本文選題：丙烯酰胺 + 生物標(biāo)志物��； 參考：《山東大學(xué)》2012年碩士論文

【摘要】：丙烯酰胺(Acrylamide,ACR)是一種重要的化工原料,主要用于生產(chǎn)其聚合物—聚丙烯酰胺(PACR)。后者則廣泛用于飲水凈化、污水處理、造紙、采礦等行業(yè)[1]。另外,高溫烹飪(120℃以上)富含淀粉的食物(如土豆和谷物等)時(shí)也會(huì)產(chǎn)生ACR。ACR可通過呼吸道、消化道、皮膚黏膜等多種途徑吸收,其中經(jīng)消化道吸收最快,以皮膚接觸導(dǎo)致職業(yè)中毒最常見。此外,哺乳動(dòng)物細(xì)胞體外培養(yǎng)和體內(nèi)試驗(yàn)均表明ACR具有神經(jīng)毒性、生殖毒性、胚胎發(fā)育毒性和遺傳毒性,但是由于缺乏流行病學(xué)方面的資料,至今能在人體內(nèi)得到驗(yàn)證的只有神經(jīng)毒性[2]。因職業(yè)接觸而導(dǎo)致ACR中毒的情況近年來時(shí)有發(fā)生。尋找ACR特異的生物標(biāo)志物,有助于評(píng)價(jià)ACR作業(yè)工人的暴露水平,預(yù)防ACR作業(yè)工人急慢性中毒的發(fā)生,為更好地保護(hù)職業(yè)暴露人群提供依據(jù)。本課題研究ACR暴露與生物材料指標(biāo)之間的相關(guān)性,以尋找早期生物標(biāo)志物,為ACR的接觸評(píng)價(jià)和效應(yīng)評(píng)價(jià)提供依據(jù)。 目的 通過測(cè)定ACR接觸工人尿中乙酰半胱氨酸-S-丙酰胺(APC)、血清中鈣離子(Ca2+)和蛋白激酶C(PKC)的含量,結(jié)合工人ACR個(gè)體暴露水平、職業(yè)相關(guān)因素和個(gè)人基本情況,探討ACR接觸工人暴露負(fù)荷與生物指標(biāo)之間的關(guān)系以尋找早期生物學(xué)監(jiān)測(cè)指標(biāo)。 研究對(duì)象與方法 1.研究對(duì)象的選擇：本研究選擇某市ACR生產(chǎn)企業(yè)接觸ACR的暴露組工人及管理、后勤崗位不接觸ACR的對(duì)照組工人共138人,其中暴露組101人,對(duì)照組37人。 2.現(xiàn)場(chǎng)職業(yè)衛(wèi)生調(diào)查：以查閱工廠資料和問卷調(diào)查相結(jié)合的形式了解工廠的生產(chǎn)工藝、工人的職業(yè)相關(guān)因素和個(gè)人基本情況。 3.樣品的采集：空氣中ACR濃度的樣品采集按照國家標(biāo)準(zhǔn)《工作場(chǎng)所空氣中有害物質(zhì)監(jiān)測(cè)的采樣規(guī)范》(GBZ159-2004)中規(guī)定的方法,使用個(gè)體空氣采樣器,連接硅膠采樣管進(jìn)行采樣；皮膚ACR污染量的樣品采集用經(jīng)蒸餾水潤(rùn)濕的醫(yī)用棉球擦拭皮膚；尿樣的采集使用聚乙烯塑料瓶收集工人班后尿；血樣的采集使用一次性無添加劑真空采血管采集班后工人前臂靜脈血。 4.樣品的測(cè)定：空氣中ACR濃度和工人皮膚ACR污染量的測(cè)定采用氣相色譜法；工人尿中APC濃度的測(cè)定采用可見分光光度法；血清中Ca2+濃度的測(cè)定應(yīng)用Ca2+試劑盒法(甲基百里香酚藍(lán)法)；血清中PKC濃度的測(cè)定應(yīng)用ELISA檢測(cè)試劑盒法。 5.數(shù)據(jù)處理與統(tǒng)計(jì)分析：建立Word文本資料數(shù)據(jù)庫和Excel數(shù)據(jù)資料數(shù)據(jù)庫,應(yīng)用SPSS16.0軟件進(jìn)行統(tǒng)計(jì)分析。對(duì)數(shù)據(jù)進(jìn)行正態(tài)性檢驗(yàn)和方差齊性檢驗(yàn),應(yīng)用單因素方差分析、簡(jiǎn)單線性相關(guān)分析和多元回歸分析對(duì)各指標(biāo)之間的關(guān)系進(jìn)行研究。 結(jié)果 1.空氣ACR濃度和皮膚ACR污染量測(cè)定所用氣相色譜法的線性范圍為0.10～2.0μg/ml,回歸方程為y=3186918.408x+4388.271,相關(guān)系數(shù)為0.9997,檢出限為1.4×10-5μg/ml,保留時(shí)間為1.633min。 2.工人尿APC測(cè)定所用分光光度法,游離巰基化合物的標(biāo)準(zhǔn)曲線y=2.375x-0.003,相關(guān)系數(shù)為0.998,游離巰基化合物的檢出下限為0.002mmol/L:總巰基化合物的標(biāo)準(zhǔn)曲線為y=1.611x+0.000,相關(guān)系數(shù)為0.999,游離毓基化合物的檢出下限為0.01mmol/L。 3.ACR暴露組工人空氣ACR濃度為O.040±O.015mg/m3,對(duì)照組工人空氣ACR未檢出；ACR暴露組工人皮膚ACR污染量為2.003±0.961μg,對(duì)照組工人皮膚ACR未檢出。 4.ACR暴露組和對(duì)照組工人尿中APC濃度分別為0.65±0.22mmol/L和0.54±0.18mmol/L,暴露組高于對(duì)照組(P0.05)；暴露組和對(duì)照組工人血清中Ca2+濃度分別為2.18±0.15mmol/L和2.32±0.33mmol/L,暴露組低于對(duì)照組(P0.05)；暴露組和對(duì)照組工人血清中PKC濃度分別為235.06±93.08nmol/L和194.73±98.98nmol/L暴露組高于對(duì)照組工人(P0.05)。 