本文選題：環(huán)境雌激素　切入點(diǎn)：重組基因酵母　出處：《華中科技大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：隨著工農(nóng)業(yè)的發(fā)展，環(huán)境雌激素在我們的生活中被廣泛應(yīng)用，經(jīng)研究發(fā)現(xiàn)，它們可以干擾人類和動(dòng)物的內(nèi)分泌系統(tǒng)，引起發(fā)育、行為和生殖等各方面的異常變化，這嚴(yán)重影響了人類的健康。食品是人類生存和發(fā)展的物質(zhì)基礎(chǔ)，是人類暴露于環(huán)境雌激素的主要途徑。因此，，開(kāi)展食品中環(huán)境雌激素的監(jiān)測(cè)，對(duì)于發(fā)現(xiàn)和消除對(duì)人類健康的潛在危害具有重要意義。 方法：本研究采用重組基因酵母篩檢法（YES）對(duì)中國(guó)四個(gè)城市居民經(jīng)常食用的16種142個(gè)動(dòng)物性食品的雌激素活性進(jìn)行評(píng)價(jià)，并結(jié)合高效液相色譜-串聯(lián)質(zhì)譜法（LC-MS/MS）分析樣品中常見(jiàn)8種環(huán)境雌激素的含量。 結(jié)果：主要內(nèi)容包括兩部分： 第一部分：動(dòng)物性食品中總的雌激素活性測(cè)定 本部分采用該方法對(duì)動(dòng)物性食品中總的雌激素活性進(jìn)行測(cè)定。結(jié)果表明所有樣品均檢測(cè)到一定的雌激素活性，生物雌二醇當(dāng)量濃度（EEQbio）在8.29~118.32ng/g之間。四大類動(dòng)物性食品中，家畜樣品測(cè)得的雌激素活性值最高，EEQbio為34.28~118.32ng/g，比雞蛋測(cè)得值高4~5倍，雞蛋樣品測(cè)得的雌激素活性值最低，EEQbio為8.29~20.91ng/g；家禽、水產(chǎn)品測(cè)得的EEQbio的范圍在12.82~110.94ng/g之間。 第二部分：動(dòng)物性食品中8種常見(jiàn)環(huán)境雌激素含量的測(cè)定 采用高效液相色譜-串聯(lián)質(zhì)譜方法（LC/MS/MS）檢測(cè)了樣品中雌酮（E1）、17β-雌二醇（E2）、雌三醇（E3）、乙炔基雌二醇（EE2）、己烯雌酚（DES）、雙酚A（BPA）、壬基酚（4-NP）、辛基酚（4-t-OP）等8種常見(jiàn)的環(huán)境雌激素。結(jié)果表明，天然雌激素E1、E3在動(dòng)物性食品中均未檢出，所有的樣品中都檢測(cè)到E2，檢出率達(dá)100%，其濃度范圍在0.44~15.04ng/g之間，中值濃度為7.93ng/g。人工合成雌激素EE2在部分樣品中有檢出，總體檢出率為33.1%，在樣品中檢測(cè)到的最高濃度為2.8ng/g，DES在任何樣品中均未檢測(cè)到。酚類環(huán)境雌激素BPA和4-NP有較高的檢出率約83%，4-t-OP的檢測(cè)值均低于檢測(cè)下限。BPA和4-NP的濃度范圍在nd~35.76ng/g之間。 結(jié)論：重組基因酵母篩檢法是篩選和定量分析樣品總雌激素活性的一種快速、有效的方法，重組基因酵母對(duì)雌激素類化合物有較高的靈敏度。所有樣品測(cè)得的EEQbio（8.29-118.32ng/g）均高于化學(xué)雌二醇當(dāng)量（EEQchem，0.44-15.91ng/g），兩種方法測(cè)得的EEQ值有一定的相關(guān)性（r=0.84）。
[Abstract]:Objective: with the development of industry and agriculture, environmental estrogens are widely used in our daily life. It has been found that they can disturb the endocrine system of humans and animals and cause abnormal changes in development, behavior and reproduction. Food is the material foundation of human survival and development and the main way of human exposure to environmental estrogens. Therefore, the monitoring of environmental estrogens in food is carried out. It is of great significance to discover and eliminate the potential harm to human health. Methods: in this study, the estrogenic activity of 16 kinds of 142 kinds of animal food frequently consumed by four cities in China was evaluated by recombinant gene yeast screening method (YESs). The contents of 8 common environmental estrogens in the samples were analyzed by HPLC-MS-MS. Results: the main content consists of two parts:. Part one: determination of total estrogen activity in animal foods. In this part, the total estrogenic activity in animal food was determined by this method. The results showed that all samples had certain estrogenic activity, and the biological equivalent concentration of estradiol was between 8.29 and 118.32 ng / g. The estradiol activity of domestic animal samples was 34.28 ~ 118.32ng / g, which was 4 ~ 5 times higher than that of eggs. The lowest estrogenic activity of egg samples was 8.29 ~ 20.91ng / g, and the range of EEQbio measured by poultry and aquatic products was 12.82110.94ng / g. Part II: determination of eight common environmental estrogens in animal foods. Eight common environmental estrogens were detected by HPLC / MS / MS / MS / MS / MS, including estradiol E1O17 尾 -estradiol (E2N), estriol (E3N), ethylestradiol (E2N), diethylstilbestrol (DESN), bisphenol A (BPAA), nonylphenol 4-NPand octylphenol (4-t-OPP), and so on. No natural estrogen E _ 1C _ 3 was detected in animal food, and E _ 2 was detected in all samples, and the detection rate was 100%. The concentration range was between 0.44 ~ 15.04 ng / g, and the median concentration was 7.93 ng / g. Synthetic estrogen EE2 was detected in some samples. The total detectable rate was 33.1. The highest concentration of 2.8ng / g des was not detected in any sample. The detection rates of phenolic environmental estrogens BPA and 4-NP were higher than those of the lower limit of detection. BPA and 4-NP were in the range of nd~35.76ng/g. Conclusion: the screening method of recombinant gene yeast is a rapid and effective method for screening and quantitative analysis of total estrogen activity of samples. The sensitivity of recombinant gene yeast to estrogen compounds was higher than that of chemical estradiol equivalent EEQbiotin (8.29-118.32ng / g), and the EEQ values measured by the two methods had a certain correlation with 0.84 ng / g ~ (-1) of estradiol equivalent (Eestradiol) = 0.44-15.91ng / g ~ (-1) 路g ~ (-1) 路g ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1)).
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R155.5

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