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紫甘薯Z103花色苷粗提液的組分分析Z103紫甘薯花色苷粗提液的液相色譜及總正離子色譜圖分別如圖 及圖 所示:加了這一句圖21 紫甘薯Z103花色苷HPLC的液相色譜圖 HPLC chromatogram of anthocyanin from purple sweet potato Z103圖22 紫甘薯Z103花色苷的總正離子色譜 Total positive iron chromatogram of anthocyanin from purple sweet potato Z103對紫甘薯Z103花色苷提取液的紫外可見光譜掃描顯示,所有供試紫甘薯前后描述不一致花色苷在紫外光區(qū)280315 nm和320340 nm處均有較強的吸收峰。 實驗方法 色譜條件Agilent ZORBAX Eclipse XDBC18液相色譜柱,100 mm, μm。(4) 紫甘薯花色苷的熱穩(wěn)定性和體外抗氧化研究。本課題的目的是以生長于沿海灘涂的耐鹽紫甘薯為原料,采用液質(zhì)聯(lián)用技術(shù)初步分析原料的花色苷組成,在次基礎(chǔ)上采用超聲輔助醇水溶劑提取技術(shù)提取原料中的花色苷,對得到的提取液采用大孔吸附樹脂進行分離純化,對所得到的產(chǎn)物進一步研究其熱穩(wěn)定性和抗氧化性。為此,花色苷給人類帶來了鮮艷的顏色和促進人體健康。在同一項研究中,給大鼠喂食葡萄籽花色苷以后,可在血漿中檢測到花色苷,但在脂蛋白(LDL加VLDL)中不能測得。Facino等(1996)[68]證明了用葡萄籽花色苷100~200g/L灌注心臟,可劑量依賴性地降低局部缺血期心室攣縮,降低冠狀動脈灌注壓改善再灌注時心肌機械功能,增加灌注前及再灌注期間心肌向灌注液釋放6酮PGF1A1,而嚴重缺血時則可抑制心律失常的發(fā)生。Matsui(2002)[63]等用從紫色甘薯的塊根中分離出來的一種二酸基花色苷,芍藥色素3O[2O(6OE阿魏酰βD吡喃葡糖基6OE咖啡酰βD吡喃葡糖苷]5βD吡喃葡糖苷,給約兩月大的雄性小鼠口服2 g/kg麥芽糖和100 mg/kg二?;ㄉ?。Tsuda等(1994)[59]用富含花色苷的葡萄、越桔提取物, mg/ml,處理人類結(jié)腸癌細胞HT29 24 h后,可抑制增殖,并認為花色苷組分影響花色苷生物活性。Yoshimoto等人(1999)[54]用鼠傷害桿菌TA 98(Salmonella typhimurium)為材料,評價了4種甘薯塊根水提取物的抗突變活性。O2應(yīng)該是O2-過量的自由基產(chǎn)生可以超過抗氧化酶如谷胱甘肽過氧化物酶、過氧化氫酶和超氧化物歧化酶及抗氧化物質(zhì)如谷胱甘肽、生育酚或抗壞血酸提供的生理抗氧化能力,造成氧化傷害,是人體疾病的發(fā)生和發(fā)展的誘因。Manhita等人(2006)[47]將此技術(shù)應(yīng)用到了黑醋栗粉以及葡萄、草莓中花青素的提取、純化研究中。膜分離技術(shù)具有電耗低,是傳統(tǒng)蒸發(fā)濃縮的1/10,操作溫度低,產(chǎn)品質(zhì)量好不是一句完整的話。目前純化的方法有:(1) 常用的洗脫純化方法%HCl水溶液洗脫,花青素等酚類物質(zhì)被吸附,糖、酸和其它水溶性物質(zhì)隨鹽酸洗掉,用乙酸乙酯洗脫其它酚類物質(zhì),最后用酸化乙醇或甲醇(%HCl,v/v)洗脫花色苷[912]。④微生物發(fā)酵或酶解法此法是利用微生物或酶的作用將細胞壁成分降解,讓胞內(nèi)的花色苷成分迅速滲透擴散出來,以利于提取。脈沖電場輔助提取法是一種新型非熱技術(shù),它具有提取溫度低、速率快、時間短、提取率高、產(chǎn)品品質(zhì)好的特點。(5) 微波輔助提取( microwave assisted extraction,MAE)微波是頻率介于300 MHz30 GHz(波長在1 cml m,介于紅外和無線電波之間)的電磁波。Corrales等人(2008)[30]采用不同的提取方式對葡萄中花青素的提取效率進行了比較研究,發(fā)現(xiàn)相同條件下,與熱(70℃)提取相比,高壓(600MPa)輔助提取花青素等多酚類的效率可以提高近50%,且其產(chǎn)物的抗氧化活性約為熱浸提物3倍。酸化甲醇不適宜工業(yè)化生產(chǎn)花色苷色素,主要用于花色苷的組成分析和結(jié)構(gòu)鑒定?;ㄉ丈卮嬖谟谥参锛毎囊号葜衃23],被細胞壁、細胞膜包裹,常規(guī)的提取方法為溶劑提取,為提高色素得率,常采用機械破碎、加熱、冷凍、酶制劑(如果膠酶、纖維素酶和蛋白酶等)、超聲波、微波和脈沖電場等技術(shù)破壞細胞壁和細胞膜,提高組織細胞的滲透性,縮短提取時間,提高色素得率,改善產(chǎn)品的質(zhì)量。