高速逆流色谱结合制备型高效液相色谱法分离制备大豆皂苷单体

doi:10.7506/spkx1002-6630-201306006

食品科学 ›› 2013, Vol. 34 ›› Issue (6): 27-32.doi: 10.7506/spkx1002-6630-201306006

高速逆流色谱结合制备型高效液相色谱法分离制备大豆皂苷单体

黄玉艾赵大云严明霞

上海交通大学农业与生物学院，上海 200240

收稿日期:2011-10-07 修回日期:2013-01-15 出版日期:2013-03-25 发布日期:2013-03-01
通讯作者: 赵大云 E-mail:dyzhao@sjtu.edu.cn
基金资助:
国家自然科学基金

Large-Scale Isolation and Preparation of Soybean Saponin by High-Speed Countercurrent Chromatography Combined with Preparative HPLC

HUANG Yu-ai，YAN Ming-xia，ZHAO Da-yun

School of Agriculture and Biology,Shanghai Jiao Tong University, Shanghai 200240, China

Received:2011-10-07 Revised:2013-01-15 Online:2013-03-25 Published:2013-03-01

摘要/Abstract

摘要： 为建立一种有效的同时分离制备多种大豆皂苷单体的方法，先采用高速逆流色谱分离去除大豆皂苷粗提物中的大豆异黄酮，然后对得到大豆总皂苷，优化制备型高效液相的流动相酸(乙酸或三氟乙酸)的含量和梯度洗脱程序，以制备高纯度的大豆皂苷单体。结果表明：流速10mL/min，流动相乙腈(含0.01%三氟乙酸)-水按梯度程序Ⅲ洗脱，出峰数量多、峰形好、分离度高。经高效液相分析与光谱特性、质谱和核磁共振鉴定了11个峰的大豆皂苷种类，包括5种A组、2种B组以及3种2,3-2H-2,5-二羟基-6-甲基-4H-吡喃-4-(DDMP)大豆皂苷。研究结果提供了从含量相对丰富的大豆胚芽中大量分离制备部分或完全乙酰化的A组以及非DDMP和完整DDMP存在的B组大豆皂苷的有效方法。

关键词: 大豆皂苷单体, 高速逆流色谱, 制备型高效液相色谱, 质谱, 核磁共振

Abstract: The objective of this study is to isolate and purify soybean saponin from soybean germs by high-speed countercurrent chromatography (HSCCC) combined with preparative HPLC. The isoflavonoids were removed from crude extract by HSCCC first. The pure soybean saponin was then obtained by preparative HPLC with acetic acid or trifluoroacetic acid (TFA) as mobile phase in a gradient elution mode. The optimal conditions for preparative HPLC were dual mobile phases composed of acetonitrile containing 0.01% (V/V) TFA and millipore water at a flow rate of 10 mL/min for gradient elution. Five compounds in group A and two compounds in group B and 3 kinds of 2,3-dihydro-2,5-dinydroxy-6-methyl-4H-pyran-4-one (DDMP)- conjugated soybean saponins with high purity were prepared and their structures were elucidated by mass spectrum (MS) and nuclear magnetic resonance (NMR). Therefore, the developed method can be used to separate partially and fully acetylated soybean saponins as well as DDMP-conjugated and unconjugated soybean saponins in the original sample.

Key words: soybean saponin, high-speed countercurrent chromatography (HSCCC), preparative high performance liquid chromatography, mass spectroscopy (MS), nuclear magnetic resonance (NMR)

中图分类号:

TS207.3

黄玉艾赵大云严明霞. 高速逆流色谱结合制备型高效液相色谱法分离制备大豆皂苷单体[J]. 食品科学, 2013, 34(6): 27-32.

参考文献

[1] Hong-Xiang Sun, Yong Xie, Yi-Ping Ye. Advances in saponin-based adjuvants[J].Vaccine, Volume 27, Issue 12, 13 March 2009, Pages 1787-1796.
[2] 杜丽洁. 大豆皂甙对高脂血症大鼠血脂及抗血栓作用的影响研究[D]山西医科大学, 2011.
[3] 郑奇志,吴家祥,肖纫秋,王心蕊,石博,卫永第. 大豆皂甙对胰岛素下的平滑肌细胞脂质过氧化反应的影响[J]白求恩医科大学学报, 1998,(05) .
[4] 靖雪妍. 大豆复合物和大豆皂苷的免疫调节作用研究[D]. 吉林大学, 2007,(03) .
[5] 袁晓洁. 大豆异黄酮和大豆皂苷缓解体力疲劳作用的研究[D]. 吉林大学, 2007,(02) .
[6] Yoshiki Y, Okubo K. Active oxygen scavenging activity of DDMP(2, 3-dihydro-2, 5-dihydroxy-6-methyl-4H-pyran-4-one) saponin in soybean seed .Biosci Biotechnol Biochem, 1995,59, 59 :1556-1557.
[7] 马利刚.大豆异黄酮和大豆皂苷对酒精肝损伤的干预作用[D]. 吉林大学,2009.
[8] Mark A Berhow, Elizabeth D Wagner, Steven F Vaughn, Michael J Plewa. Characterization and antimutagenic activity of soybean saponins[J]. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Volume 448, Issue 1, 14 March 2000, Pages 11-22.
[9] 高学敏,靳莉,汪锦邦,徐燕玲,叶恩赐. 大豆总皂甙抗衰老作用的研究 I.生命实验的研究[J]食品工业科技, 1999,(S1) .
[10] Shiraiwa M.Composition and structure of“group A saponin”in soybean seed[J].Agri Biol Chem,1991,55(2):315～322.
[11] Shiraiwa M.Composition and structure of“group B saponin”in soybean seed[J].Agri Biol Chem,1991,55(4):911～917.
[12] 王东冬.大豆皂苷的提取纯化与分离检测[D]. 上海交通大学,2007.
[13] 吴永慧,贾竞夫,唐波,孙向军,赵大云. 高速逆流色谱分离大豆皂苷和异黄酮[J]中国粮油学报, 2010,(12) .
[14] Decroos K,Vincken J-P,Heng L,et al. Simultaneous quanti-fication of differently glycosylated,acetylated,and 2,3-di-hydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one-conjugated soyasaponins using reversed phase high per-formance liquid chromatography with evaporative light scatter-ing detection[J] .Journal of Chromatography A, 2005,1072, 1072 :185-193 .
[15] 马杰. 高含量大豆皂苷制备和分离工艺研究[D]浙江大学, 2007 .
[16] 全吉淑,尹学哲,田中真实,金泽武道. 大豆胚轴中B类、E类和DDMP大豆皂甙的分离纯化[J]食品科学, 2003, (02) .
[17] 徐龙权,田晶. 大豆皂苷Ⅱ的提取及HPLC-MS分析[J]大连工业大学学报, 2009,(03) .
[18] K Decroos, J Vincken, G Vankoningsveld, H Gruppen, W Verstraete. Preparative chromatographic puri?cation and surfactant properties of individual soyasaponins from soy hypocotyls[J]. Food Chemistry (2007) , Volume: 101, Issue: 1, Pages: 324-333.

高速逆流色谱结合制备型高效液相色谱法分离制备大豆皂苷单体

Large-Scale Isolation and Preparation of Soybean Saponin by High-Speed Countercurrent Chromatography Combined with Preparative HPLC

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