本文關(guān)鍵詞：綠豆芽富硒特性及HPLC-MS聯(lián)用法測定綠豆芽中硒代氨基酸　出處：《遼寧大學》2016年碩士論文　論文類型：學位論文

  更多相關(guān)文章： 硒 綠豆芽 高效液相色譜-質(zhì)譜聯(lián)用 硒代氨基酸

【摘要】：硒(Selenium, Se)是人體必需的微量元素,對人體有重要意義。它有增強免疫力、抗癌、抗氧化等重要的生理功能,在植物生長過程中向環(huán)境中添加無機硒可以通過植物自身的生物代謝將環(huán)境中的無機硒轉(zhuǎn)化為植物組織中的有機含硒化合物,通過含硒植物補硒是缺硒地區(qū)提高人體硒水平的有效方法。植物體中的硒主要以有機硒和無機硒的形式存在,而相對于無機硒化合物,有機形態(tài)的硒化合物通常具有較高的生理功能及較低的毒性,如硒蛋白和硒代氨基酸。綠豆芽的蛋白質(zhì)含量較高,且營養(yǎng)豐富,在綠豆的發(fā)芽過程中能更好地將環(huán)境中的無機硒轉(zhuǎn)化為自身的有機硒。本文以自培富硒綠豆芽為對象研究硒對綠豆芽生長過程中生理特性及其含硒量的影響,同時測定綠豆芽可溶性蛋白質(zhì)中的硒含量,并采用液相色譜-質(zhì)譜聯(lián)用技術(shù)對富硒綠豆芽中硒甲基硒代半胱氨酸、硒代蛋氨酸和硒代胱氨酸進行定性定量分析,得出結(jié)果如下：1、低濃度的硒對于綠豆芽的生長有一定的促進作用,當施硒濃度小于5mg/L時,綠豆芽的發(fā)芽率和長度與對照組比均有提高；當施硒濃度大于5 mg/L時,綠豆芽發(fā)芽率開始降低,在12 mg/L時較對照組降低30.3%,下胚軸及根的長度較對照組分別降低36.7%和31.6%。用考馬斯亮藍法測定綠豆芽中可溶性蛋白質(zhì)含量表明綠豆芽中蛋白質(zhì)含量在14%-19%范圍內(nèi),且硒有利于增加綠豆芽中蛋白質(zhì)的含量,即在一定程度上促進綠豆芽營養(yǎng)成分的合成。2、以HN03為消解劑,采用微波消解-石墨爐原子吸收光譜法對富硒綠豆芽中總硒含量進行消化及測定,研究不同消解劑量的使用對樣品消化效果的影響,結(jié)果表明,在1-12 mg/L施硒范圍內(nèi),綠豆芽中總硒含量隨著外源施硒濃度的增加而增大,在施硒濃度為12mg/L時綠豆芽中總硒含量達到最大值7275μg/kg,但高濃度的硒會對綠豆芽的生長產(chǎn)生毒害,抑制綠豆芽的生長。3、以超純水、HCl、aOH、Tris-HCl為浸提劑,對富硒綠豆芽中溶解態(tài)硒進行提取并測定其含量,結(jié)果表明,綠豆芽中可溶態(tài)硒的分布規(guī)律為：酸溶態(tài)堿溶態(tài)Tris-HCl溶解態(tài)水溶態(tài)。各個溶解態(tài)硒含量均隨施硒濃度增加而增大,酸溶態(tài)、堿溶態(tài)、Tris-HCl溶解態(tài)和水溶態(tài)中硒含量分別占總硒量的50%-55%、35%-45%、30%-35%和20%-30%。說明在酸性介質(zhì)中,硒更容易通過綠豆芽進行遷移轉(zhuǎn)化,從而更能被人體吸收利用。采用堿溶酸沉等電點法對富硒綠豆芽中可溶性蛋白質(zhì)進行提取,研究堿液濃度、料液比、浸提時間、浸提溫度及等電點對蛋白質(zhì)提取率的影響,最佳的提取條件為：提取液濃度0.20 mol/L、料液比1：50、浸提時間90 min、浸提溫度40℃及等電點pH為4.8,在此條件下提取綠豆芽可溶性蛋白質(zhì),使用微波消解儀對蛋白質(zhì)沉淀進行消化,并使用石墨爐原子吸收光譜法測定蛋白質(zhì)中的硒含量。結(jié)果表明,綠豆芽蛋白質(zhì)中硒含量隨著施硒濃度的增加而增大,最高濃度為1710μg/kg,占總硒含量的23.5%。高濃度的硒不僅能增加蛋白質(zhì)中硒含量,且能增大蛋白質(zhì)中硒占總硒的比例。4、采用PLC-MS聯(lián)用法對富硒綠豆芽樣品中硒代胱氨酸、硒代蛋氨酸和硒甲基硒代半胱氨酸進行定性定量分析,對每種氨基酸進行裂解電壓和碰撞能的優(yōu)化,選擇最佳提取條件,采用MRM模式進行監(jiān)測。結(jié)果表明,硒甲基硒代半胱氨酸是富硒綠豆芽中的主要硒代氨基酸；而硒代蛋氨酸和硒代胱氨酸的含量相對較小,僅占總硒含量的5%以下。因此,綠豆芽可以通過其生長代謝過程將無機硒有效地轉(zhuǎn)化成有機硒,作為營養(yǎng)、安全、高效的功能性補硒食品,是提高低硒地區(qū)人體內(nèi)含硒水平的有效途徑。
[Abstract]:Se (Selenium, Se) is an essential trace element, has important significance to the human body. It has enhanced immunity, anticancer, antioxidant and other important physiological functions, adding inorganic selenium to the environment in the process of plant growth through biological metabolism of plants in the environment of inorganic selenium into organic plant tissues selenium compounds, the selenium selenium selenium deficiency in plant is an effective method to improve the level of selenium in plants. Selenium mainly exists in the form of organic and inorganic selenium, rather than inorganic selenium compounds, physiological function of selenium compounds in organic form usually have higher and lower toxicity, such as protein and seleno amino acids. The protein content of mung bean is high, and rich nutrition, during the germination of mung bean can better inorganic selenium into organic selenium in the environment itself. Based on the self training of selenium enriched mung bean Discusses the effect of selenium on bud growth and physiological characteristics of mung bean sprouts in the process and the content of selenium, and selenium content of soluble protein in mung bean sprouts, and using liquid chromatography-mass spectrometry of Se methylselenocysteine enriched mung bean sprouts, qualitative and quantitative analysis of selenomethionine and selenocystine the results were as follows: 1, the low concentration of selenium for a certain role in promoting the growth of mung bean sprouts, when the selenium concentration is less than 5mg/L, the germination rate and the length of mung bean sprouts than in the control group were increased; when the concentration of selenium was greater than 5 mg/L, mung bean germination rate began to decrease at 12 mg/L when compared with the control group decreased 30.3%, hypocotyl and