从元宝枫叶中提取、分离纯化绿原酸的工艺研究
|
摘要
绿原酸是一种天然的抗氧化剂,具有免疫调节、抗癌抗衰老等作用,同时还被认为是很有希望的抗艾滋病毒先导化合物。目前我国绿原酸产业不发达,仍需进口部分绿原酸产品特别是高纯度制品才能满足市场需求,因此急需一种低成本、高得率的绿原酸生产工艺,开展此方面相关研究具有较高的经济价值和广阔的市场前景。
我国传统植物元宝枫中富含绿原酸,本文即以元宝枫叶为提取原料,开发一种能降低绿原酸生产成本、提高产物纯度与得率,并减少废弃物污染的工艺,为发展完善绿原酸产业提供理论依据,使其进一步市场化。研究结果如下：
1分别使用石油醚和乙酸乙酯对元宝枫叶原料进行热回流提取预处理,结果证明其可优化提取工艺流程效率,有利于提高绿原酸产品纯度。
2对比数种提取方法之间的效率,根据结果确认使用超声波法提取元宝枫叶中绿原酸的。针对其设计单因素考察试验与正交试验,优选得出超声提取元宝枫叶中绿原酸的最佳参数：超声功率150W,提取时间15 min,料液比1：12(m／v),提取2次。使用此方法绿原酸得率达3.14%。
3采用大孔树脂层析法分离纯化元宝枫叶提取液中的绿原酸,通过对4种不同类型树脂的筛选发现LSA-21树脂具有较高的吸附率和解吸率,用Langmuir方程分析得出其对绿原酸的理论最大吸附量可达90.91 mg/g,4h内可基本吸附完全。进一步研究确定最佳分离纯化工艺条件为：pH 3.0,以8BV(柱体积)绿原酸含量约为2 mg／ml的元宝枫叶提取液上柱吸附,6BV70%乙醇进行洗脱。
4本实验选用反相高效液相色谱法进行测定,绿原酸的保留时间在7 min左右,与杂质色谱峰之间分离良好,且峰面积与绿原酸含量的线性关系符合定量分析原则,能较好满足精确测定的要求。经检测,按试验确定最佳工艺路线处理所得终产物中绿原酸纯度为78.14%,得率2.81%,绿原酸总回收率为73.48%。
此方法效果优于传统工艺,缩短了生产周期,降低了生产成本,符合绿原酸产业化要求,值得大力推广。
Chlorogenic acid, a kind of natural anti-oxidant, has an immuno-modulatory, anti-aging and anti-cancer effect. Moreover, it was considered as promising anti-HIV leading compounds. Recently, Chinese Chlorogenic acid industry hasn't developed well. It is still essential to import some Chlorogenic acid products to meet market demands, particularly products in high purity. So there is an urgent need for a low-cost product process with high yield of Chlorogenic acid. Carrying out research in this area has high economic value and broad marketing prospects.
Traditional Chinese plant Acer truncatum Bunge is rich in Chlorogenic acid. This paper is about developing a new product process of Chlorogenic acid with the maple leaves as the raw materials. It is hoped to reduce product costs, improve purity and yield of the product, reduce pollution by the waste. The aim of the paper is providing a theoretical basis for the industry of Chlorogenic acid to make the market of it more perfect. The results are as follows:
1 Petroleum ether and ethyl acetate were used in heatened reflux extraction for pretreatment of the maple leaves. The results proved that it could optimize the efficiency of extraction process, improve the purity of Chlorogenic acid.
2 According to the contrasting test results of some extraction methods, the ultrasonic method is chosen for the extracting of Chlorogenic acid. According to the result of single-factor and orthogonal test, the best program is:ultrasonic power is 150W, extracting time is 15 minutes, solid to liquid ratio is 1:12 (m/v), and extracting for 2 times. The yield of Chlorogenic acid with this method could reach 3.14%.
3 Using macroporous resin chromatography to purify Chlorogenic acid from crude extracts of the maple leaves. Through the chosen test of 4 different types of resins, it is found that LSA-21 resin performed best. The analysis with the Langmuir equation proved its theoretical maximum of adsorption of capacity Chlorogenic acid is up to 90.91 mg/g, achieving in complete absorption in 4 hours. Further research confirmed the optimum conditions for separation and purification is:pH 3.0, using 8 BV (column volume) of crude extraction from maple leaves, with a content of Chlorogenic acid at about 2 mg/ml, for adsorption,6 BV of 70%ethanol for desorption.
