1，4-二棕榈酰羟脯氨酸的合成工艺研究
|
摘要
1,4-二棕榈酰羟脯氨酸(1,4-Dipalmitoyl Hydroxyproline),作为胶原蛋白主要组成成分之一的羟脯氨酸衍生物,是一种化妆品中有效的抗皱抗衰老添加剂。本文以肖顿·鲍曼(Schotten·Banmamm)缩合反应法对酰氯进行氨解和醇解制备1,4-二棕榈酰羟脯氨酸,并探索1,4-二棕榈酰羟脯氨酸的非水相酶催化反应。
1.首先制备了用于酯化和酰胺化的棕榈酰氯,通过对反应物摩尔比、反应时间和反应温度因素进行了考察,得到制备棕榈酰氯反应的适宜条件为:反应摩尔比n(酸):n(三氯化磷)=1:2、反应温度70-80℃、反应时间3 h。
2.用肖顿·鲍曼缩合反应对棕榈酰氯进行了氨解和醇解,制备得到了1,4-二棕榈酰羟脯氨酸。通过单因素实验考察了各因素对制备二棕榈酰羟脯氨酸的影响。得到了反应条件为:以丙酮为有机溶剂,反应pH为8~9,有机溶剂(指丙酮)体积分数为50%,反应温度为15℃,原料摩尔比n(羟脯氨酸) : n(棕榈酰氯)=1: 2.4,反应时间为4 h。此条件下,反应转化率可达98.13%,产率可达60%。为了得到最佳的反应条件,运用了正交设计和响应面分析手段对实验进行优化。结果表明,对于二棕榈酰羟脯氨酸的产率,影响因素顺序为:原料摩尔比>反应温度>丙酮体积分数>反应时间。得到的最佳反应条件为有机溶剂体积分数为55%,反应温度为15℃,原料摩尔比为n(羟脯氨酸) : n(棕榈酰氯)=1:2.5,反应时间为5 h。采用三因素三水平的响应面分析法,建立了反应的二次多项式数学模型,并以1,4-二棕榈酰羟脯氨酸的产率为响应值作响应面和等高线,考察了反应温度、有机溶剂体积分数和原料摩尔比对产率的影响。得出了反应拟合方程,其反应的最佳工艺条件为:有机溶剂体积分数为55%,反应温度为15℃,原料摩尔比为n(羟脯氨酸) : n(棕榈酰氯)=1:2.5。此条件下,产率可达58.26%。
3.考察了1,4-二棕榈酰羟脯氨酸的非水相酶合成反应。对酶种类、有机溶剂、加酶量和加水量等因素进行的比较实验,可知Lipolase脂肪酶和Savinase蛋白酶对催化效果较好;在疏水性溶剂异辛烷中转化率较大;加酶量增加转化率有适当的增加;必要加水量在1-2%下较为合适,在水量过少或过多的情况下都容易导致酶失活而使转化率降低。
1,4-dipalmitoyl hydroxyproline, plant-derived lipid agent consisting of hydroxyproline (natural amino acid) & palmitic acid, is anti-aging agent which can be applied to cosmetics. This experiment focused on the synthesis of 1,4-dipalmitoyl hydroxyproline by Schotten Banmamm condensation reactions, and studied the non-aqueous enzyme catalysis.
1.Firstly, palmitoyl chloride used in esterification and amidation, By studying of the influencing facts such as molar ratio of materials, the time, temperature in detail, the optimum reaction conditions were obtained as follows: molar ratio of materials was 1:2, reaction temperature was 70-80℃, reaction time was 3 h.
2.1,4-dipalmitoyl hydroxyproline was obtained by Schotten Banmamm condensation reactions. The different influencing factors of reaction were studied. Then the reaction conditions were obtained as follows: volume ratio of acetone was 50%, molar ratio of materials was 1:2.5, pH=8~9, reaction temperature was 15℃, reaction time was 4 h. The conversion of reaction had reached to 98.13 % under the optimal conditions, and the yield 60%. Orthogonal design and Response surface methodology were used in the experiment for optimum condition. The experimental results show that the molar ratio of materials had the largest effect, the second was reaction temperature, third was volume ratio of acetone and reaction time had the least effect. And the optimum reaction conditions were obtained as follows: volume ratio of acetone was 55%, molar ratio of materials was 1:2.5, reaction temperature was 15℃, reaction time was 5 h. By using the three-level-three-factor response surface methodology, a quadratic polynomial mathematical model was established. The influences of the reaction temperature, volume ratio of organic agent and the molar ratio of materials were investigated by determining the yield as the response. The optimum reaction conditions were obtained as follows: volume ratio of acetone was 55%, reaction temperature was 15℃, molar ratio of materials was 2.5. And in these conditions, the yield reaches 58.26%.
