利用高速逆流色谱（HSCCC）对千层塔提取物废液中含量较高的2个同系物进行初步纯化分离,再利用高效液相色谱（HPLC）对收集到的样品进行同系物分离,得到8mg化合物A,7mg化合物B,纯度均在95%以上.经核磁共振仪（NMR）鉴定2个组分的化学结构,结果证明,千层塔提取物废液所含的主要成分即2个同系物分别为3,4-二羟基肉桂酸和3,4-二甲氧基肉桂酸.结论也同时证实高速逆流色谱与高效液相色谱联用能够有效分离样品所含同系物,且工业提取千层塔有效成分后的废液中仍含有价值物质.

High-speed counter-current chromatography(HSCCC) is first used preliminarily to purify two homologues with higher content in waste from extracts of Huperzia serrata, and high performance liquid chromatography(HPLC) is used to seperate the two components. 8 mg compound A and 7 mg compound B are obtained, and purity of both compounds reaches more than 95%. Then chemical structure of the two components is identified using the nuclear magnetic resonance apparatus(NMR). The results show that the two components are 3,4-dihydroxy cinnamic acid and 3,4-dimethoxy cinnamic acid. Conclusion can be made that the homologues can be purified by combination of high-speed counter-current chromatography and high performance liquid chromatography, and there still are valuable substances in waste from extracts of Huperzia serrata.

利用高速逆流色谱法从100 mg诃子醇提物中一次性分离制备得到8.6 mg没食子酸。通过分析型高速逆流色谱对5种溶剂系统进行筛选,确定以正己烷-乙酸乙酯-甲醇-水（体积比为1∶5∶1∶5）为两相溶剂体系并放大到制备型上,以上相为固定相,下相为流动相,在主机转速850 r/min、流动相流速2 mL/min、检测波长254 nm的条件下进行分离制备,获得4个分离峰（组分Ⅰ、Ⅱ、Ⅲ、Ⅳ）。经高效液相色谱检测,按照面积归一法计算,其中组分Ⅲ的纯度达96.40%。经电喷雾电离质谱分析,并结合与没食子酸标准品的高效液相色谱测定结果的对比,确定组分Ⅲ为没食子酸。该方法简便、快速、重复性好,适合于诃子中没食子酸的分离制备。

A separation method based on high-speed counter-current chromatography ( HSCCC)has been established for the isolation and preparation of gallic acid from the ethanol extract of Terminalia chebula Retz. After comparing five kinds of solvent protocols of HSCCC, the two-phase system of n-hexane-ethyl acetate-methanol-water(1:5:1:5,v/v/v/v)was final-ly chosen as the operating solvent of HSCCC for the separation of gallic acid,in which the low-er phase was used as the mobile phase and the upper phase as stationary phase. The detection in the experiments was performed with an ultraviolet detector at 254 nm. Under the conditions of rotation speed of 850 r/min,lower phase flow rate of 2 mL/min,four peaks(Ⅰ/Ⅱ/Ⅲ/Ⅳ) were displayed on HSCCC chromatogram. Among them,only peak Ⅲ contained a large amount of gallic acid( about 96. 40%),which was confirmed by electrospray ionization mass spectrom-etry( ESI-MS)and high performance liquid chromatographic( HPLC)analysis. As much as 8. 6 mg

花色苷是高等植物中最重要的水溶性色素。因其种类繁多、来源广泛、安全无毒并有一定的营养和保健功效而引起国内外的广泛关注,具有十分重要的开发价值和广阔的应用前景。文中介绍了国内外花色苷分离纯化(层析法、高速逆流色谱、膜分离法、固相萃取)、以及花色苷鉴定(高效液相色谱-串联质谱法,核磁共振法)的研究方法,并对各种方法进行了分析评价。对全面认识和开发利用花色苷具有一定的参考价值。

Anthocyanins are the most important water-soluble pigment in plants.It caused widespread concern at home and abroad because of its variety and wide range of sources,safety and rich in nutrition and health effects,it has a very important development value and broad application prospects.The present paper mentioned some methods for anthocyanin separation and purification (chromatography,high-speed countercur-rent chromatography,membrane separation,solid phase extraction),and identification (high performance liq-uid chromatography-tandem mass spectrometry,nuclear magnetic resonance spectroscopy).

