高效液相色谱法同时检测粮食中常见8 种真菌毒素的含量

建立免疫亲和柱净化-高效液相色谱法同时测定粮食中黄曲霉毒素B1（aflatoxins,AFB1）、黄曲霉毒素B2（AFB2）、黄曲霉毒素G1（AFG1）、黄曲霉毒素G2（AFG2）、赭曲霉毒素A（ochratoxin A,OTA）、玉米赤霉烯酮（zearalenone,ZEN）、呕吐毒素（deoxynivalenol,DON）和T-2毒素的检测方法。样品经乙腈-水提取后,用免疫亲和柱净化,Agilent Elipse Plus C18（100 mm×4 mm,3.5 μm）色谱柱分离,以甲醇-乙腈-水-乙酸为流动相,流速1 mL/min,柱温35 ℃,进样量20 μL,检测系统为可变波长检测器串联光化学衍生器串联荧光检测器。根据信噪比为3的峰响应值,确定各真菌毒素的检出限为：AFB1 0.446 ng/mL、AFB2 0.152 ng/mL、AFG1 0.523 ng/mL、AFG2 0.334 ng/mL、ZEN 7 ng/mL、OTA 0.7 ng/mL、DON 200 ng/mL、T-2毒素100 ng/mL。样品中各真菌毒素的平均加标回收率,玉米为80.0%～104.5%,小麦为83.2%～102.8%,方法精密度为2.6%～10.2%。从样品前处理到分析整个过程耗时约2 h。本方法简便、快速、灵敏度高,适用于粮食中多种真菌毒素的快速测定。     

3种真菌毒素(ZEN、DON及T2)完全抗原的合成与鉴定

目的:合成玉米赤霉烯酮(ZEN)、脱氧雪腐镰刀菌烯醇(DON)和T2 3种真菌毒素的完全抗原。方法:将ZEN半抗原、DON及T2与阳离子化的载体蛋白(BSA和OVA)偶联,制备高特异性的完全抗原。用高性能基质辅助激光解吸电离-飞行时间质谱仪(MALDI-TOF)检测偶联物及载体蛋白的分子质量,用商品化的ZEN、DON和T2单克隆抗体对偶联物进行免疫滴定验证。结果:平均每个BSA偶联上的ZEN、DON和T2的分子个数分别为20.81,6.03,4.92个,平均每个OVA偶联上的ZEN、DON和T2分子个数分别为7.17,3.05,2.80个,偶联物与ZEN、DON及T2的抗体呈阳性反应。结论:合成的ZEN、DON及T2完全抗原为3种毒素抗体的制备及免疫学方法的建立奠定了基础。     

基于噬菌体展示纳米抗体的绿色免疫PCR检测脱氧雪腐镰刀菌烯醇

目的：在获得脱氧雪腐镰刀菌烯醇（deoxynivalenol,DON）抗独特型纳米抗体的前期基础上,将纳米抗体作为酶标抗原的替代物,应用于荧光定量免疫聚合酶链式反应（polymerase chain reaction,PCR）体系,实现DON的高灵敏、绿色免疫分析。方法：将特异性结合DON抗体的噬菌体展示纳米抗体（P-28）作为竞争抗原,以编码P-28纳米抗体的DNA为靶标,设计特异性PCR扩增引物,优化荧光定量免疫PCR退火温度、抗DON抗体浓度、P-28投入量等参数,建立基于间接竞争模式的荧光定量免疫PCR检测DON的方法。结果：本研究建立的荧光定量免疫PCR方法线性检测范围为0.1～1 000 ng/mL,IC50值为（3.96±2.21）ng/mL,最低检出限为0.048 ng/mL,并与其他真菌毒素无交叉反应。结论：该方法直接使用噬菌体展示纳米抗体作为竞争抗原的替代物,应用于免疫PCR体系,避免了使用传统的化学合成酶标抗原所带来的环境污染、操作毒性等缺陷,并具有良好的特异性及灵敏度。     