3.In order to study the non-aqueous enzyme catalysis, the influencing factors were investigated. The results show that the catalysis effects of Lipolase lipase and Savinase albuminuria are better than others; Conversion of reaction, much bigger in isooctane, will increase by adding of enzyme. And the optimum water content is 1~2%.
1.首先制备了用于酯化和酰胺化的棕榈酰氯,通过对反应物摩尔比、反应时间和反应温度因素进行了考察,得到制备棕榈酰氯反应的适宜条件为:反应摩尔比n(酸):n(三氯化磷)=1:2、反应温度70-80℃、反应时间3 h。
2.用肖顿·鲍曼缩合反应对棕榈酰氯进行了氨解和醇解,制备得到了1,4-二棕榈酰羟脯氨酸。通过单因素实验考察了各因素对制备二棕榈酰羟脯氨酸的影响。得到了反应条件为:以丙酮为有机溶剂,反应pH为8~9,有机溶剂(指丙酮)体积分数为50%,反应温度为15℃,原料摩尔比n(羟脯氨酸) : n(棕榈酰氯)=1: 2.4,反应时间为4 h。此条件下,反应转化率可达98.13%,产率可达60%。为了得到最佳的反应条件,运用了正交设计和响应面分析手段对实验进行优化。结果表明,对于二棕榈酰羟脯氨酸的产率,影响因素顺序为:原料摩尔比>反应温度>丙酮体积分数>反应时间。得到的最佳反应条件为有机溶剂体积分数为55%,反应温度为15℃,原料摩尔比为n(羟脯氨酸) : n(棕榈酰氯)=1:2.5,反应时间为5 h。采用三因素三水平的响应面分析法,建立了反应的二次多项式数学模型,并以1,4-二棕榈酰羟脯氨酸的产率为响应值作响应面和等高线,考察了反应温度、有机溶剂体积分数和原料摩尔比对产率的影响。得出了反应拟合方程,其反应的最佳工艺条件为:有机溶剂体积分数为55%,反应温度为15℃,原料摩尔比为n(羟脯氨酸) : n(棕榈酰氯)=1:2.5。此条件下,产率可达58.26%。
3.考察了1,4-二棕榈酰羟脯氨酸的非水相酶合成反应。对酶种类、有机溶剂、加酶量和加水量等因素进行的比较实验,可知Lipolase脂肪酶和Savinase蛋白酶对催化效果较好;在疏水性溶剂异辛烷中转化率较大;加酶量增加转化率有适当的增加;必要加水量在1-2%下较为合适,在水量过少或过多的情况下都容易导致酶失活而使转化率降低。
1,4-dipalmitoyl hydroxyproline, plant-derived lipid agent consisting of hydroxyproline (natural amino acid) & palmitic acid, is anti-aging agent which can be applied to cosmetics. This experiment focused on the synthesis of 1,4-dipalmitoyl hydroxyproline by Schotten Banmamm condensation reactions, and studied the non-aqueous enzyme catalysis.
1.Firstly, palmitoyl chloride used in esterification and amidation, By studying of the influencing facts such as molar ratio of materials, the time, temperature in detail, the optimum reaction conditions were obtained as follows: molar ratio of materials was 1:2, reaction temperature was 70-80℃, reaction time was 3 h.
2.1,4-dipalmitoyl hydroxyproline was obtained by Schotten Banmamm condensation reactions. The different influencing factors of reaction were studied. Then the reaction conditions were obtained as follows: volume ratio of acetone was 50%, molar ratio of materials was 1:2.5, pH=8~9, reaction temperature was 15℃, reaction time was 4 h. The conversion of reaction had reached to 98.13 % under the optimal conditions, and the yield 60%. Orthogonal design and Response surface methodology were used in the experiment for optimum condition. The experimental results show that the molar ratio of materials had the largest effect, the second was reaction temperature, third was volume ratio of acetone and reaction time had the least effect. And the optimum reaction conditions were obtained as follows: volume ratio of acetone was 55%, molar ratio of materials was 1:2.5, reaction temperature was 15℃, reaction time was 5 h. By using the three-level-three-factor response surface methodology, a quadratic polynomial mathematical model was established. The influences of the reaction temperature, volume ratio of organic agent and the molar ratio of materials were investigated by determining the yield as the response. The optimum reaction conditions were obtained as follows: volume ratio of acetone was 55%, reaction temperature was 15℃, molar ratio of materials was 2.5. And in these conditions, the yield reaches 58.26%.
