银杏叶聚戊烯醇的分离纯化及其亲水衍生物的合成
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摘要
首先对银杏叶聚戊烯醇含量测定的方法及方法学进行了考察,结果表明:采用高效液相色谱,以C18为色谱柱,以异丙醇/甲醇(体积比32∶18)为流动相,在210 nm下测定聚戊烯醇具有较高的精密度(RSD为0.89%)、较好的稳定性(RSD为3.26%),平均加标回收率为97.4%(RSD为1.75%)。然后通过正己烷提取、皂化、乙醇和丙酮脱蜡,以及硅胶柱层析(V(乙醚)∶V(正己烷)=3∶97)获得纯度为98.6%的聚戊烯醇。在此基础上,对高纯度聚戊烯醇末端羟基进行亲水改性,先与邻苯二甲酰亚胺进行光延反应(mitsunobu反应),再与水合肼进行还原反应,合成了氨基聚戊烯醇,并通过红外和核磁共振氢谱对其结构进行了表征,确认了氨基聚戊烯醇的成功合成。
The analytical methodology and method performance for the determination of polyprenol in Ginkgo biloba leaves were investigated for the first time. The results showed that of the high performance liquid chromatography with C18 as chromatographic column, isopropanol methanol(volume ratio, 32∶18) as mobile phase, 210 nm as detection wavelength showed high precision(RSD=0.89%) and good stability(RSD=3.26%). In addition, the average standard recovery and RSD value of polyprenol were 97.4% and 1.75%, respectively. Then, polyprenol with the purity of 98.6% was obtained by extraction with n-hexane, saponification, de-waxing with ethanol and acetone, and silica gel column chromatography(V(diethyl ether)∶V(n-hexane)=3∶97). On this basis, the terminal hydroxyl group of high-purity polyprenol was modified to synthesize aminopolyprenol. Firstly, by mitsunobu reaction with phthalimide, followed by reduction reaction with hydrazine hydrate, to obtain aminopolyprenol. The structure of amino polyprenol was characterized by infrared and nuclear magnetic resonance spectroscopy, which confirmed the aminopolyprenol was successfully synthesized.
The analytical methodology and method performance for the determination of polyprenol in Ginkgo biloba leaves were investigated for the first time. The results showed that of the high performance liquid chromatography with C18 as chromatographic column, isopropanol methanol(volume ratio, 32∶18) as mobile phase, 210 nm as detection wavelength showed high precision(RSD=0.89%) and good stability(RSD=3.26%). In addition, the average standard recovery and RSD value of polyprenol were 97.4% and 1.75%, respectively. Then, polyprenol with the purity of 98.6% was obtained by extraction with n-hexane, saponification, de-waxing with ethanol and acetone, and silica gel column chromatography(V(diethyl ether)∶V(n-hexane)=3∶97). On this basis, the terminal hydroxyl group of high-purity polyprenol was modified to synthesize aminopolyprenol. Firstly, by mitsunobu reaction with phthalimide, followed by reduction reaction with hydrazine hydrate, to obtain aminopolyprenol. The structure of amino polyprenol was characterized by infrared and nuclear magnetic resonance spectroscopy, which confirmed the aminopolyprenol was successfully synthesized.
引文
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[11]YANG L,WANG C Z,YE J Z,et al. Hepatoprotective effects of polyprenols from GBL on CCl4-induced hepato toxicity in rats[J]. Fitoterapia,2011,82(6):834-840.
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[13]高凌,亢寿海,张琴芬,等. 银杏叶聚戊烯醇抗癌作用的初步研究[J]. 药学与临床研究,2003,11(1):1-3. GAO L,KANG S H,ZHANG Q F,et al. A preliminary study of antitumor effects of polyprenols extracted from fofium gingko[J]. Pharmaceutical and Clinical Research,2003,11(1):1-3.
[14]王成章,沈兆邦,郑光耀,等. 银杏叶聚戊烯基磷酸酯体内外抗肿瘤的生物活性研究[J]. 林产化学与工业,2006,26(3):13-16. WANG C Z,SHEN Z B,ZHENG G Y,et al. Antitumor biological activity polyprenyl phosphate from leaves of Ginkgo biloba L. in vivo and vitro[J]. Chemistry and Industry of Forest Products,2006,26(3):13-16.
[15]王成章,沈兆邦. 银杏叶聚戊烯醇联合化疗药对Heps和Ec荷瘤小鼠的抗癌作用[J]. 中国生化药物杂志,2003,24(3):113-115. WANG C Z,SHEN Z B. The effects on transplanted Heps and EC tumors in mouse for polyprenols in combination with chemotherapeutic drugs[J]. Chinese Journal of Biochemical Pharmaceutics,2003,24(3):113-115.
[16]李春斌,孙磊,李黎,等. 银杏叶聚戊烯醇分离分析方法的研究进展[J]. 大连民族学院学报,2007,9(1):41-43. LI C B,SUN L,LI L,et al. Research progress of the separation and analysis methods of polyprenols in Ginkgo biboba leaves[J]. Chinese Journal of Dalian Nationalities University,2007,9(1):41-43.
