本文選題：游離棉酚　切入點(diǎn)：棉酚酮　出處：《新疆醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:1.本實(shí)驗(yàn)通過對實(shí)驗(yàn)大鼠、鯽魚、雞喂食含有棉酚的自制棉籽飼料,模擬動物進(jìn)食過程,實(shí)驗(yàn)周期內(nèi)定期取實(shí)驗(yàn)動物各組織、器官,建立HPLC檢測實(shí)驗(yàn)樣品中游離棉酚及其代謝產(chǎn)物(棉酚酮)殘留的方法。并采用高靈敏度的UPLC-MS/MS分析游離棉酚及其代謝產(chǎn)物在大鼠體內(nèi)的組織分布。2.建立牛乳中游離棉酚及其旋光異構(gòu)體的HPLC檢測方法。并根據(jù)上述方法對市售的動物源性食品進(jìn)行游離棉酚及其代謝產(chǎn)物殘留的檢測。方法:1.用自制的含定量棉酚的棉籽飼料飼養(yǎng)實(shí)驗(yàn)動物一定周期,定期取實(shí)驗(yàn)動物各組織器官(心、肝、脾、肺、腎、肉、血漿),將各樣品勻漿,加入提取劑(0.2%甲酸乙腈溶液)及甘草次酸內(nèi)標(biāo)液,經(jīng)除蛋白、離心等前處理方法后,采用HPLC及UPLC-MS/MS法確定動物體內(nèi)各個組織器官中游離棉酚及其代謝產(chǎn)物的殘留和組織分布。2.采用以(R)-(-)-2-胺基-1-丙醇為手性拆分試劑、甘草次酸為內(nèi)標(biāo)物的柱前衍生高效液相色譜法,對牛乳中游離棉酚及其旋光異構(gòu)體含量進(jìn)行檢測。結(jié)果:1.HPLC及UPLC-MS/MS測定動物體內(nèi)游離棉酚及其代謝產(chǎn)物的分析方法良好。HPLC法測定大鼠和雞體內(nèi)各組織器官中游離棉酚及棉酚酮的線性范圍為0.5-20μg/mL,鯽魚為0.1-10μg/mL,檢出限0.03μg/mL。UPLC-MS/MS法檢測大鼠體內(nèi)各組織器官中游離棉酚及棉酚酮的線性范圍均為0.01-10μg/mL,檢出限為0.002μg/mL。UPLC-MS/MS分析表明大鼠體內(nèi)游離棉酚及棉酚酮的含量隨飼養(yǎng)時(shí)間延長而蓄積增加,體內(nèi)棉酚含量依次為:肝臟、血漿、脾臟、肺臟、心臟、睪丸(雄性)、腎臟、肌肉,且肝臟中蓄積最多,動物體內(nèi)代謝物棉酚酮僅在肝臟中發(fā)現(xiàn)。通過對雌、雄大鼠進(jìn)行對比分析,結(jié)果顯示棉酚在雌性大鼠體內(nèi)的蓄積更高。2.牛乳中棉酚及其旋光異構(gòu)體濃度線性范圍均為2-10μg/mL。3.運(yùn)用所建立的檢測方法,對市售動物源性食品中游離棉酚及其代謝產(chǎn)物進(jìn)行檢測,包括魚、雞、豬、羊(肝、肉)共58份樣品,檢出率為51.7%。結(jié)論:本研究建立了動物體內(nèi)和乳制品中游離棉酚及棉酚酮HPLC檢測方法,為市場上動物源性食品中游離棉酚及棉酚酮的檢測提供了數(shù)據(jù)支持。同時(shí)通過UPLC-MS/MS法提高了高靈敏度,進(jìn)一步證實(shí)了棉酚在動物體內(nèi)的代謝產(chǎn)物為棉酚酮,為體內(nèi)棉酚代謝產(chǎn)物的研究提供實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Objective 1. In this experiment, the experimental rats, crucian carp and chickens were fed with self-made cottonseed feed containing gossypol to simulate the feeding process of the animals. The tissues and organs of the experimental animals were taken regularly during the experiment cycle. A HPLC method for the determination of free gossypol and its metabolites (gossypol ketone) in experimental samples was established. The tissue distribution of free gossypol and its metabolites in rats was analyzed by high sensitivity UPLC-MS/MS. 2. Free gossypol and its metabolites in milk were established. HPLC method for detection of gossypol and its optical isomers. Determination of free gossypol and its metabolite residues in animal-derived foods sold on the market based on the above method. Methods: 1. Feeding with cotton seed feed containing quantitative gossypol. Experimental animals have a certain period, The tissues and organs (heart, liver, spleen, lung, kidney, meat, plasma) of experimental animals were regularly taken. The samples were homogenized and added with the extract of 0.2% acetonitrile formate solution, and the internal standard solution of glycyrrhetinic acid was added. The residual and tissue distribution of free gossypol and its metabolites in animal tissues and organs were determined by HPLC and UPLC-MS/MS methods. A precolumn derivatization high performance liquid chromatography (HPLC) was developed, which was used as chiral resolution reagent and glycyrrhetinic acid as internal standard. The content of free gossypol and its rotatory isomers in milk was determined. Results 1. The determination of free gossypol and its metabolites in animals by HPLC and UPLC-MS/MS. The method of HPLC was good for the determination of free gossypol in tissues and organs of rats and chickens. The linear range of gossypol and gossypol was 0.5-20 渭 g / mL and 0.1-10 渭 g / mL, respectively. The detection limit of 0.03 渭 g/mL.UPLC-MS/MS was 0.01-10 渭 g / mL for free gossypol and gossypol in various tissues and organs of rats. The detection limit of free gossypol and gossypol was 0.002 渭 g/mL.UPLC-MS/MS. The accumulation increased with the increase of feeding time. The contents of gossypol in vivo are as follows: liver, plasma, spleen, lung, heart, testis (male, kidney, muscle, and liver). The metabolite gossypol is found only in the liver. The results of comparative analysis in male rats showed that gossypol accumulated higher in female rats. The linear range of gossypol and its optical isomers in milk was 2-10 渭 g / mL 路3.Using the established method, A total of 58 samples of free gossypol and its metabolites, including fish, chicken, pig, sheep (liver, meat), were detected in animal food. Conclusion: this study established a HPLC method for the detection of free gossypol and gossypol in animal and dairy products. It provides data support for the detection of free gossypol and gossypol in animal food on the market. At the same time, the high sensitivity of gossypol in animal body is improved by UPLC-MS/MS method, which further proves that the metabolite of gossypol in animal body is gossypol. To provide experimental basis for the study of gossypol metabolites in vivo.
【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】：R99

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李東雨;張亞剛;孫蓮;姚軍;;牛乳中游離棉酚及其旋光異構(gòu)體含量高效液相色譜法測定[J];食品科技;2016年07期

2 梁建華;;對我國食用棉籽油質(zhì)量及安全的探究[J];食品安全導(dǎo)刊;2016年18期

3 馬駿;羅華明;張玲茜;;液相色譜串聯(lián)質(zhì)譜測定水產(chǎn)品中硝基呋喃類代謝物殘留[J];浙江農(nóng)業(yè)科學(xué);2016年04期

4 趙曉亞;付曉芳;李晶;王鵬;葉誠;尚吟竹;鄭茜s，

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