【摘要】：目的：建立食品中交鏈孢菌酮酸(TeA)、交鏈孢酚(AOH)、交鏈孢酚單甲醚(AME)和騰毒素(TEN)的UPLC-MS/MS協(xié)同測(cè)定方法,并利用所建方法對(duì)國產(chǎn)小麥粉及其制品、番茄及其制品、柑橘及其制品中的4種交鏈孢毒素污染狀況進(jìn)行研究；體外建模,探討食品中交鏈孢毒素經(jīng)口的生物可及性和生物利用率；利用毒理學(xué)關(guān)注閾值(TTC)方法,對(duì)我國居民通過小麥及其制品暴露TeA、AOH、AME和TEN對(duì)其健康的影響風(fēng)險(xiǎn)進(jìn)行評(píng)估。方法：(1)通過對(duì)質(zhì)譜、色譜條件條件的優(yōu)化,確定各種方法學(xué)參數(shù),建立協(xié)同檢測(cè)食品中TeA、AOH、TEN和AME的UPLC-MS/MS方法。(2)利用所建立的方法對(duì)采自我國河南、山東、安徽、吉林和北京5個(gè)省(市)的323份小麥粉及其制品、70份番茄及其制品和86份柑橘及其制品中的4種交鏈孢毒素污染水平進(jìn)行測(cè)定。(3)體外條件下,模擬胃腸道生理消化過程,構(gòu)建小麥粉制品中交鏈孢毒素在口腔、胃和小腸中生物可及性評(píng)價(jià)模型,并用真菌毒素吸附劑對(duì)模型進(jìn)行驗(yàn)證；以人克隆結(jié)腸腺癌細(xì)胞(Caco-2)單層細(xì)胞系為工具,構(gòu)建Caco-2細(xì)胞轉(zhuǎn)運(yùn)TeA、AOH和AME 3種毒素的腸轉(zhuǎn)運(yùn)體外模型并進(jìn)行了驗(yàn)證,結(jié)合3種交鏈孢毒素的經(jīng)口生物可及性,對(duì)3種交鏈孢毒素的生物利用率進(jìn)行研究。(4)運(yùn)用TTC方法對(duì)我國居民通過小麥粉及其制品暴露4種交鏈孢毒素對(duì)其健康的影響風(fēng)險(xiǎn)進(jìn)行評(píng)估,結(jié)合研究獲得的生物利用數(shù)據(jù)對(duì)其膳食內(nèi)暴露量進(jìn)行初步評(píng)估。結(jié)果：(1)建立的UPLC-MS/MS檢測(cè)四種交鏈孢毒素方法的檢出限在0.1-8 μg/kg之間,加標(biāo)回收率在58.9-116.0%之間,等同或高于國際同類方法的回收率,相對(duì)標(biāo)準(zhǔn)偏差(RSD)在3.0-12.4%之間,RSD低于20%,可滿足實(shí)際檢測(cè)要求。(2)TeA為污染三類食品的主要交鏈孢毒素,在小麥粉中的檢出率為99.4%,小麥粉中TeA污染水平范圍為1.76-520 μg/kg(均值：88.4μg/kg)。小麥粉中AOH、 AME和TEN的檢出率分別為6.1%、91.2%和97.2%,污染水平范圍分別為16.0-98.7 μg/kg(均值：30.2 μg/kg)、0.320-61.8 μg/kg(均值：3.77 μg/kg)和2.25-129 μg/kg(均值：27.1 μg/kg)。小麥粉中TeA的污染水平存在地域差異,含量由高到低依次為：河南、安徽、北京、山東、吉林。小麥粉及其制品樣品中四種毒素含量呈正相關(guān),分別為：AOH與AME(r=0.904, P0.05)、TeA與AME(r=0.757, P0.01)、TeA與AOH+AME (r=0.860, P0.01)、TEN與TeA(r=0.747,P0.01。番茄醬和番茄汁中TeA的檢出率均為100%,含量范圍分別為10.2-1787 μg/kg均值：338μg/kg)和7.4-278 μg/kg(均值：87 μg/kg)。進(jìn)口番茄醬樣品中TeA含量顯著高于國產(chǎn)樣品(P0.05)。番茄醬中AOH和AME的含量范圍分別為2.5-300μg/kg(均值：80 μg/kg)和0.32-38 μg/kg(均值：8.0 μg/kg); TEN的含量范圍1.53-15.8 μg/kg(均值：4.9 μg/kg)。番茄汁中AME含量范圍為0.20-5.8 μg/kg (均值：1.42μg/kg)。柑橘汁樣品中TeA檢出率為25.0%,污染水平范圍為1.21-4.3μg/kg(均值：1.80 μg/kg),其他毒素均未檢出。(3)建立了評(píng)價(jià)交鏈孢毒素生物可及性和生物利用率體外消化模型。饅頭樣品中TeA、AOH和AME的經(jīng)口生物可及性分別為72.1±5.4%、7.0±0.8%和10.2±1.0%。成功構(gòu)建了體外評(píng)價(jià)交鏈孢毒素腸轉(zhuǎn)運(yùn)的Caco-2細(xì)胞膜型。AOH、AME和TeA在不同濃度和轉(zhuǎn)運(yùn)時(shí)間的吸收率范圍為1.5%和63.0%,TeA為易于吸收的毒素。(4)全人群和各年齡組人群通過小麥及其制品暴露AOH和AME量的均值和P97.5位數(shù)均超過TTC值,其中以2-6歲人群的暴露量最高,其次是7-17歲年齡組人群；全人群和各年齡組人群中高食物量消費(fèi)者AOH的內(nèi)暴露上限(UB)也超過TTC,應(yīng)引起關(guān)注。全人群和各年齡組人群通過小麥及其制品暴露TeA和TEN的均值和P97.5位數(shù)均低于TTC值,對(duì)健康影響的風(fēng)險(xiǎn)較低。結(jié)論：建立的協(xié)同檢測(cè)4種交鏈孢毒素的方法靈敏、特異,可滿足不同基質(zhì)中4種毒素分析需要；我國小麥及其制品、番茄制品和柑橘制品被交鏈孢毒素污染,以TeA污染最重；構(gòu)建的生物可及性體外消化模型穩(wěn)定、重復(fù)性好,完全可以滿足對(duì)食品中各類交鏈孢毒素生物可及性的研究；所建3種毒素Caco-2單層細(xì)胞模型穩(wěn)定,可用于3種毒素體外生物利用率研究中。我國居民高通過小麥及其制品中暴露AOH和AME對(duì)其健康影響的風(fēng)險(xiǎn)較高,應(yīng)引起關(guān)注。在充分獲取數(shù)據(jù)的基礎(chǔ)上,有必要對(duì)污染水平和污染率較高的TeA做進(jìn)一步的評(píng)估。
[Abstract]:OBJECTIVE: To establish a UPLC-MS/MS method for the simultaneous determination of alternosporin ketoacid (TeA), alternosporin (AOH), alternosporin monomethyl ether (AME) and tenotoxin (TEN) in food, and to study the contamination status of four kinds of alternosporin in wheat flour and its products, tomato and its products, citrus and its products. To investigate the oral bioavailability and bioavailability of Alternaria toxins in foods, and to assess the health risk of TeA, AOH, AME and TEN exposure to wheat and its products by toxicological concern threshold (TTC) method. A UPLC-MS/MS method for the simultaneous determination of TeA, AOH, TEN and AME in food was developed. (2) The contamination levels of 4 cross-linked sporotoxins in 323 wheat flour and its products, 70 tomatoes and their products and 86 citrus and their products from Henan, Shandong, Anhui, Jilin and Beijing provinces (cities) were determined in vitro. The bioavailability of Alternaria toxin in wheat flour products in oral, stomach and small intestine was evaluated by simulating gastrointestinal physiological digestion process, and the model was validated by mycotoxin adsorbent. Human