One step purification of corilagin and ellagic acid from Phyllanthus urinaria using high-speed countercurrent chromatography

High-speed countercurrent chromatography (HSCCC) has been successfully applied to the preparative separation of corilagin and ellagic acid in one step from the Chinese medicinal plant Phyllanthus urinaria L. by use of direct and successive injections of a crude methanolic extract. Some aspects concerning the practical use of this technique in the described application are considered.     

高速逆流色谱法同时分离制备头花蓼中没食子酸和原儿茶酸

对蓼科蓼属头状蓼组植物头花蓼进行化学成分的研究。本研究建立了HPLC测定中药头花蓼水提喷雾干燥粉末中化学成分含量的方法,然后应用高速逆流色谱法对头花蓼水提喷雾干燥粉末的乙酸乙酯粗提物的化学成分进行了半制备性分离研究,通过对分离方法和溶剂系统的筛选,寻找到最佳的溶剂系统(正己烷∶乙酸乙酯∶甲醇∶水=1∶5∶1∶5),上相为固定相,转速840 r/min,流速2.0 mL/min,进样量756 mg,检测波长272 nm。结果显示,在该条件下经一步分离可同时得到质量为148.5 mg和9.2 mg的两种产物,纯度为99.8%和97.4%,经紫外、红外、质谱及核磁共振等方法进行结构分析,确定分别为没食子酸和原儿茶酸。     

高速逆流色谱分离制备川西獐牙菜中两种苷类化合物

采用高速逆流色谱从川西獐牙菜中分离制备了两种高纯度苷类化合物.以正丁醇-氯仿-甲醇-水(3.4∶8∶5∶6,v/v)为溶剂系统,主机转速为800 rpm,流速:O～210 min,1.5mL/min;210 ～360m in,2.5 mL/min,检测波长254 nm的条件下进行分离制备,在360 min内从100 mg样品中一步分离制备得到1-O-樱草糖-3,7,8-三甲氧基(口山)酮(Ⅰ,11 mg)和异荭草苷(Ⅱ,24 mg).经HPLC检测,两个化合物的纯度均在99％以上,结构由UV、1H和13C NMR鉴定.     

Separation and purification of echinacoside from Penstemon barbatus (Can.) Roth by recycling high-speed counter-current chromatography

Echinacoside is an important bioactive compound extracted from Cistanche tubulosa which was endangered by overexploitation. It is imperative to find an alternative source. Echinacoside was isolated from Penstemon barbatus (Can.) Roth for the first time. The peak contents of echinacoside are 9.09 ± 0.32 mg/g and 7.25 ± 0.36 mg/g respectively in the leaves and roots annually. The methanolic extracts from 20 g of dried powder of the roots of P. barbatus were pre-purified by AB-8 resin and the fraction containing echinacoside was further purified by conventional high-speed counter-current chromatography (HSCCC) and recycling HSCCC with the solvent system n-butanol–water (1:1, v/v). Totally 42.0 mg echinacoside with a purity of 96.3% was recovered. The recovery rate of echinacoside by recycling HSCCC reached 91.0%. The structure of our echinacoside confirmed by IR, ¹H NMR and ¹³C NMR is identical to the standard sample. This indicates that P. barbatus might be ideal source for preparation of large scale of echinacoside.     

高速逆流色谱法分离纯化丹参并尝试制订中药指纹图谱

用国产高速逆流色谱（HSCCC）分离纯化中草药——丹参,选用正己烷乙醇水体系,固定相保留率达到788%。采用分步洗脱,3个产地丹参各分离得到12个洗脱组分,经高效液相色谱仪和紫外光谱仪检测证明3张HSCCC洗脱图谱中对应洗脱峰为同一组分。HSCCC洗脱图谱不包含非共有峰,并且对应洗脱峰保留时间的相对标准偏差RSD<3%,符合国家标准关于制订指纹图谱方法学考察资料的技术参数。因此,HSCCC作为制订指纹图谱的方法之一具有可行性。     

高速逆流色谱法分离制备丹酚酸B

采用高速逆流色谱法分离纯化丹参水溶性成分丹酚酸类物质,制备丹酚酸B化学对照品。分离采用的溶剂系统为正己烷-乙酸乙酯-水-甲醇(1.5:5:5:1.5),上相做固定相,下相做流动相,流速为1.7 mL/min,仪器转速850 rpm,进样量80 mg,纯度用HPLC方法测定。结果表明:一次分离可制备63.4 mg丹酚酸B,其纯度为98.6%。该方法操作简单,可作为高纯度丹酚酸B化学对照品的制备分离方法。     

Preparative isolation and purification of four flavonoids from the petals of nelumbo nucifera by high‐speed counter‐current chromatography

Introduction – Flavonoids, the primary constituents of the petals of Nelumbo nucifera, are known to have antioxidant properties and antibacterial bioactivities. However, efficient methods for the preparative isolation and purification of flavonoids from this plant are not currently available. Objective – To develop an efficient method for the preparative isolation and purification of flavonoids from the petals of N. nucifera by high‐speed counter‐current chromatography (HSCCC). Methodology – Following an initial clean‐up step on a polyamide column, HSCCC was utilised to separate and purify flavonoids. Purities and identities of the isolated compounds were established by HPLC‐PAD, ESI‐MS, ¹H‐NMR and ¹³C‐NMR. Results – The separation was performed using a two‐phase solvent system composed of ethyl acetate–methanol–water–acetic acid (4 : 1 : 5 : 0.1, by volume), in which the upper phase was used as the stationary phase and the lower phase was used as the mobile phase at a flow‐rate of 1.0 mL/min in the head‐to‐tail elution mode. Ultimately, 5.0 mg syringetin‐3‐O‐β‐d‐glucoside, 6.5 mg quercetin‐3‐O‐β‐d‐glucoside, 12.8 mg isorhamnetin‐3‐O‐β‐d‐glucoside and 32.5 mg kaempferol‐3‐O‐β‐d‐glucoside were obtained from 125 mg crude sample. Conclusion – The combination of HSCCC with a polyamide column is an efficient method for the preparative separation and purification of flavonoids from the petals of N. nucifera. Copyright © 2009 John Wiley & Sons, Ltd.