膜分离茶多酚工艺初步研究 论文正文

毕业论文（设计）题目膜分离茶多酚工艺初步研究指导老师专业班级生物技术及应用0701姓名学号2010年5月30日A1A0A2A3A4A5A6A0A7A8A10A11A9A13A12摘要研究膜分离技术制备低咖啡因高表没食子儿茶素没食子酸酯（EGCG）含量茶多酚生产工艺。主要采用高速离心、微滤、超滤、纳滤技术，高效液相色谱检测，结果表明，高速离心具有较好的澄清效果，微孔膜在孔径为5M,压力01MPA，流速为20ML/MIN分离咖啡因去除率023，EGCG损失率018。超滤膜5KDAL（膜单位道尔顿）在压力05MPA，流速7ML/MIN分离咖啡因去除率018，EGCG损失率011；超滤5KDAL跟1KDAL分离咖啡因去除率028，EGCG损失率004。纳滤膜浓缩压力05MPA，流速5ML/MIN咖啡因去除率023，EGCG损失率025。膜的使用及选择结合木质纤维素树脂分离技术，在一定程度上降低咖啡因的含量使其达到标准04，提高EGCG的含量达到标准50。所得产品得率为73，咖啡因的纯度为036，EGCG纯度为5950。关键词膜分离；微滤；超滤；纳滤；咖啡因；EGCGA14A15A16A17A18A19A20A15A21A22A23A24A25A26A27A27目录引言11材料与仪器211实验材料212实验试剂213试验仪器22实验方法321分离液制备3211提取液制备3212提取液离心分离322微孔膜分离323超滤膜分离324树脂分离4241上G7691液G16855G182094242G7621G5191G15925及上G7691G8939G14085425纳滤膜浓缩526分G7524方法5261HPLC分G7524G7477G12265262标准G7366G13459制G13285263茶多酚含量G16757G12651方法53结果与分G7524631标准G7366G13459制G1328632微孔膜分离效果733超滤膜分离效果与G8616较834纳滤膜浓缩效果94G5647结与G16764G1678210G2454G13783G7003G1049811A28A29A30A31A32A33A34A29A35A36A37A38A39A40A41A42A43引言膜分离技术G717121G1002G13438G7380具有G2469G4649G9520力的高G7044技术，分离G17819程G1209选择G5627G17891G17819膜为分离G1183质，在压力G5058浓度G5058G12573G6524G2172力G991,G2419料G1403G13464分选择G5627G3332G17891G17819膜,G1209达到分离、提纯的G11458的1。G11013G1122其G1872有分离、浓缩、纯G2282的G11458的，选择合G17878的膜与G6817G1328G2454G6980，G2499得到较高的G3250G6922率，G3800G10714G13007G13491G2499G4506G19393G5502G10627，G19462G8502G3818G7481G8757G7591，G993G3818G2164G2282G4410G10301质，G17891G17819液G2499G5502G10627使用，降低G6116G7424，G1955G4581G4557G10627G3671的G8757G7591。在G2520G12193工G1006生产G1025得到G5203G8879的G5224用。G5062G5332G2469G5224用的膜分离技术有G2465G9195G17891RO、纳滤NF、超滤UF、微滤MFG3247G121932。G3835多G6980膜分离G17819程G1025G10301质G993G2469生相G2476G2282,分离G13007G6980较G3835,G6817G1328G9213度在G4472G9213G5050G2503。微滤用G1122G5760G9026液（G12902子G12902径为0110M）的G17819滤；超滤G1209低G14033G13803、G7092G9921效G5224、G7092相G2476G2282、结G7512G12628单、G2356G3332G4579、G6817G1328G12628G1427、使用的压力低、产G8712量较G3835G7143G1122实G10628G14270G2172G2282的G10317G9869G13792G5203G8879G3332G5224用G1122多G12193G10301质的分离、浓缩、G1940G2282G12573G20058G3507；纳滤膜G1183G1122超滤与G2465G9195G17891G1055G19400，G14033G6142G11053分子量为3001000的G4579分子G10301质，其G19610浓缩与G17891G7524为一G1319，G2499使溶质的损失达到G7380G4579，G2465G9195G17891膜G14033将G8712与低分子量G10301质分离，其典型G5224用G7171海G8712G14085盐34。将膜分离技术G5224用G1122茶多酚制品的生产工艺G1025具有深远意义。G11013G1122浸提G17819程G1025的富G19610，一般茶提取G10301G1025咖啡因的含量都远远高G1122鲜叶，将茶提取G10301G1328为功G14033G6116分在食品和药G10301上利用时，G14085去咖啡因G7171提高产品附G2164值的前提基础G7477G122656。