root length were decreased by 36.7% and 31.6%. by Coomassie brilliant blue method for the determination of soluble protein content in mung bean sprouts that mung bean protein content in the range of 14%-19% compared with the control group, and selenium To increase the protein content of mung bean sprouts, which promote the.2 synthesis of mung bean nutritional ingredients in a certain extent, with HN03 as the digesting agent, used to digest and determination of total selenium content in mung bean sprouts with Microwave Digestion Graphite furnace atomic absorption spectrometry, study the different effects of fire, use solution dosage on the effect sample digestion results showed that in the 1-12 mg/L range of selenium, the total selenium content in mung bean sprouts increased with increasing exogenous selenium concentration, the concentration of selenium is 12mg/L total selenium content in mung bean sprouts reached a maximum value of 7275 g/kg, but high concentration of selenium on growth of mung bean sprouts poison the growth inhibition of.3, mung bean sprouts, with ultra pure water, HCl, aOH, Tris-HCl as extraction agent, the dissolved selenium enriched mung bean sprout extract and determine the content, the results show that the distribution of soluble selenium in mung bean sprouts are: acid soluble alkali soluble Tris-H Cl dissolved water soluble selenium content. All were dissolved with the selenium concentration increased, acid soluble, alkali soluble Tris-HCl, soluble and water soluble selenium content in selenium respectively of the total amount of 50%-55%, 35%-45%, 30%-35% and 20%-30%. in acidic medium, selenium is easier to transfer the green bean sprouts, so it can be absorbed by human body. By alkali solution and acid isoelectric point method for soluble protein enriched in mung bean sprouts were extracted on alkali concentration solid-liquid ratio, extraction time, effects of extraction temperature and the isoelectric point of the protein extraction rate, the best extraction conditions are: the concentration of 0.20 mol/L, solid-liquid ratio 1:50, extraction time 90 min, extraction temperature of 40 DEG C and isoelectric point of pH was 4.8, the extraction of soluble protein in mung bean under this condition, the precipitated proteins were digested using microwave digestion instrument, and using graphite furnace atomic absorption spectrometry measurement Set the selenium content in protein. The results showed that the selenium content of mung bean protein increases with the concentration of selenium, the highest concentration was 1710 g/kg, the total selenium content in high concentration of 23.5%. can not only increase the selenium content of selenium in protein, and can increase the total protein of selenium in selenium by ratio of.4. PLC-MS method for generation of selenium enriched mung bean samples of cystine, selenomethionine and Se methylselenocysteine in qualitative and quantitative analysis, optimize the cracking voltage and collision energy of each amino acid, choose the best extraction conditions, using MRM mode monitoring. The results showed that Se methylselenocysteine is the main se selenium in mung bean sprouts on behalf of amino acids; and selenomethionine and selenium content of cystine is relatively small, only the total selenium content is below 5%. Therefore, mung bean sprouts can be through the growth and metabolism of inorganic selenium effectively Transforming into organic selenium, as a nutritive, safe and efficient functional selenium supplement, is an effective way to improve the level of selenium in the human body in the low selenium area.

【學位授予單位】：遼寧大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S649;O657.63

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