colon adenocarcinoma cell (Caco-2) monolayer cell line was used to construct the intestine of Caco-2 cells transporting TeA, AOH and AME toxins. The in vitro transport model was validated, and the bioavailability of three Alternaria toxins was studied in combination with the bioavailability of three Alternaria toxins. (4) The risk of exposure to four Alternaria toxins from wheat flour and its products was assessed by TTC method, and the bioavailability of the three Alternaria toxins was also evaluated. Results: (1) The detection limits of the four methods were 0.1-8 ug/kg, and the recovery rates were 58.9-116.0%. The recoveries were equivalent to or higher than those of the international methods. The RSD ranged from 3.0-12.4%, and RSD was lower than 20%. (2) TeA was the main cross-linked spore toxin in three kinds of food. The detection rate of TeA in wheat flour was 99.4%. The contamination range of TeA in wheat flour was 1.76-520 ug/kg (mean: 88.4 ug/kg). The detection rates of AOH, AME and TEN in wheat flour were 6.1%, 91.2% and 97.2% respectively, and the contamination range was 16.0-98.7 ug/kg (mean: 30.2%). The contamination levels of TeA in wheat flour varied from high to low in order: Henan, Anhui, Beijing, Shandong, Jilin. The contents of four toxins in wheat flour and its products were positively correlated, respectively: AOH and AME (r = 0.904, P 0.05). TeA and AME (r = 0.757, P 0.01), TeA and AOH + AME (r = 0.860, P 0.01), TEN and TeA (r = 0.747, P 0.01). The detection rates of TeA in tomato sauce and tomato juice were 100%, and the contents ranged from 10.2 to 1787 ug/kg with an average value of 338 ug/kg and 7.4 to 278 ug/kg with an average value of 87 ug/kg. The TeA content in imported tomato sauce samples was significantly higher than that in domestic samples (P 0.05). The contents of AOH and AME in sauce ranged from 2.5 to 300 ug/kg (mean: 80 ug/kg) and 0.32 to 38 ug/kg (mean: 8.0 ug/kg), TEN ranged from 1.53 to 15.8 ug/kg (mean: 4.9 ug/kg), AME ranged from 0.20 to 5.8 ug/kg (mean: 1.42 ug/kg) in tomato juice, TeA detection rate in citrus juice was 25.0%, and pollution level ranged from 1.2%. 1-4.3 ug/kg (mean: 1.80 ug/kg) and no other toxins were detected. (3) An in vitro digestion model was established to evaluate the bioavailability and bioavailability of Alternaria toxin. The oral bioavailability of TeA, AOH and AME in steamed bread samples were 72.1 (+ 5.4%), 7.0 (+ 0.8%) and 10.2 (+ 1.0%) respectively. The absorption rates of AOH, AME and TeA were 1.5% and 63.0% at different concentrations and transport times, and TeA was easily absorbed toxin. (4) The mean and P97.5 digits of AOH and AME exposures to wheat and its products exceeded TTC values in the whole population and all age groups, with the highest exposure in the 2-6 year-old group, followed by the 7-17 year-old group. The upper limit of internal exposure (UB) of AOH in high-food-intake consumers was also higher than that of TTC in the whole population and all age groups. The mean and P97.5 digits of TEA and TEN exposure to wheat and its products were lower than that of TTC in the whole population and all age groups, and the risk of health effects was lower. The method is sensitive and specific, and can meet the needs of four toxins analysis in different substrates; wheat and its products, tomato products and citrus products are polluted by Alternaria toxin, TeA pollution is the most serious; the bioavailability in vitro digestion model is stable and reproducible, and it can completely meet the requirements of Alternaria toxin bioavailability in food. The single-layer cell model of three toxins Caco-2 is stable and can be used to study the bioavailability of the three toxins in vitro. TeA for further evaluation.
【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R155.5