G11458前膜分离浓缩茶多酚的研究报道较G4581，浙江一企G1006需要咖啡因低G112204,EGCG高G112250，在G1055前的研究G1025，咖啡因的含量061，EGCG的含量4550达G993到所需要求。因此膜的使用研究G2476的尤为重要，G7424G7003就G4557微孔膜、超滤膜分离，纳滤浓缩G1328了一些研究，膜使用让结果达到了所需要求，并G14033扩G3835G1025试生产。A28A29A30A31A32A33A34A29A35A36A37A38A39A40A41A44A431材料与仪器11实验材料茶叶（炒青G3835宗茶沫，购G1122金华浦江）表没食子儿茶素没食子酸酯EGCG（实验G4472提供）12实验试剂95乙醇（食用级），柠檬酸、酒石酸亚铁、磷酸氢钠、磷酸二氢钾G12573均为分G7524纯试剂，乙醇（色谱纯），冰乙酸，G8712为蒸馏G8712。13实验仪器核酸蛋白G14270G2172测定仪（SHQT6A上海琪G10317分G7524仪器有限公司）；N2000色谱G6980据工G1328站（浙江G3835G4410智达信息工程有限公司）；分光光度G16757（752N上海精G4506科G4410仪器有限公司）；分G7524天G5191（SPS202F梅G10317勒托利多称重设备G13007G13491有限公司）；喷雾干燥器（SY6000上海G1002远生G10301设备工程有限公司）；冷冻干燥仪（12LD德国ALPHA）；超声波清G8939器（KQ500DA昆山市超声仪器有限公司）；层G7524G7621（1640MM，40800MM上海亚荣生G2282仪器厂）；旋转蒸G2469仪（RE52AA上海亚荣生G2282仪器厂）；真空泵（SHBIII郑州长城科工贸有限公司）；微孔G17819滤膜上海市G7044亚G1940G2282器G1226厂；超滤膜G4579试G5191板超滤膜设备FLOWMEM0250厦门G1002达膜科技有限公司；卷式膜G4579试设备（RNF0460厦门G1002达膜科技有限公司）；高速离心机DL5型上海安亭科G4410仪器厂。A45A46A47A48A49A50A51A46A52A53A54A55A56A57A58A59A602方法21分离液的制备211提取液制备称取10000G绿茶沫，G1209112G8616例G216430酒精浓度的溶剂，65G8712浴超声G2164G9921提取，30MIN每15MING15842MIN，G13445G5079G17819滤提取液、重G3809一G8437，合并滤液，G2375得提取液17L。212提取液离心分离得到提取液，高速离心机离心，离心料液G186117L，转速10000转/MIN，流速10M3/H，压力05MPA，流速5L/MIN，所得澄清溶液16LG5050G2503，取16LG17839G15904分离G2375为G5465分离的分离液。22微孔膜分离微滤膜主要G5224用G1122G6142G11053G20075G12902G10301,液G1319的澄清G1209及G3835G18108分G13466G14752的去除,并G1328为超滤G17819程的前G3800G10714。取分离液16L溶液G17902G17819孔径为5M滤G14467，压力为01MPA，流速20ML/MIN，G6922G19610滤液，G8712G8939浓缩液，G1863机G2530G6922G19610G13007G13491G8543G11053液，G6289取G17891G17819液和G6142G11053液的溶液G17819滤分G2047取滤液G1582检测分G7524。试验工艺G328221G3926G991G726图21微孔膜试验流程示意图23超滤膜分离超滤膜G14033G6142G11053分子量在上G2327G14279G6980G2325G987的G3835分子,除G14033G4448G6116微滤的除G20075G12902、除G14752和澄清G1328用G3818,G17836G14033除去微滤膜G993G14033G1998去的G11161G14752、G9921G9316、G14026G1319和蛋白质G12573G3835分子G22821G2419料液G81452G17767液泵3压力表4微滤G13464G12265G5502G10627G19412G3A61A62A63A64A65A66A67A62A68A69A70A71A72A73A74A75A76合G10301。超滤G1209G1852G3250流方式G17839G15904，在超滤时一般G5785G1929G991，膜压力为05MPA，流速7ML/MING13475微孔膜G17819G2530取15LG17891G17819液G1820G17902G17819G6142G11053分子量5KDAL的膜G20293超滤，G1889G17902G17819G6142G11053分子量1KDAL的膜G20293超滤。G5332机G17839料液并G6922G19610滤液，G8712G8939浓缩液，G1863机G2530G6922G19610G13007G13491G8543G11053液。G6289取G17891G17819液和浓缩液的溶液G17819滤分G2047取滤液G1582检测分G7524。