氧化铝柱层析分离纯化磷脂的工艺优化
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摘要
高纯度磷脂作为一种优良的表面活性剂和乳化剂已经被广泛应用于食品、医药、化妆品等行业。目前国内高纯度磷脂产品尚依赖进口，对其需求也在逐年增加。因此，迫切需要开发出适合我国国情的高纯度磷脂的制备方法。针对我国鸡蛋资源非常丰富但是利用率低下的现状，本课题组一直致力于高纯度蛋黄磷脂的制备研究并且取得了较大的进展。
本文在课题组已有研究的基础上，以“氧化铝柱层析分离纯化磷脂的工艺优化”为目标，考察了不同洗脱条件对磷脂在氧化铝柱上的洗脱特性的影响，并进一步进行了氧化铝再生以及工艺连续上样的实验研究，建立并且优化了氧化铝柱层析分离纯化卵磷脂(Phosphatidylcholine，简称PC)和脑磷脂(Phosphatidylethanolamine，简称PE)的完整工艺。
系统考察了甲醇—氯仿；甲醇—氯仿—三乙胺；甲醇—氨水；和甲醇—氯仿—氨水洗脱体系对氧化铝分离纯化磷脂工艺的影响。得到最佳的洗脱液组成为甲醇：氯仿：氨水=80：120：10(v／v)。在此条件下，PC和PE的回收率均达到98％以上，理论产量分别为0.31g／g吸附剂和0.04g／g吸附剂。
对不同洗脱条件下氧化铝的再生以及工艺的连续上样进行了实验研究。结果表明：在洗脱液为甲醇：氯仿=1：4(v／v)的条件下，PC和PE回收率分别达到99.8％和86.8％，理论产量分别达到0.32g／g吸附剂和0.03g／g吸附剂，再次上样量和三次上样量分别达到首次上样量的80.2％和79.7％，并可同时得到纯度大于95％的PC产品和纯度大于80％的PE产品。在有机溶剂和碱组成的洗脱液体系中，再生和连续上样效果最佳的是甲醇：氯仿：氨水=80：120：10(v／v)的洗脱液。在此条件下，再次上样量为10.85g，达到首次上样量的94.7％。
对磷脂在不同溶剂中的稳定性进行了研究。研究结果表明，磷脂在氨水或者三乙胺存在的有机溶剂中是不稳定的。
本文的研究建立了氧化铝柱层析分离纯化高纯度磷脂的完整工艺，该工艺具有分离效果好、工艺简单、投资少、处理量较大等优点。研究结果为高纯度卵磷脂和脑磷脂的制备工艺工业化提供了重要的基础数据。
High-purity phosphohpids are widely used in the fields of food, pharmacy, cosmetic and so on because of the special surface activity and emulsification. .At present most of this production is imported in China and the requirement enhances every year. So it is necessary to develop the preparation of high-purity phospholipids. The research has been doing and great process has been made in our lab.
Aiming at the process optimization for simultaneous purification of PC and PE by alumina column chromatography, the effect of different elution phase on the elution characteristic of PC (Phosphatidylcholine) and PE (Phosphatidyl-ethanolamine) on aluminium oxide column was studied. The regeneration of alumina oxide and the contiunous loading of the process were experimented. The process was developed and optimized absolutely.
The elution effect of chloroform-methanol, methanol-chloroform-triethylamine, methanol-ammonia and methanol-chloroform-ammonia on the process for simultaneous purification of PC and PE by alumina column chromatography was studied. The optimized proportion of elution phase is methanol: chloroform:
ammonia = 80 ∶120∶10 (v/v) and under this condition the recoveries of PC and PE
were both beyond 98% and the theoretical output of PC and PE were 0.31g/g sorbent and 0.04 g/g sorbent.