5.簡(jiǎn)單線性相關(guān)分析：ACR暴露工人尿中APC濃度、血清PKC濃度與皮膚ACR污染量呈正相關(guān)關(guān)系(P0.01)；血清中Ca2+濃度與皮膚ACR污染量呈負(fù)相關(guān)關(guān)系(P0.01)。 6.多元回歸分析：對(duì)工人尿中APC、血清中Ca2+和PKC濃度的影響因素中,進(jìn)入方程的因素為皮膚ACR污染量。 結(jié)論 1.ACR暴露能夠引起工人尿中APC、血清中Ca2+和PKC濃度的改變。 2.工人尿中APC、血清中Ca2+和PKC的濃度變化主要是通過皮膚吸收ACR引起的。 3.工人尿中APC、血清中Ca2+和PKC可能為ACR暴露的生物標(biāo)志物。 4.空氣ACR濃度和皮膚ACR污染量使用毛細(xì)管柱氣相色譜儀進(jìn)行測(cè)定,分離效果好、高效、快速；個(gè)體空氣采樣使用硅膠采樣管,符合現(xiàn)場(chǎng)采樣要求。
[Abstract]:Acrylamide (ACR) is an important chemical raw material, mainly used in the production of polymer polyacrylamide (PACR). The latter is widely used in drinking water purification, sewage treatment, paper making, mining and other industries [1].. The high temperature cooking (above 120 degrees C) rich in flour (such as potatoes and grains) will also produce ACR.ACR through the call. Absorption of the channels, digestive tract, skin and mucous membrane, including the fastest absorption of the digestive tract, and the most common cause of occupational poisoning by skin contact. In addition, mammalian cells in vitro culture and in vivo tests indicate that ACR has neurotoxicity, reproductive toxicity, embryonic development toxicity and hereditary toxicity, but because of lack of epidemiology. Materials, only neurotoxic [2]. caused by occupational exposure to ACR poisoning, have occurred in recent years. Looking for ACR specific biomarkers will help to evaluate the exposure level of ACR workers, prevent the occurrence of acute and chronic poisoning in ACR workers, and provide a better protection for the occupational exposure population. According to this topic, we study the correlation between ACR exposure and biomaterial indicators, in order to find early biomarkers, and provide basis for ACR contact evaluation and effect evaluation.
objective
By measuring the content of acetylcysteine -S- propanamide (APC) in urine of ACR workers, serum calcium ion (Ca2+) and protein kinase C (PKC), the relationship between exposure load and biological index of workers in ACR contact workers was explored to find the early biological monitoring index by combining the exposure level of ACR individual, occupational related factors and individual basic conditions.
