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。     

直接稀释-超高效液相色谱-串联质谱法快速测定谷物及其制品中16种真菌毒素

目的建立谷物及其制品中黄曲霉毒素、赭曲霉毒素、单端孢霉烯族类毒素、伏马菌素等16种真菌毒素的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。方法 5 g样品加入20 ml乙腈-水-甲酸(79∶20∶1,V/V)溶液提取,10 000 r/min离心5 min后稀释,Cortecs C_(18)色谱柱(100 mm×3.0 mm,1.6μm)分离,以乙腈-0.1%甲酸水作为流动相梯度洗脱,采用四极杆串联质谱仪电喷雾模式检测,同位素稀释内标法定量。结果 16种真菌毒素在线性范围内(0.05~200 ng/ml)具有良好的线性关系,相关系数(r~2)0.99,方法定量限为0.1~50μg/kg,加标回收率为72.67%~126.92%之间,相对标准偏差(RSD)为0.15%~16.83%。应用本方法分析英国弗帕斯检测技术研究所(FAPAS)质控样品,测定结果均在标示范围内,方法准确度良好。结论本方法具有良好的灵敏度、回收率和重复性,前处理方法简单,适用于常规实验室对谷物及其制品中真菌毒素的检测,满足日常真菌毒素监测工作的需要。     

杂质吸附固相萃取–液相色谱串联质谱法同时测定粮食中15种真菌毒素

建立了同步检测粮食中15种真菌毒素的杂质吸附固相萃取–超高效液相色谱串联质谱方法。样品经0.1%酸化84%乙腈水溶液提取,杂质吸附柱Prime-HLB净化后,加入稳定同位素内标补偿基质效应干扰,选择Phenomenon Kinetex Bipheny l100A作为分离柱,采用梯度洗脱,超高效液相色谱–串联质谱（UPLC-MS/MS）检测,内标法定量。结果表明,15种真菌毒素的线性相关系数均大于0.996,检出限为0.2~15 μg/kg,定量限为0.8~30 μg/kg;三种基质3个添加水平的回收率为76.9%~116.1%,相对标准偏差（RSD）为1.4%~10%;对玉米基质标准物质进行检测验证,表明8种真菌毒素的检测值均在标示范围内;采用本方法检测了市售粮食135批次,共有97批样品检出真菌毒素,检出率为71%,多毒素同时污染现象较为普遍。方法可用于粮食及其制品中真菌毒素的快速检测。     

基于复合萃取技术-超高效液相色谱-串联三重四极杆质谱法同时测定谷物及其制品中10种真菌毒素

目的 建立超高效液相色谱串联三重四极杆质谱法(ultra high performance liquid chromatography tandem with triple quadrupole mass spectrometry, UPLC-MS/MS)同时测定谷物及其制品中的10种真菌毒素的检测方法。方法 以0.1%甲酸-80%乙腈水为提取溶剂,经过涡旋、振荡提取后,加入Supel QuE z-sep+作为净化剂进行初级净化,离心后的上清液过PRiME HLB固相萃取柱二次净化,经UPLC-MS/MS测定,质谱采用多反应监测模式(multiple reaction monitoring mode, MRM)定性,基质匹配标准溶液外标法定量。结果10种真菌毒素在谷物中的检出限为0.2~10.0 μg/kg,定量限为0.6~20 μg/kg。在玉米、花生、大米3种谷物基质中的回收率为73.2%~98.1%,相对标准偏差为1.3%~9.3%。结论 该方法的灵敏度满足国家标准对真菌毒素的限量要求,实验操作简便快速,成本较低,定性定量分析可靠,适合谷物中真菌毒素的多组分高通量快速分析测定。     

QuEChERS/超高效液相色谱-串联质谱法测定茶叶中28种农药残留

建立了QuEChERS/超高效液相色谱-串联质谱（UPLC-MS/MS）测定绿茶中28种农药残留的分析方法。样品经1%乙酸乙腈提取,QuEChERS方法净化后进行UPLC-MS/MS测定。采用电喷雾离子源（ESI）,正离子模式扫描,多反应检测（MRM）模式采集,基质匹配工作曲线法定量。结果表明:28种目标化合物在各自浓度范围内线性良好,相关系数（r）为0.990~0.998,检出限（LOD,S/N≥3）为0.08~1.26 μg/kg,定量限（LOQ,S/N≥10）为0.28~4.20 μg/kg;在10、50、80 μg/kg 3个加标水平下平均回收率为72.5%~118.5%,相对标准偏差（n=6）为0.8%~16.3%。该方法简单、快速、重现性好、稳定性高,可满足茶叶中28种农药残留的检测要求。     

