高效液相色谱柱后衍生法检测啤酒中黄曲霉毒素的研究

现在大多数检测黄曲霉毒素的方法,如酶联免疫测试盒、免疫亲和层析净化荧光光度法等,都只能检测单一的黄曲霉毒素B₁或黄曲霉毒素总量,而且检测限高,重复性和稳定性不好,难以得到理想的测试结果。本文主要研究啤酒样品经pH7.0磷酸盐缓冲溶液调节pH值后,用含有黄曲霉毒素特异抗体的免疫亲和层析柱净化富集,黄曲霉毒素交联在层析介质的抗体上,用吐温-20/PBS将免疫亲和柱上杂质除去,再以甲醇通过免疫亲和层析柱洗脱,洗脱液经C₁₈柱分离,然后用液相色谱柱后衍生法,用荧光检测器进行检测。本方法可同时分离检测出黄曲霉毒素B₁、B₂、G₁和G₂,检测限可达0.2μg/L。     

免疫亲和层析净化荧光光度法快速检测鸡饲料中黄曲霉毒素

基于食品中采用免疫亲和层析净化荧光光度法测定黄曲霉毒素含量,建立了免疫亲和层析净化荧光光度法测定鸡饲料中黄曲霉毒素B1、B2、G1、G2含量。该方法的检出限2.0 μg/kg,相对标准偏差低于10%,平均回收率85%~90%。     

免疫亲和层析净化荧光光度法快速测定酱油及醋中黄曲霉毒素

为检测酱油和醋中黄由霉毒素的含量，建立了免疫亲和层析净化荧光光度法。试样由甲醇一水提取，提取液被过滤、稀释后，上交联着黄曲霉毒素B1、B2、G1、G2特异抗体的免疫亲和层析柱净化。以甲醇通过免疫亲和层析柱洗脱，溴溶液衍生，荧光光度计测定衍生物的黄曲霉毒素(B1 B2 G1 G2)含量。酱油和醋中检出限分别为2．5μgkg和1μg／kg。添加回收率在85％以上。该方法准确、简单、快速、安全。     

免疫亲和层析荧光光度法测定枸杞中黄曲霉毒素

利用免疫亲和层析荧光光度法测定枸杞中的黄曲霉毒素(AFT),样品由甲醇-水提取,提取液经过滤、稀释,用黄曲霉毒素免疫亲和柱净化。以甲醇洗脱,溴溶液显色,荧光光度计测定黄曲霉毒素(B1+B2+G1+G2)含量。枸杞的最低检出限为1μg/kg,5μg/kg的加标回收率为99.6%,10μg/kg的加标回收率为114.0%。该方法准确、快速、安全。     

免疫亲和层析净化高效液相色谱法和胶体金快速定量法测定小麦中呕吐毒素含量的比较

采用免疫亲和层析净化高效液相色谱法和胶体金快速定量法测定小麦中呕吐毒素含量。结果表明:用这两种方法测定添加不同浓度呕吐毒素的小麦,测定结果的相对标准偏差分别为1.59%~3.08%和7.28%~11.50%,且两种方法测定呕吐毒素呈阳性的小麦样品的测定结果相对偏差小,相关系数为0.995,相关性好。这两种方法各有优缺点,免疫亲和层析净化高效液相色谱法精密度好,能够对呕吐毒素进行精准定量,但试验步骤较为复杂,耗时长;胶体金快速定量法检测简便快速,能用于大量样品的快速筛查,预期在粮食中真菌毒素检测方面具有很好的应用前景。     

免疫亲和层析净化高效液相色谱法测定呕吐毒素的应用与分析

阐述免疫亲和层析净化高效液相色谱法测定呕吐毒素的原理、试样制备、样品提取、仪器调试等操作过程和技术要领,开展检验质量控制,确保了免疫亲和层析净化高效液相色谱法呕吐毒素测定的准确应用。     