试验工艺G328222G3926G991G726图22超滤、纳滤试验流程示意图24树脂分离241上样液调配将精滤液G6365G8616例G16855G18209G6116PH4，酒精度为10的绿茶溶液，为上G7691液。242柱平衡及上样洗脱G7621G5191G15925G726用10酒精，柠檬酸G16855PH4G5191G15925树脂G7621，使G6984G1022树脂G7621G1319G13007G3800G1122酒精度为10，PH4。上G7691量取500ML，上G7691G726G1209V15L/MIN流速上G7691，上G7691G4448G6116G2530G19757G1363410MIN。10G8939G14085G726G12095BV（层G7524G7621G5214程）G1319G12227的10酒精PH4G8939G14085，流速G6523制在16L/MINV18L/MIN。40G8939G14085G726G12093BVG1319G12227的40酒精G8939G14085，流速G6523制在17L/MINV19L/MIN。70G8939G14085G726G12092BVG1319G12227的70酒精G8939G14085，流速G6523制在17L/MINV19L/MIN。1G2419料液G81452G17767液泵3压力表4G2465G9195G17891（超滤、纳滤G13464G1226）5流量G167576G5502G10627G194127浓液G194128流量G16757G19412G3A61A62A63A64A65A66A67A62A68A69A70A71A72A73A74A77A7625纳滤膜浓缩采用NF200卷式纳滤G13464G1226，在压力050MPA、流速5ML/MING4472G9213G7477G1226G991G6817G1328G17828G15904，试验流程G3926G32822所G12046。取40G8939G14085液G17839G15904纳滤G6817G1328，G5332机G17839料液并G6922G19610浓缩液，G1863机G2530G6922G19610并G8939G1998G13007G13491G8543G11053浓缩液，浓缩液分G2047G1582检测分G7524。浓缩液G2375得茶多酚G6116品。26分析方法261HPLC分析条件1）流G2172相G18209制G726A相G72605冰G18271酸3乙G14108G8712定G4493B相G72605冰G18271酸30乙G14108G8712定G44932）G6152安PH680高效液相色谱仪，色谱G7621G726C18G7621250MM46MM,5UM，V1ML/MIN，波长280NM，G17839G7691量20UL，G7621G921330，检测器G726UVD170UG13055G3818检测器流G2172相G2476G2282G3926表21G726表21HPLC流动相配比随时间变化表检测时G19400（MIN）A相B相045MIN10004550MIN01005060MIN1000262标准曲线制作G18209制标准溶液G726准G11842称取表没食子儿茶素没食子酸酯（EGCG）0250G,准G11842称取咖啡因0125G,G18209G6116G8609液5MG/ML,G18209G6116EGCG为05，1，15，2，25MG/ML的浓度，用G1122制G1328标准G7366G13459。263茶多酚含量计算方法茶多酚得率G16757G12651G13475验公式G3926G991G726AA1315N1V1/A0315N0V0公式21公式G1025G2476量含义A得率；A0G726G2419液茶多酚检测G2572光度；A78A79A80A81A82A83A84A79A85A86A87A88A89A90A91A92A93A1G726茶多酚检测G2572光度；315G726G13475验值（企G1006检测G13475验值）；N0G726茶多酚检测G17819程G2419液G12244G18334的G1505G6980；N1G726茶多酚检测G17819程G12544二G8437G12244G18334的G1505G6980；V0G726G2419溶液G1319G12227；V1G726G12544二G8437检测溶液G1319G12227；3结果与分析31标准曲线的制作将0、05、1、15、2，25MG/ML6G1022G993G2528浓度的EGCG和咖啡因标准品，G6365G10043261检测方法，得G7691品浓度与G7691品G1998G4804G19766G12227相G5224值G1393G8437为G7260、185938、376523、531483、7143、856011（EGCG）,0、192747、371133、501032、650444、767510（咖啡因）；将此浓度与G1998G4804G12227分G19766G12227值在EXCELG1025G17839G15904G13459G5627G3250G5414，制得标准G7366G13459G3926G328231、32所G12046G726A94A95A96A95A97A98A99A100A101A102A103A104A104A105A106A107A108A109A107A104A105A106