建立高速逆流色谱分离纯化杭白菊总黄酮结晶中芹菜素-7-O-芸香糖、木犀草素-7-O-葡萄糖、芹菜素-7-O-葡萄糖以及金合欢素-7-O-葡萄糖4种黄酮类化合物。高速逆流分离过程分为两步,分别采用乙酸乙酯-乙醇-水-乙酸（体积比4：1：5：0.2）和氯仿-甲醇-水（体积比4：3：2）两个体系。在第一步中,100 mg的总黄酮结晶分离得到了11.2 mg的芹菜素-7-O-芸香糖、15.3 mg的木犀草素-7-O-葡萄糖、28.2 mg的芹菜素-7-O-葡萄糖。然后收集到尾吹液,旋蒸至干得到35 mg的浸膏。在第二步中,当采用氯仿-甲醇-水（体积比4：3：2）体系时,从35 mg的浸膏中分离纯化得到14.5 mg的金合欢素-7-O-葡萄糖。4个化合物的纯度分别为99.4%、93.6%、99.1%和99.5%,电喷雾电离质谱和氢、碳核磁共振波谱鉴定化合物的结构。

High-speed counter-current chromatography ( HSCCC) method for isolation and purification of flavonoids crystalloid, i. e. apigenin-7-O-rutinoside( I) , luteolin-7-O-glucoside( II) , apigenin-7-O-glucoside( III) and acacetin-7-O-glucoside( IV) from Chrysanthemum morifolium Ramat. was established successfully. The separation was performed in two steps with two different types of solvent systems: ethyl acetate-ethanol-water-acetic acid (4:1:5:0. 2, v/v) and chloroform-methanol-water (4:3:2, v/v). In the first separation step, 100 mg of the flavonoids crystalloid yielded 11. 2 mg of apigenin-7-O-rutinoside, 15. 3 mg of luteolin-7-O-glucoside, and 28. 2 mg of apigenin-7-O-glucoside. Then we collected tail blowing fluid and obtained 35 mg of extract by evaporation to dryness. In the second step, when we used [ chloroform-methanol-water (4:3:2, v/v)] solvent system to separate the 35 mg of extract, 14. 5 mg of acacetin-7-O-glucoside could be obtained. Their four compounds purities were 99. 4

采用高速逆流色谱(HSCCC)技术从铁棒锤根氯仿提取物中分离制备了一种高纯度咪唑类生物碱1H-imidazole-2-carboxylic acid,butyl ester(ICABE)。采用高效液相色谱(HPLC)测定目标化合物在两相溶剂中的分配系数,优化HSCCC分离ICABE的溶剂体系,确定了以正己烷-氯仿-乙醇-水(10∶1∶13∶2,v/v/v/v)为HSCCC的两相溶剂系统,以上相为固定相,下相为流动相,流动相流速为1.8 mL/min,主机转速850 r/min,检测波长为230 nm条件下进行分离制备,在350 min内从100 mg粗样品中一步分离得到7.5 mg ICABE,经HPLC检测其纯度达98%以上(峰面积归一化法),结构由UV、1H-NMR和13C-NMR得以鉴定。该方法简便、快速,所得产物纯度高,适合于铁棒锤中ICABE的制备分离。

Aconitum pendulum Busch is rich C19 diterpenoid alkaloids,but there is no report of imidazole alkaloid in Aconitum pendulum Busch. In this study,an imidazole alkaloid named 1H-imidazole-2-carboxylic acid,butyl ester( ICABE)was successfully separated from Aconi-tum pendulum Busch with semi-preparative high-speed counter-current chromatography( HSC-CC). The partition coefficient was measured by HPLC to select the solvent systems for ICABE separation by HSCCC. The separation was performed with a two-phase solvent system com-posed of n-hexane-chloroform-ethanol-water(10:1:13:2,v/v/v/v). The upper phase was used as the stationary phase and the lower phase as the mobile phase. It was operated at a flow rate of 1. 8 mL/min. The apparatus was rotated at 850 r/min,and the detection wavelength was set at 230 nm. Under the selected conditions,a high efficiency separation of HSCCC was achieved, and 7. 5 mg of ICABE was obtained from 100 mg of the crude sample of Aconitum pendulum in one-step separ