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 周兵;強(qiáng)勝;;鏈格孢菌毒素細(xì)交鏈孢菌酮酸對(duì)蠶豆根尖細(xì)胞微核和有絲分裂的影響[J];應(yīng)用與環(huán)境生物學(xué)報(bào);2007年06期

2 周啟霆，徐儀寶，康愛麗，白東魯;交鏈孢醇和交鏈孢醇單甲醚的制備[J];中國藥物化學(xué)雜志;1996年01期

3 周良模;黃明賢;王光華;朱道乾;;極性PEG20M交鏈柱的研制[J];分析化學(xué);1987年06期

4 顧侃英;王昌時(shí);陸婉珍;;交鏈SE-54彈性石英開管柱[J];分析化學(xué);1985年02期

5 李樹正;岳東霞;曾強(qiáng);張素華;楊昭;張志銘;;若干種化合物鈍化交鏈孢酸的研究[J];植物保護(hù)學(xué)報(bào);1997年03期

6 石聞華,苗健,婁振嶺,裴留城,甄應(yīng)中,劉桂亭;應(yīng)用薄層層析法檢測(cè)互隔交鏈孢C_(12)-b_3主要代謝產(chǎn)物的分析結(jié)果報(bào)告(摘要)[J];河南醫(yī)科大學(xué)學(xué)報(bào);1987年03期

7 陳真光，李琦，李金祥;直接交鏈染料的應(yīng)用研究[J];紡織導(dǎo)報(bào);1996年05期

8 楊星星;劉細(xì)霞;王弘;徐振林;沈玉棟;孫遠(yuǎn)明;;細(xì)交鏈孢菌酮酸酶聯(lián)免疫吸附分析方法研究[J];分析化學(xué);2012年09期

9 黃宏度;顧文娟;閔恩澤;;幾種交鏈蒙脫石的裂化活性與水蒸汽穩(wěn)定性[J];石油煉制與化工;1983年07期

10 董志芳;陸瓊;;新型交鏈染色體系的研究[J];染料工業(yè);1987年02期

相關(guān)會(huì)議論文 前1條

1 李蓉;陳同強(qiáng);劉志剛;;交鏈孢毒抗原成份的分析[A];第一屆全國變態(tài)反應(yīng)學(xué)術(shù)研討會(huì)論文匯編[C];2001年

相關(guān)博士學(xué)位論文 前2條

1 趙凱;食品中交鏈孢毒素污染、生物利用及風(fēng)險(xiǎn)評(píng)估研究[D];中國疾病預(yù)防控制中心;2015年

2 趙利輝;交鏈孢菌激活蛋白激發(fā)水稻的應(yīng)答基因與抗病性分析[D];中國農(nóng)業(yè)科學(xué)院;2006年

相關(guān)碩士學(xué)位論文 前2條

1 崔娜;細(xì)交鏈孢菌的抗藥性分化研究[D];廣西大學(xué);2004年

2 張寧;交鏈孢菌cDNA文庫構(gòu)建及激活蛋白基因克隆與融合表達(dá)[D];中國農(nóng)業(yè)科學(xué)院;2006年

，

本文編號(hào)：2216839