4 In this experiment, HPLC is used for determination of Chlorogenic acid. The retention time of it is about 7 minutes, with a good separation of the peaks of targeting products and impurities. The linear relationship between quantity of Chlorogenic acid and peak area has a consistency with the principles of quantitative analysis and can satisfy the requirements of precise determination. The test of experiment with the optimum producting process is as follows:purity of Chlorogenic acid product was 78.14%, with a yield 2.81%, a total recovery of 73.48%.
This method is better than the traditional process, reducing the production cycle and cost, meeting the industrial requirements of chlorogenic acid, so worth promoting vigorously.
我国传统植物元宝枫中富含绿原酸,本文即以元宝枫叶为提取原料,开发一种能降低绿原酸生产成本、提高产物纯度与得率,并减少废弃物污染的工艺,为发展完善绿原酸产业提供理论依据,使其进一步市场化。研究结果如下：
1分别使用石油醚和乙酸乙酯对元宝枫叶原料进行热回流提取预处理,结果证明其可优化提取工艺流程效率,有利于提高绿原酸产品纯度。
2对比数种提取方法之间的效率,根据结果确认使用超声波法提取元宝枫叶中绿原酸的。针对其设计单因素考察试验与正交试验,优选得出超声提取元宝枫叶中绿原酸的最佳参数：超声功率150W,提取时间15 min,料液比1：12(m／v),提取2次。使用此方法绿原酸得率达3.14%。
3采用大孔树脂层析法分离纯化元宝枫叶提取液中的绿原酸,通过对4种不同类型树脂的筛选发现LSA-21树脂具有较高的吸附率和解吸率,用Langmuir方程分析得出其对绿原酸的理论最大吸附量可达90.91 mg/g,4h内可基本吸附完全。进一步研究确定最佳分离纯化工艺条件为：pH 3.0,以8BV(柱体积)绿原酸含量约为2 mg／ml的元宝枫叶提取液上柱吸附,6BV70%乙醇进行洗脱。
4本实验选用反相高效液相色谱法进行测定,绿原酸的保留时间在7 min左右,与杂质色谱峰之间分离良好,且峰面积与绿原酸含量的线性关系符合定量分析原则,能较好满足精确测定的要求。经检测,按试验确定最佳工艺路线处理所得终产物中绿原酸纯度为78.14%,得率2.81%,绿原酸总回收率为73.48%。
此方法效果优于传统工艺,缩短了生产周期,降低了生产成本,符合绿原酸产业化要求,值得大力推广。
Chlorogenic acid, a kind of natural anti-oxidant, has an immuno-modulatory, anti-aging and anti-cancer effect. Moreover, it was considered as promising anti-HIV leading compounds. Recently, Chinese Chlorogenic acid industry hasn't developed well. It is still essential to import some Chlorogenic acid products to meet market demands, particularly products in high purity. So there is an urgent need for a low-cost product process with high yield of Chlorogenic acid. Carrying out research in this area has high economic value and broad marketing prospects.
Traditional Chinese plant Acer truncatum Bunge is rich in Chlorogenic acid. This paper is about developing a new product process of Chlorogenic acid with the maple leaves as the raw materials. It is hoped to reduce product costs, improve purity and yield of the product, reduce pollution by the waste. The aim of the paper is providing a theoretical basis for the industry of Chlorogenic acid to make the market of it more perfect. The results are as follows:
1 Petroleum ether and ethyl acetate were used in heatened reflux extraction for pretreatment of the maple leaves. The results proved that it could optimize the efficiency of extraction process, improve the purity of Chlorogenic acid.
2 According to the contrasting test results of some extraction methods, the ultrasonic method is chosen for the extracting of Chlorogenic acid. According to the result of single-factor and orthogonal test, the best program is:ultrasonic power is 150W, extracting time is 15 minutes, solid to liquid ratio is 1:12 (m/v), and extracting for 2 times. The yield of Chlorogenic acid with this method could reach 3.14%.