The regeneration and continous loading were experimented and under the condition of elution phase 80% chloroform-methanol the recoveries of PC and PE were 99.8% and 86.8%; the theoretical outputs were 0.32 g/g sorbent and 0.03 g/g sorbent, the second and third loading amount were as much as the 80.2% and 79.7% of the firet loading amount. PC and PE with purities over 95% and 80% could be
obtained. Under the elution condition of methanol: chloroform: ammonia =80 ∶120 ∶
10 (v/v), the second loading amount was 10.85g which was 94.7% of the fist loading amount.
Based on the former research, the effect of the different elution phases on lecition stability was studied. Experimental results showed that PC and PE were unstable in the solvent with ammonia or triethylamine. The result was valuable for the following work and industrial practice.
The process for simultaneous purification of PC and PE by alumina column chromatography was developed and optimized which has satisfied effect, simple techniques, less investment and large amount loading. The work will provide important basic data for industrial preparation of high-purity phospholipids with alumina column chromatography.
本文在课题组已有研究的基础上，以“氧化铝柱层析分离纯化磷脂的工艺优化”为目标，考察了不同洗脱条件对磷脂在氧化铝柱上的洗脱特性的影响，并进一步进行了氧化铝再生以及工艺连续上样的实验研究，建立并且优化了氧化铝柱层析分离纯化卵磷脂(Phosphatidylcholine，简称PC)和脑磷脂(Phosphatidylethanolamine，简称PE)的完整工艺。
系统考察了甲醇—氯仿；甲醇—氯仿—三乙胺；甲醇—氨水；和甲醇—氯仿—氨水洗脱体系对氧化铝分离纯化磷脂工艺的影响。得到最佳的洗脱液组成为甲醇：氯仿：氨水=80：120：10(v／v)。在此条件下，PC和PE的回收率均达到98％以上，理论产量分别为0.31g／g吸附剂和0.04g／g吸附剂。
对不同洗脱条件下氧化铝的再生以及工艺的连续上样进行了实验研究。结果表明：在洗脱液为甲醇：氯仿=1：4(v／v)的条件下，PC和PE回收率分别达到99.8％和86.8％，理论产量分别达到0.32g／g吸附剂和0.03g／g吸附剂，再次上样量和三次上样量分别达到首次上样量的80.2％和79.7％，并可同时得到纯度大于95％的PC产品和纯度大于80％的PE产品。在有机溶剂和碱组成的洗脱液体系中，再生和连续上样效果最佳的是甲醇：氯仿：氨水=80：120：10(v／v)的洗脱液。在此条件下，再次上样量为10.85g，达到首次上样量的94.7％。
对磷脂在不同溶剂中的稳定性进行了研究。研究结果表明，磷脂在氨水或者三乙胺存在的有机溶剂中是不稳定的。
本文的研究建立了氧化铝柱层析分离纯化高纯度磷脂的完整工艺，该工艺具有分离效果好、工艺简单、投资少、处理量较大等优点。研究结果为高纯度卵磷脂和脑磷脂的制备工艺工业化提供了重要的基础数据。
High-purity phosphohpids are widely used in the fields of food, pharmacy, cosmetic and so on because of the special surface activity and emulsification. .At present most of this production is imported in China and the requirement enhances every year. So it is necessary to develop the preparation of high-purity phospholipids. The research has been doing and great process has been made in our lab.
Aiming at the process optimization for simultaneous purification of PC and PE by alumina column chromatography, the effect of different elution phase on the elution characteristic of PC (Phosphatidylcholine) and PE (Phosphatidyl-ethanolamine) on aluminium oxide column was studied. The regeneration of alumina oxide and the contiunous loading of the process were experimented. The process was developed and optimized absolutely.
The elution effect of chloroform-methanol, methanol-chloroform-triethylamine, methanol-ammonia and methanol-chloroform-ammonia on the process for simultaneous purification of PC and PE by alumina column chromatography was studied. The optimized proportion of elution phase is methanol: chloroform:
ammonia = 80 ∶120∶10 (v/v) and under this condition the recoveries of PC and PE
were both beyond 98% and the theoretical output of PC and PE were 0.31g/g sorbent and 0.04 g/g sorbent.