柱后衍生-高效液相色谱法检测核桃粉中黄曲霉毒素B1含量的研究

建立了一种免疫亲和层析净化-高效液相色谱法测定核桃粉中黄曲霉毒素B．含量的办法。将核佻粉样品用甲醇-水提取、稀释后直接过免疫亲和枉净化,经甲醇洗脱后用HPLCNIJ定。该方法检测黄曲霉毒素B．的回收率在84．0～92．7％之间,相对标准偏差在1．51～4．38％之间,最低检出限为0．5μg／kg,且检测结果准确可靠,重复性好,可进行核挑粉中黄曲霉毒素B1含量的检测。     

SPE法低成本快速检测乳制品中黄曲霉毒素M1

黄曲霉毒素,1993年被世界卫生组织的癌症研究机构划定为1类致癌物.其中黄曲霉毒素B1毒性和致癌性最强,而黄曲霉毒素M1是黄曲霉毒素B1的代谢物,主要存在于牛奶中.卫生部官网一份有关食物中毒的说明文件指出,黄曲霉毒素主要损伤肝脏,致癌性很强,我国乳及乳制品中规定黄曲霉毒素M1限量为0.5μg/kg. 目前检测黄曲霉毒素的方法通常为免疫亲和层析净化高效液相色谱法和酶联免疫吸附法(ELISA)这两种方法.酶联免疫吸附法(ELISA)操作简单,快速,但灵敏度低且会出现假阳性,需使用免疫亲和柱法进行精确定性、定量,而免疫亲和柱又存在使用繁琐、价格昂贵、储存条件苛刻、保存期短等缺点.     

食品中赭曲霉毒素样品前处理及其检测方法研究进展

赭曲霉毒素广泛存在于谷物、葡萄和咖啡等众多食品中,具有极强的肝肾毒性和致畸、致癌作用,可经食物链传播效应累积在人体血液及内脏中,对人类健康造成巨大威胁。因此有必要建立食品中快速、灵敏的赭曲霉毒素分析方法,保障食品安全、维护人类的健康。食品基质复杂多样,且赭曲霉毒素在样品中含量极低,检测前需要高效的前处理技术对目标物进行分离和富集。本文总结了基于分子印迹材料、纳米石墨烯、生物材料、磁性纳米材料等新型分离介质的液相萃取、QuEChERS、固相萃取、免疫亲和层析和场辅助提取(微波辅助提取、磁性固相萃取、超声辅助提取)等前处理技术,综述了色谱法、免疫分析法和生物传感器在食品中赭曲霉毒素分析中的应用,并对其发展趋势进行了展望。     

免疫亲和层析净化-超高效液相色谱法测定生鲜牦牛乳中黄曲霉毒素M_1及M_2

使用免疫亲和层析净化-超高效液相色谱法同时测定了生鲜牦牛乳中的黄曲霉毒素M_1(Aflatoxin M_1,AFM_1)及黄曲霉毒素M_2(Aflatoxin M_2,AFM_2)。黄曲霉毒素M_1仪器线性范围0.400-100μg/mL,仪器定量限0.400ng/mL,仪器检出限0.100ng/mL,方法检出限0.005μg/kg,回收率85.2-95.5%;黄曲霉毒素M_2仪器线性范围0.100-100μg/mL,仪器定量限0.100ng/mL,仪器检出限0.03ng/mL,方法检出限0.0015μg/kg,回收率82.1-93.8%。使用该方法对来自若尔盖地区的10个生鲜牦牛乳进行了测定,结果表明生鲜牦牛乳中黄曲霉毒素M_1和M_2安全可控。     

Survey of Canadian retail coffees for ochratoxin A

One-hundred and one specimens of coffee were gathered from retail outlets across Canada and analysed for ochratoxin A. Seventy-one specimens were roasted beans or roasted ground coffee, and 30 were instant (or 'soluble') coffees. All samples were extracted with methanol-sodium bicarbonate. The extracts were cleaned up either by immunoaffinity column chromatography or by a combination of solid-phase extraction and immunoaffinity column chromatography. Ochratoxin A was quantified by liquid chromatography (LC) with fluorescence detection. The minimum quantifiable level was 0.1 ng g^-1. Ochratoxin A was present, above the minimum quantifiable level, in 42 (59%) of 71 beans and ground coffee and in 20 (67%) of 30 instant coffees. The mean ochratoxin A level in the positive samples of beans and ground coffee was 0.6 ng g^-1, and the mean level in the positive samples of instant coffee was 1.1 ng g^-1.     

