番茄红素的提取纯化工艺及稳定性研究
|
摘要
番茄红素是一种天然类胡萝卜素，它具有防癌抗癌，提高免疫力，延缓衰老等多种保健功能。本文主要研究了从天然番茄中提取番茄红素粗产品，并进一步将粗产品纯化的工艺路线和工艺条件。
采用皂化法对番茄进行预处理不仅可除去脂肪酸甘油酯及各种游离脂肪酸，而且在番茄细胞组织的保护下，番茄红素不容易在碱性条件下被破坏。番茄皂化的最佳工艺条件为：温度70℃，KOH浓度0.5mol/L，皂化时间0.5h。从皂化后的番茄中提取的番茄红素产品有番茄红素晶体出现，产品的色价由原来的21.2提高到66.7。
分别采用固液浸取法、索氏提取法和微波萃取法用石油醚从皂化后干燥番茄粉末中提取番茄红素，考察了温度、时间、液固比、功率等各种提取条件对番茄红素提取量的影响。固液浸取法最佳工艺条件：温度50～60℃，提取时间1h，液固比40:1，番茄红素产量为22.54mg/g(干燥番茄粉末)；索氏提取的最佳工艺条件：温度45℃，提取时间4h，液固比100:1，番茄红素产量为22.69mg/g(干燥番茄粉末)；微波萃取的最佳工艺条件：功率400w，萃取时间50s，番茄红素提取量23.25mg/g(干燥番茄粉末)。三种方法对比表明微波萃取的提取量最大，而且提取速度最快。
用柱色谱法分离纯化提取阶段得到的产品—番茄红素油树脂。根据番茄红素的性质和薄层色谱的试验结果，色谱系统的固定相选用硅胶，流动相选用V(正庚烷):V(丙酮)=6:1的混合液，洗脱方式为梯度洗脱。为了提高混合物的分离度，对色谱操作条件进行了优化，确定出最佳流动相流速为5ml/min，高径比30:1。纯化后的产品经HPLC检验，番茄红素质量百分含量达到18.9％。另外，从番茄红素油树脂中还得到β-胡萝卜素和叶黄素。
影响番茄红素稳定性的因素主要有光、空气、温度、和pH值，这些因素作用程度的大小取决于番茄红素的存在状态，空气对番茄粉末的稳定性影响最大，光对提取液的稳定性影响最大。
Lycopene is a kind of natural carotenoid, it has the function of various health cares, such as resisting cancer, increasing the immunity and deferring decrepitude etc. The paper mainly studies the technologic processes and conditions by which oleoresin be extracted from natural tomatoes then be purified into lycopene crystal.
Alkaline saponification can remove the glyceride and fatty acid, and release lycopene. Under the protection of tomato cell tissue lycopene will not denature. The best conditions of tomato saponification are temperature 70 C, concentration of KOH 0.5 mol/L and time 0.5h.The lycopene oleoresin extracted from sapnification tomato included some crystal, and its color value increased to 66.7
The methods including solid-liquid leaching, soxholet extraction and microwave extraction have been utilized to gain crude product from dry tomato powder with petroleum benzene. The factors such as temperature, time, ratio of liquid-solid, microwave power will affect the amount of oleoresin. The result shows that the best conditions of solid-liquid leaching are temperature 50-60 C, time Ih, ratio of liquid-solid 40:1,and the amount of oleoresin is 22.54mg/g (tomato powder); the best conditions of soxholet extraction are temperature 45 C, time 4h, ratio of liquid-solid 100:1,and the amount of oleoresin is 22.69mg/g (tomato powder); the best conditions of microwave extraction are power 400w, time 60s, and the amount of oleoresin is 22.54mg/g (tomato powder). The comparation of the three methods proves that microwave extraction can be used in industrial production of lycopene because of its high efficiency and output.