A103A110A111A112A102A106A105A113A113A114A106A115A106A106A104A106A106A116A106A106A114A106A106A107A106A106A106A106A107A115A103A117A118A119A120A121A122A120A123A124A125A126A127A128A129A130A131A132A133A134A129A130A131A124图31EGCG标准曲线图A135A136A137A138A139A140A141A136A142A143A144A145A146A147A148A149A150A151A152A153A154A155A156A157A158A159A160A161A162A163A164A165A166A167A160A168A163A160A169A169A170A171A159A161A163A172A172A168A168A161A168A161A161A169A161A161A173A161A161A174A161A161A164A161A161A161A161A164A168A160A175A176A177A178A179A180A178A181A182A183A184A185A186A187A188A164A157A189A190A187A188A164A182图32咖啡因标准曲线图G11013G328231、32G2499得EGCG标准G7366G13459为Y34401X14035，R20998咖啡因标准G7366G13459为Y30517X32344，R209922。32微孔分离效果微孔膜分离取分离G2419液16LG17902G17819孔径为5M滤G14467，压力为01MPA，流速20ML/MIN，G6922G19610G6142G11053液2L、G17891G17819液14L检测结果G3926G328233所G12046G726A191A192A193A194A195A196A193A197A191A195A195A194A196A195A196A196A191A195A195A194A193A198A197A199A191A195A200A194A195A200A201A193A191A195A200A194A197A199A193A192A201A191A195A197A194A197A202A202A192A192A191A203A204A205A206A207A208A209A210A211A204A208A205A207A212A205A213A212A204A208A208A207A209A210A203A209A204A208A212A207A209A208A205A213A204A208A209A207A209A208A206A214A204A203A203A207A210A213A206A204A203A211A215A216A217A218A219A220A221A222A223A216A220A217A219A223A223A222A222A216A220A220A219A222A215A223A221A216A220A223A219A222A223A222A224A216A220A222A219A222A222A221A218A216A220A224A219A218A217A220A217A225A216A226A220A219A226A225A222A217A217A216A226A226A219A217A220A222A217A220A216A226A215A217A226A216图33微孔膜分离原液、微孔截留液、微孔透过液液相检测分析图A227A228A2291A230A231A232A233A234A235A2362A230EGCGA234A235A236A237A238A239HPLCA240A241A242A243A244A245A246A247结合茶多酚含量检测与G328233检测分G7524结果，得G1998分离液16LG12244G183345G1505A0649茶多酚含量227，咖啡因含量1643、EGCG含量1903。微孔G6142G11053液2LG12244G20G3G21G3G20G3G21G3G20G3G21G3G2419液G3G6142G11053液G3G17891G17819液G3A248A249A250A251A252A253A254A249A255A1A10A11A0A13A3A12A5G183345G1505A10607茶多酚含量2123。咖啡因含量1050、EGCG含量1033。微孔G17891G17819液14LG12244G183345G1505A20650茶多酚含量2273，咖啡因含量1259、EGCG含量1554，茶多酚得率8763。G11013G328233得G1998，微孔膜的使用降低咖啡因含量，G2528时EGCG含量G1075相G5224的G1955G4581了。