每一種平衡形式的花色苷含量取決于溶液的pH值和花色苷的結(jié)構(gòu)。(2) 花色苷的光譜特性在酸性溶液中,花色苷及其糖苷配基顯示出兩個最大的特征吸收:一個位于可見光區(qū)的465~550 nm之間,另一個較小的位于紫外275 nm[13,17];%鹽酸的甲醇中,在可見光區(qū)520~560 nm處存在特征吸收峰;在紫外區(qū)270~280 nm處有特征吸收峰,如果花色苷存在?;孔⒁庹Z句,其在310~320 nm處存在特征吸收峰[18]。圖 13 紫甘薯花色苷的結(jié)構(gòu) The structure of anthocyanin in purple sweet potatoHaborne(1955)[13]曾鑒定紫甘薯塊根部分花色苷為帶咖啡酸的矢車菊素3槐糖苷5葡糖苷;Shi等(1966)[14]鑒定出紫甘薯中兩種主要的花色苷為帶三個分子阿魏酸和一分子咖啡酸的芍藥素3二槐糖苷5葡糖苷和含二分子阿魏酸及一分子咖啡酸的芍藥素3槐糖苷5葡糖苷。與花青素以糖苷鍵結(jié)合的主要糖類有,αD葡萄糖,βD葡萄糖,αD半乳糖,αD甘露糖,βD木糖,βL阿拉伯糖,αD鼠李糖以及由這些單糖構(gòu)成的二糖和三糖等[8],其中,3單糖苷、5雙糖苷、3,5二糖苷和3,7二糖苷是最常見的糖苷。 花色苷的一般結(jié)構(gòu)圖 11 花青素的結(jié)構(gòu) The structure of anthocyanidin花色苷是以黃酮核為基礎(chǔ)的一類糖苷,是花青素(配體)與糖相結(jié)合形成的配糖體。)的清除實驗 103 本章小結(jié) 104第六章 結(jié)論與展望 106 結(jié)論 106 展望 108參考文獻 109發(fā)表文章 115致 謝 116第一章 文獻綜述 花色苷的去掉概述花色苷(anthocyanins)是自然界中一類廣泛存在于植物中的水溶性天然色素[1],是花青素嚴格一點的話,應(yīng)該是花色素,花青素多數(shù)情況下是矢車菊素的別名。自由基的清除實驗 93(1)[DPPH scavenging capacity were anthocyanins from purple sweet potato (PSP) tea polyphenols (TP) vitamin E (VE) pigments from red cabbage (RC), with IC50 being mg/mL, mg/mL, mg/mL, mg/mL respectively. For O2ˉ(7) 在四個體外抗氧化性體系研究中,脂質(zhì)體體系, mg/mL的紫甘薯花色苷(%)與同等什么意思?紫甘藍紅色素(%)的抗氧化能力相近,且高于茶多酚(%)與維生素E(%);各抗氧化劑對DPPH%; g/100g,%;對不同提取方法所得的提取液進行LCMS分析比較。耐鹽紫甘薯原料易得、價格低廉,能夠適應(yīng)工業(yè)化生產(chǎn)的需要,紫甘薯花色苷在食品、醫(yī)藥及化妝品等領(lǐng)域具有很大的應(yīng)用前景。主要研究內(nèi)容如下:(1) 紫甘薯花色苷粗提液的HPLCDADESI/MS圖譜分析表明,紫甘薯花色苷主要為被咖啡酸、阿魏酸、對羥基苯甲酸等芳香酸?;氖杠嚲哲蘸蜕炙庈眨蛔细适鞿103含有15種具有花色苷特征的組分,其中有7種是以矢車菊花色素為苷元,8種是以芍藥花色素為苷元;單糖苷的有1種,其他為雙糖苷,形成花色苷的糖苷為葡萄糖苷和槐糖苷;非?;幕ㄉ沼?種,單?;挠?種,雙?;挠?種,?;鶠榭Х人?、阿魏酸、對羥基苯甲酸;紫甘薯Z10Z110和Z103三個品種所含花色苷的種類基本相同,含量不同?;ㄉ战M分在大孔吸附樹脂HPD300上的吸附行為為放熱過程,低溫有利于吸附容量的提高。關(guān)鍵詞:紫甘薯花色苷;組分分析;提?。患兓?;抗氧化性 ABSTRACTSalttolerant purple sweet potato is a Convolvulaceae annual herb, which is planted in the coastal beach. The anthocyanins from purple sweet potato are a class of highquality acylated natural colorants with highcolored capability, watersolubility and better stability. It also has the vital signifieance to treatments and