Oleoresin can be purified by column-chromatography. The immobile phase is silica-gel and the mobile phase is the mixture of V (hexane): V (acetone)=1.5:1. The result of optimization for column-chromatography condition proves that the best flow speed of immobile phase is 5ml/min, and the ratio of height-diameter is 30:1. The needle crystal can be gained from the concentrated eluting liquid, and the measurement by HPLC shows that the mass
percentage of lycopene is 18.9%. Other ingredients separated from oleoresin are B-carotene and lutehi.
Light, oxide, temperature and acid salt tolerance will affect the stability of lycopene. The degree of lycopene denaturalization is often different from the state in which lycopene exists. Oxide denatures tomato powder more evidently and light affects the stability of lycopene solvent more distinctly.
采用皂化法对番茄进行预处理不仅可除去脂肪酸甘油酯及各种游离脂肪酸，而且在番茄细胞组织的保护下，番茄红素不容易在碱性条件下被破坏。番茄皂化的最佳工艺条件为：温度70℃，KOH浓度0.5mol/L，皂化时间0.5h。从皂化后的番茄中提取的番茄红素产品有番茄红素晶体出现，产品的色价由原来的21.2提高到66.7。
分别采用固液浸取法、索氏提取法和微波萃取法用石油醚从皂化后干燥番茄粉末中提取番茄红素，考察了温度、时间、液固比、功率等各种提取条件对番茄红素提取量的影响。固液浸取法最佳工艺条件：温度50～60℃，提取时间1h，液固比40:1，番茄红素产量为22.54mg/g(干燥番茄粉末)；索氏提取的最佳工艺条件：温度45℃，提取时间4h，液固比100:1，番茄红素产量为22.69mg/g(干燥番茄粉末)；微波萃取的最佳工艺条件：功率400w，萃取时间50s，番茄红素提取量23.25mg/g(干燥番茄粉末)。三种方法对比表明微波萃取的提取量最大，而且提取速度最快。
用柱色谱法分离纯化提取阶段得到的产品—番茄红素油树脂。根据番茄红素的性质和薄层色谱的试验结果，色谱系统的固定相选用硅胶，流动相选用V(正庚烷):V(丙酮)=6:1的混合液，洗脱方式为梯度洗脱。为了提高混合物的分离度，对色谱操作条件进行了优化，确定出最佳流动相流速为5ml/min，高径比30:1。纯化后的产品经HPLC检验，番茄红素质量百分含量达到18.9％。另外，从番茄红素油树脂中还得到β-胡萝卜素和叶黄素。
影响番茄红素稳定性的因素主要有光、空气、温度、和pH值，这些因素作用程度的大小取决于番茄红素的存在状态，空气对番茄粉末的稳定性影响最大，光对提取液的稳定性影响最大。
Lycopene is a kind of natural carotenoid, it has the function of various health cares, such as resisting cancer, increasing the immunity and deferring decrepitude etc. The paper mainly studies the technologic processes and conditions by which oleoresin be extracted from natural tomatoes then be purified into lycopene crystal.
Alkaline saponification can remove the glyceride and fatty acid, and release lycopene. Under the protection of tomato cell tissue lycopene will not denature. The best conditions of tomato saponification are temperature 70 C, concentration of KOH 0.5 mol/L and time 0.5h.The lycopene oleoresin extracted from sapnification tomato included some crystal, and its color value increased to 66.7
The methods including solid-liquid leaching, soxholet extraction and microwave extraction have been utilized to gain crude product from dry tomato powder with petroleum benzene. The factors such as temperature, time, ratio of liquid-solid, microwave power will affect the amount of oleoresin. The result shows that the best conditions of solid-liquid leaching are temperature 50-60 C, time Ih, ratio of liquid-solid 40:1,and the amount of oleoresin is 22.54mg/g (tomato powder); the best conditions of soxholet extraction are temperature 45 C, time 4h, ratio of liquid-solid 100:1,and the amount of oleoresin is 22.69mg/g (tomato powder); the best conditions of microwave extraction are power 400w, time 60s, and the amount of oleoresin is 22.54mg/g (tomato powder). The comparation of the three methods proves that microwave extraction can be used in industrial production of lycopene because of its high efficiency and output.