The regeneration and continous loading were experimented and under the condition of elution phase 80% chloroform-methanol the recoveries of PC and PE were 99.8% and 86.8%; the theoretical outputs were 0.32 g/g sorbent and 0.03 g/g sorbent, the second and third loading amount were as much as the 80.2% and 79.7% of the firet loading amount. PC and PE with purities over 95% and 80% could be
obtained. Under the elution condition of methanol: chloroform: ammonia =80 ∶120 ∶
10 (v/v), the second loading amount was 10.85g which was 94.7% of the fist loading amount.
Based on the former research, the effect of the different elution phases on lecition stability was studied. Experimental results showed that PC and PE were unstable in the solvent with ammonia or triethylamine. The result was valuable for the following work and industrial practice.
The process for simultaneous purification of PC and PE by alumina column chromatography was developed and optimized which has satisfied effect, simple techniques, less investment and large amount loading. The work will provide important basic data for industrial preparation of high-purity phospholipids with alumina column chromatography.
引文
[1] Ansell G B, Hawthone J N. Phospholipid's chemistry, metabolism and function [M]. New York: Elsevier Publishing Company New, 1964.3.
[2] 凌关庭．从卵磷脂商品看中国保健食品市场培育和发展[J]．粮食与油脂，2000，(3)．15-16．
[3] Weber E J, JAOCS. 1981, 58: 898.
[4] 迟玉杰，，林淑英．卵黄卵磷脂提取与应用的研究进展[J]．食品与发酵工业，Vol．28 No．5．
[5] 李涛，王天志．卵磷脂的研究进展，中药材[J]，2002，25：10．
[6] 曹栋，裘爱泳，曹兴国．磷脂结构、性质、功能及研究现状[J]．粮食与油脂，2004，5：3-6．
[7] 陈红．神经鞘磷脂的研究[J]．食品科技，2000，1：3-5．
[8] 徐生庚，裘爱泳主译，贝雷．油脂化学与工艺学[M]．第五版，北京：中国轻工出版社，2001．
[9] Thomas. The component fatty acids of some vegetable seed phosphatides [J]. Biochem., 1942, 36: 815-821.
[10] 蔡秋声．蛋黄磷脂[J]．粮食与油脂，1999，3：12-15．
[11] 崔小明．大豆磷脂的开发和应用前景[J]，甘肃化工，2001，1：10-12．
[12] 钮竹安．大豆磷脂国内外市场和开发[J]．粮食与油脂，1999，1：33-37．
[13] 胡兴中，徐振山．我国大豆磷脂的开发利用状况与产业发展设想[J]．中国油脂，1999，3：54-56．
[14] 汪勇．大豆磷脂酰胆碱分离，纯化和检测方法[J]，粮食与油脂，2003．2．
[15] 彭一鸣，蒋笃孝．PTLC法纯化大豆卵磷脂及鉴定[J]．中国油脂，2002，27，6：62-64．
[16] Lendrath G. Behavior of vegetable phosphlipids in thin-layer chromatography: optimization of mobile phase, detection and direct evaluation [J]. J. Chromatography, 1990, 502: 385.
[17] Gabriels M, Camu F, Plaizier-Vercammen J. Improved thin-layer chromato- graphic method for the separation of cholesterol, egg phosphatidylcholine, and their degradation products [J]. AOAC International, 2002, 85, 6: 1273-1287.
[18] Nzai J M, Proctor A. Phospholipids determination in vegetable oil by thin-layer chromatography and imaging densitometry [J]. Food chemistry, 1998, 63, 4: 571-576.
[19] AOCS official Method Ja7-86, 1997.
[20] 王智华．反相离子对高效液相色谱法分离不同种类的磷脂酰胆碱[J]，色谱，2002，20，3：249-252．
[21] Amari J V, Brown P R, Isolation and purification of lecithin by preparative high performance liquid chromatography[J], J Chromatogr, 1990, 517: 219.