3 Using macroporous resin chromatography to purify Chlorogenic acid from crude extracts of the maple leaves. Through the chosen test of 4 different types of resins, it is found that LSA-21 resin performed best. The analysis with the Langmuir equation proved its theoretical maximum of adsorption of capacity Chlorogenic acid is up to 90.91 mg/g, achieving in complete absorption in 4 hours. Further research confirmed the optimum conditions for separation and purification is:pH 3.0, using 8 BV (column volume) of crude extraction from maple leaves, with a content of Chlorogenic acid at about 2 mg/ml, for adsorption,6 BV of 70%ethanol for desorption.
4 In this experiment, HPLC is used for determination of Chlorogenic acid. The retention time of it is about 7 minutes, with a good separation of the peaks of targeting products and impurities. The linear relationship between quantity of Chlorogenic acid and peak area has a consistency with the principles of quantitative analysis and can satisfy the requirements of precise determination. The test of experiment with the optimum producting process is as follows:purity of Chlorogenic acid product was 78.14%, with a yield 2.81%, a total recovery of 73.48%.
This method is better than the traditional process, reducing the production cycle and cost, meeting the industrial requirements of chlorogenic acid, so worth promoting vigorously.
引文
[1]吴龙奇,朱文学,张玉先,等.杜仲中绿原酸含量及提取检测方法分析[J].食品科学,2005,26(1)：187-192.
[2]汪萌,张翠,刘泉.元宝枫的药用植物化学成分及药理作用研究进展[J].黑龙江医药,2008,21(1)：70-73.
[3]李鹏,彭修娟,杨新杰.大孔吸附树脂技术在中药化学研究中的应用[J].亚太传统医药,2010,6(2)：117-120.
[4]、傅立国,陈潭清,郎楷永,等.中国高等植物(第八卷)[M].青岛：青岛出版社,2001.319.
[5]尉芹,王兰珍,马希汗,等.元宝枫开发利用研究[M].西安：陕西科学技术出版社,1996.22-24.
[6]吴卫中,李映丽,许走,等.元宝枫化学成分的研究概况[J].中国药事,2008,22(7)：603-606
[7]苏建荣,罗香,杨文云,等.元宝枫叶内黄酮、绿原酸含量动态变化研究[J].林业科学研究,2004,17(4)：496-499.
[8]黄相中,李聪,古昆,等.元宝枫叶提取物的稳定性及抗氧化活性研究[J].云南大学学报(自然科学版),2003,25(5)：442-445.
[9]杨连利,李仲瑾,苏秀霞,等.元宝枫单宁类化合物的提取研究[J].皮革化工,2004,21(5)：1-3.
[10]魏希颖,梁健.有潜在药用价值的元宝枫等槭属植物[J].中药材,2005,28(3)：176-177.
[11]刘祥义,付惠,张加研.云南元宝枫种子含油量及其脂肪酸成分分析[J].天然产物研究与开发,2003,15(1)：38-39.
[12]张元,侯相林.元宝枫油中神经酸乙酯的分离提纯[J].中国油脂,2010,35(1)：28-31.
[13]王性炎,王妹清.神经酸新资源—元宝枫油[J].中国油脂,2005,30(9)：62-64.
[14]Michael Sefkow. First efficient synthesis of Chlorogenic acid[J]. European Journal of Organic Chemistry,2001,6:1137-1141.
[15]马丽,邱业先,杨进军,等.元宝枫蛋白酶的分离纯化及其生化性质[J].植物资源与环境学报,2005,14(1)：6-9.
[16]王兰珍,马希汉,王姝清,等.元宝枫叶营养成分分析[J].西北林学院学报,1997,12(4)：61-63.
[17]陈绍华,王亚琴,罗立新.天然产物绿原酸的研究进展[J].食品科技,2008,33(2)：195-198.
[18]宁正祥,赵谋明.食品生物化学(第二版)[M].广州：华南理工大学出社,2006：356.
[19]Xiaoxia Wang, Jingwu Wang, Nianjun Yang. Chemiluminescent determination of Chlorogenic acid in fruits[J]. Food Chemistry,2006,95(4):574-578.