Oleoresin can be purified by column-chromatography. The immobile phase is silica-gel and the mobile phase is the mixture of V (hexane): V (acetone)=1.5:1. The result of optimization for column-chromatography condition proves that the best flow speed of immobile phase is 5ml/min, and the ratio of height-diameter is 30:1. The needle crystal can be gained from the concentrated eluting liquid, and the measurement by HPLC shows that the mass
percentage of lycopene is 18.9%. Other ingredients separated from oleoresin are B-carotene and lutehi.
Light, oxide, temperature and acid salt tolerance will affect the stability of lycopene. The degree of lycopene denaturalization is often different from the state in which lycopene exists. Oxide denatures tomato powder more evidently and light affects the stability of lycopene solvent more distinctly.
引文
1. Beecher G R. Nutrient content of tomatoes and tomato products.Pro Soc Exp Biol Med. 1998, 218(2):98-100
2. Weisburger J H. Evaluation of the evidence on the role of tomato products in disease prevention. Pro Soc Exp Biol Med. 1998,218(2):140-143
3．王业勤，李勤生．天然类胡萝卜素，北京，中国医药科技出版社．1997，245-292
4. Yoav Sharoni, Michael Danilenko. Molecular mechanisms for the anticancer activity of the carotenoid lycopene. Drug Development Research. 1999, 53(2):38-45
5. Shi. J, Maguer. M. I, Kakuda. Y. Lycopene Degradation and Lsomefization in Tomato Dehydration. Food Res. Intl. 1999.32:15-21
6．李增光，吴骥陶．番茄浆中番茄红素的测定．食品与发酵工业，1991，16(2)：82-84
7．孙庆杰，丁霄霖．番茄红素的保健作用与开发．食品与发酵工业，1997，23(4)：72-75。
8. Joseph Schierle. Content and Isomeric ratio of lycopene in food and human blood plasma, Food Technology. 1997, 59 (3):459-465.
9. Di Mascio P, Kaisert S, Sies H, et al. Lycopene as the most Efficient Biological Carotenid Single Oxygen Quencher. Arch Bichern Biophys. 1989, 274(1): 1-7
10. Sharma S. K et al. Kinetic of Lycopene degradation in tomato pulp solids under different and storage conditions. Food Research International. 1996, 29(3): 309～315.
11. T. Snguelova, J. Warthesen. Lycopene stability in tomato powers. J. Food Sci. 2000, 65:67-70
12．马柏林．食用调和油中的番茄红素的稳定性研究．中国油脂．2000(6)：132-133．
13. Mario G. Ferruzzi, Minhthy L. Nguyen. Analysis of lycopene geometrical isomers in biological microsamples by liquid chromatography with coulometric array detection. Journal of chromatography. B.bimedical applications. 1992 (9): 373-378
14. Anocha K. et al. Analysis of Carotenoid. Food chemistry. 1998, 63(4):577-584
15．张连富丁宵霖．番茄红素简便测定方法的建立．食品与发酵工业．2001，27(3)：51-55
16. O. Mikes. Laboratory Handbook of chromatographic. New York. John Wiley & Sons. 1979
17．果秀敏，翟彤宇．薄层色谱法分离菠菜叶片中色素的研究．河北农业大学学报．
18．成都科技大学分析化学教研组合编．分析化学：华东化工学院高等教育出版社．1988，5
19．奥·米克斯(捷克)．杨文澜译．色谱及有关方法的试验手册．机械工业出版社．1986，9
20. H. Pfander, R. Riesen et al. Carotenoids in Human Health. Appl. Chem. 1994, 66:947-955
21．陆冰真，翟永信．薄层层析在食品中的应用．北京：北京大学出版社．1991，6
22. Teodor hodisan, Carmen Soxaciu, et al. Carotenoid composition of Rosa Canina fruits determined by
thin-layer Chromatography. Journal of Pharmaceutical and Biomedical Analysis. 1997, (16):521-528
23．高崑玉．色谱法在精细化工中的应用．中国石化出版社．1997，11
