RP-HPLC法制备高纯度天然辣椒红色素的研究
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摘要
辣椒红色素是从茄科植物辣椒(Capsicum annuum L.)中提取出的一类含有多种成分的脂溶性四萜类化合物。作为一种天然的食用色素,它不仅着色力强,安全无毒,而且具有很高的营养价值,是世界卫生组织和许多国家允许使用的天然色素之一。并广泛应用于食品、医药、化妆品等行业中。
现在市场上的辣椒红色素产品大多是粗品,辣椒红色素的纯度低。含有辣椒碱,使辣椒红色素含有辣味,限制了辣椒红色素在食品等行业的应用,另外辣椒红色素是混合物,含有成分的不确定性,增加了辣椒红色素的安全隐患。
1本论文首先对用于分离辣椒红色素的分离介质进行了研究。在研究中以硅胶为原料,键合十八烷基三甲基氯硅烷,合成C_(18)分离介质,并对合成的分离介质进行红外光谱表征。用合成的C_(18)作为分离介质进行装填制备柱。
2对皂化辣椒红色素粗品的温度、碱浓度、醇浓度、用量、时间、皂化次数进行了研究。最后得到皂化辣椒红色素粗品的条件为:采用10%的氢氧化钠甲醇溶液(甲醇浓度为50%),20mL的皂化溶液皂化1mL的色素,在50℃下,每次2h,皂化3次。
3对皂化后辣椒红色素产物进行RP—HPLC法制备分离。考察了制备色谱的洗脱剂配比、流速、检测波长等因素。得到制备条件为:制备柱(C18,φ300mm×4.6mm,10μm);洗脱剂V(甲醇-水=95-5):V(乙酸乙酯)=85:15;检测波长460nm:流速1.0mL/mim;进样量20μL。基于此最后所得到的辣椒红素、β-胡萝卜素产品的纯度均达到了97%以上。
4在实验室级制备高纯度辣椒红色素数据的基础上,进行了工业级放大设计,得到了放大设计的相关参数:放大后工业级设计的洗脱剂流速、上样量、压力及制备塔所需填料的量等,最后对工业级放大设计后的成本和辣椒红色素的产出效益进行了核算和经济可行性分析。通过参数的计算和经济可行性分析,为今后工业化RP—HPLC法制备高纯度辣椒红色素提供了参考。
The hot pepper erythrophyll, extracted from Capsicum annuum L., contains many kinds of terpene compounds soluble in grease Capsanthin is allowed and encouraged to be used as one of the medical natural pigments by World Health Organization and some developed countries. T As one kind of edible natural pigment, the hot pepper erythrophyll has the tinting strength strong, safe non-toxic and the very strong nutritional value . Widely applies in the filed of food, medicine, cosmetics and so on.
Nowadays, the hot pepper product mostly is thick in the market. The purity of the hot pepper erythrophyll is low , and contains the capsicine, that make the hot pepper erythrophyll contain piquancy, Has limited the hot pepper erythrophyll applications in food and so on . Moreover the hot pepper erythrophyll purity is low, ie mixture,and includes the ingredient uncertainty, that increased the security hidden danger.
1 At first,this thesis has studied the separation medium which separated hot pepper erythrophyll .In the research take the silica gel as raw material, responded with the octadecyl three methyl chloride silicon hydride, synthesizes C18 to separate the medium, and carries on the infrared spectrum attribute to the synthesis separation medium. Carries on the filling preparation column with synthesis C18 as the separation medium .
2 Has studied the saponified hot pepper erythrophyll thick's temperature, alkaline solution density, the mellow density, tSaponified fluid amount used, saponified time, saponified number of times . Saponified condition: Be treated with 10% methanol sodium hydroxidie solution (the density of methanol is 50%), Add the saponification solution in the ratio of 1g pigment 20mL, at 50℃, 2h each time, 3 times .
3 After saponifying the raw materail then carries on the RP-HPLC preparation separation. Inspected preparation factors and so on chromatograph eluent allocated proportion, speed of flow, examination wave length. Preparation chromatograph purification condition : Reversed-phase column(C_(18),300mm×4.6mm,φ10μm), Chromatograms were monitored at 450nm; the mobile phase was methanol water ethyl acetate(80.75: 4.25:15, v/v); the flow rate was 1mL/min;injection volume 20μL. Finally obtains Capsanthin element andβ-carrot element pure purification has respectively achieved more than 97%.