[20]陈育如,唐刚,刘虎,等.烟草废料中绿原酸的提取工艺研究[J].生物加工过程,2009,7(6)：55-58.
[21]刘军海,裘爱泳.绿原酸及其提取纯化和应用前景[J].粮食与油脂,2003,(9)：44-46.
[22]田士林,李莉.杜仲叶、皮内绿原酸含量的测定与比较[J].安徽农业科学,2006,34(13)：3084-3086.
[23]Ohnishi M, Morishita H, Iwahashi H, et al. Inhibitory effects of Chlorogenic acids on linoleic acid peroxidation and haemolysis[J]. Phytochemistry,1994,36(3):579-583.
[24]Schindler P W, Below P, Hemmerle H, et al. Identification of two new inhibitors of the hepatic glucose-6-phosphatase system[J]. Drug Development Research,1994,44(1):34-40.
[25]胡宗福,于文利,赵亚平.绿原酸清除活性氧和抗脂质过氧化的研究[J].食品科学,2006,27(2)：128-130.
[26]Kasai H. Action of Chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesismodel[J]. Food and Chemical Toxicology,2000,38(5):467-461.
[27]Y Jiang K Satoh, K Kusama, et al. Interaction between Chlorogenic acid and antioxidants [J]. AnticancerRes,2000,20(4):2473-2476.
[28]Rodriguez de Sotillo. Chlorogenic acid modifies plasma and liver concentrationsof:cholesterol, triacylglycerol, and minerals in (fa/fa) Zucker rats[J]. Journal of Nutritional Biochemistry,2002, 13(12):717-726.
[29]Hulme A C. The isolation of Chlorogenic acid from the apple fruit[J]. Biochem,1953,53(3):337-40.
[30]刘军海,裘爱泳.绿原酸的提取分离和含量测定[J].中国油脂,2005,30(3)：54-56.
[31]吴红,李稳宏,吴道澄.杜仲叶水提醇沉工艺对绿原酸含量的影响[J].第四军医大学学报,2002,23(10)：931.
[32]薛焰,郭立玮,沈静,等.超临界萃取与溶剂法联用提取滁菊有效成分的工艺研究[J].南京中医药大学学报,2004,20(2)：102-103.
[33]张玲,尚立霞.金银花提取工艺研究[J].中药新药与临床药理,2003,14(1)：59-61.
[34]周贵新.用超临界二氧化碳从杜仲叶中提取有效成分的方法[P].中国专利：CN124560,A.
[35]刘祥义,傅惠.超声波提取元宝枫叶中绿原酸的研究[J].云南化工,2003,30(2)：23-25.
[36]韦藤幼,赵群莉,阮莉姣,等.微波预处理法提取金银花中的绿原酸[J].中成药,2003,25(7)：534-537.
[37]刘佳佳,赵国玲,王晖,等.金银花绿原酸酶法提取新工艺研究[J].中成药,2002,24(6)：416-418.
[38]胡青平,徐建国,李琪,等.酶法提取元宝枫叶绿原酸的新工艺研究[J].食品科学,2006,27(7)：
159-162.
[39]姜忠义,吴洪.膜技术在中药有效部位和有效成分提取分离中的应用[J].离子交换与吸附,2002,18(2)：155-192.
[40]孙波.杜仲叶制备绿原酸工艺[P].中国专利：CN1273964,A.
[41]刘军海.杜仲叶中高纯绿原酸提取纯化工艺研究[D].江苏无锡：江南大学.2005.
[42]杨祖金,江燕斌,葛发欢,等.超滤膜技术分离杜仲叶绿原酸的研究[J].中药材,2008,31(4)：585-588.
[43]郭立纬,金万勤.无机陶瓷膜分离技术对中药药效物质基础研究的意义[J].膜科学与技术,2002,22(4)：48-49.
[44]彭小文,王金,廉志清.膜除杂浓缩对绿原酸提取液醇沉的影响[J].化学与生物工程.2009,26(9)：49-51.
[45]江邦和,胡晓忠.离子交换与吸附树脂在中药有效成分提取中的应用[J].离子交换与吸附,2001,1 7(40)：379-384.
