硅胶柱层析和半制备液相色谱法从曲克芦丁母液中分离制备高纯度曲克芦丁及有关物质

doi:10.5246/jcps.2020.07.046

中国药学(英文版) ›› 2020, Vol. 29 ›› Issue (7): 487-493.DOI: 10.5246/jcps.2020.07.046

硅胶柱层析和半制备液相色谱法从曲克芦丁母液中分离制备高纯度曲克芦丁及有关物质

刘少静^1,2,3, 秦蓓^1,2*, 韩红芳¹, 李立^1,2, 余丽丽^1,2, 徐小静^1,2

1. 西安医学院药学院, 陕西西安 710021
2. 西安医学院药物研究所, 陕西西安 710021
3. 西北大学化工学院制药工程专业, 陕西西安 710069

收稿日期:2020-01-02 修回日期:2020-02-15 出版日期:2020-07-31 发布日期:2020-04-12
通讯作者: Tel.: +86-29-86177536, E-mail: liushaojingbmgw@163.com
基金资助:
Shaanxi science and technology hall project (Grant No. 2018JM7060); Provincial Key Discipline Construction Project of pharmacy of Xi’an Medical University (Grant No. 2016YXXK08).

Preparative separation of high-purity troxerutin and related substances from mother liquor of troxerutin by silica gel column chromatography and semi-preparative liquid chromatography

Shaojing Liu^1,2,3, Bei Qin^1,2*, Hongfang Han¹, Li Li^1,2, Lili Yu^1,2, Xiaojing Xu^1,2

1. College of pharmacy, Xi’an Medical University, Shaanxi 710021, China
2. Institute of Medicine, Xi’an Medical University, Shaanxi 710021, China
3. College of Chemical Engineering, Department of harmaceutical Engineering, Northwest University, Shaanxi 710069, China

Received:2020-01-02 Revised:2020-02-15 Online:2020-07-31 Published:2020-04-12
Contact: Tel.: +86-29-86177536, E-mail: liushaojingbmgw@163.com
Supported by:
Shaanxi science and technology hall project (Grant No. 2018JM7060); Provincial Key Discipline Construction Project of pharmacy of Xi’an Medical University (Grant No. 2016YXXK08).

摘要/Abstract

摘要：

曲克芦丁(Troxerutin, TRO)是由芦丁羟乙基化制备的半合成黄酮类化合物的混合物,常用于治疗脑血管疾病,主要活性成分为三羟乙基芦丁(亦称曲克芦丁)。曲克芦丁生产母液中含有大量的三羟乙基芦丁及其它羟乙基芦丁衍生物,该研究采用硅胶柱层析和半制备液相色谱相结合的方法,从TRO母液中回收制备高纯度的TRO。在硅胶柱色谱分离中:考察了硅胶与样品比例及洗脱液组成,以获得最佳的分离效果。结果表明,当硅胶与样品的比为50:1,丙酮–乙酸乙酯–水–冰醋酸(10:10:3:1, v/v/v/v)为洗脱剂时, TRO与相邻杂质的分离效果较好。用硅胶柱层析法, 从1 kg TRO母液中制备得150 g (纯度80%) TRO。进一步采用半制备HPLC纯化,制备得纯度高达98% (w/w)的TRO 100 g,同时得到纯度大于98%的四羟乙基芦丁和四羟乙基槲皮素。本文首次提出从TRO生产废液中分离制备高纯度TRO,具有一定的环境效益和经济效益。

关键词: 曲克芦丁母液, 曲克芦丁, 硅胶柱层析, 半制备液相色谱法, NMR

Abstract:

Troxerutin (TRO) is a mixture of semi-synthetic flavonoids prepared by hydroxyethylation of rutin, and it is commonly used for the treatment of cerebrovascular diseases. The main active ingredient is trishydroxyethyl rutin. The mother liquor of TROcontains a lot of TRO and other derivatives of hydroxyethylated rutin. In order to make full use of the mother liquor of TRO, an efficient method was developed for recovering high-purity TRO from mother liquor of TRO by combining silica gel column chromatography with semi-preparative liquid chromatography. In the silica gel column chromatographic separation, the ratio of silica gel to sample and eluent composition were investigated to obtain optimum separation effect. The results showed that when the ratio of silica gel to sample was 50, and acetone–ethyl acetate–water–glacial acetic acid (10:10:3:1, v/v/v/v) was used as the eluent, the separation effect of TRO and adjacent impurities was good. Moreover, 150 g ofTRO with a purity of 80% could be obtained from 1 kg of mother liquor of TRO by the silica gel column chromatographic separation, and the results were consistent with the quality standard of TRO raw material. Subsequently, the semi-preparative HPLC was performed, and 100 g TRO with a purity of up to 98% (w/w) was obtained. Meanwhile, tetrahydroxyethylrutin and tetrahydroxyethylquercetin with purity greater than 98% were obtained. This work proposed the separation and preparation of TRO with high-purity from the production waste of TRO for the first time, which had certain environmental benefits and economic benefits.

Key words: Mother liquor of troxerutin, Troxerutin, Silica-gel column chromatography, Semi-preparative HPLC, NMR

中图分类号:

R917

Supporting:

刘少静, 秦蓓, 韩红芳, 李立, 余丽丽, 徐小静. 硅胶柱层析和半制备液相色谱法从曲克芦丁母液中分离制备高纯度曲克芦丁及有关物质[J]. 中国药学(英文版), 2020, 29(7): 487-493.

Shaojing Liu, Bei Qin, Hongfang Han, Li Li, Lili Yu, Xiaojing Xu. Preparative separation of high-purity troxerutin and related substances from mother liquor of troxerutin by silica gel column chromatography and semi-preparative liquid chromatography[J]. Journal of Chinese Pharmaceutical Sciences, 2020, 29(7): 487-493.

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硅胶柱层析和半制备液相色谱法从曲克芦丁母液中分离制备高纯度曲克芦丁及有关物质

Preparative separation of high-purity troxerutin and related substances from mother liquor of troxerutin by silica gel column chromatography and semi-preparative liquid chromatography

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