LAURENCE BISHOP MEMORIAL LECTURE THE CURRENT STATUS OF ANALYTICAL METHODS IN THE BREWING INDUSTRY*

Progress concerning the presentation of analytical methods in terms of format and precision is reviewed. Methods are described in which instrumental techniques such as gas chromatography, high performance liquid chromatography and near infrared spectroscopy are used to estimate a range of analytes in brewing raw materials and final products. Methods relying on immunoaffinity columns are described for the detection and determination of mycotoxins.     

粮谷中农药残留前处理技术的研究进展

样品前处理技术是保证样品检测结果准确性的重要因素。随着农产品检测技术的发展,粮谷中农药残留前处理技术也从传统的前处理技术阶段发展到新型高效的前处理技术阶段。本文通过检索文献,按照时间顺序以及各国的发展历程,对固相萃取、固相微萃取、超临界流体萃取、微波辅助萃取、凝胶渗透色谱、基质固相分散萃取、加速溶剂萃取、免疫亲和色谱和分散固相萃取一系列样品前处理技术在检测粮谷中农药残留分析方法中的应用进行了简单介绍和概述。归纳了测定粮谷中农药残留的常用提取溶剂、净化方法和分析仪器,并对日常检验工作中如何利用每种前处理技术提出了建议和展望。本文可为食品安全检测领域的科技人员进行进一步的研究和应用提供借鉴和参考。     

粮食加工品中玉米赤霉烯酮和呕吐毒素污染 情况调查

目的初步了解粮食加工品中玉米赤霉烯酮和呕吐毒素的污染情况。方法对北京地区和网购的200批次样品进行随机抽检,分别采用液相色谱法、免疫亲和层析净化高效液相色谱法对粮食加工品中的玉米赤霉烯酮和呕吐毒素进行了检测。结果初步调查发现玉米碾磨加工品中的玉米赤霉烯酮和小麦粉中的呕吐毒素检出率较高,分别达34%和74%;散装样品中玉米赤霉烯酮含量不合格率和检出率明显高于预包装样品。结论建议监管部门加强对散装玉米碾磨加工品和小麦粉等粮食加工品中毒素指标的监控力度和监测频次,同时针对我国特殊敏感人群加快设定合理限值。     

Natural occurrence of ochratoxin A in wolfberry fruit wine marketed in China

Wolfberry fruit wine (WFW) is widely used as a global functional food to improve the immune system and prevent human disease. A total of 36 bottled WFWs were randomly collected in China between 2005 and 2010. Samples were analysed for the presence of ochratoxin A (OTA) using immunoaffinity column (IAC) clean-up and high-performance liquid chromatography with fluorescence detection (HPLC-FLD). Positive results were confirmed by liquid chromatography–electrospray ionisation–tandem mass spectrometry (LC-ESI-MS/MS). The limit of detection (LOD), based on a signal-to-noise ratio of 3, was 0.05?ng?mL^–1. Recoveries ranged from 78.3% to 94.7% and relative standard deviations from 1.1% to 4.3% within the spiking range of 0.2–20?ng?mL^–1. OTA was detected in one sample, below the maximum allowable limit as established by the European community.     