[22] Toshihiko Yamagishi. Hiroshi Akiyama. Shinichi Kimura and Masashi Toyoda. Quantitative Determination of phosphatidylcholine by an HPLC-RI system. JAOCS Vol.66.no. 12 (December 1689)
[23] 孔秀阁，阎丽波．浅析浓缩大豆磷脂生产工艺[J]，中国油脂，2001，26，2．
[24] 剂凯洋，高纯度大豆粉末磷脂的制取新工艺研究[J]，中国油脂，2000，25，5．
[25] 李卫，邵友元．柱层析法分离脑磷脂和卵磷脂[J]，湖北工学院学报，2001，16，2：63-65．
[26] Lea C H, Rhodes D N. The chromatographic separation of glycerophospholipids [J]. Biochem J, 1955, 60:353.
[27] Singleton W S, Gray M S, Brown M L. White Chromatographically homogeneous lecithin from egg phospholipids. JAOCS, 1965, 42, 1: 53-56.
[28] Nielsen J R, A simple chromatographic method for purification of egg lecithin [J]. Lipids, 1980, 15, 6: 481-484.
[29] Bandi Z L, Moslen M T, Reynolds E S. Ion-exchange chromatography of phosphatides and glycolipids on quaternary triethylammonium glycophase covalently bound to controlled pore glass[J]. J Chromatogr, 1982, 249: 93-101.
[30] Un Hoi do, Shi-lung Lo. Separation of phospholipids using a diethylaminoethyl-silica gel column and thin-layer chromatography [J]. J Chromatogr, 1986, 381:233-240.
[31] Juneja L R, Sugino H, Fujiki M. Preaparation of Pure Phospholipids from Egg Yolk [J]. Egg Uses Process Technology. Chapter twelve, 139-149.
[32] 曹栋，裘爱泳，超临界流体分离磷脂酰胆碱[J]，中国油脂，2002，27，3：72-74．
[33] Juneja L R, Yamane T. Shoichi Shimizu.Enzymatic Method of Increasing Phosphatidylcholine content of lecithin. JAOCS, 1989, 66, 5: 714-717.
[34] 顾霞敏．高含量大豆卵磷脂的制备工艺研究．浙江大学硕士论文，2006．
[35] Niki E, Yamamoto Y, Kamiya Y. Oxidation of phosphatidylcholine and its inhibition by vitamin E and vitamin C [A]. Edited by Bors W, Saran M, Tait D. Oxgen Radicals Chem. Biol. [C]. Berlin: de Gruyter. 1984. 273—280.
[36] 北川浩，铃木谦一郎．吸附的基础与设计[M]．1983，北京：化学工业出版社．
[37] 叶振华．吸着分离过程基础[M]．1988，北京：化学工业出版社．
[38] 中国市政工程华北设计院，中国工程建渗标准化协会标准饮用水除氟设计规程[M]1993．
[39] 张燕，谢虹，贾文波，吴志刚．活性氧化铝除氟剂的再生性能研究[J]．武汉大学学报(理学版)，2005，51，4：457-460
[40] 韩金勇，宣启波，于传娥，郭绍山．双氧水生产中废氧化铝的再生利用研究[J]．中国资源综合利用，2000．04：12-13．
[41] 成慧梅．小型水厂强碱再生活性氧化铝除氟滤料初探[J]．内蒙古科技与经济，2004．20：50-51
[42] 吴奕．活性炭的再生方法[J]．化工生产与技术，2005，12，1：20-22
[43] Racz R.Adsorbent Usage for VOC Removal in Sythetic Latices Macromot. Symp.2000.155:171-180