4 Based on the data of prepares the high-purity hot pepper erythrophyll in the laboratory level, Has carried on the technical grade enlargement design variable computation, After obtained the enlargement technical grade design correlation parameter. And has carried on the economical feasibility analysis after the technical grade enlargement design cost. Through the parameter computation and economical feasibility analysis ,Will prepare the high-purity hot pepper erythrophyll for the RP-HPLC industrialization to provide the reference
现在市场上的辣椒红色素产品大多是粗品,辣椒红色素的纯度低。含有辣椒碱,使辣椒红色素含有辣味,限制了辣椒红色素在食品等行业的应用,另外辣椒红色素是混合物,含有成分的不确定性,增加了辣椒红色素的安全隐患。
1本论文首先对用于分离辣椒红色素的分离介质进行了研究。在研究中以硅胶为原料,键合十八烷基三甲基氯硅烷,合成C_(18)分离介质,并对合成的分离介质进行红外光谱表征。用合成的C_(18)作为分离介质进行装填制备柱。
2对皂化辣椒红色素粗品的温度、碱浓度、醇浓度、用量、时间、皂化次数进行了研究。最后得到皂化辣椒红色素粗品的条件为:采用10%的氢氧化钠甲醇溶液(甲醇浓度为50%),20mL的皂化溶液皂化1mL的色素,在50℃下,每次2h,皂化3次。
3对皂化后辣椒红色素产物进行RP—HPLC法制备分离。考察了制备色谱的洗脱剂配比、流速、检测波长等因素。得到制备条件为:制备柱(C18,φ300mm×4.6mm,10μm);洗脱剂V(甲醇-水=95-5):V(乙酸乙酯)=85:15;检测波长460nm:流速1.0mL/mim;进样量20μL。基于此最后所得到的辣椒红素、β-胡萝卜素产品的纯度均达到了97%以上。
4在实验室级制备高纯度辣椒红色素数据的基础上,进行了工业级放大设计,得到了放大设计的相关参数:放大后工业级设计的洗脱剂流速、上样量、压力及制备塔所需填料的量等,最后对工业级放大设计后的成本和辣椒红色素的产出效益进行了核算和经济可行性分析。通过参数的计算和经济可行性分析,为今后工业化RP—HPLC法制备高纯度辣椒红色素提供了参考。
The hot pepper erythrophyll, extracted from Capsicum annuum L., contains many kinds of terpene compounds soluble in grease Capsanthin is allowed and encouraged to be used as one of the medical natural pigments by World Health Organization and some developed countries. T As one kind of edible natural pigment, the hot pepper erythrophyll has the tinting strength strong, safe non-toxic and the very strong nutritional value . Widely applies in the filed of food, medicine, cosmetics and so on.
Nowadays, the hot pepper product mostly is thick in the market. The purity of the hot pepper erythrophyll is low , and contains the capsicine, that make the hot pepper erythrophyll contain piquancy, Has limited the hot pepper erythrophyll applications in food and so on . Moreover the hot pepper erythrophyll purity is low, ie mixture,and includes the ingredient uncertainty, that increased the security hidden danger.
1 At first,this thesis has studied the separation medium which separated hot pepper erythrophyll .In the research take the silica gel as raw material, responded with the octadecyl three methyl chloride silicon hydride, synthesizes C18 to separate the medium, and carries on the infrared spectrum attribute to the synthesis separation medium. Carries on the filling preparation column with synthesis C18 as the separation medium .
2 Has studied the saponified hot pepper erythrophyll thick's temperature, alkaline solution density, the mellow density, tSaponified fluid amount used, saponified time, saponified number of times . Saponified condition: Be treated with 10% methanol sodium hydroxidie solution (the density of methanol is 50%), Add the saponification solution in the ratio of 1g pigment 20mL, at 50℃, 2h each time, 3 times .
3 After saponifying the raw materail then carries on the RP-HPLC preparation separation. Inspected preparation factors and so on chromatograph eluent allocated proportion, speed of flow, examination wave length. Preparation chromatograph purification condition : Reversed-phase column(C_(18),300mm×4.6mm,φ10μm), Chromatograms were monitored at 450nm; the mobile phase was methanol water ethyl acetate(80.75: 4.25:15, v/v); the flow rate was 1mL/min;injection volume 20μL. Finally obtains Capsanthin element andβ-carrot element pure purification has respectively achieved more than 97%.