北京地区部分市售食用植物油中玉米赤霉烯酮和脱氧雪腐镰刀菌烯醇的污染状况分析

目的分析北京市售植物油中玉米赤霉烯酮和脱氧雪腐镰刀菌烯醇的污染状况。方法实验采用同位素稀释-超高效液相色谱-串联质谱法,在多反应监测模式下对所采集玉米油、花生油、调和油等食用油样中的玉米赤霉烯酮(ZEN)、脱氧雪腐镰刀菌烯醇(DON)进行了检测,并对其污染现状进行分析。结果 ZEN和DON在3~500 ng/mL浓度范围内线性良好,相关系数为0.999,加标回收率为95.05%~107.10%,ZEN和DON方法的检出限分别为0.03μg/kg和0.9μg/kg;ZEN和DON方法的定量限分别为0.1μg/kg和3μg/kg。对北京市售30个油样检测结果表明,DON为未检出,ZEN检出率为100%,最高含量为333μg/kg,最低含量为1.95μg/kg,平均含量为67.7μg/kg,低于欧盟规定的限量标准400μg/kg。结论通过分析食用油中真菌毒素的含量状况,初步了解了北京市售植物油中玉米赤霉烯酮和脱氧雪腐镰刀碱烯醇的污染状况。     

免疫快速同步检测玉米中五种真菌毒素胶体金试纸条的构建和应用

玉米作为重要的粮食和饲料来源，容易受到多种真菌毒素的污染，给居民健康和养殖业造成严重的危害。本文优化了适合五种真菌毒素（黄曲霉毒素B1、伏马毒素、T-2毒素、脱氧雪腐镰刀菌烯醇和玉米赤霉烯酮）同时提取的样品前处理方法，并构建了五检测线胶体金试纸条，检测结果可以通过肉眼定性，结合Image J软件可以进行定量分析，并应用于实际玉米样品检测。结果表明，最佳提取条件为90%乙腈/水，涡旋20 min，样品的添加回收率为71.9%-113.3%，相对标准偏差为0.9%-7.5%；测定基质添加样品建立标准曲线，通过肉眼判定得到五种真菌毒素的消线值分别为80.0、1000.0、100.0、400.0和200.0 μg/kg；结合Image J 软件提取多重检测试纸条的检测线灰度值，结果表明当AFB1、FB1、T-2、DON和ZEN的浓度为8.0、20.0、10.0、40.0、20.0 μg/kg时，各真菌毒素浓度对应的检测线显色值比空白对照显色值低。最后，通过检测11个实际玉米样品，结果表明部分玉米样品受到了不同程度的AFB1、DON和ZEN的污染。因此，本研究开发的免疫快速同步检测玉米中五种真菌毒素胶体金试纸条可以用于玉米中真菌毒素的现场快速筛查。     

电子束辐照降解玉米中玉米赤霉烯酮和呕吐毒素

利用电子束辐照(electron beam irradiation,EBI)降解玉米赤霉烯酮(zearalenone,ZEN)和呕吐毒素(deoxynivalenol,DON),考察EBI对单一及混合毒素的降解效果,分析毒素降解及相互作用规律。结果表明,在0～10 k Gy范围内,随着剂量的增加,标准品溶液中单一毒素降解率升高,且毒素初始浓度越低,降解率越高。10k Gy时,1μg/m L的ZEN和DON降解率分别可达76. 04%和89. 31%。以EBI处理单一毒素为对照,DON与ZEN混合后同时经EBI处理,标准品和玉米浆中ZEN降解率均呈显著降低,DON降解率无明显趋势变化。14. 20%(质量分数)水分含量的玉米粉中混合毒素经EBI处理后,ZEN和DON降解率均低于单一毒素对照组;水分含量为19. 80%(质量分数)时,ZEN降解率低于对照组而DON高于对照组。该研究阐明了EBI同时降解玉米中ZEN和DON的效果,为EBI在粮食真菌毒素降解中的应用提供了理论参考。     