[44] Hajime Tamon, M.O. Influence of Surface Oxides on Ethanol Regeneration of Spent Carbonaceous Adsorbents. Journal of Colloid and Interface Science [J], 1997. 196: 120-122
[45] 童少平，魏红，刘维．臭氧氧化法再生活性炭的研究[J]，工业水处理，2005．25，2：31-33
[46] 郑德海，奚风涛．氮气再生硅胶吸附水分在溶解乙炔生产上的应用[J]，深冷技术，1994．4：27-29
[47] 熊德琴，李春晖，林培喜．吸水硅胶的快速再生试验[J]．茂名学院学报， 2005. 15, 4:20-21
[48] Wei-Nong Zhang, Zhi-Xiong Hua, Yu-Qi Feng, Shi-Lu Da. Optimization of parameters of high-performance displacement chromatography for separation of soybean phosphatidylcholine and phosphatidylethanolamine[J], Journal of Chromatography A, 2005, 1068:269-278
[49] 杨莹琴，马稳，马万山．多孔质天然沸石颗粒吸附剂对铜离子的吸附及再生[J]．非金属矿，2004．27，2：44-48
[50] 李彬，宁平，陈玉保，邱琳．除磷稀土吸附剂的再生实验研究．昆明理工大学学报(理工版)，2005．30，4：88-91
[51] 苗青，许家鸾，王京霞．D101吸附树脂再生工艺研究研究：Ⅰ：再生液酸、碱、醇浓度及浸泡时间对再生效果的影响[J]．中国现代应用药学杂志，2004．21，2：152-154．
[52] 曾骆，戴静．吸附树脂再生效果的实验研究[J]．时珍过医国药，2005．16，4：279-280．
[53] 韩珍琼，刘期凯．啤酒生产中硅藻土再生处理方法的研究[J]．食品工业科技，2004．25，12：77-79．
[54] 王扬，郑红，李海云，吴祖义，梁树平．三价铬离子改性膨润土除酚效果及再生研究[J]．非金属矿，2005．28，4：44-47．
[55] Salgin U, Y.N., Calimli A, Desorption of salicylic acid from modified bentonite by using supercritical fluids in packed bed column [J]. Separation Sciebce and Technology, 2004.39, 11: 2677-2694.
[56] A. Tejeda-Mansir, R.E., R.M. Montesinos, R. Guzman, Modelling regeneration effects on protein an affinity chromatography [J]. Bioprocess Engineering, 1997. 17: 39-44.
[57] 陈南羽，封禄田．卵磷脂的应用与开发[J]．化学工业与工程技术，1999，20，2：21-23．
[58] Bernard F.Szuhaj. Lectihins, Sources, Manufacture&Use.AOCS, 1989:10-11
[59] 李卫，邵友元，黄光斗．卵磷脂的提取及热稳定性研究[J]．甘肃化工，2001，2：71-74．
[60] 何新霞，徐丽珊，杨玲．大豆磷脂胶丸稳定性的初步考察[J]．中国药学杂志，1999，34，8：538-540．
[61] 张世宏．大豆磷脂湿胶的稳定性研究[J]．武汉工业学院学报，2000，2：4-6．
[62] 黄莉．光照条件下粉末磷脂储藏稳定性初探[J]．粮食储藏．1997，26，5：29-33．
[63] 徐丽珊，何新霞，周虞灿．磷脂复合物的稳定性研究[J]．食品科学，2000，21，1：21-22．
[64] 周景春，孟冬娅，石富成．大豆油、水分和铁对大豆磷脂酸值和碘值的影响[J]．中国生化药物杂志，2001，22，2：90-92．
[65] Renkonen O. Breakdown of lecithin on aluminum oxide columns [J]. Lipid Research, 1962, 3, 2: 181-183.
[66] Kazumasa Y, Koji U, Tetsuo N. Autoxidation of Phosphatidyl- ethanolamine and Phosphatidylcholine in Organic Solvents[J]. Yukagaku, 1992, 41, 7:582-585.
[67] Niki E, Yamamoto Y, Kamiya Y. Oxidation ofphosphatidylcholine and its inhibition by vitamin E and vitamin C[A]. Edited by Bors W, Saran M, Tait D. Oxgen Radicals Chem. Biol. [C]. Berlin: de Gruyter. 1984. 273—280.
