大孔树脂-高速逆流色谱法分离纯化地黄中毛蕊花糖苷（英文）

该文建立了大孔树脂-高速逆流色谱分离中药材地黄中有效成分毛蕊花糖苷的方法。考察了4种大孔树脂对地黄粗提物中毛蕊花糖苷的静态吸附与解吸情况,其中D101大孔树脂对目标成分的吸附率与解吸率最理想,实验结果表明体积分数为10%的乙醇洗脱得到的毛蕊花糖苷含量最高,目标成分含量从4.9%提高到32.6%。最后,部分纯化的样品(165 mg)采用高速逆流色谱进一步纯化,两相溶剂系统由乙酸乙酯-正丁醇-水(1∶4∶5,v/v/v)组成,分离得到45 mg纯度为96%的毛蕊花糖苷。     

大孔树脂-高速逆流色谱分离纯化薇甘菊中的黄酮类化合物

该文建立了大孔树脂-高速逆流色谱分离薇甘菊中黄酮类物质的方法。分离条件为:采用大孔树脂AB-8,洗脱液为50%（v/v）乙醇水溶液,高速逆流色谱溶剂体系为正丁醇-乙酸-水（4：1：5,v/v）。从薇甘菊中分离到4种黄酮类物质:槲皮素-3-O-芸香糖苷（纯度90.2%）、山奈酚-3-O-芸香糖苷（纯度98.55%）、木犀草苷（纯度98.33%）和紫云英苷（纯度99.23%）。建立的大孔树脂-高速逆流色谱方法简单、高效,可扩展应用于从其他植物中分离黄酮类物质。     

大孔树脂-高速逆流色谱分离纯化薇甘菊中的黄酮类化合物（英文）

该文建立了大孔树脂-高速逆流色谱分离薇甘菊中黄酮类物质的方法。分离条件为:采用大孔树脂AB-8,洗脱液为50%(v/v)乙醇水溶液,高速逆流色谱溶剂体系为正丁醇-乙酸-水(4∶1∶5,v/v)。从薇甘菊中分离到4种黄酮类物质:槲皮素-3-O-芸香糖苷(纯度90.2%)、山奈酚-3-O-芸香糖苷(纯度98.55%)、木犀草苷(纯度98.33%)和紫云英苷(纯度99.23%)。建立的大孔树脂-高速逆流色谱方法简单、高效,可扩展应用于从其他植物中分离黄酮类物质。     

大孔吸附树脂结合酶解法分离纯化虎杖中白藜芦醇的研究

研究了大孔吸附树脂结合酶解法提取和纯化虎杖中白藜芦醇的方法,采用HPLC法测定虎杖中白藜芦醇的含量,考查了β-糖苷酶对虎杖药材酶解前后白藜芦醇含量的变化,并经静态吸附考察了4种树脂,最后确定以H1020作为提取分离白藜芦醇的树脂.此树脂吸附量较高,脱附容易,有利于得到质量较好的白藜芦醇产品,经该树脂吸附解吸,饱和吸附量可达51.4mg/g,解吸率达92.5%.大孔树脂分离纯化白藜芦醇的含量可达71.5%,而上柱前粗提物中白藜芦醇含量为8.71%,说明采用本法分离纯化虎杖中白藜芦醇是可行的.     

大孔阴离子树脂分离纯化微生物转化液中5-氟尿苷的研究

探寻从微生物转化液中分离纯化5-氟尿苷(5-FUR)的工艺条件。先后尝试了大孔吸附树脂、大孔弱碱性阴离子交换树脂以及硅胶柱色谱的分离方法,首先利用大孔弱碱性阴离子交换树脂吸附,然后用1mol/L NaCl动态洗脱得到含量较高的两种含氟化合物,最终通过硅胶柱色谱,采用正己烷-乙酸乙酯(含10%醋酸)为流动相进行洗脱,得到了纯度为100%的纯品。在目标化合物和杂质理化性质极其相似,含量极其悬殊的条件下,成功去除含量极高的杂质,实现了5-FUR的分离纯化,圆满地达到了预期的目的。     

