鹰嘴豆文献中的图片

高效液相色谱法测定19个鹰嘴豆品种中寡糖的含量Abstract

Theextractionprocedureofoligosaccharidesfromchickpeaseedswasoptimizedandsufficientextractionwasachievedwith50%ofethanol-waterinaratioof10:1forsolventtodefattedchickpeamealat50°Cfor30min.Undertheoptimalextractionconditions,thecontentsofoligosaccharidesin19cultivarsofchickpeaseedsweredeterminedbyHPLC.Theresultsshowedthatthemainα-galactooligosaccharides(α-GOS)inchickpeaseedswereraffinose,stachyose,verbascoseandanunknownglycoside,whichwasisolated,purified,andidentifiedasciceritol.Ciceritol,themainsugarinallthechickpeasamples,accountedforabout50%ofthetotalα-GOS.

Therewereconsiderablevariationsinthelevelsofα-GOSandsucrosebetweenspecies,especiallyverbascosewhichcouldonlybedetectedin7samples.Withthehighestamountofα-GOSandalowamountofsucrose,the171wasthebestchoiceforobtainingα-GOSforuseasaprebioticinfunctionalfoods.Keywords:Chickpea(CicerarietinumL)seed;α-Galactooligosaccharide;Sucrose;Optimization;Extractionmethod;HPLC通过实验对鹰嘴豆种子中寡糖的提取进行优化，最终确定了寡糖的提取条件为石油醚脱脂，料液比1：10，提取溶剂为50%乙醇，50℃水浴加热30分钟，并运用HPLC对19个鹰嘴豆品种中低聚糖的含量进行了测定。结果表明：鹰嘴豆种子中主要含有的α-低聚半乳糖有棉子糖（三糖）,水苏糖（四糖），毛蕊花糖（五糖），和一个未知的糖，并对其进行了分离，鉴定，确定为Ciceritol。并且Ciceritol是鹰嘴豆中含量最多的低聚糖，约占α-GOS的50%。α-GOS和蔗糖（二糖）的含量在不同品种间存在较大差异，尤其是毛蕊花糖，只在7个品种中检测到。通过含量测定，找到了19个鹰嘴豆品种中，171鹰嘴豆中α-低聚半乳糖的含量最高，同时蔗糖含量相对较低，是制作功能性食品的最佳选择。Abstract

EthanolconcentrationTempera-tureExtractiontimeRatio(m/v)10%20℃15min1：5230%50℃30min1：10350%70℃60min1：15470%90℃1：20580%

Opitimizationofextractionconditionsforoligosachharides寡糖的提取与制备1.先将种子在85℃下干燥2小时，粉碎过6号筛。2.取500g干粉，用2L石油醚脱脂，得到脱脂鹰嘴豆。3.取脱脂鹰嘴豆，一式三份，每份1g，用10ml50%乙醇50℃温浸3次，每次30min，合并上清，减压浓缩成浸膏。4.浸膏复溶于5.0ml的流动相(乙腈:水=75:25)Fig.1Chromatographicprofileoffreesugarsinextractionofchickpeaseeds.1,Ribose;2,Fructose;3,Sucrose;4,Unknowncompound;5,Maltose;6,Raffinose;7,Ciceritol;8,Stachyose;9,Vebascose.Agilent1100seriesSugar-DColum(4.6*250mm)Ace:Water=75:25Flowrate=1ml/minInjvolume=20ul核糖果糖蔗糖(二糖）未知麦芽糖Ciceritol（三糖）水苏糖（四糖）毛蕊花糖（五糖）棉子糖（三糖）ExtractionandIdentificationofCiceritol

Theextractofchickpeaseedswasloadedontoacolumn(3×50cm)ofcharcoal–Celite(1:1,w/w)andsuccessivelyelutedwithwaterandincreasingconcentrationsofethanol(5–30%).ThefractionswerescreenedbyreactionwithnaphthoresorcinolandthosewithsugarswerefurtherdetectedbyHPLC.Thefractionscontainingciceritolwerecombined,concentratedinvacuo,andpurifiedbygelpermeationonacolumn(1.5×90cm)ofBiogelP-2.Degassedwaterwasusedastheeluentataflowrateof20ml/h.Thefractionscontainingpureciceritol,analyzedbyHPLC,werecombined,concentrated,andfreeze-dried.ThestructurewasconfirmedbyESI-TOF-MS(AppliedBiosystemsmassspectrometer)andNMR(BrukerDRX500)withD2Oasthesolvent.该级分通过间二萘酚反应进行筛选并用HPLC进一步检测。将含ciceritol的馏分进行合并，减压浓缩，并通过BiogelP-2凝胶色谱柱（1.5×90厘米）进行纯化，并以20ml/h的流速用脱气水进行洗脱。通过HPLC分析该级分，合并，浓缩，冷冻干燥得到纯的Ciceritol。其结构经ESI-TOF-MS仪（AppliedBiosystems质谱仪）和NMR（Bruker公司DRX500）进行鉴定。将鹰嘴豆种子的提取物装载到木炭硅藻土柱（3×50厘米）上，并依次用水及浓度不断增加的乙醇（5-30％）进行梯度洗脱。Fig.2Thechemicalstructureofciceritol.α-D-吡喃半乳糖基–(1→6)​-α-D-吡喃半乳糖基–(1→2)-4-O-甲基-手性肌醇O-α-

D-galactopyranosyl-(1→6)-O-α-D-galatopyranosyl-(1→2)-1D-4-O-

methyl-chiro-inositol123456123456123456被甲基化的肌醇（环己六醇）M/Z

541[M+的Na]

+=518α-1,6糖苷键Fig.3Theeffectsofextractionconditionsonthecontentsofα-GOSandsucroseinachickpeaseeds.Concentrationsof