酒醅中31种真菌毒素的液相色谱-串联质谱检测方法。样品使用乙腈与1%的甲酸水混合溶液（85∶15,V/V）提取,经QuEChERS法净化后,采用多反应监测模式（MRM）检测,可同时对31种真菌毒素进行定性定量分析。结果表明,该方法的相关系数R2为0.991～0.999,线性关系良好,检出限和定量限分别为0.1～5.0 μg/kg和0.4～16.5 μg/kg,平均回收率为83.1%～108.7%,回收率实验结果的相对标准偏差（RSD）为2.5%～9.6%。对10个随机抽取的酒醅样品进行检测,检出的真菌毒素分别有：黄曲霉毒素、脱氧雪腐镰刀菌烯醇及其衍生物、T-2毒素、新茄病镰刀菌烯醇、玉米赤霉烯酮、赭曲霉毒素、杂色曲霉素。该方法的灵敏度、准确度和精密度良好,可满足白酒生产原料中31种真菌毒素的检测。     

QuEChERS-超高效液相色谱串联质谱法测定发酵黑茶中的10种真菌毒素

建立了云南普洱茶、湖南黑茶、广西六堡茶和湖北老青茶等发酵黑茶黄曲霉毒素B1（AFB1）、黄曲霉毒素B2（AFB2）、黄曲霉毒素G1（AFG1）、黄曲霉毒素G2（AFG2）、脱氧雪腐镰刀菌烯醇（DON）、3-乙酰基脱氧雪腐镰刀菌烯醇（3-AcDON）、玉米赤霉烯酮（ZEN）、T-2毒素（T-2）、HT-2毒素（HT-2）、赭曲霉毒素A（OTA）等10种真菌毒素的简单、快速、高灵敏度超高效液相色谱-串联质谱检测方法。样品采用乙腈/水溶液（84+16）提取试样、QuEChERS净化,试样以C18柱分离,多反应监测（MRM）模式进行定量与定性分析,采用外标法定量。结果表明,在优化的条件下,10种真菌毒素在各自的线性响应范围内线性关系良好,相关系数（r）均不小于0.9995,定量限（LOQ）为0.1~10 μg/kg,在三个不同加标水平下平均回收率为61.9%~120.3%,相对标准偏差（RSD）为3.2%~16.1%。采用建立的方法对市面上销售的黑茶中的10种真菌毒素进行了筛查,数批产品检出真菌毒素。该方法简单、准确、可靠,可适用于发酵黑茶中多种真菌毒素的快速分析。     

硫色镰刀菌（Fusarium sulphureum）体外产毒条件的筛选

对影响硫色镰刀菌（Fusarium sulphureum）体外产生真菌粗毒素的温度、时间、pH值、培养方式及培养基种类进行筛选。结果表明：F. sulphureum的最佳产毒条件是Richard培养基、25 ℃、pH 4.5、振荡培养10 d。在单因素筛选的基础上,以温度、pH值和培养时间为自变量,绿豆种子胚根平均长度为响应值,利用响应面分析法对该菌产毒条件的优化结果表明,22.2 ℃、pH 5.1、振荡培养12 d最有利于体外产毒。     

Quantitative Analysis of 10 Mycotoxins in Wheat Flour by Ultrahigh Performance Liquid Chromatography‐Tandem Mass Spectrometry with a Modified QuEChERS Strategy

A simple and sensitive analytical method for quantitative analysis of 10 mycotoxins was developed and validated by a combination of modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure with ultrahigh performance liquid chromatography‐tandem mass spectrometry (UHPLC‐MS/MS). Sample preparation involved QuEChERS with dispersive solid phase extraction for clean‐up, and analysis was performed by reversed‐phase UHPLC‐MS/MS using electrospray negative ionization and multiple reaction monitoring. Under optimized conditions, the calibration curves displayed good linear relationships with all coefficients of determinations (r²) higher than 0.998. The limits of quantification for all target mycotoxins were lower than 7 μg/kg. Trueness and precision for the analytes were 70% to 116% average recoveries and 2% to 13% relative standard deviations (RSDs). The validated method was used to analyze 46 wheat flour samples for the targeted mycotoxins. The method can be used as a rapid and robust tool for screening mycotoxin in cereal products.     