4 Based on the data of prepares the high-purity hot pepper erythrophyll in the laboratory level, Has carried on the technical grade enlargement design variable computation, After obtained the enlargement technical grade design correlation parameter. And has carried on the economical feasibility analysis after the technical grade enlargement design cost. Through the parameter computation and economical feasibility analysis ,Will prepare the high-purity hot pepper erythrophyll for the RP-HPLC industrialization to provide the reference
引文
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[4]程绍玲.辣椒的分析与综合利用[J].粮食加工,2002,(4):52-55.
[5]张继民,胡林华.辣椒色素提取工艺及稳定性[J].安徽机电学院学报,1999,14(1):21-25.
[6]张亚军.干红辣椒深度开发综合加工价值高[J].市场博览,43.
[7]陈洁,陈庶来,陆道礼,等.超临界C02萃取辣椒红色素的研究初探[J].农业工程学报,1998,9(2):89-94.
[8]李涛.天然类胡萝卜素(叶黄素)的分离纯化与分子修饰[M].2002
[9]Btitton G,Structurue and Properties of Carotenoids in Relation to Function.FASEB J.,1995,(9):1551-1558
[10]B.H.Davies,S.Matthew,J.T.O.Kint.The nature and Biosynthesis of the carotenoids of Different Color Varieties of Capsicum annuum.Phytochemisty,1970,9:797-805
[11]B.Camara,R.Moneger.Free and esterified Carotenoids in gree and red fruits of Capsicum annuum.Phytochemisty,1987,(17):91-93.
[12]Zechmeister L,Cholonoky L.Paprika coloringmatters.I Ann Chen.,1927,454(1):54-71.
[13]Simpson K.L.Relative Value of Carotenoids as Precursors of Vitamin A.Proc.Nutr.Soc,1983,42:7-17.
[14]M.Isabel.Minguez-Mosquera,Damaso,Homero-Mendez.Formation and Transformation of Pigments during the Fruit Ripening of capsicum annuum Cv.Bloa and Agridulce J.Agdc.Food Sci.,1971,(36):38-44.
[15]T.Philip,W.W.Nawar,F.J.Francis,The nature of Fatty Acids and Capsanthin Esters in Paprika J.food Sci.,1971,(36):98-100.
[16]M.I.Minguez-Mosquera,D.Homero-Mendez.Changes in Carotenoid Esterification during the Fruit Ripening of Capsanthin annuum Cv.Bola.J.Agric.Food Chem.,1994b,(42):640-644.
[17]谢碧霞,杜红岩.绿色食品开发利用[M].中国中医药出版社,2003.
[18]Pierre P.Mouly,Emile M.GAydou,Josiane Corsetti。Characterization of Paprika(Capsicum annuum)Extract in Orang Juices by Liquid Chromatography of Carotenoid Profiles.J.Agric.Food Chem.,1999,(47):968-976.
[19]韦来红,刘怀珍,陆应林等.辣椒中辣椒红色素与蛋黄着色功能的初探.中国畜产与食品。 1998.6:263-264.
[20]刘成梅,游海主编.天然产物有效成分的分离与应用.化学工业出版社.2003:459-467.
[21]施飞群.红辣椒中辣椒素和辣椒红素的提取[J].化学世界,1994(3):158-160.
[22]陈里,丁来欣,包训祥.辣椒红素提取工艺的研究[J].南京理工大学学报.1995,19(3):255-257.
[23]贺文智,索全伶,王宏伟,等.辣椒红色素提取精制方法[J].内蒙古工业大学报,2001,20(1):10-13.
[24]张继民,胡林华.辣椒色素提取工艺及稳定性,安徽机电学院学报,1999,14(1):21-25.
[25]蔡立志,朱文利.天然色素辣椒红提取工艺研究,中国粮油学报,1994,9(2):23-25.
[26]姚祖风,姜洪杰,张世文.关于辣椒红色素的研究[J].吉首大学学报(自然科学版),2000,21f(2):48-52.
[27]郭湘泗.从辣椒中提取红色素的方法[J].CN 1 082570A,1994.
[28]陈勇.辣椒红色素的制备方法[J].CN 1 297965A,2001.
[29]张万轩,钟建华,陈家威.从辣椒中提取无辣味红色素[J].湖北大学学报,1995,17(2):177-179.
[30]李新岗,刘毅,黄蕴慧,等.辣椒中辣椒红素的简便分离方法[J].南京军医学院学报.2000,22(3):189-190.
[31]袁庆云.用硅胶柱层析分离辣椒红色素[J].贵州农业科学,1999,27(6):56-57.
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