淡碱蒸胚对玉米胚及其毛油中真菌毒素的降解消除作用

利用湿热和碱性条件对真菌毒素的破坏作用,以不同质量分数淡碱(NaOH)溶液调节玉米胚达到适宜蒸胚水分质量分数(14%),用115℃过热蒸汽蒸胚30 min,对蒸胚前后玉米胚及所制取玉米毛油中黄曲霉毒素B_1(aflatoxin,AFB_1)、玉米赤霉烯酮(zearalenone,ZEN)和呕吐毒素(deoxynivalenol,DON)含量进行检测,研究淡碱蒸胚对玉米胚及其毛油中真菌毒素的降解消除效果及碱液质量分数对降解消除效果的影响。结果表明:碱液质量分数为5.29%时,淡碱蒸胚对玉米胚中ZEN和DON的降解消除效果最好,ZEN含量从697.41μg/kg降至246.25μg/kg(降解率64.69%),DON含量由2 417.07μg/kg降至802.83μg/kg(降解率66.78%),所制取玉米毛油中ZEN含量从266.44μg/kg减少至140.02μg/kg,DON含量从150.76μg/kg降低至108.47μg/kg;碱液质量分数为6.55%时,淡碱蒸胚对玉米胚中AFB_1的降解消除效果最好,AFB_1含量从7.91μg/kg降至0.77μg/kg(降解率90.30%),所制取玉米毛油中AFB_1含量从2.11μg/kg减少至0.51μg/kg。随碱液质量分数的增大,淡碱蒸胚所得玉米胚及其毛油的色泽有所加深,毛油酸价和过氧化值明显降低。采用淡碱蒸胚不仅能对玉米胚中真菌毒素进行有效降解,大幅降低玉米胚及其毛油中真菌毒素含量,同时也能降低玉米粕中真菌毒素含量,实现玉米油食用安全和玉米粕饲用安全的同步提升。     

固相萃取柱净化-液相色谱-串联质谱法测定糕点中脱氧雪腐镰刀菌烯醇及其衍生物和玉米赤霉烯酮

建立使用超高效液相色谱-串联质谱仪测定糕点中玉米赤霉烯酮(ZEN)、脱氧雪腐镰刀菌烯醇(DON)及其衍生物3-乙酰基-脱氧雪腐镰刀菌烯醇(3-Ac DON)、15-乙酰基-脱氧雪腐镰刀菌烯醇(15-Ac DON)4种真菌毒素的方法。该方法在4 min内完成4种真菌毒素的分析,线性范围5~1000μg/kg,标准曲线的相关系数均在0.997以上,DON、ZEN的最低检出限为1.0μg/kg,3-Ac DON、15-Ac DON的最低检出限为1.5μg/kg。通过对134份市售预包装糕点样品进行分析发现,4种真菌毒素均有一定检出,其中ZEN、DON、3-Ac DON和15-Ac DON的检出率分别为2.2%、78.4%、3.0%和5.2%,所有样品中4种真菌毒素含量水平均未超过食品安全国家标准限量要求。     

时间分辨荧光免疫层析法定量检测谷物中黄曲霉毒素B1和玉米赤霉烯酮残留

建立了一种快速定量检测谷物产品中黄曲霉毒素(Aflatoxin B1,AFB1)和玉米赤霉烯酮(Zearalenone,ZEN)的时间分辨荧光免疫层析方法。采用时间分辨荧光微球标记黄曲霉毒素B1抗体和玉米赤霉烯酮抗体,研究了如p H值、标记抗体浓度、荧光探针使用量、检测T线包被原浓度、质控C线羊抗鼠Ig G浓度、样品前处理方法等因素对时间分辨荧光免疫层析方法灵敏度的影响。结果表明:AFB1的检出限为0.80 ng/mL,线性范围(IC20~IC80)为0.81~5.67 ng/mL,半抑制浓度(IC50)为2.15 ng/mL。在ZEN检出限为4.58 ng/mL,线性范围(IC20~IC80)为4.76~85.60 ng/mL,半抑制浓度(IC50)为20.19 ng/mL。方法特异性良好,与T-2毒素、脱氧雪腐镰刀菌烯醇、伏马毒素、赭曲霉毒素A多种真菌毒素交叉率小于10%。通过选择玉米、麦麸、大豆、小麦进行添加回收试验,AFB1的添加回收率在97.1%~108.7%之间,ZEN的添加回收率在92.8%~109.1%之间,相对标准偏差小于15%。选取经HPLC-MS/MS检测过的FAPAS标准质控样本进行测试,检测结果与其结果一致。在实际产品检测对比中,与市售胶体金免疫层析卡,ELISA试剂盒的检测结果基本一致。本方法操作简单快速、可定量,检测过程约25 min,适用于谷物样品中黄曲霉毒素B1和玉米赤霉烯酮的现场快速筛。     