Occurrence of emerging mycotoxins in cereals and cereal-based products from the Korean market using LC-MS/MS

The present study aimed to investigate the occurrence of emerging mycotoxins in cereals (n = 61) and cereal-based products (n = 36) collected from Korean market. First of all, using the quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction method, and ultrahigh-pressure liquid chromatography (UPLC) with triple quadruple tandem mass spectrometry (MS/MS), we developed a simple and fast method for quantitative determination of eight emerging mycotoxins including alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), beauvericin (BEA) and enniatins (ENs; ENA, ENA1, ENB and ENB1). The developed analytical method was validated in parameters of linearity, precision and accuracy. For UPLC-MS/MS analysis, the recoveries of emerging mycotoxins from spiked samples at three concentration levels ranged from 82.7% to 108.8% with RSDs between 0.4% and 14.7%. Analytical methods were applied to determine the contamination of mycotoxins in cereal and cereal-based product samples. Sixty-three of the total 97 samples were contaminated with at least one emerging mycotoxin. The maximum number of emerging mycotoxins observed in a single sample was six out of eight analytes. The highest level of contamination was detected in cereal at 70.9 μg/kg for alternariol monomethyl ether (AME). However, currently there is no international standard for emerging mycotoxins in food. Accordingly, it is necessary to establish a database of emerging mycotoxins contamination through continuous monitoring.     

Multiclass mycotoxin analysis in edible oils using a simple solvent extraction method and liquid chromatography with tandem mass spectrometry

ABSTRACT

A simple and rapid method for the simultaneous determination of 11 mycotoxins – aflatoxins B₁, B₂, G₁ and G₂; fumonisins B₁, B₂ and B₃; ochratoxin A; zearalenone; deoxynivalenol; and T-2 toxin – in edible oils was established using liquid chromatography tandem mass spectrometry (LC-MS/MS). In this study, QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe), QuEChERS with dispersive liquid–liquid microextraction, and solvent extraction were examined for sample preparation. Among these methods, solvent extraction with a mixture of formic acid/acetonitrile (5/95, v/v) successfully extracted all target mycotoxins. Subsequently, a defatting process using n-hexane was employed to remove the fats present in the edible oil samples. Mass spectrometry was carried out using electrospray ionisation in polarity switching mode with multiple reaction monitoring. The developed LC-MS/MS method was validated by assessing the specificity, linearity, recovery, limit of quantification (LOQ), accuracy and precision with reference to Commission Regulation (EC) 401/2006. Mycotoxin recoveries of 51.6–82.8% were achieved in addition to LOQs ranging from 0.025 ng/g to 1 ng/g. The edible oils proved to be relatively uncomplicated matrices and the developed method was applied to 9 edible oil samples, including soybean oil, corn oil and rice bran oil, to evaluate potential mycotoxin contamination. The levels of detection were significantly lower than the international regulatory standards. Therefore, we expect that our developed method, based on simple, two-step sample preparation process, will be suitable for the large-scale screening of mycotoxin contamination in edible oils.     

Risk Assessment of Mycotoxins and Predictive Mycology in Sri Lankan Spices: Chilli and Pepper