32超滤分离效果及比较超滤膜分离膜压力为05MPA，流速7ML/MING13475微孔膜G17819G2530取14LG17891G17819液结果检测G3926G328234G991G726A2A4A6A4A2A7A6A4A8A4A6A4A8A7A6A7A4A4A4A9A14A15A2A16A2A2A6A17A4A18A8A16A2A7A6A19A19A20A21A16A2A19A6A17A18A8A7A16A22A23A23A23A24A25A26A27A28A29A30A31A32A23A32A22A28A33A29A31A22A34A29A35A28A33A33A31A35A34A29A34A28A33A22A31A35A34A22A36A28A33A35A31A35A36A23A30A28A32A27A29A23A28A37A38A39A40A39A41A40A42A38A43A44A43A45A41A39A40A38A46A43A40A47A41A39A39A38A45A46A45A37A41A39A48A38A45A43A44A42A41A39A46A38A45A48A39A40A44A41A39A45A38A45A47A42A40A40A41A43A39A38A39A42A42A40A39A41A43A43A38A44A42A42A40A43A41A43A48A40A48A41A49A50A51A52A53A54A55A56A57A50A54A54A53A58A58A57A59A50A54A59A53A58A49A58A58A50A54A58A53A58A58A49A55A50图34超滤膜过分子量5KDAL、1KDAL截留液、微孔透过液液相检测分析图G1186G328234G1025得G19985KDALG6142G11053液2LG12244G183345G1505A30650茶多酚含量227。咖啡因含量1290，EGCG含量1035。5KDALG17891G17819液12LG12244G183345G1505A40400茶多酚含量1399，咖啡因含量1034，EGCG含量1377。茶多酚得率5275。1KDALG6142G11053液2LG12244G183345G1505A50485茶多酚含量1696咖啡因含量1098，EGCG含量1160。1KDALG17891G17819液12LG12244G183345G1505A60358茶多酚含量1347，咖啡因含量908，EGCG含量1491。茶多酚得率4721。G1186上G17860G3282谱G2499G1209G11487G1998，G993G12661G7171G178195KDAL膜G17836G71711KDAL膜G6142G11053液G1025咖啡因含量都较高，G16840明二G12193膜孔径都G14033有效的去除一定的咖啡因，G1306EGCG含量G1075有所降低。G20G3G21G3G20G3G21G3G20G3G21G3G20G3G21G3G24G78G71G68G79G6142G11053液G3G24G78G71G68G79G17891G17819液G3G20G78G71G68G79G6142G11053液G3G20G78G71G68G79G17891G17819液G3A60A61A62A63A64A65A66A61A67A68A69A70A71A72A73A74A7533纳滤膜浓缩效果将40G8939G14085液溶液G17902G17819蒸馏法检测含G3278率，G2528时结合高效液相色谱法测定咖啡因、EGCG含量得G726咖啡因含量0607、EGCG含量6077，检测HPLCG3282谱G3926G328235所G12046G726A76A77A76A78A77A76A79A76A77A76A79A78A77A76A80A76A77A76A80A78A77A76A81A76A77A76A81A78A77A76A82A76A77A76A82A78A77A76A78A76A77A76A78A78A77A76A83A76A77A76A84A78A76A76A76A78A76A76A79A85A76A76A76A79A85A78A76A76A80A85A76A76A76A80A85A78A76A76A81A85A76A76A76A86A87A88A89A90A91A79A80A82A76A92A93A94A95A96A97A98A97A99A100A98A79A101A102A96A101A103A104A79A84A80A79A77A76A76A83A80A84A80A79A77A82A79A81A81A84A80A80A77A78A105A76A82A84A80A81A77A81A83A80A78A84A80A82A77A76A80A106A83A84A80A105A77A81A76A79A105A84A80A105A77A78A83A82A107A108A109A107A110A111A112A113A112A108A109A107A110A114A115A112A116A117A108A118A116A110A117A107A114A116A116A108A118A116A110A115A117A112A116A109A108A118A109A110A115A107A107A116A118A108A118A118A110A114A118A114A116A115A108A118A111A110A117A117A109A116A113A108A118A114A110A118A107A111A116A111A108A113A118A110A116A109A117A119A120A121A122A109A107A117A123A124图3540乙醇洗脱液HPLC检测图谱将40G8939G14085液纳滤浓缩，G6984G1022浓缩时G19400G17340度为24G4579时，浓缩冷冻干燥G2530测得G726茶多酚