3.In order to study the non-aqueous enzyme catalysis, the influencing factors were investigated. The results show that the catalysis effects of Lipolase lipase and Savinase albuminuria are better than others; Conversion of reaction, much bigger in isooctane, will increase by adding of enzyme. And the optimum water content is 1~2%.
引文
[1]李卫年.抗衰老化妆品发展趋势[J].日用化学品科学,2006,29(7):14-18.
[2]彭园,黄小琴.中医药延缓皮肤老化的研究[J].湖北中医杂志,2006,28(10):18-20.
[3]王春霖,郭芳,王永利.自由基与衰老[J].河北医科大学学报,2005,26(4):308-311.
[4] Levine RL,Stadtman ER.Oxidative modification of protein during aging[J].Exp Gerontal,2001,36(9):1495-1502.
[5] Sohal RS,Mockett RJ,Orr WC.Mechanisms of aging:In appraisal of t he oxidative stress hypothesis[J].Free Radic Biol Med,2002,23(5):575-586.
[6]刘仲荣,杨军,杨慧兰等.抗皮肤老化化妆品活性成分的研究进展[J].中国美容医学,2005, 14(3):362-365.
[7]虞瑞尧.保湿化妆品与皮肤科学[J].中国美容医学,2002,11(4):393-395.
[8]冯文帅.透明质酸及其发展应用前景[J].四川食品与发酵,2001,1:4-6.
[9]林春梅,张小东,管洪义等.美白化妆品原料的开发及其祛斑美白机制[J].职业与健康,2003,19(8):98-99.
[10]陈燕妮,卢云,吴昊等.N-酰基氨基酸系列表面活性剂的合成和应用[J].化学研究与应用,2001,13(2):192-195.
[11]王峰,张兆国.钯催化的酰胺羰基化反应[J].有机化学,2002,22(8):536-542.
[12]黄涛,郭华,黄开勋.硬酯酰甘氨酸的合成工艺研究[J].化学世界,2003,7:376- 342.
[13]刘筱虹,张春霞,王佛华等.混合氨基酸月桂酰胺的合成[J].天然产物研究与开发,1997,9(4):79-81.
[14]唐晓彬.N-酰基肌氨酸钠合成及在防水加脂剂中的应用[J].皮革化工,2000, 17(2):38-40.
[15]李红,黎四芳.N-月桂酰肌氨酸钠的合成研究[J].厦门大学学报,2005,44(3): 386-389.
[16]祝阳,蒋立建.十一烯酰-L-谷氨酸的制备[J].广东化工,2007,34(11):37-39.
[17]郭雪飞,廖发盆,黄小华.吡哆醇二棕榈酸酯的合成[J].应用化工,2006,35(7): 532-534.
[18] A.B.Madhan Kumar,K.Panduranga Rao.Poly(palmitoyl-L-hydroxyproline ester) microspheres as potential oral controlled drug delivery system[J].International journal of pharmaceutics,1997,149:107-114.
[19] Shen Zong-xuan,Liu Yan-hua,CHEN Wu-hong etc.Synthesis of 4-Substituted Prolines from(4R)-Hydroxy-(s)-Proline[J].CHenmiCAL RESEARCH,2005,16(1): 15-18.
[20] R·P·乌布赖,R·R·高德特,F·D·乌德.碱金属氨基酸的制备[P].中国专利, 96194561.3.1996-5-31.
[21]于伟民,薛永强,梁西良等.乙酸乙酯的精制方法[J].化学与粘合,2005,27(2): 108-111.
[22]黄志良,何壁如,莫驰.L-抗坏血酸苯甲酸酯的合成与性能研究[J].食品科学,2004,25(4):87-90.
[23]刘长波,高瑞昶.L-抗坏血酸棕榈酸酯的合成研究进展[J].中国油脂,2003, 28(10):46-49.
[24]谢文磊,李魁,王宏雁.酯交换法合成L-抗坏血酸硬脂酸酯及其抗氧化性能[J].高校化学工程学报,2002,4(16):441-445.
[25]施志浩,魏运洋,丁亚明等.酯交换法合成L-天冬酰胺叔丁酯[J].化学研究与应用,2006,18(7):856-859.
[26]常雁红,王化军.酯交换法合成碳酸二钾酯研究进展[J].化工进展,2007,26(5): 642-646
[27]姜绍通,李岩,邵平.酯交换法生产生物柴油的研究进展[J].农产品加工学刊, 2006,10:5-8.
[28]王文魁,包宗宏,王本庭等.酯交换法生产生物柴油研究进展[J].拖拉机和农用运输车,2008,35(1):3-5.
[29]陆豫,甘利军,陈葆仁.L-抗坏血酸酸棕榈酸酯的合成[J].精细化工,1996,13(3): 17-18.
[30]李德江,龙德清,付和清.肉桂酸苄酯的合成[J].合成化学,2004,12(4):378-380.
[31]粟晖,郑燕升,李军生等.维生素E油酸酯的合成研究[J].食品科学,2005,26(8): 135-138.
[32]芮斌,陶文沂，蒋惠亮.曲酸双棕搁醋的制备和性能[J]旧用化学工业，2003，33(2): 124-126.
[33]孙建军.月桂酰柠檬酸酯的合成[J].郑州粮食学院学报,2000,21,(1):29-31.
[34] Klibanov A M.Enzymes that work in organic solvents[J].Chem Tech,1986,6:354-359.
[35] Lipase-catalyzed enatioselective esterification of (S)-naproxen hydroxyalkyl ester in organic media[J].Biotechnology Letters,2003,25:413-416.
[36] EELIN SOO,ABU BAKAR SALLEH,MAHIRAN BASRI.etal.Optimization of the enzyme catalysed synthesis of amino acid-based surfactants from palm oil fractions[J].J.Biosci Bioeng,2003,95(4):361- 367.
[37]姜雯虹,韩萍芳,徐宁等.酶法合成果糖棕榈酸酯[J].化学反应工程与工艺,2006,22(6): 565-569.
[38]李祖义,陈倩.酶法合成表面活性剂[J].工业微生物,2001,31(2):42-48.
[39]王福海.硬脂酸及脂肪酸衍生物生产工艺[M].北京:轻工业出版社,1991.291-293.
[40]许志勤,高兰兴.组织羟脯氨酸测定方法的改进[J].解放军预防医学杂志,1990,8(1): 40-42.
[41]关静,叶萍,武继民.胶原海绵的羟脯氨酸含量测定[J].氨基酸和生物资源,2000,22(1): 52-54.
[42]葛洪,张之强,王军等.N-十四酰基甘氨酸钠的合成和性能[J].精细石油化工,2006,23(5): 34-36.
[43]吴有炜.试验设计与数据处理[M].江苏:苏州大学出版社,2002.135-143.
[2]彭园,黄小琴.中医药延缓皮肤老化的研究[J].湖北中医杂志,2006,28(10):18-20.
[3]王春霖,郭芳,王永利.自由基与衰老[J].河北医科大学学报,2005,26(4):308-311.
[4] Levine RL,Stadtman ER.Oxidative modification of protein during aging[J].Exp Gerontal,2001,36(9):1495-1502.
[5] Sohal RS,Mockett RJ,Orr WC.Mechanisms of aging:In appraisal of t he oxidative stress hypothesis[J].Free Radic Biol Med,2002,23(5):575-586.
[6]刘仲荣,杨军,杨慧兰等.抗皮肤老化化妆品活性成分的研究进展[J].中国美容医学,2005, 14(3):362-365.
[7]虞瑞尧.保湿化妆品与皮肤科学[J].中国美容医学,2002,11(4):393-395.
[8]冯文帅.透明质酸及其发展应用前景[J].四川食品与发酵,2001,1:4-6.
[9]林春梅,张小东,管洪义等.美白化妆品原料的开发及其祛斑美白机制[J].职业与健康,2003,19(8):98-99.
[10]陈燕妮,卢云,吴昊等.N-酰基氨基酸系列表面活性剂的合成和应用[J].化学研究与应用,2001,13(2):192-195.
[11]王峰,张兆国.钯催化的酰胺羰基化反应[J].有机化学,2002,22(8):536-542.
[12]黄涛,郭华,黄开勋.硬酯酰甘氨酸的合成工艺研究[J].化学世界,2003,7:376- 342.
[13]刘筱虹,张春霞,王佛华等.混合氨基酸月桂酰胺的合成[J].天然产物研究与开发,1997,9(4):79-81.
[14]唐晓彬.N-酰基肌氨酸钠合成及在防水加脂剂中的应用[J].皮革化工,2000, 17(2):38-40.
[15]李红,黎四芳.N-月桂酰肌氨酸钠的合成研究[J].厦门大学学报,2005,44(3): 386-389.
[16]祝阳,蒋立建.十一烯酰-L-谷氨酸的制备[J].广东化工,2007,34(11):37-39.
[17]郭雪飞,廖发盆,黄小华.吡哆醇二棕榈酸酯的合成[J].应用化工,2006,35(7): 532-534.
[18] A.B.Madhan Kumar,K.Panduranga Rao.Poly(palmitoyl-L-hydroxyproline ester) microspheres as potential oral controlled drug delivery system[J].International journal of pharmaceutics,1997,149:107-114.
[19] Shen Zong-xuan,Liu Yan-hua,CHEN Wu-hong etc.Synthesis of 4-Substituted Prolines from(4R)-Hydroxy-(s)-Proline[J].CHenmiCAL RESEARCH,2005,16(1): 15-18.
[20] R·P·乌布赖,R·R·高德特,F·D·乌德.碱金属氨基酸的制备[P].中国专利, 96194561.3.1996-5-31.
[21]于伟民,薛永强,梁西良等.乙酸乙酯的精制方法[J].化学与粘合,2005,27(2): 108-111.
[22]黄志良,何壁如,莫驰.L-抗坏血酸苯甲酸酯的合成与性能研究[J].食品科学,2004,25(4):87-90.
[23]刘长波,高瑞昶.L-抗坏血酸棕榈酸酯的合成研究进展[J].中国油脂,2003, 28(10):46-49.
[24]谢文磊,李魁,王宏雁.酯交换法合成L-抗坏血酸硬脂酸酯及其抗氧化性能[J].高校化学工程学报,2002,4(16):441-445.
[25]施志浩,魏运洋,丁亚明等.酯交换法合成L-天冬酰胺叔丁酯[J].化学研究与应用,2006,18(7):856-859.
[26]常雁红,王化军.酯交换法合成碳酸二钾酯研究进展[J].化工进展,2007,26(5): 642-646
[27]姜绍通,李岩,邵平.酯交换法生产生物柴油的研究进展[J].农产品加工学刊, 2006,10:5-8.
[28]王文魁,包宗宏,王本庭等.酯交换法生产生物柴油研究进展[J].拖拉机和农用运输车,2008,35(1):3-5.
[29]陆豫,甘利军,陈葆仁.L-抗坏血酸酸棕榈酸酯的合成[J].精细化工,1996,13(3): 17-18.
[30]李德江,龙德清,付和清.肉桂酸苄酯的合成[J].合成化学,2004,12(4):378-380.
[31]粟晖,郑燕升,李军生等.维生素E油酸酯的合成研究[J].食品科学,2005,26(8): 135-138.
[32]芮斌,陶文沂，蒋惠亮.曲酸双棕搁醋的制备和性能[J]旧用化学工业，2003，33(2): 124-126.
[33]孙建军.月桂酰柠檬酸酯的合成[J].郑州粮食学院学报,2000,21,(1):29-31.
[34] Klibanov A M.Enzymes that work in organic solvents[J].Chem Tech,1986,6:354-359.
[35] Lipase-catalyzed enatioselective esterification of (S)-naproxen hydroxyalkyl ester in organic media[J].Biotechnology Letters,2003,25:413-416.
[36] EELIN SOO,ABU BAKAR SALLEH,MAHIRAN BASRI.etal.Optimization of the enzyme catalysed synthesis of amino acid-based surfactants from palm oil fractions[J].J.Biosci Bioeng,2003,95(4):361- 367.
[37]姜雯虹,韩萍芳,徐宁等.酶法合成果糖棕榈酸酯[J].化学反应工程与工艺,2006,22(6): 565-569.
[38]李祖义,陈倩.酶法合成表面活性剂[J].工业微生物,2001,31(2):42-48.
[39]王福海.硬脂酸及脂肪酸衍生物生产工艺[M].北京:轻工业出版社,1991.291-293.
[40]许志勤,高兰兴.组织羟脯氨酸测定方法的改进[J].解放军预防医学杂志,1990,8(1): 40-42.
[41]关静,叶萍,武继民.胶原海绵的羟脯氨酸含量测定[J].氨基酸和生物资源,2000,22(1): 52-54.
[42]葛洪,张之强,王军等.N-十四酰基甘氨酸钠的合成和性能[J].精细石油化工,2006,23(5): 34-36.
[43]吴有炜.试验设计与数据处理[M].江苏:苏州大学出版社,2002.135-143.