磁控双色上转换荧光适配体传感器同时检测玉米和燕麦粉中的赭曲霉毒素A与玉米赤霉烯酮

目的 建立上转换荧光法同时检测食品中赭曲霉毒素A（ochratoxin A,OTA）与玉米赤霉烯酮（zearalenone,ZEN）的含量 方法 利用溶剂热法合成了两种油酸封端的核壳型上转换纳米材料,通过表面改性法和戊二醛法制备了表面分别偶联OTA、ZEN适配体的核壳型上转换荧光探针。同时制备了表面原位生长四氧化三铁纳米颗粒的二硫化钼纳米片,作为淬灭剂。OTA、ZEN和适配体特异性结合后,通过磁分离后检测溶液的荧光强度值,从而实现OTA和ZEN浓度的检测。结果 该方法在最佳检测条件下,OTA与ZEN的浓度在0.05~500.00 ng/mL的线性检测范围内,与两种上转换荧光探针的荧光强度的对数值呈良好的线性关系,相关系数分别为0.9949和0.9972,对OTA的检测限为3.97×10-2 ng/mL,对ZEN的检测限为3.11×10-2 ng/mL,应用于玉米粉和燕麦粉中OTA和ZEN的检测,加标回收率为91.7%~109.4%。结论 该方法成功检测灵敏度较高,并具有较好的特异性,可用于食品中OTA和ZEN的高灵敏检测。     

Natural occurrence of Fusarium mycotoxins of the 1990 barley crop in Korea.

During the barley harvest in June 1990, there was a great deal of rainfall and high humidity in the southern part of Korea, and natural occurrence of Fusarium mycotoxins was suspected in barley samples. The samples of undergrade barley were obtained from four provinces and analysed for the presence of deoxynivalenol (DON) and nivalenol (NIV) by gas chromatography and zearalenone (ZEN) by high performance liquid chromatography. Of 37 samples, 33, 37 and 10 were positive for DON, NIV and ZEN, respectively. The husked barley contained 29-677 ng/g for DON, 114-1546 ng/g for NIV and 183-1416 ng/g for ZEN. The naked barley contained 38-645 ng/g for DON, 85-4569 ng/g for NIV and 40-1081 ng/g for ZEN. The average concentration of NIV in naked barley was higher than that in husked barley, but the average concentration of DON in husked barley was higher than that in naked barley. The survey indicates that the 1990 barley crop in Korea was heavily contaminated with Fusarium mycotoxins.     

超导体包被免疫荧光试纸法同时测定玉米中脱氧雪腐镰刀菌烯醇和玉米赤霉烯酮适用性研究

为了能同时快速测定玉米中脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)和玉米赤霉烯酮(Zearalenone,ZEN)两种毒素,采用基于超导体包被的免疫荧光快速定量检测技术,检测了多个玉米样品,结合加标回收率、稳定性、检出限、精密度等指标,并与液相色谱法的检测结果进行比较分析,评估方法的适用性。结果表明,超导体包被的免疫荧光试纸法检测DON含量在100～2 500μg/kg,检测ZEN含量在5～200μg/kg的范围内线性良好。DON在500～2 000μg/kg添加水平范围内,回收率为103.72%～108.17%,ZEN在50～150μg/kg添加水平范围内,回收率为97.81%～111.27%。该方法于液相色谱法检测结果对比,DON相对偏差在3.72%～8.17%,ZEN相对偏差在2.19%～11.27%,均低于液相色谱法允许偏差23%和15%,超导体包被的免疫荧光试纸法是一种有效、实用、快速、定量的分析方法,能满足同时检测玉米中DON和ZEN的要求。     

Simultaneous detection of deoxynivalenol and zearalenone by dual-label time-resolved fluorescence immunoassay

BACKGROUND: The health risks of deoxynivalenol (DON) and zearalenone (ZEN) necessitate the development of analytical methods for widespread food and feed screening. We sought to establish a rapid, economic and sensitive dual‐label time‐resolved fluoroimmunoassay (TRFIA) to detect DON and ZEN simultaneously. Eu³⁺‐ and Sm³⁺‐labelled antibodies were used, as lanthanides are more stable and have narrower emission spectra than most fluorescent dyes. RESULTS: The limit of detection was 0.0194 ng mL^?1 for DON (range: 0.0194–100 ng mL^?1) and 0.37 ng mL^?1 for ZEN (range: 0.37–50 ng mL^?1). DON recovery in spiked cereal samples was 88–107%, and for ZEN was 83–108%, with both intra‐ and inter‐assay coefficients of variation (CVs) less than 5%. The dual‐label TRFIA results correlated well with ELISA results (correlation coefficients: 0.9733 for DON and 0.9784 for ZEN). CONCLUSION: The dual‐label TRFIA is a simple, fast and sensitive method for high‐throughput screening of DON and ZEN in food and feedstuff. Copyright © 2010 Society of Chemical Industry     

