本文選題：大豆 + 老化��； 參考：《南昌大學(xué)》2017年碩士論文

【摘要】：大豆(Glycine max(L.)Merr)是一種富含油脂和蛋白質(zhì)的豆科植物,是世界重要糧食作物之一。然而,大豆種子屬于短命種子,隨著儲藏時間的延長,大豆種子發(fā)生劣變,種子活力下降,發(fā)芽率和發(fā)芽勢降低,從而導(dǎo)致大豆的利用價值和種用價值降低。因此,尋找有效的方法檢測大豆種子活力具有積極的生產(chǎn)實踐意義。本文采用電噴霧萃取電離質(zhì)譜(Extractive electrospray ionization mass spectrometry,EESI-MS)技術(shù)直接檢測不同活力大豆種子水提液,獲得其正離子模式下指紋圖譜,結(jié)合主成分分析(Principal component analysis,PCA)、聚類分析(Cluster analysis,CA)和判別分析(Discriminant analysis,DA)等多變量分析方法對其進(jìn)行區(qū)分,實現(xiàn)不同活力大豆種子的快速評價。同時,采用高效液相色譜(High performance liquid chromatography,HPLC)和EESI-MS技術(shù)分析大豆種子老化過程中酚類化合物的動態(tài)變化。主要研究內(nèi)容及結(jié)果如下:1、采用人工加速老化法對種子進(jìn)行處理,獲得不同活力水平的大豆種子,老化后的種子分別進(jìn)行標(biāo)準(zhǔn)發(fā)芽試驗,以未經(jīng)老化處理的種子為對照,獲得大豆種子老化過程中各項發(fā)芽指標(biāo)和種子活力的變化規(guī)律。結(jié)果表明,未經(jīng)老化處理的大豆種子的發(fā)芽指標(biāo)和活力指數(shù)與經(jīng)老化處理后的大豆種子之間的差異極顯著(P0.01)。隨著老化處理時間的延長,種子的各項發(fā)芽指標(biāo)和活力指數(shù)均呈下降趨勢,老化至第8 d時,種子的各項發(fā)芽指標(biāo)和活力指數(shù)均為0,說明種子活力隨著老化處理時間的延長而下降,直至完全喪失。種子活力指標(biāo)與老化處理時間相關(guān)性分析表明,活力指標(biāo)與老化時間都呈極顯著的負(fù)相關(guān)關(guān)系。2、為了對不同活力的大豆種子進(jìn)行快速鑒別,本文采用EESI-MS,在無需復(fù)雜樣品預(yù)處理前提下,對不同老化處理時間的大豆種子水提液進(jìn)行直接質(zhì)譜分析,獲得EESI-MS指紋圖譜。PCA的前3個主成分因子的累計貢獻(xiàn)率為81.50%。CA和DA的正確率都為100%,對外部驗證樣本進(jìn)行DA,正確率為90%。通過質(zhì)譜鑒定的物質(zhì)有γ-氨基丁酸、谷氨酰胺和乙�；玖夏拒�,實驗結(jié)果表明,隨著大豆種子老化處理時間的延長,γ-氨基丁酸含量逐漸增加,谷氨酰胺與大豆異黃酮中的乙酰基染料木苷含量逐漸減少。EESI-MS結(jié)合多變量分析能對不同活力的大豆種子進(jìn)行有效區(qū)分。3、本文采用HPLC對經(jīng)老化處理過程中大豆種子的19種酚類化合物進(jìn)行了定性定量分析,結(jié)果表明,大豆種子在老化處理過程中19種酚類化合物的含量均發(fā)生了明顯的變化。其中,原兒茶酸、蘆丁和桑黃素的含量隨老化處理時間的增加而減少,分別從183.60μg/g、14.03μg/g和55.03μg/g降至138.10μg/g、12.37μg/g和38.60μg/g;黃豆苷元、黃豆黃素、染料木素和黃芩黃素的含量隨著老化處理時間的增加而增加,分別從12.57μg/g、痕量(Trace;Tr)、9.10μg/g和0.83μg/g增加至91.97μg/g、6.60μg/g、76.10μg/g和3.53μg/g;咖啡酸、表兒茶素、阿魏酸、黃豆苷、染料木苷和白藜蘆醇的含量隨老化處理時間的增加呈先上升后下降的趨勢,其中阿魏酸的含量在老化處理2 d時最大,其余5種酚類化合物的含量均在老化處理4 d時最大;綠原酸、楊梅素、肉桂酸、山奈酚、對-硝基苯甲酸和鷹嘴豆芽素A的含量隨著老化處理時間的增加呈現(xiàn)出不規(guī)則的變化,其中綠原酸、楊梅素和山奈酚的變化趨勢相同,肉桂酸、對-硝基苯甲酸和鷹嘴豆芽素A的變化趨勢一致。19種酚類化合物的總含量隨老化處理時間的增加呈先上升后下降的趨勢,在老化處理第4 d時達(dá)到最大值6830.88μg/g。4、進(jìn)一步采用EESI-MS對不同老化時間的大豆種子醇提取物進(jìn)行直接質(zhì)譜分析,獲得其正離子模式下指紋圖譜,并對四種典型異黃酮進(jìn)行定性定量分析。結(jié)果表明,不同老化處理時間的大豆種子黃豆苷元、染料木素、黃豆苷和染料木苷的含量呈極顯著差異。黃豆苷元和染料木素含量隨老化時間的增加而增加;黃豆苷和染料木苷含量則隨老化時間的增加呈現(xiàn)先上升后下降的趨勢,在老化處理4 d時達(dá)到最大值。此結(jié)果與HPLC實驗結(jié)果中黃豆苷元、染料木素、黃豆苷和染料木苷含量變化趨勢相同。同時,對不同老化處理時間大豆種子的醇提物指紋譜圖進(jìn)行PCA分析,結(jié)果表明,不同活力大豆種子醇提物中的物質(zhì)差異顯著,能夠被明顯區(qū)分在PCA二維空間的不同的區(qū)域。本文采用EESI-MS與HPLC作為研究平臺,對不同活力大豆種子進(jìn)行快速鑒定,并對大豆種子老化過程中酚類物質(zhì)的動態(tài)變化作了較系統(tǒng)的研究,建立了快速、靈敏、有效大豆種子樣品質(zhì)譜檢測新方法,為大豆種子的貯藏及種質(zhì)資源的研究與利用提供理論依據(jù)。
[Abstract]:Glycine max (L.) Merr is a kind of bean plant rich in oil and protein. It is one of the most important food crops in the world. However, soybean seeds belong to the short life seeds. With the prolongation of storage time, the seeds of soybean are deteriorated, the vigor of the seeds descends, the germination rate and the germinating potential are reduced, thus the use value and the value of the soybean are reduced. Therefore, it is of great practical significance to find effective methods to detect the vigor of soybean seed. This paper uses electrospray ionization mass spectrometry (Extractive electrospray ionization mass spectrometry, EESI-MS) to directly detect the water extract of soybean seeds with different vigor, and obtain the fingerprint of its positive ion mode and combine the principal component. Principal component analysis (PCA), cluster analysis (Cluster analysis, CA) and discriminant analysis (Discriminant analysis, DA) are used to distinguish them and realize the rapid evaluation of different vigor soybean seeds. The main research contents and results are as follows: 1, the seeds were treated with artificial accelerated aging method, and the soybean seeds with different vigor were obtained. The aged seeds were tested for standard germination respectively, and the aging of the seeds was compared to the aging of soybean seeds. The results showed that the difference between the germination index and the vigor index of the untreated soybean seeds was very significant (P0.01). The germination index and the vigor index of the seeds decreased with the prolongation of the aging treatment time. When aging to eighth D, the germination index and