超高效液相色谱-串联质谱法测定水产品中50种兽药残留

目的 建立QuEChERS-超高效液相色谱-质谱/质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定水产品中50种兽药残留(三苯甲烷类、酰胺醇类、硝基咪唑类、磺胺类、喹诺酮类)的方法。方法 样品经过含1%甲酸的乙腈溶液提取, 采用QuEChERS方法净化, 通过相同C18色谱柱分离, 经2个不同流动相系统(0.1%甲酸水溶液-甲醇、5 mmol/L乙酸铵水溶液-甲醇)梯度洗脱后, 采用UPLC-MS/MS电喷雾正离子模式、正负切换模式分别分析测定, 基质添加标准曲线外标或内标法定量。结果 50种药物标准曲线相关系数均≥0.997, 检出限为0.0319~0.356 418 μg/kg, 加标回收率为60.7%~114%, 相对标准偏差为1.06%~14.1%。结论 本方法快速、高效, 可应用于水产品中多种兽药残留的快速准确筛查和检测。     

QuEChERS前处理结合超高效液相色谱-串联质谱法同时检测茶叶中14种农药残留

建立了QuEChERS（Quick,Easy,Cheap,Effective,Rugged,Safe）前处理结合超高效液相色谱-串联质谱法（UPLC-MS/MS）测定茶叶中14种农药残留的方法。茶叶样品经含1%乙酸的乙腈提取,QuEChERS净化柱净化后直接进行UPLC-MS/MS测定。比较了4种QuEChERS净化柱对茶叶提取液的净化效果和农药残留的回收率,选取净化效果和回收率最好的净化柱进一步考察方法的基质效应、线性关系、回收率等。结果表明,方法的检出限为0.001~2.000 μg/L,定量限为0.003~5.000 μg/L;14种农药在相应浓度范围内呈良好的线性关系,决定系数R2为0.9926~0.9989。在100、500、800 μg/kg 3个添加水平下14种农药的回收率为71.2%~117.4%,相对标准偏差为1.1%~14.2%。该方法具有操作简单快速、灵敏度高、回收率好等优点,能够满足茶叶中多农残快速检测要求。     

超高效液相色谱-串联质谱法同时检测饲料原料、饲料成品中18种真菌毒素含量

目的 建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)同时测定饲料原料和饲料成品中18种真菌毒素的方法。方法 样品经50%乙腈水溶液提取后, 经MycoSpin TM 400净化柱净化, 同位素内标稀释后, 上机测定。结果 在低、中、高3个添加水平下, 18种毒素的空白样品平均加标回收率为62.6%~112.53%(预混料除外), 相对标准偏差(relative standard deviation, RSD)为1.82%~13.83%。18种真菌毒素在各自的线性范围内线性关系良好, 线性系数均大于0.995, 检出限(limit of detection, LOD, S/N=3)和定量限(limit of quantity, LOQ, S/N=10)分别为0.15~10 μg/L和0.5~30 μg/L。对2018年初的饲料原料以及饲料成品近900个样品进行了18种真菌毒素的普查工作, 结果显示呕吐毒素和伏马菌素的阳性检出率最高, 分别为93%和91%。结论 此方法操作简单、灵敏度高且应用范围广, 可以用于饲料中真菌毒素的大规模普查。     