the prevention disease, such as tumor, senile, cardiovascular of humanity. Purple sweet potatoes are so facile and cheap that they can meet the expectation of industry. There are great potential applications of anthocyanins from purple sweet potato in food pharmaceutical and cosmetic industries. With the salttolerant purple sweet potatoes as raw material, the anthocyanins were extracted via conventional solvent method and ultrasonicassisted method respectively, the effect of ultrosonicassistant treatment was studied under the optimal conditions of solvent method. To obtain a high content of anthocyanins and better quality products, macroporous resins were used in purple sweet potato pigments purification. Reversedphase HPLC with electrospray ionization mass spectrometry were also used to rapidly identify the main anthocyaninsins in indifferent varieties of purple sweet potato, extracts with different methods and eluent with gradientelution. At last, the heat stability and antioxidant activity was studied. Details are concluded as follows:(1) Spectrometry analysis of HPLCDADESI/MS for crude extracts showed that most of anthocyanins in salttolerant purple sweet potato are cyanin and peonin acylated by caffeic acid, ferulic acid, phydroxybenzoic acid and other aromatic acid. Purple sweet potato Z103 has 15 constituents of anthocyanins, of which seven cyanin and eight peonin, one glucoside and the other are diglucosidase, three nonacylated, six acylated and six diacylated anthocyanins, acyl were caffeic acid, ferulic acid, phydroxybenzoic. There was no significant difference in the ponents in different purple sweet potato varieties Z103, Z109, Z109, but the contents are different. (2) The results showed that under the condition of pH , ethanol concentration 20% (volume fraction), liquidsolid ratio 1:30 (g:mL), extraction temperature 50℃, and extraction time 120 min, the extraction rate of anthocyanins was % with conventional solvent method , the content of starch in the residue was g/100g and the loss rati