[46]稻波治.Production opurified Chlorogenic acid[P].日本专利：JP4145048,1992.
[47]张虹,柳正良,王洪泉.大孔吸附树脂在药学领域的应用[J].中国医药工业杂志,2001,32(1)：41-44.
[48]高春荣,胡锦蓉,孙君社.金银花中绿原酸的提取工艺[J].中国农业大学学报,2003,8(4)：5-8.
[49]钱骅,赵伯涛.杜仲叶绿原酸的提取分离[J].中国野生植物资源,1999,20(4)：15-16.
[50]李进飞,黄可龙,李春华.LSA-2 1型树脂对绿原酸吸附分离性能的研究[J].华西药学杂志,2004,19(1)：1-4.
[5 1]彭密军,周春山,钟世安.制备型高效液相色谱法分离纯化绿原酸[J].中南大学学报,2004,35(3)：408-412.
[52]Ian A. Sutherland, Derek Fisher. Role of counter-current chromatography in the modernisation of Chinese herbal medicines[J]. Journal of Chromatography A,2009,1216(4):740-753.
[53]Hai-Tao Lu, Yue Jiang. Feng.Application of preparative high-speed counter-current chromatography for separation of Chlorogenic acid from Flos Lonicerae[J]. Journal of Chromatography A,2004, 1026(1-2):185-190.
[54]张毅敏,谭天伟.环糊精键合凝胶介质分离纯化金银花中绿原酸过程工程学报[J].2009,9(5)：969-974.
[55]张广强,姜玉风.三波长分光光度法测定清热解毒注射剂中绿原酸的含量[J].中国中医杂志,1 990,15(8)：33-34.
[56]谭晓杰,景丹,陈晓辉,等.RP-HPLC法测定野菊花中绿原酸的含量[J].中药材,2004,27(4)：256-257.
[57]袁华,邓良,孙炎彬,等.可见分光光度法测定杜仲叶中的绿原酸[J].生物加工过程,2007,5(1)
：71-73.
[58]张平,苏小飞,方桐辉,等.石油醚、乙酸乙酯预处理在金银花绿原酸萃取中的应用研究[J].安徽科技,2009,2：51-52
[59]梅林.金银花中绿原酸的酶法提取工艺优化[J].广州化学,2007,32(4)：30-34.
[60]Bin Zhanga, Ruiyuan Yangb, Yan Zhaoa, et al. Separation of Chlorogenic acid from honeysuckle crude extracts by macroporous resins[J]. Journal of Chromatography B,2008,867 (2):253-258.
[61]沈奇,赵厚民,张卫明.蒲公英绿原酸提取分离工艺的研究[J].食品科学,2006,27(7)：140-144.
[62]刘斌,石任兵,余超.影响大孔吸附树脂吸附分离中药化学成分的因素[J].中草药,2002,33(5)：475-477.
[2]汪萌,张翠,刘泉.元宝枫的药用植物化学成分及药理作用研究进展[J].黑龙江医药,2008,21(1)：70-73.
[3]李鹏,彭修娟,杨新杰.大孔吸附树脂技术在中药化学研究中的应用[J].亚太传统医药,2010,6(2)：117-120.
[4]、傅立国,陈潭清,郎楷永,等.中国高等植物(第八卷)[M].青岛：青岛出版社,2001.319.
[5]尉芹,王兰珍,马希汗,等.元宝枫开发利用研究[M].西安：陕西科学技术出版社,1996.22-24.
[6]吴卫中,李映丽,许走,等.元宝枫化学成分的研究概况[J].中国药事,2008,22(7)：603-606
[7]苏建荣,罗香,杨文云,等.元宝枫叶内黄酮、绿原酸含量动态变化研究[J].林业科学研究,2004,17(4)：496-499.
[8]黄相中,李聪,古昆,等.元宝枫叶提取物的稳定性及抗氧化活性研究[J].云南大学学报(自然科学版),2003,25(5)：442-445.
[9]杨连利,李仲瑾,苏秀霞,等.元宝枫单宁类化合物的提取研究[J].皮革化工,2004,21(5)：1-3.
[10]魏希颖,梁健.有潜在药用价值的元宝枫等槭属植物[J].中药材,2005,28(3)：176-177.