固相萃取-超高效液相色谱-串联质谱法测定禽蛋中的67种禁用药物

摘要：目的 建立固相萃取-超高效液相色谱-三重四极杆串联质谱法（Solid Phase Extraction-ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry, SPE-UPLC-QqQ-MS）测定禽蛋中的67种禁用药物残留的分析方法。方法 禽蛋样品用Na2EDTA-Mcllvaine缓冲溶液提取,加入钨酸钠沉淀蛋白,经Anavo HLB固相萃取柱富集净化后采用 ACQUITY UPLC HSS T3柱(2.1×100 mm,1.8 μm)分离,正离子模式以0.1%甲酸水溶液-甲醇为流动相、负离子模式以水-乙腈为流动相进行梯度洗脱,在UPLC-MS/MS多反应监测模式下进行定性定量分析,同位素内标法或基质配外标法定量。结果67种禁用药物在0.1~100 μg/kg范围内均呈现良好的线性关系,相关系数R2≥0.99,方法的定量限范围为0.1~0.5 μg/kg。平均加标回收率为56.3%~110.1%,相对标准偏差为1.5% ~16.8%。结论 本方法操作简单、准确性高、成本较低,可实现禽蛋中禁用药物残留的高通量分析,为禽蛋产品质量监管提供技术支持。     

UPLC-MS/MS与HPLC测定茶叶中10种氨基甲酸酯类农药残留的比较

比较了超高效液相色谱-串联质谱(UPLC-MS/MS)与高效液相色谱(HPLC)法测定茶叶中10种氨基甲酸酯类农药残留的分析方法。样品经乙腈提取,氨基固相萃取柱净化,HPLC和UPLC-MS/MS测定。结果表明,在相同的前处理条件下,两种方法均具有较好的准确度和精密度,UPLC-MS/MS法在0.01~0.1mg/kg添加水平下回收率为75.2%~115.6%,相对标准偏差(RSD)为4.4%~16.7%。HPLC法在0.05~1.0mg/kg添加水平下回收率为82.7%~120.7%,相对标准偏差(RSD)为0.8%~12.9%。UPLC-MS/MS法相对于HPLC法有更高的灵敏度,10种氨基甲酸酯类农药在UPLC-MS/MS上检出限为0.02~0.3μg/kg,HPLC法检出限为3~7.5μg/kg;其高灵敏度和选择性,使其更适于多农药残留同时检测和微量残留检测。     

婴幼儿谷类辅助食品加工工艺对其脱氧雪腐镰刀菌烯醇及衍生物影响

脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)是由镰刀菌在侵染小麦等禾谷类作物过程中产生的一种有毒次级代谢产物,主要污染小麦、大麦、燕麦、黑麦和玉米等谷物。DON在植物和微生物作用下会转化形成各种衍生物,且这些衍生物可与DON原型同时存在,增加了谷物及其相关制品的安全风险。谷类辅食是婴幼儿膳食中的重要组成部分,且婴幼儿对毒素的敏感性更强,应严格限制DON及其衍生物在谷类辅食中被检出。已有研究表明谷物类食品的制作加工方式会对DON及其衍生物的含量水平产生影响,但专门针对婴幼儿消费人群的谷类辅助食品的加工过程如何影响DON及其衍生物水平还鲜有报道。目前婴幼儿谷类辅助食品中DON及其衍生物含量存在超标问题,严重影响婴幼儿这类特殊人群的生长发育和健康。本文从婴幼儿谷类辅食中DON及其衍生物的污染状况及其在加工过程中的含量变化等方面进行较为详细的阐述,以期为婴幼儿谷类辅食中DON的风险评估及防控策略的制定提供有利参考。     

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.     

我国市售婴幼儿谷类辅助食品营养成分含量水平研究

了解我国市售婴幼儿谷类辅助食品营养成分含量水平,为GB 10769-2010《食品安全国家标准 婴幼儿谷类辅助食品》标准修订提供参考。收集我国市售婴幼儿谷物辅助食品、婴幼儿高蛋白谷物辅助食品、婴幼儿生制类谷物辅助食品和婴幼儿饼干类辅助食品,对GB 10769-2010中规定的基本营养成分和可选择营养成分含量进行检测,对检测结果进行统计分析。共收集婴幼儿谷类2004份,其中婴幼儿谷物辅助食品1850份,婴幼儿高蛋白辅助食品50份,婴幼儿生制类食品51份,婴幼儿饼干53份。婴幼儿高蛋白谷物辅助食品蛋白质平均含量高于其他3类谷物辅助食品蛋白质含量,为0.88 g/100 kJ,差异具有统计学意义(P<0.05)。婴幼儿饼干类辅助食品能量和脂肪平均含量高于其他3类谷物辅助食品能量和脂肪含量,分别为1777 kJ/100 g和0.55 g/100 kJ,差异具有统计学意义(P<0.05)。GB 10769-2010有下限值和上限值范围值的营养素,维生素A、维生素D、钙、铁、锌、维生素E、磷、碘、钾等含量平均值分别接近各自理论平均值。GB 10769-2010无上限值范围值的营养素,市售产品其含量范围的最大值是国标下限值的2.83~67.04,其含量范围的95百分位数是国标下限值的2.05~16.35。GB 10769-2010中未制定上限值的营养成分出现离群值。建议综合考虑我国市售婴幼儿谷类辅助食品营养素含量分布情况,对无上限值的营养素进行修订。     

Kavut, a traditional Turkish cereal product: production method and some chemical and sensorial properties

Kavut is a cereal‐based product made from wholemeal wheat and barley flour, milk or milk powder, fat and sugar. In this study, the best processing method and formulation were investigated by analysing different formulations and processing methods for the kavut. Four different flour combinations (100% wheat without barley, 75% wheat + 25% barley, 50% wheat + 50% barley, 25% wheat + 75% barley), two shortening (butter and margarine) and three different roasting periods (1, 1.5, 2 min) at 250 °C were used in the study. Significant changes were observed in the physical and chemical properties of the cereal by heat treatment in the kavut production. Increase of barley flour in the mixture decreased protein content, softness, altered L colour values, appearance, texture, mouth‐feel, and general acceptance and increased ash content, +a and the absorbance value of kavut. While the longer roasting period resulted in reduced softness and colour (L‐value) of kavut, roasting period did not significantly affect the general acceptance of kavut. Kavut made from only whole wheatflour was most preferred by the panellists.     

Evaluation of enniatins A,A1, B,B1 and beauvericin in Portuguese cereal-based foods

Sixty-one samples of Portuguese cereal-based foods were analysed for the occurrence of emerging mycotoxins called enniatins (A, A1, B and B1) and beauvericin. Samples were extracted with a mixture of acetonitrile/water (85/15, v/v) using an Ultra-Turrax homogeniser, and mycotoxins were detected with liquid chromatography coupled to a mass spectrometer. This method was validated and adequate values of recovery (70–103%) and relative standard deviation (<15%) were obtained. Signal suppression/enhancement was studied and matrix-matched calibration used to minimise this effect, but no additional clean-up step was necessary. The mass spectrometer was operated in selected reaction monitoring (SRM) mode and with selected transitions for each compound to quantify and to qualify them. Fifty-nine per cent of samples were contaminated. The percentages of enniatins were 53%, 49%, 44% and 16% for A1, B, B1, and A, respectively, and for beauvericin it was 1.6%. For the total samples, the mean contamination was 30, 24, 15, 2.1 and 0.1?ng?g^?1 for enniatins A1, B, B1 and A, and beauvericin, respectively. The wheat-based samples showed higher levels and greater prevalence than any other cereals monitored. These results were used to estimate the daily intake of ENs from wheat-based cereal by the Portuguese population. At the same time, the usefulness of this method in the analysis of other important mycotoxins (aflatoxin B₁, ochratoxin A, deoxynivalenol, T-2 toxin, fumonisin B₁ and zearalenone) was evaluated.     

谷物及制品中真菌毒素前处理及检测技术研究进展

谷物及其制品在生产、贮藏、运输的各个环节均易受到真菌毒素的污染,且真菌毒素种类多、浓度低、毒性强、性质差异大,防治困难。文章综述了谷物及其制品中真菌毒素的前处理技术(液液萃取技术、固相萃取技术、QuEChERS技术、免疫亲和层析技术)和检测技术(免疫层析技术、光谱技术、液相色谱技术、液质联用技术),并对真菌毒素检测技术的发展趋势进行了展望。     