Co-occurrence of mycotoxins in maize and maize-derived food in China and estimation of dietary intake

ABSTRACT

A total of 576 samples marketed in China, including maize, maize flour, maize grits and maize meal, was determined for the simultaneous presence of 12 mycotoxins (FB₁, FB₂, FB₃, DON, 3-DON, 15-DON, ZEN, AFB₁, AFB₂, AFG₁, AFG₂ and OTA) using a validated UPLC-MS/MS for multi-mycotoxin method. DON were the most widespread mycotoxins (63%), followed by FB₁ (57%) and ZEN (46%). 78% of the samples was contaminated with at least one of these mycotoxins. Risk assessment indicated that maize and maize-derived food intake does not pose a potential risk for general adult population with respect to individual mycotoxin. However, two or more mycotoxins were detected in 60% of all samples, and a combination of up to seven different mycotoxins was found. A particular attention should be paid to the combined exposure of mycotoxins, in this cases the estimated daily intake might increase greatly due to the high frequency of co-occurrence.     

Development and application of analytical methods for the determination of mycotoxins in organic and conventional wheat.

The aim of this study was to develop a multicomponent analytical method for the determination of deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEN), nivalenol (NIV), 3-acetyl-DON (3-acDON), 15-acetyl-DON (15-acDON), zearalenol (ZOL) and citrinin (CIT) in wheat. It also aimed to survey the presence and amounts of DON, OTA and ZEN in Belgian conventionally and organically produced wheat grain and in wholemeal wheat flours. After solvent extraction, an anion-exchange column (SAX) was used to fix the acidic mycotoxins (OTA, CIT), whilst the neutral mycotoxins flowing through the SAX column were further purified by filtration on a MycoSep cartridge. OTA and CIT were then analysed by high-performance liquid chromatography (HPLC) using an isocratic flow and fluorescence detection, while the neutral mycotoxins were separated by a linear gradient and detected by double-mode (ultraviolet light fluorescence) detection. The average DON, ZEN and OTA recovery rates from spiked blank wheat flour were 92, 83 and 73% (RSDR = 12, 10 and 9%), respectively. Moreover, this method offered the respective detection limits of 50, 1.5 and 0.05 microg kg-1 and good agreement with reference methods and inter-laboratory comparison exercises. Organic and conventional wheat samples harvested in 2002 and 2003 in Belgium were analysed for DON, OTA and ZEN, while wholemeal wheat flour samples were taken from Belgian retail shops and analysed for OTA and DON. Conventional wheat tended to be more frequently contaminated with DON and ZEN than organic samples, the difference being more significant for ZEN in samples harvested in 2002. The mean OTA, DON and ZEA concentrations were 0.067, 675 and 75 microg kg-1 in conventional samples against 0.063, 285 and 19 microg kg-1 in organically produced wheat in 2002, respectively. Wheat samples collected in 2003 were less affected by DON and ZEN than the 2002 harvest. Organic wholemeal wheat flours were more frequently contaminated by OTA than conventional samples (p < 0.10). The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours (p < 0.10).     

Deoxynivalenol,zearalenone and T-2 in grain based swine feed in Hungary

Fusarium genera can produce trichothecenes like deoxynivalenol (DON), zearalenone (ZEN) and T-2 toxin, which can occur in feed cereal grains. Enzyme-linked immunosorbent assays (ELISA) tests of different Hungarian swine feedstuff proved that these mycotoxins were present. In this survey, 45 feed samples from 3 significant Hungarian swine feedstuff manufacturers were tested. ELISA methodology validation showed mean recovery rates in ranges from 85.3% to 98.1%, with intermediate precision of 86.9-96.9% and variation coefficients of 3.4–5.7% and 5.9–7.1%, respectively. The results showed that among Fusarium toxins, generally DON was present in the highest concentration, followed by T-2 and finally ZEN in all tested swine feeds. Each of the mycotoxins was found above the limit of detection in all swine feedstuffs. Boars feed’s DON (average ± standard deviation was 872 ± 139 µg kg^?1) and ZEN (172 ± 18 µg kg^?1) results of one of the manufacturers were above the guidance values. It indicates the necessity for efficient monitoring of DON, ZEN and T-2 mycotoxins in swine feeds.     