[68] 徐小红．高纯度蛋黄卵磷脂制备方法的研究，浙江大学硕士论文，2002．
[69] 许晓慧．蛋黄卵磷脂的提取及卵磷脂的纯化工艺研究．西北大学硕士论文，2002．
[70] 许海丹．蛋黄磷脂的精制工艺及其稳定性研究．浙江大学硕士论文，2004．
[71] Stephen H, Stephen T. solubilities of Inorganic and Organic Compounds [M], Volume Ⅰ Part Ⅰ. New York: Pergamon Press, 1963.370.
[2] 凌关庭．从卵磷脂商品看中国保健食品市场培育和发展[J]．粮食与油脂，2000，(3)．15-16．
[3] Weber E J, JAOCS. 1981, 58: 898.
[4] 迟玉杰，，林淑英．卵黄卵磷脂提取与应用的研究进展[J]．食品与发酵工业，Vol．28 No．5．
[5] 李涛，王天志．卵磷脂的研究进展，中药材[J]，2002，25：10．
[6] 曹栋，裘爱泳，曹兴国．磷脂结构、性质、功能及研究现状[J]．粮食与油脂，2004，5：3-6．
[7] 陈红．神经鞘磷脂的研究[J]．食品科技，2000，1：3-5．
[8] 徐生庚，裘爱泳主译，贝雷．油脂化学与工艺学[M]．第五版，北京：中国轻工出版社，2001．
[9] Thomas. The component fatty acids of some vegetable seed phosphatides [J]. Biochem., 1942, 36: 815-821.
[10] 蔡秋声．蛋黄磷脂[J]．粮食与油脂，1999，3：12-15．
[11] 崔小明．大豆磷脂的开发和应用前景[J]，甘肃化工，2001，1：10-12．
[12] 钮竹安．大豆磷脂国内外市场和开发[J]．粮食与油脂，1999，1：33-37．
[13] 胡兴中，徐振山．我国大豆磷脂的开发利用状况与产业发展设想[J]．中国油脂，1999，3：54-56．
[14] 汪勇．大豆磷脂酰胆碱分离，纯化和检测方法[J]，粮食与油脂，2003．2．
[15] 彭一鸣，蒋笃孝．PTLC法纯化大豆卵磷脂及鉴定[J]．中国油脂，2002，27，6：62-64．
[16] Lendrath G. Behavior of vegetable phosphlipids in thin-layer chromatography: optimization of mobile phase, detection and direct evaluation [J]. J. Chromatography, 1990, 502: 385.
[17] Gabriels M, Camu F, Plaizier-Vercammen J. Improved thin-layer chromato- graphic method for the separation of cholesterol, egg phosphatidylcholine, and their degradation products [J]. AOAC International, 2002, 85, 6: 1273-1287.
[18] Nzai J M, Proctor A. Phospholipids determination in vegetable oil by thin-layer chromatography and imaging densitometry [J]. Food chemistry, 1998, 63, 4: 571-576.
[19] AOCS official Method Ja7-86, 1997.
[20] 王智华．反相离子对高效液相色谱法分离不同种类的磷脂酰胆碱[J]，色谱，2002，20，3：249-252．
[21] Amari J V, Brown P R, Isolation and purification of lecithin by preparative high performance liquid chromatography[J], J Chromatogr, 1990, 517: 219.
[22] Toshihiko Yamagishi. Hiroshi Akiyama. Shinichi Kimura and Masashi Toyoda. Quantitative Determination of phosphatidylcholine by an HPLC-RI system. JAOCS Vol.66.no. 12 (December 1689)
[23] 孔秀阁，阎丽波．浅析浓缩大豆磷脂生产工艺[J]，中国油脂，2001，26，2．
[24] 剂凯洋，高纯度大豆粉末磷脂的制取新工艺研究[J]，中国油脂，2000，25，5．
[25] 李卫，邵友元．柱层析法分离脑磷脂和卵磷脂[J]，湖北工学院学报，2001，16，2：63-65．
[26] Lea C H, Rhodes D N. The chromatographic separation of glycerophospholipids [J]. Biochem J, 1955, 60:353.