This research contributes to a better understanding of the mycotoxin problem associated with two important spices in world trade;, chilli and pepper by a multidisciplinary approach including analytical chemistry, risk assessment, food chemistry and predictive mycology. More specifically, this work provides important insights in mycotoxin contamination of these spices and associated risks in Sri Lanka. Firstly, a simple extraction method based on the QuEChERS approach was developed and successfully validated for the simultaneous determination of multiple mycotoxins using an advanced chromatographic technique, LC-MS/MS. The method was applied on complex spices for quantitative screening of seventeen mycotoxins. In addition to the classical aflatoxins and ochratoxin A, the spices were also found to be contaminated with several other toxicologically significant mycotoxins. Chilli samples (87%) were more frequently contaminated with mycotoxins than peppers (65%). Subsequently, the mycotoxins screening results and the collected consumption data were integrated in a quantitative risk assessment study. The results showed that AFB1 exposure via chilli consumption is of a public health concern in Sri Lanka, pepper is of lesser extent a risk due to the lower consumption. The toxigenic mould characterization in black peppers showed that Aspergillus flavus and/or Aspergillus parasiticus were the predominant moulds (73%) found, with considerable contamination (60%) of Penicillium spp. and A. niger. Furthermore, predictive mould growth models on peppercorns were developed at three temperatures and seven water activity levels for both A. flavus and A. parasiticus isolates. Based on the research, suitable storage conditions for black peppercorns were suggested and the way forward in managing the risk towards mycotoxins posed by the consumption of these two spices in Sri Lanka.     

UPLC-ESI-MS-MS结合QuEChERS同时测定柑橘中的4 种真菌毒素

建立同时检测柑橘中链格孢酚甲基乙醚、交链孢酚、腾毒素和橘青霉素4 种真菌毒素的QuEChERS-超高效液相色谱-质谱分析方法。待测样品经乙腈提取,无水MgSO4和NaCl脱水盐析,C18键合相分散萃取净化,以ACQUITY UPLC BEH C18液相色谱柱为分离柱,以0.1%甲酸溶液和乙腈作为流动相梯度洗脱,在电喷雾离子源电离、多反应监测模式条件下进行测定,外标法定量。该方法在5.0～200 μg/L范围内线性关系良好,相关系数（R2）不小于0.992 7,检出限为0.2～0.7 μg/kg。3 个不同加标水平的平均回收率为78.0%～103.3%,相对标准偏差为2.6%～10.6%。结果表明,该方法快速、准确、灵敏,适用于柑橘中4 种真菌毒素残留的快速确证检测。     

生物识别元件功能化微纳米磁性材料在真菌毒素检测中的应用

真菌毒素是真菌产生的一类强毒性次级代谢产物,可通过食品或食物链危害人类的健康。建立快速、准确、灵敏的检测方法是控制真菌毒素危害的有效手段之一。功能化微纳米磁性材料因具有超顺磁性、比表面积大等特性,近年来在建立样品前处理新方法及生物传感器分析等领域的应用日益增多。本文在概述真菌毒素传统检测方法及微纳米磁性材料特点的基础上,综述近期生物识别元件功能化微纳米磁性材料在样品前处理及利用生物传感器高灵敏快速检测真菌毒素中的应用,并对其发展进行展望。     

Development and Comparison of Sample Preparation Techniques for Chromatographic Analysis of Sulfonamide Residues in Bovine Milk

Most studies to determine sulfonamide residues in milk samples have used solid-phase extraction as the sample preparation technique. However, the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method, introduced in 2003, has been used in the extraction of various compounds in food matrices. This study aimed to evaluate two sample preparation techniques: solid-phase extraction and QuEChERS, for chromatographic analysis of sulfonamides (sulfathiazole, sulfamethazine, and sulfadimethoxine) in bovine milk. The chromatographic parameters and the QuEChERS extraction procedure were developed by using different experimental designs, obtaining good peak resolution, recovery, precision, accuracy, linearity, selectivity, and limits of detection and quantification. In contrast, using solid-phase extraction, acceptable recoveries and selectivity were not achieved, despite the number of articles published that have applied this sample preparation technique for sulfonamide analysis. As a result of the experiments performed, probably sulfonamides are retained together with other components of the matrix in the sample pretreatment step (prior to its addition in the cartridge containing solid phase), which is an important part of solid-phase extraction with raw whole milk. Therefore, QuEChERS is a better method than solid-phase extraction for the analysis of sulfonamide residues in milk. Validation tests demonstrated that the method is appropriate, within the maximum residue limit (0.1 mg kg^?1). Moreover, it was possible to use a lower amount of solvent compared with previously published articles (6 mL against 10 or 15 mL).