[11]刘祥义,付惠,张加研.云南元宝枫种子含油量及其脂肪酸成分分析[J].天然产物研究与开发,2003,15(1)：38-39.
[12]张元,侯相林.元宝枫油中神经酸乙酯的分离提纯[J].中国油脂,2010,35(1)：28-31.
[13]王性炎,王妹清.神经酸新资源—元宝枫油[J].中国油脂,2005,30(9)：62-64.
[14]Michael Sefkow. First efficient synthesis of Chlorogenic acid[J]. European Journal of Organic Chemistry,2001,6:1137-1141.
[15]马丽,邱业先,杨进军,等.元宝枫蛋白酶的分离纯化及其生化性质[J].植物资源与环境学报,2005,14(1)：6-9.
[16]王兰珍,马希汉,王姝清,等.元宝枫叶营养成分分析[J].西北林学院学报,1997,12(4)：61-63.
[17]陈绍华,王亚琴,罗立新.天然产物绿原酸的研究进展[J].食品科技,2008,33(2)：195-198.
[18]宁正祥,赵谋明.食品生物化学(第二版)[M].广州：华南理工大学出社,2006：356.
[19]Xiaoxia Wang, Jingwu Wang, Nianjun Yang. Chemiluminescent determination of Chlorogenic acid in fruits[J]. Food Chemistry,2006,95(4):574-578.
[20]陈育如,唐刚,刘虎,等.烟草废料中绿原酸的提取工艺研究[J].生物加工过程,2009,7(6)：55-58.
[21]刘军海,裘爱泳.绿原酸及其提取纯化和应用前景[J].粮食与油脂,2003,(9)：44-46.
[22]田士林,李莉.杜仲叶、皮内绿原酸含量的测定与比较[J].安徽农业科学,2006,34(13)：3084-3086.
[23]Ohnishi M, Morishita H, Iwahashi H, et al. Inhibitory effects of Chlorogenic acids on linoleic acid peroxidation and haemolysis[J]. Phytochemistry,1994,36(3):579-583.
[24]Schindler P W, Below P, Hemmerle H, et al. Identification of two new inhibitors of the hepatic glucose-6-phosphatase system[J]. Drug Development Research,1994,44(1):34-40.
[25]胡宗福,于文利,赵亚平.绿原酸清除活性氧和抗脂质过氧化的研究[J].食品科学,2006,27(2)：128-130.
[26]Kasai H. Action of Chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesismodel[J]. Food and Chemical Toxicology,2000,38(5):467-461.
[27]Y Jiang K Satoh, K Kusama, et al. Interaction between Chlorogenic acid and antioxidants [J]. AnticancerRes,2000,20(4):2473-2476.
[28]Rodriguez de Sotillo. Chlorogenic acid modifies plasma and liver concentrationsof:cholesterol, triacylglycerol, and minerals in (fa/fa) Zucker rats[J]. Journal of Nutritional Biochemistry,2002, 13(12):717-726.
[29]Hulme A C. The isolation of Chlorogenic acid from the apple fruit[J]. Biochem,1953,53(3):337-40.
[30]刘军海,裘爱泳.绿原酸的提取分离和含量测定[J].中国油脂,2005,30(3)：54-56.
[31]吴红,李稳宏,吴道澄.杜仲叶水提醇沉工艺对绿原酸含量的影响[J].第四军医大学学报,2002,23(10)：931.
[32]薛焰,郭立玮,沈静,等.超临界萃取与溶剂法联用提取滁菊有效成分的工艺研究[J].南京中医药大学学报,2004,20(2)：102-103.
[33]张玲,尚立霞.金银花提取工艺研究[J].中药新药与临床药理,2003,14(1)：59-61.
[34]周贵新.用超临界二氧化碳从杜仲叶中提取有效成分的方法[P].中国专利：CN124560,A.
[35]刘祥义,傅惠.超声波提取元宝枫叶中绿原酸的研究[J].云南化工,2003,30(2)：23-25.
[36]韦藤幼,赵群莉,阮莉姣,等.微波预处理法提取金银花中的绿原酸[J].中成药,2003,25(7)：534-537.