[27] Singleton W S, Gray M S, Brown M L. White Chromatographically homogeneous lecithin from egg phospholipids. JAOCS, 1965, 42, 1: 53-56.
[28] Nielsen J R, A simple chromatographic method for purification of egg lecithin [J]. Lipids, 1980, 15, 6: 481-484.
[29] Bandi Z L, Moslen M T, Reynolds E S. Ion-exchange chromatography of phosphatides and glycolipids on quaternary triethylammonium glycophase covalently bound to controlled pore glass[J]. J Chromatogr, 1982, 249: 93-101.
[30] Un Hoi do, Shi-lung Lo. Separation of phospholipids using a diethylaminoethyl-silica gel column and thin-layer chromatography [J]. J Chromatogr, 1986, 381:233-240.
[31] Juneja L R, Sugino H, Fujiki M. Preaparation of Pure Phospholipids from Egg Yolk [J]. Egg Uses Process Technology. Chapter twelve, 139-149.
[32] 曹栋，裘爱泳，超临界流体分离磷脂酰胆碱[J]，中国油脂，2002，27，3：72-74．
[33] Juneja L R, Yamane T. Shoichi Shimizu.Enzymatic Method of Increasing Phosphatidylcholine content of lecithin. JAOCS, 1989, 66, 5: 714-717.
[34] 顾霞敏．高含量大豆卵磷脂的制备工艺研究．浙江大学硕士论文，2006．
[35] Niki E, Yamamoto Y, Kamiya Y. Oxidation of phosphatidylcholine and its inhibition by vitamin E and vitamin C [A]. Edited by Bors W, Saran M, Tait D. Oxgen Radicals Chem. Biol. [C]. Berlin: de Gruyter. 1984. 273—280.
[36] 北川浩，铃木谦一郎．吸附的基础与设计[M]．1983，北京：化学工业出版社．
[37] 叶振华．吸着分离过程基础[M]．1988，北京：化学工业出版社．
[38] 中国市政工程华北设计院，中国工程建渗标准化协会标准饮用水除氟设计规程[M]1993．
[39] 张燕，谢虹，贾文波，吴志刚．活性氧化铝除氟剂的再生性能研究[J]．武汉大学学报(理学版)，2005，51，4：457-460
[40] 韩金勇，宣启波，于传娥，郭绍山．双氧水生产中废氧化铝的再生利用研究[J]．中国资源综合利用，2000．04：12-13．
[41] 成慧梅．小型水厂强碱再生活性氧化铝除氟滤料初探[J]．内蒙古科技与经济，2004．20：50-51
[42] 吴奕．活性炭的再生方法[J]．化工生产与技术，2005，12，1：20-22
[43] Racz R.Adsorbent Usage for VOC Removal in Sythetic Latices Macromot. Symp.2000.155:171-180
[44] Hajime Tamon, M.O. Influence of Surface Oxides on Ethanol Regeneration of Spent Carbonaceous Adsorbents. Journal of Colloid and Interface Science [J], 1997. 196: 120-122
[45] 童少平，魏红，刘维．臭氧氧化法再生活性炭的研究[J]，工业水处理，2005．25，2：31-33
[46] 郑德海，奚风涛．氮气再生硅胶吸附水分在溶解乙炔生产上的应用[J]，深冷技术，1994．4：27-29
[47] 熊德琴，李春晖，林培喜．吸水硅胶的快速再生试验[J]．茂名学院学报， 2005. 15, 4:20-21