[2]陶冉,王成章,孔振武. 银杏叶类脂的化学成分研究[J]. 林产化学与工业,2014,34(4):71-76. TAO R,WANG C Z,KONG Z W. Chemical constituents of Ginkgo biloba L. leaves lipids[J]. Chemistry and Industry of Forest Products,2014,34(4):71-76.
[3]王成章,沈兆邦,陈祥. 五种针叶中聚戊烯醇的化学结构[J]. 林产化学与工业,1994,14(3):1-8. WANG C Z,SHEN Z B,CHEN X. Structure of polyprenols from five conifer needles[J]. Chemistry and Industry of Forest Products,1994,14(3):1-8.
[4]王成章,沈兆邦,谭卫红,等. 银杏叶聚戊烯醇的化学、纯化和药效研究[J]. 天然产物研究与开发,2001,13(2):43-45. WANG C Z,SHEN Z B,TAN W H. The studies on the chemistry,purification and pharmacological of polyprenols from Ginkgo biloba L. [J]. Natural Product Research and Development,2001,13(2):43-45.
[5]王成章. 银杏叶聚戊烯醇的分离和衍生物合成机理及生物活性研究[D]. 北京:中国林业科学研究院,2007. WANG C Z. Study of separation and derivant synthesis and bioactives of polyprenols from Ginkgo biloba L.(GP)[D]. Beijing:Chinese Academy of Forestry,2007.
[6]王成章,沈兆邦,高凌. 银杏叶聚戊烯醇抗肿瘤的生物活性研究[J]. 天然产物研究与开发,2002,14(5):18-20. WANG C Z,SHEN Z B,GAO L. The biological activeness of polyprenols from Ginkgo biloba L. on antitumor[J]. Natural Product Research and Development,2002,14(5):18-20.
[7]王成章,沈兆邦,刘妤婵,等. 银杏叶聚戊烯醇对小鼠免疫功能和诱导肿瘤细胞凋亡的研究[J]. 林产化学与工业,2005,25(3):9-12. WANG C Z,SHEN Z B,LIU Y C. Studies of Ginkgo biloba L. polyprenols on mmunological function and inducing apoptosis of tumor in mouse[J]. Chemistry and Industry of Forest Products,2005,25(3):9-12.
[8]DEFEUDIS F V,DRIEU K. "Stress ealleviating" and "vigilancee enhancing" actions of Ginkgo biloba extract(EGb761)[J]. Drug Development Research,2004,62(1):1-25.
[9]MAHADY G B. Ginkgo biloba for the prevention and treatment of cardiovascular disease:A review of the literature[J]. Journal of Cardiovascular Nursing,2002,16(4):21-32.
[10]SMITH J V,LUO Y. Studies on molecular mechanisms of Ginkgo biloba extract[J]. Applied Microbiology and Biotechnology,2004,64(4):465-472.
[11]YANG L,WANG C Z,YE J Z,et al. Hepatoprotective effects of polyprenols from GBL on CCl4-induced hepato toxicity in rats[J]. Fitoterapia,2011,82(6):834-840.
[12]TAO R,WANG C Z,KONG Z W. Antibacterial/antifungal activity and synergistic interactions between polyprenols and other lipid isolated from Ginkgo biloba L. leaves[J]. Molecules,2013,18(2):2166-2182.
[13]高凌,亢寿海,张琴芬,等. 银杏叶聚戊烯醇抗癌作用的初步研究[J]. 药学与临床研究,2003,11(1):1-3. GAO L,KANG S H,ZHANG Q F,et al. A preliminary study of antitumor effects of polyprenols extracted from fofium gingko[J]. Pharmaceutical and Clinical Research,2003,11(1):1-3.
[14]王成章,沈兆邦,郑光耀,等. 银杏叶聚戊烯基磷酸酯体内外抗肿瘤的生物活性研究[J]. 林产化学与工业,2006,26(3):13-16. WANG C Z,SHEN Z B,ZHENG G Y,et al. Antitumor biological activity polyprenyl phosphate from leaves of Ginkgo biloba L. in vivo and vitro[J]. Chemistry and Industry of Forest Products,2006,26(3):13-16.
[15]王成章,沈兆邦. 银杏叶聚戊烯醇联合化疗药对Heps和Ec荷瘤小鼠的抗癌作用[J]. 中国生化药物杂志,2003,24(3):113-115. WANG C Z,SHEN Z B. The effects on transplanted Heps and EC tumors in mouse for polyprenols in combination with chemotherapeutic drugs[J]. Chinese Journal of Biochemical Pharmaceutics,2003,24(3):113-115.
[16]李春斌,孙磊,李黎,等. 银杏叶聚戊烯醇分离分析方法的研究进展[J]. 大连民族学院学报,2007,9(1):41-43. LI C B,SUN L,LI L,et al. Research progress of the separation and analysis methods of polyprenols in Ginkgo biboba leaves[J]. Chinese Journal of Dalian Nationalities University,2007,9(1):41-43.