vigor index of the seeds were 0, indicating that the seed vigor decreased with the prolonging of the aging treatment time, until completely lost. The correlation analysis between the vigor index and the aging time showed that the vigor index and the aging time had a very significant negative correlation of.2, in order to be different vitality. The rapid identification of soybean seeds was made by using EESI-MS. Under the precondition of no complex sample pretreatment, the soybean seed water extract with different aging time was analyzed by direct mass spectrometry. The cumulative contribution rate of the first 3 principal component factors of the EESI-MS fingerprint.PCA was 81.50%.CA and DA, both of which were 100%, and the external verification samples were verified. DA, the correct rate of 90%. was identified by mass spectrometry: gamma aminobutyric acid, glutamine and acetyldyestuside. The experimental results showed that the content of gamma aminobutyric acid increased gradually with the prolongation of aging treatment time of soybean seeds, and the content of glutamine and acetyl dye in soybean isoflavones gradually decreased by.EESI-MS binding. Multivariate analysis can effectively distinguish.3 from soybean seeds with different vitality. In this paper, 19 kinds of phenolic compounds in soybean seeds were analyzed qualitatively and quantitatively by HPLC. The results showed that the content of 19 phenolic compounds in the aging process of soybean seeds all changed obviously. The content of acid, rutin and saoflavin decreased with the increase of aging treatment time, from 183.60 mu g/g, 14.03 mu g/g and 55.03 mu g/g to 138.10 mu g/g, 12.37 mu g/g and 38.60 mu g/g, and the content of daioside, flavin, genistein and baicalein increased with the increase of aging treatment, from 12.57 Mu g/g, trace (Trace; Tr), respectively. 9.10 mu g/g and 0.83 mu g/g increased to 91.97 mu g/g, 6.60 mu g/g, 76.10 mu g/g and 3.53 mu g/g; caffeic acid, epicatechin, ferulic acid, daidzein, geniside and resveratrol increased first and then decreased with the aging treatment time, and ferulic acid content was the largest in aging treatment 2 D, and the other 5 phenolic compounds contained The amount of chlorogenic acid, myricetin, cinnamic acid, kaempferol, and the content of A with chlorogenic acid, myricetin and kaempferol were the same, and the changes of chlorogenic acid, myricetin and kaempferol were the same, cinnamic acid, P - Nitrobenzoic Acid and olecranon A of A were changed. The total content of.19 phenols increased first and then decreased with the increase of aging treatment time, reaching the maximum value of 6830.88 mu g/g.4 at fourth D aging treatment, and further using EESI-MS to analyze the alcohol extracts of soybean seeds with different aging time, and obtain the fingerprint of their positive ion mode. The qualitative and quantitative analysis of four typical isoflavones was carried out. The results showed that the content of Huang Dougan, genistein, geniside and genistein increased with the aging time, while the content of daidzein and genistein increased with the aging time. The trend of increasing first and then decreasing, reaching the maximum when the aging treatment was 4 d. This result was the same as the content of daidzein, genistein, daidzein and genistein in the experimental results of HPLC. At the same time, the fingerprint spectrum of the alcohol extracts of soybean seeds with different aging treatment time was analyzed by PCA. The results showed that the different vitality was great. The substance difference in soybean seed alcohol extracts is significant and can be clearly distinguished in different regions of PCA two-dimensional space. This paper uses EESI-MS and HPLC as the research platform for rapid identification of different vigor soybean seeds, and systematically studies the dynamic transformation of phenolic substances in the aging process of soybean seeds, which is rapid and sensitive. A new method for mass spectrometry of effective soybean seed samples is provided to provide theoretical basis for soybean seed storage and research and utilization of germplasm resources.