[48] Wei-Nong Zhang, Zhi-Xiong Hua, Yu-Qi Feng, Shi-Lu Da. Optimization of parameters of high-performance displacement chromatography for separation of soybean phosphatidylcholine and phosphatidylethanolamine[J], Journal of Chromatography A, 2005, 1068:269-278
[49] 杨莹琴，马稳，马万山．多孔质天然沸石颗粒吸附剂对铜离子的吸附及再生[J]．非金属矿，2004．27，2：44-48
[50] 李彬，宁平，陈玉保，邱琳．除磷稀土吸附剂的再生实验研究．昆明理工大学学报(理工版)，2005．30，4：88-91
[51] 苗青，许家鸾，王京霞．D101吸附树脂再生工艺研究研究：Ⅰ：再生液酸、碱、醇浓度及浸泡时间对再生效果的影响[J]．中国现代应用药学杂志，2004．21，2：152-154．
[52] 曾骆，戴静．吸附树脂再生效果的实验研究[J]．时珍过医国药，2005．16，4：279-280．
[53] 韩珍琼，刘期凯．啤酒生产中硅藻土再生处理方法的研究[J]．食品工业科技，2004．25，12：77-79．
[54] 王扬，郑红，李海云，吴祖义，梁树平．三价铬离子改性膨润土除酚效果及再生研究[J]．非金属矿，2005．28，4：44-47．
[55] Salgin U, Y.N., Calimli A, Desorption of salicylic acid from modified bentonite by using supercritical fluids in packed bed column [J]. Separation Sciebce and Technology, 2004.39, 11: 2677-2694.
[56] A. Tejeda-Mansir, R.E., R.M. Montesinos, R. Guzman, Modelling regeneration effects on protein an affinity chromatography [J]. Bioprocess Engineering, 1997. 17: 39-44.
[57] 陈南羽，封禄田．卵磷脂的应用与开发[J]．化学工业与工程技术，1999，20，2：21-23．
[58] Bernard F.Szuhaj. Lectihins, Sources, Manufacture&Use.AOCS, 1989:10-11
[59] 李卫，邵友元，黄光斗．卵磷脂的提取及热稳定性研究[J]．甘肃化工，2001，2：71-74．
[60] 何新霞，徐丽珊，杨玲．大豆磷脂胶丸稳定性的初步考察[J]．中国药学杂志，1999，34，8：538-540．
[61] 张世宏．大豆磷脂湿胶的稳定性研究[J]．武汉工业学院学报，2000，2：4-6．
[62] 黄莉．光照条件下粉末磷脂储藏稳定性初探[J]．粮食储藏．1997，26，5：29-33．
[63] 徐丽珊，何新霞，周虞灿．磷脂复合物的稳定性研究[J]．食品科学，2000，21，1：21-22．
[64] 周景春，孟冬娅，石富成．大豆油、水分和铁对大豆磷脂酸值和碘值的影响[J]．中国生化药物杂志，2001，22，2：90-92．
[65] Renkonen O. Breakdown of lecithin on aluminum oxide columns [J]. Lipid Research, 1962, 3, 2: 181-183.
[66] Kazumasa Y, Koji U, Tetsuo N. Autoxidation of Phosphatidyl- ethanolamine and Phosphatidylcholine in Organic Solvents[J]. Yukagaku, 1992, 41, 7:582-585.
[67] Niki E, Yamamoto Y, Kamiya Y. Oxidation ofphosphatidylcholine and its inhibition by vitamin E and vitamin C[A]. Edited by Bors W, Saran M, Tait D. Oxgen Radicals Chem. Biol. [C]. Berlin: de Gruyter. 1984. 273—280.
[68] 徐小红．高纯度蛋黄卵磷脂制备方法的研究，浙江大学硕士论文，2002．
[69] 许晓慧．蛋黄卵磷脂的提取及卵磷脂的纯化工艺研究．西北大学硕士论文，2002．
[70] 许海丹．蛋黄磷脂的精制工艺及其稳定性研究．浙江大学硕士论文，2004．
[71] Stephen H, Stephen T. solubilities of Inorganic and Organic Compounds [M], Volume Ⅰ Part Ⅰ. New York: Pergamon Press, 1963.370.