24. K. Jinno, Y.lin. Separation of carotenoids by High-performance chromatography with Polymeric and Monomeric Octadecylsitica Station Phase. Chroma. 1995, 41 (6):311-317
25．王强等．反相高效液相色谱法同时测定番茄中5中类胡萝卜素．色谱．1997，15(6)：534-538
26. Curt Enaeehiser, Nada Simunovit, et al. Separation of Geometrical Carotenoid Ismers in Biological Extracts Using a Polymeric C30 Column in Reversed-phase Liquid Chromatography. J. Agric. Food Chem. 1996, 44:3887-3893
27. Katherine S. Epler, et al. Evaluation of reversed-phase liquid chromatographic columns for recovery and selectivity of selected Carotenoids. J. Chroma. 1992, 595:89-101
28. T. Lacker, S. strohschein. Separation and identification of various carotenoids by C30 reversed-phase high-performance liquid chromatography coupled to UV and atmospheric pressure chemical ionization mass spectrometric detectim. J. Chroma. 1999, 854:37-44
29. Emenhiser C., Sander L. et al. J. Capability of polymeric C_(30)stationary phase to resolve cis-tran carotenoid isomers in reverse liquid chromagraphy. Chroma. 1995, 707:205-216
30. Di Mascio P, Kaisert S, Sies H, et al. Lycopene as the most Efficient Biological Carotenid Single Oxygen Quencher. Arch Bichern Biophys. 1989, 274(1): 1-7
31. Van KH, Brouwer IA, West CE, et al. Bioacailability of Lutein from Vegetables is 5 times high than that of beta-carotene. Am J Clin Nutr. 1999.70(2): 179-182
32. Shi. J, Maguer. M. I, Kakuda. Y. Lycopene Degradation and Isomerization in Tomato Dehydration. Food Res. Intl. 1999, 32:15-21
33. Rao A V. Agarwal S. Baioavailability and in vivo antioxidant properities of lycopene from tomato products and their possible role in the prevention of cancer. Nutr Cancer. 1998, 31 (3):199-203
34. Rao A V. Agarwal S. Tomato Lycopene and low density lipoprotein oxidation:a human dietary intervention study. Lipids. 1998, 33(10):981-984
35. Campbell L, Mi Kyung Kim, Sei Hyun Ahn. Relationship of serum α-tocopherol, carotenoids and retinol with the risk of breast cancer. Nutrition Research. 1991, 108:45～48
36. Giovannucci E, Ascherio A et al. Carotenoid and Cancer. Food Technology. 1995, 87:1767～1776
37. Nir Z, et al. Lycopene from tomatoes[J]. IFI. 1993, 6:45-51
38. P. Lindner et al. Distribution of protein, lycopene and elements Ca, Mg, P and N among carious fractions of tomato juice. J. Food Sci. 1984,49:1214-1218
39．姜文候．β-胡萝卜素的应用、市场和天然型产品的发酵法生产．食品与发酵工业，1994 (3)：65-71．
40．周静，徐以玲．提取番茄皮色素的初步研究．新疆农垦科技．1992，4：28-29
41．蔡俊，邱雁临，谈小兰等．番茄红素提取工艺的研究．食品与发酵工业．2000，26(2)：50-53
42．D．哈提尔，Y．拉维．稳定的番茄红素浓缩物及制备方法．CN 1176577．1998，3
43. Rodeny L, Des Moines et al. Process for the isolation and purification of lycopene crystal. WO 98/43620.1998,6
44．文刚，胡光，李新．番茄酱制取结晶番茄红素及番茄红素油树脂的方法．CN1298904A．2001，6
45. PCT/US98/02335, WO 98/43620(美国专利)
46. Dubodel N P, et al. Piscchchevaya Promyshlennost, Moscow, 1995,(1), 16
47．安本光政等，日本食品科学工学会第42回大会讲演集，1995，3
48. Wo 96/13178, 1996, 5
49．孙庆杰，丁霄霖．超临界CO_2萃取番茄红素的初步研究。食品与发酵工业．1998，24(1)：3-6
50. Inakuma T et al. Effect of drying methods on extraction of lycopene in tomato skin with supercritical carbon dioxide. J. of the Japanese Soceity for Food Science and Technology. 1996,32 (5):740-743.
51. Gavrilov A. S et al. Industrial production of a microbiological method. Applied Biochemistryand Microbiology. 1996, 32(5), 492-494
52. Minura Y et al. Production of lycopene by the food yeast, Candida utilis that dose not naturally synthesize carotenoid. Biotechnology and Bioengineering, 1998, 58(2/3):306-308
53．岑宁，王杰．柑橘果皮番茄红素诱导合成研究冲国南方果树．1996，25 (3)：6-7
54. Sari H. Hakila I. Marina Heinoen. Chromatographic Purification of Natural Lycopene. J. Agric.Food Chem, 1994 42(6):1314-1316
55．何碧烟，欧光南．食品添加剂对番茄红素稳定性的影响．集美大学学报．2000，5(1)：47-51
56．李伟，孙新虎．番茄红素溶解度的测定．食品与发酵工业．2002，28(8)：54-56
57．Kingston H M，Jassic L B编．郭振库，卜玉兰，刘正阳等译．分析化学中的微波制样技术．北京：气象出版社，1992
58．郭振库，卜玉兰．微波萃取技术．色谱．1997，15(6)：499-501
59．郝金玉，黄若华，王平艳．微波萃取技术进展．云南化工．1999，3：26-29
60．古华民，张斌东，胡男．应用统计方法．暨南大学出版社．1994
61．达世禄．色谱学导论．武汉大学出版社．1999
62. Sahl W. Sies H. Persaectives in Biochemistry and Biophysics. Lycopene:A Biologically Important Carotenoid for Humans. Arch. Biochem. & Biophys. 1996, 336:1-9
63. Shi J. Maguer M. L., Kakuda Y. Lycopene Degradation and lsomerization in Tomato Dehydration. Food Rea. Intl. 1999, 32:15-21
2. Weisburger J H. Evaluation of the evidence on the role of tomato products in disease prevention. Pro Soc Exp Biol Med. 1998,218(2):140-143
3．王业勤，李勤生．天然类胡萝卜素，北京，中国医药科技出版社．1997，245-292
4. Yoav Sharoni, Michael Danilenko. Molecular mechanisms for the anticancer activity of the carotenoid lycopene. Drug Development Research. 1999, 53(2):38-45
5. Shi. J, Maguer. M. I, Kakuda. Y. Lycopene Degradation and Lsomefization in Tomato Dehydration. Food Res. Intl. 1999.32:15-21
6．李增光，吴骥陶．番茄浆中番茄红素的测定．食品与发酵工业，1991，16(2)：82-84
7．孙庆杰，丁霄霖．番茄红素的保健作用与开发．食品与发酵工业，1997，23(4)：72-75。
8. Joseph Schierle. Content and Isomeric ratio of lycopene in food and human blood plasma, Food Technology. 1997, 59 (3):459-465.
9. Di Mascio P, Kaisert S, Sies H, et al. Lycopene as the most Efficient Biological Carotenid Single Oxygen Quencher. Arch Bichern Biophys. 1989, 274(1): 1-7
10. Sharma S. K et al. Kinetic of Lycopene degradation in tomato pulp solids under different and storage conditions. Food Research International. 1996, 29(3): 309～315.
11. T. Snguelova, J. Warthesen. Lycopene stability in tomato powers. J. Food Sci. 2000, 65:67-70
12．马柏林．食用调和油中的番茄红素的稳定性研究．中国油脂．2000(6)：132-133．
13. Mario G. Ferruzzi, Minhthy L. Nguyen. Analysis of lycopene geometrical isomers in biological microsamples by liquid chromatography with coulometric array detection. Journal of chromatography. B.bimedical applications. 1992 (9): 373-378
14. Anocha K. et al. Analysis of Carotenoid. Food chemistry. 1998, 63(4):577-584
15．张连富丁宵霖．番茄红素简便测定方法的建立．食品与发酵工业．2001，27(3)：51-55
16. O. Mikes. Laboratory Handbook of chromatographic. New York. John Wiley & Sons. 1979
17．果秀敏，翟彤宇．薄层色谱法分离菠菜叶片中色素的研究．河北农业大学学报．
18．成都科技大学分析化学教研组合编．分析化学：华东化工学院高等教育出版社．1988，5
19．奥·米克斯(捷克)．杨文澜译．色谱及有关方法的试验手册．机械工业出版社．1986，9
20. H. Pfander, R. Riesen et al. Carotenoids in Human Health. Appl. Chem. 1994, 66:947-955
21．陆冰真，翟永信．薄层层析在食品中的应用．北京：北京大学出版社．1991，6
22. Teodor hodisan, Carmen Soxaciu, et al. Carotenoid composition of Rosa Canina fruits determined by
thin-layer Chromatography. Journal of Pharmaceutical and Biomedical Analysis. 1997, (16):521-528
23．高崑玉．色谱法在精细化工中的应用．中国石化出版社．1997，11
24. K. Jinno, Y.lin. Separation of carotenoids by High-performance chromatography with Polymeric and Monomeric Octadecylsitica Station Phase. Chroma. 1995, 41 (6):311-317
25．王强等．反相高效液相色谱法同时测定番茄中5中类胡萝卜素．色谱．1997，15(6)：534-538
26. Curt Enaeehiser, Nada Simunovit, et al. Separation of Geometrical Carotenoid Ismers in Biological Extracts Using a Polymeric C30 Column in Reversed-phase Liquid Chromatography. J. Agric. Food Chem. 1996, 44:3887-3893
27. Katherine S. Epler, et al. Evaluation of reversed-phase liquid chromatographic columns for recovery and selectivity of selected Carotenoids. J. Chroma. 1992, 595:89-101
28. T. Lacker, S. strohschein. Separation and identification of various carotenoids by C30 reversed-phase high-performance liquid chromatography coupled to UV and atmospheric pressure chemical ionization mass spectrometric detectim. J. Chroma. 1999, 854:37-44
29. Emenhiser C., Sander L. et al. J. Capability of polymeric C_(30)stationary phase to resolve cis-tran carotenoid isomers in reverse liquid chromagraphy. Chroma. 1995, 707:205-216
30. Di Mascio P, Kaisert S, Sies H, et al. Lycopene as the most Efficient Biological Carotenid Single Oxygen Quencher. Arch Bichern Biophys. 1989, 274(1): 1-7
31. Van KH, Brouwer IA, West CE, et al. Bioacailability of Lutein from Vegetables is 5 times high than that of beta-carotene. Am J Clin Nutr. 1999.70(2): 179-182
32. Shi. J, Maguer. M. I, Kakuda. Y. Lycopene Degradation and Isomerization in Tomato Dehydration. Food Res. Intl. 1999, 32:15-21
33. Rao A V. Agarwal S. Baioavailability and in vivo antioxidant properities of lycopene from tomato products and their possible role in the prevention of cancer. Nutr Cancer. 1998, 31 (3):199-203
34. Rao A V. Agarwal S. Tomato Lycopene and low density lipoprotein oxidation:a human dietary intervention study. Lipids. 1998, 33(10):981-984
35. Campbell L, Mi Kyung Kim, Sei Hyun Ahn. Relationship of serum α-tocopherol, carotenoids and retinol with the risk of breast cancer. Nutrition Research. 1991, 108:45～48
36. Giovannucci E, Ascherio A et al. Carotenoid and Cancer. Food Technology. 1995, 87:1767～1776
37. Nir Z, et al. Lycopene from tomatoes[J]. IFI. 1993, 6:45-51
38. P. Lindner et al. Distribution of protein, lycopene and elements Ca, Mg, P and N among carious fractions of tomato juice. J. Food Sci. 1984,49:1214-1218
39．姜文候．β-胡萝卜素的应用、市场和天然型产品的发酵法生产．食品与发酵工业，1994 (3)：65-71．
40．周静，徐以玲．提取番茄皮色素的初步研究．新疆农垦科技．1992，4：28-29
41．蔡俊，邱雁临，谈小兰等．番茄红素提取工艺的研究．食品与发酵工业．2000，26(2)：50-53
42．D．哈提尔，Y．拉维．稳定的番茄红素浓缩物及制备方法．CN 1176577．1998，3
43. Rodeny L, Des Moines et al. Process for the isolation and purification of lycopene crystal. WO 98/43620.1998,6
44．文刚，胡光，李新．番茄酱制取结晶番茄红素及番茄红素油树脂的方法．CN1298904A．2001，6
45. PCT/US98/02335, WO 98/43620(美国专利)
46. Dubodel N P, et al. Piscchchevaya Promyshlennost, Moscow, 1995,(1), 16
47．安本光政等，日本食品科学工学会第42回大会讲演集，1995，3
48. Wo 96/13178, 1996, 5
49．孙庆杰，丁霄霖．超临界CO_2萃取番茄红素的初步研究。食品与发酵工业．1998，24(1)：3-6
50. Inakuma T et al. Effect of drying methods on extraction of lycopene in tomato skin with supercritical carbon dioxide. J. of the Japanese Soceity for Food Science and Technology. 1996,32 (5):740-743.
51. Gavrilov A. S et al. Industrial production of a microbiological method. Applied Biochemistryand Microbiology. 1996, 32(5), 492-494
52. Minura Y et al. Production of lycopene by the food yeast, Candida utilis that dose not naturally synthesize carotenoid. Biotechnology and Bioengineering, 1998, 58(2/3):306-308
53．岑宁，王杰．柑橘果皮番茄红素诱导合成研究冲国南方果树．1996，25 (3)：6-7
54. Sari H. Hakila I. Marina Heinoen. Chromatographic Purification of Natural Lycopene. J. Agric.Food Chem, 1994 42(6):1314-1316
55．何碧烟，欧光南．食品添加剂对番茄红素稳定性的影响．集美大学学报．2000，5(1)：47-51
56．李伟，孙新虎．番茄红素溶解度的测定．食品与发酵工业．2002，28(8)：54-56
57．Kingston H M，Jassic L B编．郭振库，卜玉兰，刘正阳等译．分析化学中的微波制样技术．北京：气象出版社，1992
58．郭振库，卜玉兰．微波萃取技术．色谱．1997，15(6)：499-501
59．郝金玉，黄若华，王平艳．微波萃取技术进展．云南化工．1999，3：26-29
60．古华民，张斌东，胡男．应用统计方法．暨南大学出版社．1994
61．达世禄．色谱学导论．武汉大学出版社．1999
62. Sahl W. Sies H. Persaectives in Biochemistry and Biophysics. Lycopene:A Biologically Important Carotenoid for Humans. Arch. Biochem. & Biophys. 1996, 336:1-9
63. Shi J. Maguer M. L., Kakuda Y. Lycopene Degradation and lsomerization in Tomato Dehydration. Food Rea. Intl. 1999, 32:15-21