本文選題：果蔬籽 + 巰基化合物　； 參考：《延邊大學》2017年碩士論文

【摘要】：生物體內(nèi)含有多種抗氧化劑,其中含半胱氨酸殘基的巰基化合物在維持生物體內(nèi)氧化還原氛圍中起到了不可替代的作用。據(jù)文獻報道;生物體內(nèi)的巰基化合物含量水平于與眾多疾病緊密相連,其含量高低也成為了許多病理條件的一個判斷標志。由于生活節(jié)奏的日益加速,大多數(shù)人時常處在高壓力的狀態(tài),這也刺激人體內(nèi)活性氧自由基的過量增加,從而加速機體的衰老,與此同時過量的活性氧自由基也易于誘發(fā)基因突變導(dǎo)致癌癥等惡性疾病的產(chǎn)生。另據(jù)研究表明;人體通過攝入外源性抗氧化劑,像還原型谷胱甘肽(GSH),N-乙�；�-半胱氨酸等含巰基化合物,可明顯提高人體血液內(nèi)的GSH含量水平,這對提高機體抗病能力、保健均具有重要的作用[1]。因此,開發(fā)一種準確、靈敏、經(jīng)濟又快速的測定含半胱氨酸殘基的巰基化合物含量的分析方法,并以此來篩選富含巰基化合物的各類食材作為外源性抗氧化補充劑,這對防病、治病等方面具有實際意義。本研究利用對巰基具有選擇性的、并與其定量衍生化的衍生化試劑5,5'-二硫雙-2-硝基苯甲酸(DTNB),用含過量衍生化試劑的提取液,采用細胞破壁、衍生化為一體的前處理方法,分別用HPLC法和光譜法實時測定了果蔬籽中GSH和P-SH含量,并從中篩選出了四種富含GSH和蛋白巰基的果蔬籽。HPLC測定結(jié)果發(fā)現(xiàn),柚子籽、南瓜籽、黃瓜子、冬瓜子中含較豐富的GSH和蛋白巰基(P-SH),其中柚子籽中GSH和P-SH巰基含量分別為1.7304 μmol/g(GSH),1.2028μmol/g(P-SH),南瓜籽含量分別為 1.0445 μmol/g(GSH)和 1.8878 μmol/g(P-SH)。黃瓜籽和冬瓜籽中未檢測到GSH等小分子巰基化合物,但含相對豐富的蛋白巰基(P-SH),其含量分別為4.0325 μmol/g和4.1623 μmol/g。對所選定的果蔬樣品也分別采用文獻已報道的熒光分光光度法和紫外-可見分光光度法進行了比較測定,發(fā)現(xiàn)熒光光度法測定GSH含量和紫外-可見分光光度法測定結(jié)果略微偏離色譜測定結(jié)果,這可能是由于樣品中的共存組分對GSH的測定有一定的光譜干擾。本研究中采用的HPLC測定方法的回收率在99.92%-101.32%之間,GSH和P-SH測定檢測限分別為0.1384 μmol/g和0.2184μmol/g,基本滿足準確測定的要求。由于生物樣品成分復(fù)雜,在果蔬籽中含有大量的不飽和脂肪酸、黃酮類物質(zhì)、維生素和礦物質(zhì)等。為了探討果蔬籽中其他化學成分對實驗測定結(jié)果的影響,在本研究中將柚子籽和南瓜籽作為樣品,采用高效液相色譜法和光譜法分別對黃酮類化合物和不飽和脂肪酸對色譜和光譜測定干擾進行了探究。結(jié)果表明;采用光譜法測定樣品中巰基化合物含量時受到黃酮類物質(zhì)的干擾較大,但采用高效液相色譜法測定其含量時并無顯著影響;于此同時,樣品中的不飽和脂肪酸類物質(zhì)對樣品的測定結(jié)果基本不構(gòu)成干擾或干擾相對較小。綜上所述,本研究為進一步從植物中篩選富含巰基化合物的準確分析方法、以及特定果蔬籽中的活性巰基化合物的含量提供了重要的分析數(shù)據(jù)。
[Abstract]:There are many antioxidants in the organism, and the sulfhydryl compounds containing cysteine residues play an irreplaceable role in maintaining the redox atmosphere in the organism. It is reported in the literature that the levels of sulfhydryl compounds in organisms are closely linked to many diseases, and the content of sulfhydryl compounds has also become one of the many pathological conditions. Judging sign. Because of the increasing pace of life rhythm, most people are often in high stress state, which also stimulates the excessive increase of reactive oxygen radicals in the body, and thus accelerates the aging of the body, while excessive reactive oxygen free radicals can also induce gene mutation to cause cancer and other malignant diseases. Through the intake of exogenous antioxidants, such as GSH, N- acetyl cysteine and other sulfhydryl compounds, the content of GSH in human blood can be significantly increased, which has an important role in improving the body's ability to resist disease and health care. Therefore, a accurate, sensitive, economical and rapid determination of cysteine containing cysteine is made. The analytical method of the content of sulfhydryl compounds in the residues is used to screen all kinds of edible materials rich in sulfhydryl compounds as a exogenous antioxidant supplement, which is of practical significance for the prevention and treatment of the diseases. This study uses the sulfhydryl group, which is selective, and its derivatization reagent 5,5'- two sulfur double -2- Nitrobenzoic Acid. DTNB), the content of GSH and P-SH in fruit and vegetable seeds was measured in real time by HPLC method and spectral method, and the results of four kinds of fruit and vegetable seeds rich in GSH and protein sulfhydryl were detected by HPLC method and spectral method. Rich GSH and protein sulfhydryl (P-SH) were contained in the seeds, in which the content of GSH and P-SH mercapto in grapefruit seed was 1.7304 mu mol/g (GSH), 1.2028 mol/g (P-SH), and the pumpkin seed content was 1.0445 micron (GSH) and 1.8878 mu mol/g (P-SH), and the small molecule mercapto compounds were not detected in cucumber seed and winter melon seed, but the relative rich protein was contained. The content of sulfhydryl group (P-SH) is 4.0325 mu mol/g and 4.1623 mol/g. respectively, and the selected samples of fruits and vegetables are also compared by the reported fluorescence spectrophotometry and UV visible spectrophotometric method. It is found that the determination of GSH content by fluorometric method and the UV visible spectrophotometric method are slightly deviated from the chromatographic test. This may be due to the spectral interference of the coexisting components in the samples for the determination of GSH. The recovery rate of the HPLC method used in this study is between 99.92%-101.32% and GSH and P-SH, respectively. The detection limits are 0.1384 mu mol/g and 0.2184 mu mol/g, respectively. Fruit and vegetable seeds contain a large number of unsaturated fatty acids, flavonoids, vitamins and minerals. In order to explore the effects of other chemical components in fruit and vegetable seeds on the experimental results, pomelo seeds and pumpkin seeds were used as samples in this study. The flavonoids and unsaturated fatty acids were used respectively by high performance liquid chromatography and light spectrum method. The results show that the determination of the content of sulfhydryl compounds in the samples by spectral method is greatly disturbed by the flavonoids, but there is no significant influence on the determination of the content by high performance liquid chromatography. At the same time, the determination results of the unsaturated fatty acids in the samples are basic. No interference or interference is relatively small. In summary, this study provides important analytical data for the accurate analysis of sulfhydryl compounds rich in plants, as well as the content of active sulfhydryl compounds in specific fruit and vegetable seeds.
【學位授予單位】：延邊大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：TS209;O657

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