[37]刘佳佳,赵国玲,王晖,等.金银花绿原酸酶法提取新工艺研究[J].中成药,2002,24(6)：416-418.
[38]胡青平,徐建国,李琪,等.酶法提取元宝枫叶绿原酸的新工艺研究[J].食品科学,2006,27(7)：
159-162.
[39]姜忠义,吴洪.膜技术在中药有效部位和有效成分提取分离中的应用[J].离子交换与吸附,2002,18(2)：155-192.
[40]孙波.杜仲叶制备绿原酸工艺[P].中国专利：CN1273964,A.
[41]刘军海.杜仲叶中高纯绿原酸提取纯化工艺研究[D].江苏无锡：江南大学.2005.
[42]杨祖金,江燕斌,葛发欢,等.超滤膜技术分离杜仲叶绿原酸的研究[J].中药材,2008,31(4)：585-588.
[43]郭立纬,金万勤.无机陶瓷膜分离技术对中药药效物质基础研究的意义[J].膜科学与技术,2002,22(4)：48-49.
[44]彭小文,王金,廉志清.膜除杂浓缩对绿原酸提取液醇沉的影响[J].化学与生物工程.2009,26(9)：49-51.
[45]江邦和,胡晓忠.离子交换与吸附树脂在中药有效成分提取中的应用[J].离子交换与吸附,2001,1 7(40)：379-384.
[46]稻波治.Production opurified Chlorogenic acid[P].日本专利：JP4145048,1992.
[47]张虹,柳正良,王洪泉.大孔吸附树脂在药学领域的应用[J].中国医药工业杂志,2001,32(1)：41-44.
[48]高春荣,胡锦蓉,孙君社.金银花中绿原酸的提取工艺[J].中国农业大学学报,2003,8(4)：5-8.
[49]钱骅,赵伯涛.杜仲叶绿原酸的提取分离[J].中国野生植物资源,1999,20(4)：15-16.
[50]李进飞,黄可龙,李春华.LSA-2 1型树脂对绿原酸吸附分离性能的研究[J].华西药学杂志,2004,19(1)：1-4.
[5 1]彭密军,周春山,钟世安.制备型高效液相色谱法分离纯化绿原酸[J].中南大学学报,2004,35(3)：408-412.
[52]Ian A. Sutherland, Derek Fisher. Role of counter-current chromatography in the modernisation of Chinese herbal medicines[J]. Journal of Chromatography A,2009,1216(4):740-753.
[53]Hai-Tao Lu, Yue Jiang. Feng.Application of preparative high-speed counter-current chromatography for separation of Chlorogenic acid from Flos Lonicerae[J]. Journal of Chromatography A,2004, 1026(1-2):185-190.
[54]张毅敏,谭天伟.环糊精键合凝胶介质分离纯化金银花中绿原酸过程工程学报[J].2009,9(5)：969-974.
[55]张广强,姜玉风.三波长分光光度法测定清热解毒注射剂中绿原酸的含量[J].中国中医杂志,1 990,15(8)：33-34.
[56]谭晓杰,景丹,陈晓辉,等.RP-HPLC法测定野菊花中绿原酸的含量[J].中药材,2004,27(4)：256-257.
[57]袁华,邓良,孙炎彬,等.可见分光光度法测定杜仲叶中的绿原酸[J].生物加工过程,2007,5(1)
：71-73.
[58]张平,苏小飞,方桐辉,等.石油醚、乙酸乙酯预处理在金银花绿原酸萃取中的应用研究[J].安徽科技,2009,2：51-52
[59]梅林.金银花中绿原酸的酶法提取工艺优化[J].广州化学,2007,32(4)：30-34.
[60]Bin Zhanga, Ruiyuan Yangb, Yan Zhaoa, et al. Separation of Chlorogenic acid from honeysuckle crude extracts by macroporous resins[J]. Journal of Chromatography B,2008,867 (2):253-258.
[61]沈奇,赵厚民,张卫明.蒲公英绿原酸提取分离工艺的研究[J].食品科学,2006,27(7)：140-144.
[62]刘斌,石任兵,余超.影响大孔吸附树脂吸附分离中药化学成分的因素[J].中草药,2002,33(5)：475-477.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |