大果沙棘果中亚油酸的分离纯化及降血脂功能研究
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摘要
通过超声波-超临界CO2联用法提取大果沙棘果油,对提取工艺进行研究,确定了优化的提取工艺条件为:超声强度128w,超声时间40min,料液比1:12条件下对大果沙棘果渣进行超声波辅助提油,之后用萃取压力24MPa,萃取温度33℃,萃取时间75min,分离Ⅰ压力10MPa、分离Ⅰ温度35℃、分离Ⅱ压力8MPa、分离Ⅱ温度38℃的超临界萃取设备继续提油,所提的油得率为9.24%,亚油酸得率为2.44%。用超声波-超临界CO2联用法提取大果沙棘果油是一种创新的提取方法,实验表明这种联用法具有实际意义。用尿素包合法和硝酸银硅胶柱层析法纯化大果沙棘果中亚油酸,通过响应面分析法优化尿素包合法纯化亚油酸的工艺参数,确定的纯化条件为:尿素:FFA=2.4:1;反应温度为66℃;包合时间为11.7h,纯化得到的亚油酸含量为57.66%,亚油酸得率为5.88%。再用AgNO3:硅胶=1:10,PUFA:吸附剂=1:25的柱层析条件进一步纯化亚油酸,经气相色谱分析确定可纯化得到的最高亚油酸含量为82.53%,亚油酸回收率为82.63%。实验结果表明,尿素包合法和硝酸银-硅胶柱层析法是有效的纯化亚油酸的方法。通过药效学试验研究纯化得到的大果沙棘果亚油酸对小鼠的降血脂活性,结果为大果沙棘果亚油酸对高血脂模型小鼠的TC和TG水平具有明显的降低作用(p<0.05),对HDL-C水平有明显升高作用(p<0.02),证明大果沙棘果亚油酸具有降血脂功能。
Extrated oil from fruit residue of Hippophae rhamneides by ultrasonic-SFE. The optimized extraction condition: in the condition of ultrasonic intensity 128w; ultrasonic time, 40min; the ratio between material and solvent is 1:12 using ultrasonic extract fruit oil, then pouring in the extraction department of SFE, in the condition of extraction pressure, 24MPa; extration temperature, 33℃,extraction time, 75min; separationⅠpressure, 10MPa; separationⅠtempatrature, 26℃; separationⅡpressure, 8MPa; separationⅡtemparature, 38℃, at this condition, the yield of oil is 9.24% and the yield of LA is 2.44%. The extract method of ultrasonic-SFE is an innovative method. The extraction experiment result show the method is practical meaning. Urea adduct method and silver-nitrite silica gel columniation chromatogram was adopted to pruifity LA of Hippophae rhbmneides. Study the purifitying technics parameter of urea adduct method by using response face analysis to determine the optimed purifity condition: FFA:urea= 2.4:1; reaction temperature 66℃; adduct time 11.7h. At the condition, the content of LA is 57.66% and the yield of LA is 5.88%. Using silver silica gel of silver-nitrite: silica gel =1:10 as the sorbent, in the condition of sample: sorbent=1:25 using columniation chromatogram further purify LA. By GC analyse the purifying result, the content of LA is 82.53% and the reclaim rate of LA is 82.63%. The experiment result show that Urea adduct method and silver silica gel of silver-nitrite method effectively purify LA. The lowering hyterlipemia lever’s activity was studied by medical trial. LA can significantly fall the level of TC and TG (p<0.05), can significantly hoist the level of HDL-C (p<0.02) .The result show that LA has the function of lowering hyterlipemia.
Extrated oil from fruit residue of Hippophae rhamneides by ultrasonic-SFE. The optimized extraction condition: in the condition of ultrasonic intensity 128w; ultrasonic time, 40min; the ratio between material and solvent is 1:12 using ultrasonic extract fruit oil, then pouring in the extraction department of SFE, in the condition of extraction pressure, 24MPa; extration temperature, 33℃,extraction time, 75min; separationⅠpressure, 10MPa; separationⅠtempatrature, 26℃; separationⅡpressure, 8MPa; separationⅡtemparature, 38℃, at this condition, the yield of oil is 9.24% and the yield of LA is 2.44%. The extract method of ultrasonic-SFE is an innovative method. The extraction experiment result show the method is practical meaning. Urea adduct method and silver-nitrite silica gel columniation chromatogram was adopted to pruifity LA of Hippophae rhbmneides. Study the purifitying technics parameter of urea adduct method by using response face analysis to determine the optimed purifity condition: FFA:urea= 2.4:1; reaction temperature 66℃; adduct time 11.7h. At the condition, the content of LA is 57.66% and the yield of LA is 5.88%. Using silver silica gel of silver-nitrite: silica gel =1:10 as the sorbent, in the condition of sample: sorbent=1:25 using columniation chromatogram further purify LA. By GC analyse the purifying result, the content of LA is 82.53% and the reclaim rate of LA is 82.63%. The experiment result show that Urea adduct method and silver silica gel of silver-nitrite method effectively purify LA. The lowering hyterlipemia lever’s activity was studied by medical trial. LA can significantly fall the level of TC and TG (p<0.05), can significantly hoist the level of HDL-C (p<0.02) .The result show that LA has the function of lowering hyterlipemia.
引文
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24许青媛,陈春梅.沙棘油对实验性血栓形成及凝血系统的影响.天然产物研究与开发. 1991, 3(3): 70~73
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27 M. Basu, R. Prasad, P. Jayamurthy etal. Anti-atherogenic effects of seabuckthorn (Hippophaea rhamnoides L.) seed oil. Phytomedicine. 2007
28程康华,高拥军.沙棘油的综合开发利用.林产化工通讯. 2004, 38(6): 42~46
29李忌,王尚萱,郑荣梁.沙棘提取物对癌细胞DNA、蛋白质合成及体内血浆中cAMP含量的影响.厦门大学学报. 1994, 33(4): 548~551.
30哈斯格日乐,吴岩.沙棘油对小鼠腹腔巨噬细胞功能的影响.内蒙古医学院学报. 1993, 15(1): 30~32
31刘天洁,陈运贤,钟雪云.沙棘对血液系统的作用.中药材. 2001, 24(8): 610~612
32 S. Geeth, S. Ram. Anti-oxidant and immunomodulatory properties ofseabuckthorn (hippophae rhamnoides L.) -an in vitro study. Journal of Ethnopharmacology. 2002, 79: 373~378
33程体娟,李文惠,陈卫.沙棘籽油对大鼠实验性肝纤维化的保护作用.中药药理与临床. 2002, 18(6): 15~16
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35王林杰.沙棘油与糜蛋白酶治疗小儿溃疡性口炎疗效显著.沙棘. 1992, (2): 32-35
36周张章,周才琼,阚健全.沙棘的化学成分及保健作用研究进展.粮食与食品工业. 2005, 12(2): 15~19
37刘洁,李春阳,刘志婷等.复方沙棘口服液抗衰老作用研究.长春中医学院学报. 2001, 17(1): 45~46
38徐静,李姜溶.多烯脂肪酸胶丸治疗高血脂症疗效观察.黑龙江医药, 1998, 21(4):65-67
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40顾倬云.多不饱和脂肪酸对机体免疫功能的影响.中国临床营养杂志. 2000, 8(4): 51~253
41徐贵发.ω-3和ω-6多不饱和脂肪酸的药理作用.滨州医学院学报. 2005, 28(4): 253~254
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45 J. Z. Yin, A. Q. Wang, W. Wei etal. Analysis of the operation conditions for supercritical fluid extraction of seed oil. Separation and Purification Technology. 2005, 43: 163~167
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47 T. C. Chen, Y. Hsuju. Polyunsaturated Fatty Acid Concentrates from Borage andLinseed oil Fatty Acids [J]. JAOCS. 2001, 78(5): 485~488
48 L. S. Hwang, H. L. Jer. Fractionation of Urea-Pre-treated Squid Visceral oil Ethyl Esters [J]. JAOCS. 2001, 78(5): 473~476
49邓朝霞,叶代勇,傅和清等.分子蒸馏及其在精细化工上的应用.广州化工. 2006, 34(2): 6~9
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51郑毅,郑楠,卓进锋.利用脂肪酶提高鱼油中多不饱和脂肪酸(PUFAs)甘油酯.应用与环境生物学报. 2005, 11(5): 571~574
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2任立飒.沙棘油抗突变、抗免疫抑制作用研究.沙棘. 1992, 5(4): 23~25
3李仲瑞,福凤鸣.恶性肿瘤化疗中口服沙棘油的临床观察.沙棘. 1993, 6(4): 41~43
4车锡平,霍海如,康军等.沙棘籽油的药效学试验研究.沙棘. 1998, 11(3): 34～38
5李晓花,孔令学,刘鸿章.沙棘有效成分研究进展.吉林农业大学学报. 2007, 2(10): 44~47
6程康华,高拥军.沙棘油的综合开发利用.林产化工通讯. 2004, 38(6): 42~45
7赵元藩.沙棘油的精制及其在化妆品中的应用.云南师范大学学报. 2000, 20(2): 48~49
8刘振山,史文彬.沙棘.生物学通报. 1995, 3(6): 4
9杨志刚,卢顺光,赵梅霞.沙棘属植物的生物化学和药理学研究综述.国际沙棘研究与开发. 2005, 3(4): 1~8
10吕荣森.沙棘在中国西部生态环境建设中的作用.沙棘. 2003, 16(1): 3~8
11 B. R. Yang, H. K. Kallio. Effects of harvesting time on triacylglycerols and glycerophospholipids of seabuckthorn (hippophae rhamnoides L.) berries of different origins. Journal of Food Composition and Analysis. 2002, 15: 143~157
12 S. E. Kudritskaya, L. M. Zagorodskaya, E. E. Shishkina. Carotenoids of the seabuckthorn variety obilnaya. Khim Prir Soedin. 1989, 6: 852~853
13 S. C. Thomas, H. J. Thomas and C. G. John. Phytosterol content of sea buckthorn (Hippophae rhamnoides L.) seed oil: Extraction and identification. Food Chemistry. 2007, 101: 1633~1639
14 A. Cakir. Essential oil and fatty acid composition of the fruits of Hippophae rhamnoides L. (Sea Buckthorn) and Myrtus communis L. from Turkey. Biochemical Systematic and Ecology. 2004, 32: 809~816
15张鞍灵,倪荣环.沙棘属植物化学与医疗保健功能.西部粮油科技. 2001, 26(6): 32~35
16 G. A. Loskutova, V. G. Baikov, A.V. Starkov etal. The composition of fatty acids of lipids from the fruit of Hippophae rhomnoides L. Rastit Resur, 1989, 25(1): 97~103
17赵二劳,张海容,盖青青等.沙棘黄铜的测定及氧化作用.化学研究与应用. 2003, 15(2): 284~285
18何志勇,夏文水.沙棘果汁营养成分及保健作用.食品科技. 2002, 7: 69~71
19顾清萍,韩卫平.沙棘果汁冷冻干粉的工艺研究简报.沙棘. 1999, 12(2): 32~35
20高锦明,赵明哲.沙棘挥发油化学成分研究概况.沙棘. 1995, 8(3): 2~5
21 C. J. Tian, P. Nan, J. K. Chen etal. Volatile composition of Chinese Hippophae rhamnoides and its chemotaxonomic implications. Biochemical Systematics and Ecology. 2004, 32: 431~441
22刘凤鸣,张绮丽.沙棘总黄酮对离体心脏的抗心律失常作用.中国药理学通报. 1989, 5(1):44
23 D. F. Aluokumofu. Pharmacological effects of seabuckthorn oil. Hippophae. 1992, 5: 20~25
24许青媛,陈春梅.沙棘油对实验性血栓形成及凝血系统的影响.天然产物研究与开发. 1991, 3(3): 70~73
25于云,曲树明,何跃生.沙棘叶提取物的调血脂作用.中草药. 2002, 33(9): 824~826
26 Y. Wang, Y. Lu, X. Liu etal. The protective effect of Hippohaea rhamnoides L. on hyperlipidenic serum cultured smooth muscle cells in vitro. Chung Kuo Chung Yao Tsa Chih. 1992, 10(17): 10624~6601
27 M. Basu, R. Prasad, P. Jayamurthy etal. Anti-atherogenic effects of seabuckthorn (Hippophaea rhamnoides L.) seed oil. Phytomedicine. 2007
28程康华,高拥军.沙棘油的综合开发利用.林产化工通讯. 2004, 38(6): 42~46
29李忌,王尚萱,郑荣梁.沙棘提取物对癌细胞DNA、蛋白质合成及体内血浆中cAMP含量的影响.厦门大学学报. 1994, 33(4): 548~551.
30哈斯格日乐,吴岩.沙棘油对小鼠腹腔巨噬细胞功能的影响.内蒙古医学院学报. 1993, 15(1): 30~32
31刘天洁,陈运贤,钟雪云.沙棘对血液系统的作用.中药材. 2001, 24(8): 610~612
32 S. Geeth, S. Ram. Anti-oxidant and immunomodulatory properties ofseabuckthorn (hippophae rhamnoides L.) -an in vitro study. Journal of Ethnopharmacology. 2002, 79: 373~378
33程体娟,李文惠,陈卫.沙棘籽油对大鼠实验性肝纤维化的保护作用.中药药理与临床. 2002, 18(6): 15~16
34 J. F. Xing, B. R. Yang, Y. L. Dong etal. Effects of seabuckthorn (hippophae rhamnoides L.) seed and pulp oils on experimental models of gastric ulcer in rats. Fitoterapia. 2002, 73: 644~650
35王林杰.沙棘油与糜蛋白酶治疗小儿溃疡性口炎疗效显著.沙棘. 1992, (2): 32-35
36周张章,周才琼,阚健全.沙棘的化学成分及保健作用研究进展.粮食与食品工业. 2005, 12(2): 15~19
37刘洁,李春阳,刘志婷等.复方沙棘口服液抗衰老作用研究.长春中医学院学报. 2001, 17(1): 45~46
38徐静,李姜溶.多烯脂肪酸胶丸治疗高血脂症疗效观察.黑龙江医药, 1998, 21(4):65-67
39 Kinsella J E, Lokesh B. Dietary lipids eicosanoids and immune function. Crit Care Med. 1990, 12(2): 94~113
40顾倬云.多不饱和脂肪酸对机体免疫功能的影响.中国临床营养杂志. 2000, 8(4): 51~253
41徐贵发.ω-3和ω-6多不饱和脂肪酸的药理作用.滨州医学院学报. 2005, 28(4): 253~254
42赵文斌,刘金荣,但建明.均匀设计法优化苦杏仁油超声波提取工艺.粮食与油脂. 2001, 3(11): 4~5
43 M. Perrut, J. Y. Clavier, M. Poletto etal. Mathematical modeling of sunflower seed extraction by supercritical CO2. Ind. Eng. Chem. Res. 1997, 36: 430
44 E. Reverchon, C. Marrone. Modeling and simulation of the supercritical CO2 extraction of vegetable oils. Supercrit. Fluids. 2001, 19: 161~175
45 J. Z. Yin, A. Q. Wang, W. Wei etal. Analysis of the operation conditions for supercritical fluid extraction of seed oil. Separation and Purification Technology. 2005, 43: 163~167
46 T. Yokochi. Increase in theγ-linolenic acid content by solvent xinterization of frngal oil extracted from mortierella genus [J]. JAOCS. 1990, 67(11): 846~851
47 T. C. Chen, Y. Hsuju. Polyunsaturated Fatty Acid Concentrates from Borage andLinseed oil Fatty Acids [J]. JAOCS. 2001, 78(5): 485~488
48 L. S. Hwang, H. L. Jer. Fractionation of Urea-Pre-treated Squid Visceral oil Ethyl Esters [J]. JAOCS. 2001, 78(5): 473~476
49邓朝霞,叶代勇,傅和清等.分子蒸馏及其在精细化工上的应用.广州化工. 2006, 34(2): 6~9
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