得率为73，其G1025咖啡因含量047、EGCG含量4563，检测HPLCG3282谱G3926G328236所G12046G726A125A126A125A127A126A125A128A125A126A125A128A127A126A125A129A125A126A125A129A127A126A125A130A125A126A125A130A127A126A125A131A125A126A125A131A127A126A125A127A125A126A125A127A127A126A125A132A125A126A125A133A127A125A125A125A127A125A125A128A134A125A125A125A128A134A127A125A125A129A134A125A125A125A129A134A127A125A125A135A136A137A138A139A140A128A141A125A126A131A142A143A144A145A146A145A147A148A146A128A142A149A144A142A150A151A128A133A128A152A126A130A130A141A129A133A129A125A126A153A129A132A130A133A129A130A126A125A125A125A131A133A129A130A126A152A131A125A127A133A129A132A126A152A129A130A132A133A129A153A126A131A125A132A153A133A129A141A126A127A127A153A141A133A129A152A126A153A141A152A152A133A130A128A126A129A152A131A128A125A133A130A130A126A153A152A153A128A128A133A130A131A126A130A152A129A128A129A133A130A127A126A152A153A131A128A130A133A130A132A126A127A128A127A128A131A133A130A141A126A125A153A130A128A127A133A130A141A126A141A129A141A128A132A133A131A129A126A127A132A128A154A145A155A156A129A141A125A151A142图3640乙醇洗脱液干燥物HPLC检测图谱G1186G328235、36G8616较得G1998纳滤G14033有效去除咖啡因，EGCG含量G1075降低（G11013G1122纳滤时G19400长，24G4579时），G2499G14033EGCG有G18108分G15999G8699G2282G2419因G17908G6116，G1306在生产G1025采用G3835型的纳滤设备，浓缩时G19400G3835G3835G1955G4581，G17837G1022G2476G2282G2499G1209G18003G1825，G17837G1075G8503G7171G991G8505在企G1006需要验G16789与G16311G1927的技术G1863G19202。4总结与讨论A157A158A159A160A161A162A163A158A164A165A166A167A168A169A170A171A172A173G7424G7003主要研究膜分离技术制备低咖啡因高EGCG茶多酚工艺研究，研究表明微孔膜孔径为5M滤G14467,压力01MPA，流速为20ML/MIN分离咖啡因去除率023，EGCG损失率018。超滤膜5KDAL（膜单位道尔顿）在压力05MPA，流速7ML/MIN分离咖啡因去除率018，EGCG损失率011；超滤5KDAL跟1KDAL分离咖啡因去除率028，EGCG损失率004。纳滤膜浓缩压力05MPA，流速5ML/MIN咖啡因去除率023，EGCG损失率025。G1186G17837些G6980据G11487G1998，微孔膜、超滤膜5KDAL、1KDALG2528时使用及纳滤膜咖啡因去除率相G4557较高，G13783G15397G13475G8994G6116G7424，G1177G13783G15397微滤膜跟纳滤膜的使用，研究G2469G10628微滤膜、纳滤膜的使用，所得产品得率73，咖啡因的纯度为036，EGCG纯度为5950。微滤膜用G1122茶多酚浸提液的G2033G8505分离，G17908G6116一G18108分功G14033G6116分的损失，G2499研究孔径G7368G4579的微滤膜G17839G15904分离。超滤膜G2499有效G3332澄清茶多酚浸提液，G3278G1319G7446质去除率高并G6142G11053了G3835G18108分的果G14026、G2499溶G5627蛋白质、多G12970G12573G3835分子G10301质，G13792使G13489G3835多G6980的茶多酚G17891G17819。为G1955G4579超滤膜G4557茶多酚的G6142G11053率，G6817G1328G17819程G1025在G1457G16789除G7446效果的基础上G2499G17878G5415G3698G3835膜的表G19766流速78。在G1457G16789茶多酚G17891G17819率的基础上G1075G2499G1378