《婴幼儿谷类辅助食品》(GB 10769—2010)跟踪评估研究

目的 了解我国监督部门、检验机构和生产企业对GB 10769-2010《婴幼儿谷类辅助食品》的依从性和贯彻实施情况,以期为今后标准的制修订提供资料.方法 通过现况调查,了解婴幼儿谷类辅助食品主产区的监管部门、检验机构和生产企业的基本情况、对标准条款的理解、各项指标的检测能力和标准的实施情况,收集标准执行过程中遇到的主要问题以及反馈的意见和建议.结果 向监督部门、检验机构和生产企业分别发放问卷176、90和56份,有效问卷回收率分别为95.5％、91.1％和85.7％,不同部门均认为标准的整体合理程度较高(89.3％、90.2％、95.9％),收集的反馈意见分别为235、231和207条,大型、外资企业和监督部门、检验机构中从事食品监管和检验工作时间较长以及标准使用频率较高、积极参加各项培训活动的人员反馈问题相对较多.其中产品分类、原料要求、基本营养成分、污染物和真菌毒素限量部分的反馈意见相对集中.结论 通过跟踪调查发现标准整体合理程度较高,部分指标需要跟进新的风险评估结果并及时更新指标值,某些重点营养素亟需出台配套的国标检验方法.加大监管部门、中小企业标准培训的力度和深度,并发挥媒体在标准宣贯中的积极作用,对各方正确执行标准有重要意义.     

2015~2019年度婴幼儿谷类辅助食品监督抽检结果统计与分析

目的 分析2015~2019年度婴幼儿谷类辅助食品安全整体状况,了解不合格主要指标。方法 从国家市场监督管理总局网站收集2015~2019年度婴幼儿谷类辅助食品监督抽检公告资料,对抽检结果进行统计与分析;合格率的比较采用卡方检验分析。结果 从2015～2019年度4月上旬,共抽检样品1773批次,合格率为92.39%。2015年抽检合格率最低为89.50%,其他年度抽检合理率均高于93%。中国大陆婴幼儿谷类辅助食品生产企业的产品抽检合格率为92.83%,高于其他食品生产企业的合格率87.23%。不合格项目主要为维生素类指标和钠。结论 我国市售婴幼儿谷类辅助食品合格率较高,总体呈平稳向好趋势。     

Microbiological and mycotoxicological analyses of processed cereal-based complementary foods for infants and young children from the German market

This study investigated several food safety criteria in 38 different commercial products of processed cereal-based foods (PCF) from the German market. Microbiological assessment, followed by 16S RNA gene sequencing of suspect colonies, included aerobic mesophilic bacteria, moulds, Enterobacteriaceae, Cronobacter spp., and presumptive Bacillus cereus. Mycotoxin analyses were performed by enzyme immunoassays for deoxynivalenol (DON), zearalenone (ZEN), T-2/HT-2 toxins (T-2/HT-2; oat containing products only), ergot alkaloids (EA), and alternariol (AOH). No violative result above existing European Union regulations or international guidelines was obtained. Most samples had very low aerobic mesophilic cell counts (<2.0 × 10¹ CFU/g), the maximum was 9.6 × 10² CFU/g. A few samples contained low numbers of opportunistic pathogens, most notably Cronobacter sakazakii, Acinetobacter spp., Pantoea spp., and enterotoxigenic Bacillus wiedmannii. Levels of mycotoxin contamination were very low, well below European Union maximum limits. DON was found in 10 samples, at levels of 9–35 µg/kg. T-2/HT-2 were found in all 15 oat-based products (1–8 µg/kg). All samples were negative for ZEN and EA. A high number (n = 25) of samples yielded weakly positive results for the nonregulated AOH (0.4–2 µg/kg), but just three samples exceeded a level of 1 µg/kg. No relationship between cereal composition and analytical findings for microbiological parameters and mycotoxins could be found. As long as PCF meals are freshly prepared and consumed immediately after preparation, the risk from sporadically occurring opportunistic bacteria appears to be minimal.     

Couscous, a traditional Turkish food product: production method and some applications for enrichment of nutritional value

Traditional Turkish couscous, a cereal‐based product, is made by covering bulgur with milk and flour. Traditionally Turkish couscous was produced from different flours (soyflour and oat flour) and eggs. The nutrient composition of traditional couscous was 90.6% dry matter, 11.27% protein, 2.58% fat, 71.80% carbohydrate, 42.25 mg sodium, 365.62 mg potassium, 2.73 mg iron and 48.30 mg calcium. The calorie content of traditional couscous is about 1487.41 kJ 100 g^?1. The addition of soy and oat flours increased protein content and Ca, K and Fe levels. Sensory properties were also affected. Panellists preferred traditional couscous and couscous with eggs or soyflour over couscous with oat flour.