Research objects and methods
1. the selection of the research subjects: This study selected the exposed workers and management of the ACR production enterprises in a city and the management of the exposed group of ACR, and the control group who did not contact the ACR in the logistics post was 138, of which 101 were exposed and 37 in the control group.
2. on-site occupational health survey: in the form of consulting factory data and questionnaire survey, the production process of the factory, the occupational related factors of the workers and the basic individual situation of the individual are understood.
3. sample collection: samples of ACR concentration in the air are collected in accordance with the national standard < Standard for sampling of harmful substances in the air of the workplace > (GBZ159-2004). Individual air sampler is used to connect silica sampling tube for sampling; samples of ACR pollution of skin are collected by medical cotton ball wetted with distilled water. The collection of urine samples was collected using polyethylene plastic bottles to collect urine after the workers' class.
4. determination of the samples: the determination of ACR concentration in the air and the ACR pollution of the workers' skin by gas chromatography; the determination of APC concentration in the urine of workers using visible spectrophotometry; the determination of Ca2+ concentration in serum by Ca2+ Kit (methyl thymol blue), and the determination of PKC concentration in serum by ELISA detection kit method.
5. data processing and statistical analysis: establishing Word text data database and Excel data data base, using SPSS16.0 software for statistical analysis. The normal test and variance homogeneity test of data are carried out, single factor ANOVA, simple linear correlation analysis and multiple return analysis are used to study the relationship between each index.
Result
The linear range of 1. air ACR concentration and skin ACR pollution measured by gas chromatography was 0.10 ~ 2 mu g/ml, the regression equation was y=3186918.408x+4388.271, the correlation coefficient was 0.9997, the detection limit was 1.4 * 10-5 mu g/ml, and the retention time was 1.633min.
2. the spectrophotometric method was used to determine the urine APC of workers. The standard curve of the free sulfhydryl compound was y=2.375x-0.003, the correlation coefficient was 0.998. The standard curve of the detection limit of the free sulfhydryl compound was y=1.611x+0.000, the correlation coefficient was 0.999, the detection limit of the free Yuk compound was 0.01mmol/L..
The concentration of air ACR in the exposed group of 3.ACR was O.040 + O.015mg/m3, and the air ACR in the control group was not detected, and the ACR pollution of the workers in the ACR exposed group was 2.003 + 0.961 mu g, and the skin ACR in the control group was not detected.
The urine APC concentration in the exposed group and the control group was 0.65 + 0.22mmol/L and 0.54 + 0.18mmol/L respectively, and the exposure group was higher than the control group (P0.05). The concentration of Ca2+ in the exposed group and the control group was 2.18 + 0.15mmol/L and 2.32 + 0.33mmol/L respectively. The exposure group was lower than the control group (P0.05), and the serum PKC concentration in the exposed and control groups was in the serum of the exposed and control groups. The exposure group of 235.06 + 93.08nmol/L and 194.73 + 98.98nmol/L was higher than that of the control group (P0.05).
5. simple linear correlation analysis: ACR exposed workers' urine APC concentration, serum PKC concentration was positively correlated with skin ACR pollution (P0.01), and serum Ca2+ concentration was negatively correlated with skin ACR pollution (P0.01).
6. multivariate regression analysis: the factors affecting the concentration of APC and Ca2+ and PKC in the urine of workers were skin contamination of ACR.
conclusion
1.ACR exposure can cause urinary APC and serum Ca2+ and PKC concentrations in workers.
2. the level of APC in urine and Ca2+ and PKC in serum were mainly caused by absorption of ACR through skin.
3. APC in workers urine, Ca2+ and PKC in serum may be biomarkers for ACR exposure.
4. the concentration of air ACR and the ACR pollution of skin were measured by capillary column gas chromatograph. The separation effect was good, high efficiency and rapid. The individual air sampling used silica gel tube, which was in line with the requirement of field sampling.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R131

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