本文選題：棉籽油 + 游離棉酚降解產(chǎn)物　； 參考：《新疆醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:棉籽油中游離棉酚含量檢測方法及限量已經(jīng)有國家標(biāo)準(zhǔn),但由于棉籽油中游離棉酚在常溫下極易降解(30天內(nèi)降解80%以上),因此該方法很難反映棉籽油的脫毒情況。本實驗通過自制未脫毒棉籽油樣品,考察游離棉酚在棉籽油中的降解情況,確定主要游離棉酚降解產(chǎn)物的存在并鑒定其化學(xué)結(jié)構(gòu),通過體內(nèi)、外毒性實驗考察主要游離棉酚降解產(chǎn)物的生殖毒性并建立棉籽油中主要游離棉酚降解產(chǎn)物的含量測定方法。方法:1.根據(jù)棉籽油生產(chǎn)工藝,本實驗采用正己烷溶劑浸提法提取未脫毒棉籽油樣品,室溫放置。采用HPLC測定不同放置時間的棉籽油樣品,建立游離棉酚與降解產(chǎn)物之間的線性關(guān)系。2.采用C18層析柱和半制備型高效液相色譜儀制備主要游離棉酚降解產(chǎn)物,用于質(zhì)譜結(jié)構(gòu)鑒定。3.采用液相色譜-傅立葉變換離子回旋共振質(zhì)譜儀測定主要游離棉酚降解產(chǎn)物的準(zhǔn)確分子量、推測分子式并鑒定其結(jié)構(gòu)。4.采用GC-2小鼠精母細(xì)胞毒性實驗和小鼠精子畸變實驗考察主要游離棉酚降解產(chǎn)物的體內(nèi)、外毒性。5.建立主要游離棉酚降解產(chǎn)物含量測定方法,測定24批市售棉籽油中主要游離棉酚降解產(chǎn)物含量。結(jié)果:1.棉籽油中游離棉酚與降解產(chǎn)物之間的線性關(guān)系為Y=-3.00674395X+2058258982,r=0.9900。2.經(jīng)質(zhì)譜儀檢測,主要游離棉酚降解產(chǎn)物準(zhǔn)分子離子為573.25033,推測其分子式為C34H3708,鑒定其結(jié)構(gòu)為四甲氧基棉酚。3.細(xì)胞毒性實驗結(jié)果為:應(yīng)用SPSS Statistics 17.0軟件計算,主要游離棉酚降解產(chǎn)物四甲氧基棉酚對小鼠GC-2精母細(xì)胞的半數(shù)抑制濃度IC50為6.554μg/mL。小鼠精子畸變實驗結(jié)果為:空白組小鼠精子畸形率為1.1 ±0.42%,損傷對照組小鼠精子畸形率為8.2±0.31%,高劑量組小鼠精子畸形率為4.4±0.51%,中劑量組小鼠精子畸形率為3.0±0.33%,低劑量組小鼠精子畸形率為1.8±0.44%,采用秩和檢驗法處理實驗數(shù)據(jù),陽性組與對照組精子畸形率相比P0.05,有顯著性差異,高、中、低試驗組與對照組精子畸形率相比P0.05,有顯著性差異。4.對24批市售棉籽油樣品進(jìn)行檢測,其中有20批棉籽油樣品檢測到主要游離棉酚降解產(chǎn)物,含量為0.139～0.912μg/mL。結(jié)論:通過測定棉籽油中主要游離棉酚降解產(chǎn)物的含量反映游離棉酚的脫毒情況,這種方法切實可行,為新疆地區(qū)棉籽油的生產(chǎn)和監(jiān)管提供科學(xué)有效的檢測方法。
[Abstract]:Objective: the determination method and limit of free gossypol in cottonseed oil have already had the national standard, but because the free gossypol in cottonseed oil can easily degrade more than 80% in 30 days at room temperature, it is difficult to reflect the detoxification of cottonseed oil by this method.In this experiment, the degradation of free gossypol in cottonseed oil was investigated by self-made samples of undetoxified cottonseed oil, the existence of main degradation products of free gossypol was determined and its chemical structure was identified.The reproductive toxicity of the main free gossypol degradation products was investigated and a method for determining the content of the main free gossypol degradation products in cottonseed oil was established.Method 1: 1.According to the production process of cottonseed oil, the samples of undetoxified cottonseed oil were extracted by n-hexane solvent extraction and placed at room temperature.The linear relationship between free gossypol and degradation product was established by HPLC.The main free gossypol degradation products were prepared by C18 column and semi-preparation high performance liquid chromatograph, which were used to identify the structure of gossypol by mass spectrometry.The molecular weight of the main degradation products of free gossypol was determined by liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometer. The molecular formula was deduced and its structure was identified.GC-2 mouse spermatocyte toxicity test and mouse sperm aberration test were used to investigate the in vivo and in vitro toxicity of the main free gossypol degradation products.A method was established to determine the content of major free gossypol degradation products in 24 batches of cottonseed oil.The result is 1: 1.The linear relationship between free gossypol and degradation products in cottonseed oil is YANG-3.00674395X 2058258982 ru 0.9900.2.The main free gossypol degradation product excimer ion is 573.25033, and its molecular formula is C34H3708, and its structure is identified as tetramethoxygossypol. 3.The results of cytotoxicity test showed that the median inhibitory concentration (IC50) of tetramethoxygossyl, the main degradation product of free gossypol, on mouse GC-2 spermatocytes was 6.554 渭 g 路mL ~ (-1), calculated by SPSS Statistics 17.0 software.The results of sperm aberration test in mice were as follows: sperm abnormality rate of blank group was 1.1 鹵0.42, that of injured control group was 8.2 鹵0.31, that of high dose group was 4.4 鹵0.51, that of middle dose group was 3.0 鹵0.33 and that of low dose group was small.The deformity rate of rat sperm was 1.8 鹵0.44. The experimental data were processed by rank sum test.There was significant difference in sperm malformation rate between positive group and control group (P 0.05). There was significant difference in sperm malformation rate between test group and control group (P 0.05).Twenty-four batches of cottonseed oil samples were tested, of which 20 batches of cottonseed oil samples detected the main free gossypol degradation products, the content of which was 0.139 ~ 0.912 渭 g 路mL ~ (-1).Conclusion: the detoxification of free gossypol is reflected by the determination of the main free gossypol degradation products in cottonseed oil. This method is feasible and provides a scientific and effective method for the production and supervision of cottonseed oil in Xinjiang.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TS227;R114

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