免疫亲和柱结合超高效液相色谱-串联质谱法测定牛奶中的16种真菌毒素比较研究

目的 通过比较5种不同商品化多合一免疫亲和柱的可检测目标毒素种类、回收率和稳定性,筛选性能最优的免疫亲和柱,建立免疫亲和前处理-超高效液相色谱-串联质谱法(ultraperformanceliquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定牛奶中16种真菌毒的方法。方法 牛奶样品分别经5种不同多毒素免疫亲和柱进行净化富集,用优化后的UPLC-MS/MS,在多反应监测模式下测定,同位素内标法定量,筛选覆盖牛奶中真菌毒素污染种类全面、回收率和稳定性最佳的免疫亲和柱,并进行免疫亲和柱性能评价和方法学验证,最终将方法应用于实际样品检测。结果 免疫亲和柱A对牛奶中16种真菌毒素在低、中、高3个添加水平均具有良好的准确度和精密度,加标回收率在83.6%～126.8%之间,相对标准偏差为0.2%～18.4%。柱A内各毒素残留水平低于仪器检出限,柱容量在21.8～1317.5 ng之间。使用免疫亲和柱A富集净化样品,各目标毒素在线性范围内线性良好,相关系数均大于0.99,定量限为0.0010～0.2000ng/g。应用该方法对牛奶质...     

Ochratoxin A in dried vine fruits on the Canadian retail market

Between 1998 and 2000, 151 samples of raisins and sultanas and two samples of currants were collected from retail outlets across Canada and analysed for ochratoxin A. Samples were extracted with methanol-sodium bicarbonate, and the extracts were cleaned-up by immunoaffinity column chromatography. Ochratoxin A was quantified by liquid chromatography with fluorescence detection. The minimum quantifiable level was 0.1 ng g^-1. Ochratoxin A was present, above the minimum quantifiable level, in 67 (79%) of 85 samples of raisins, in 39 (59%) of 66 samples of sultanas, and in both samples of currants. The overall mean level of ochratoxin A was 1.8 ng g^-1 in both the raisins and sultanas, and 2.8 ng g^-1 in the currants.     

赭曲霉毒素A分析方法进展

赭曲霉毒素A(OchratoxinA,OTA)是曲霉属和青霉属的某些菌种产生的一种具有致癌、致畸、免疫抑制和肝肾毒性的有毒代谢产物,世界上许多国家已制定了食品中OTA的相关法规和标准,而准确、可靠、灵敏的分析OTA方法是法规标准实施的重要依据。有机溶剂与酸(或碳酸氢钠)的混合溶液是提取食品中OTA的常用溶剂系统;含有OTA提取液的净化手段包括液液分配、固相柱萃取和免疫亲和层析等;反相液相色谱配荧光检测器、酶联免疫吸附法和液相色谱质谱联机是分析OTA的常用方法,本文就OTA的检测方法进行综述。     

Aflatoxins in nuts assayed by immunological methods

 Different kinds of raw and processed nuts available in the local retail market were investigated for aflatoxin content. Total aflatoxins and aflatoxin B₁ content were determined by immunoaffinity chromatography with fluorescence detection after reaction with bromine solution and by immunoenzymatic test kits. Of 29 investigated samples, 38% were contaminated. The total aflatoxin content in contaminated samples was between 1.20 μg/kg for peanuts and 5.50 μg/kg for walnuts. The concentration of aflatoxin B₁ found in contaminated samples was between 0.35 μg/kg for cashew nuts and 4.04 μg/kg for walnuts. The mean recovery of total aflatoxins was 95% for the Ridascreen test and 92% for immunoaffinity chromatography with fluorescence detection. For aflatoxin B₁ the mean recovery was 84%. Received: 4 March 1999 / Revised version: 17 May 1999     

免疫亲和净化-液相色谱-串联质谱法测定贝类中腹泻性贝类毒素

建立贝类中腹泻性贝类毒素的免疫亲和净化-液相色谱-串联质谱分析方法。样品采用80%甲醇溶液提取,选择磷酸盐缓冲液与提取液(4∶1,V/V)混合稀释后,免疫亲和选择专一净化,液相色谱-串联质谱分析。根据腹泻性免疫亲和柱的使用特性,对上样液、淋洗液、洗脱液等参数进行优化。质谱采用电喷雾负离子电离,多反应监测模式,外标法定量。3种分析物在1.0~100μg/L质量浓度范围内线性相关系数均大于0.996,对应的检出限和定量限均为0.3μg/kg和1.0μg/kg,平均回收率为82.7%~94.3%,相对标准偏差为0.70%~7.61%。本方法基质干扰小、净化效果强、灵敏度高,适合贝类中腹泻性贝类毒素的分析测定。