Fusarium mycotoxins in cereals harvested from Hungarian fields

The Fusarium mycotoxins deoxynivalenol (DON), zearalenone (ZEN) and T-2 frequently contaminate grain crops in Middle and Eastern Europe. In this survey, 116 cereal samples (maize, wheat, barley and oat) were examined for DON, ZEN and T-2 mycotoxins. Samples were collected from different areas in two Hungarian regions (North and South Transdanubia). The method of analysis was indirect competitive ELISA. Maize was the most contaminated grain regarding DON (86%), ZEN (41%) and T-2 (55%) toxins. The average results of the deoxynivalenol and zearalenone tests of maize proved to be significantly higher than those of barley or oat. DON was the most represented Fusarium mycotoxin followed by T-2 and ZEN. The examination of these mycotoxins would be necessary at a larger scale as to re-evaluate permissible levels, so increase of the monitoring programme would be advisable for the future.     

Development of a Simultaneous Lateral Flow Strip Test for the Rapid and Simple Detection of Deoxynivalenol and Zearalenone

The objective of this study was to develop a 1‐step simultaneous lateral flow strip test for the rapid and simple detection of deoxynivalenol (DON) and zearalenone (ZEA) in grains. Two monoclonal antibodies (MAbs) against DON and ZEA were respectively conjugated with gold nanoparticles and used to develop a lateral flow strip test for a single toxin and multiple toxins. First, individual lateral flow strips for a single toxin were optimized, and their conditions were used to develop a simultaneous lateral flow strip for multiple toxins. Limits of detection of both lateral flow strip tests for DON and ZEA were the same (DON: 50 ng/mL, ZEA: 1 ng/mL). Both methods showed cross‐reactivity for α‐zearalenol and β‐zearalenol, but no cross‐reaction to other mycotoxins. The results can be completed obtained within 15 min. The cut‐off values of the simultaneous lateral flow strip for the spiked rice and corn were 500 and 10 ng/g for DON and ZEA, respectively. The results demonstrated that the developed simultaneous lateral flow strip test offers a rapid, easy‐to‐use, and portable analytical system and can be used as a convenient qualitative tool for the on‐site detection of DON and ZEA in food and agricultural commodities.     

The coexistence of the Fusarium mycotoxins nivalenol, deoxynivalenol and zearalenone in Korean cereals harvested in 1983

A survey for the occurrence of nivalenol (NIV), deoxynivalenol (DON) and zearalenone (ZEN) in Korean cereals (totalling 53 samples) harvested in 1983, showed that 96%, 72% and 57% of the samples were contaminated with NIV, DON and ZEN, respectively. Average concentrations (micrograms/kg) in unpolished barley were 546 (NIV), 117 (DON) and 110 (ZEN), and those in polished barley were 130 (NIV) and 21 (DON). The ZEN levels were below the detection limit (1 microgram/kg). Malt, wheat and rye were also heavily contaminated with these Fusarium mycotoxins. The results of this survey show that Korean cereals harvested in 1983 were significantly contaminated with NIV, DON and ZEN, and the incidence and levels, where observed, are similar to those reported in Japan.     

呕吐毒素时间分辨荧光免疫层析法的建立与评价

构建并评价了呕吐毒素(Deoxynivalenol,DON)时间分辨荧光免疫层析法(TRFIA)。以Eu3+螯合物的纳米微球为荧光探针标记抗DON单抗,采用竞争抑制建立免疫层析定量方法,优化了微球与抗体标记量,检测的环境温度、反应时间、加样体积及缓冲体系;研究了方法的灵敏度、精密度及准确度。结果表明,每100 μL荧光微球结合纯单抗50 μg,最佳划膜浓度为1.0 mg/mL,最佳测定条件为室温(25±2)℃,10 min,加样体积100 μL,PBS (0.01 mol/L,pH7.2)的缓冲体系,方法的灵敏度为0.25 ng/mL,线性范围为0.5~25.0 ng/mL,玉米、小麦阴性样本(加标浓度200、500、1000、2000 μg/kg)平均加标回收率为91.4%~109.3%,6种典型样本经TRFIA与免疫亲和净化-高效液相色谱法(IAC-HPLC)同时测定DON含量,相关系数r达0.9754。TRFIA具有灵敏度高、速度快、定量准等技术特点,适用于谷物及制品中DON的快速定量。