【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S565.1

【參考文獻(xiàn)】

中國期刊全文數(shù)據(jù)庫 前10條

1 李水明;何曼文;王勇;;亞胺及其相關(guān)離子在區(qū)分賴氨酸和谷氨酰胺殘基中的應(yīng)用[J];質(zhì)譜學(xué)報;2014年03期

2 歐陽永中;李操;周亞飛;周振;;電噴霧萃取電離質(zhì)譜法用于摻假白酒的快速分析[J];化學(xué)學(xué)報;2013年12期

3 王鳳鵬;曾全榮;葉尚臣;尹真;;基于激光散斑的大豆活力檢測實驗研究[J];應(yīng)用激光;2013年04期

4 Nazim Hussain;Zahra Jabeen;LI Yuan-long;CHEN Ming-xun;LI Zhi-lan;GUO Wan-li;Imran Haider Shamsi;CHEN Xiao-yang;JIANG Li-xi;;Detection of Tocopherol in Oilseed Rape (Brassica napus L.) Using Gas Chromatography with Flame Ionization Detector[J];Journal of Integrative Agriculture;2013年05期

5 黃情;李云霞;魏先杰;吳輝;唐啟源;;種子劣變與修復(fù)[J];種子;2013年04期

6 瞿德敬;王俊英;王俊平;王碩;;HPLC-ESI-MSn法鑒定大豆中12種大豆異黃酮的結(jié)構(gòu)[J];現(xiàn)代食品科技;2013年04期

7 左玉;;多酚類化合物研究進(jìn)展[J];糧食與油脂;2013年04期

8 李月明;郝楠;孫麗惠;張慶芳;葉雨盛;;種子活力測定方法研究進(jìn)展[J];遼寧農(nóng)業(yè)科學(xué);2013年01期

9 郭遠(yuǎn);楊柳;王園清;薛樹鵬;張嬌;崔岳;陳學(xué)珍;謝皓;;大豆種子發(fā)育過程中異黃酮含量的動態(tài)分析[J];農(nóng)學(xué)學(xué)報;2012年11期

10 賈濱;張興磊;丁健樺;楊水平;陳煥文;;電噴霧萃取電離質(zhì)譜技術(shù)及其應(yīng)用進(jìn)展[J];科學(xué)通報;2012年20期

中國碩士學(xué)位論文全文數(shù)據(jù)庫 前2條

1 張燕慧;白沙蒿種子對人工老化的生理生化響應(yīng)研究[D];蘭州大學(xué);2016年

2 王海濤;大豆異黃酮的抑菌活性及其機(jī)制的研究[D];遼寧師范大學(xué);2009年

，

本文編號：1777914