大孔吸附树脂分离纯化昆仑雪菊总黄酮工艺的研究

筛选分离纯化昆仑雪菊总黄酮的大孔吸附树脂并建立纯化工艺条件。以大孔吸附树脂对昆仑雪菊总黄酮的吸附量、吸附率和解吸率等为指标,考察了11种型号的大孔吸附树脂进行分离纯化,并确定了该树脂分离纯化最佳工艺参数。结果显示NKA-9型大孔吸附树脂对昆仑雪菊总黄酮分离纯化效果好,其工艺条件为:以pH值为5.0的原料液上柱吸附,70%浓度的乙醇洗脱、洗脱速度3mL·min-1,在此条件下总黄酮的的静态吸附率为95.74%,解吸率为98.9%。     

在线加压溶剂微提取-湍流色谱-高效液相色谱法同时测定管花肉苁蓉中3种苯乙醇苷

建立了在线加压溶剂微提取-湍流色谱-高效液相色谱(online PLME-TFC-HPLC)法,并将其应用于管花肉苁蓉中松果菊苷、毛蕊花糖苷和异毛蕊花糖苷3种苯乙醇苷类成分含量的同时测定。微量样品粉末(0.5 mg)置于空预柱芯中并用正相硅胶填充,得到提取池后装入预柱套(Security Guard^TM)。将预柱套置于70℃柱温箱中,将一根长聚醚醚酮(PEEK)管线(1000 mm×0.13 mm)连于预柱套末端,采用0.1%(v/v)甲酸水为提取溶剂,以2.5 mL/min的速度流经PEEK管线,产生高压,实现管花肉苁蓉的在线加压溶剂微提取,通过TurboFlow cyclone色谱柱在线净化和富集。引入两个电子六通阀,将整个分析过程分为提取阶段和洗脱阶段,并在洗脱阶段将TurboFlow cyclone色谱柱中的分析物反冲至Capcell PAK C₁₈ AQ分析柱上,以0.1%(v/v)甲酸水-乙腈为流动相进行梯度洗脱,以340 nm为检测波长同时定量分析松果菊苷、毛蕊花糖苷和异毛蕊花糖苷3种苯乙醇苷类成分。结果表明,3种苯乙醇苷类在1~200 mg/L范围内线性良好,相关系数r均大于0.999,定量限分别为0.50 mg/L(松果菊苷)、0.25 mg/L(毛蕊花糖苷)和0.38 mg/L(异毛蕊花糖苷),加标回收率为83.13%~114.00%,相对标准偏差为1.89%~13.34%。该方法简便、快速、可靠,不仅节约了药材和溶剂的使用量,而且极大地简化了前处理方法,省时省力,同时显著降低了化学成分在提取过程中降解的几率,适用于管花肉苁蓉中苯乙醇苷类化合物的含量测定。     

大孔树脂对杨梅叶原花色素的纯化去糖研究

通过静态吸附与解析实验,比较了7种大孔树脂对杨梅叶原花色素的纯化效果。结果表明,HPD-500型大孔树脂的吸附效果最好。通过单因素实验,确定HPD-500型大孔树脂对杨梅叶原花色素的最佳吸附浓度为7.0mg/mL,吸附平衡时间为3h,吸附量可达到(106.61±7.83)mg/g大孔树脂;解吸液为50%乙醇水溶液,解吸时间为30min,解吸率达到96.67%。经过解吸后的产物,原花色素的含量从原来的26%提升到了45.3%,并除去了原有粗提物中的大部分糖,总糖含量从原来的24.7%下降到了4.4%。为了提高样品和大孔树脂的利用率,采用了静态吸附和动态吸附结合的方法,将固形物含量为11.83%的含有26%原花色素的粗提物溶液540mL与200g HPD-500型大孔树脂在砂芯玻璃层析柱中进行两次吸附解吸,合并两次解吸产物,基本除去了糖,原花色素含量提升至53%,原花色素的得率在80%以上,实现了原花色素的有效纯化。     

基于大孔树脂与制备液相色谱技术快速分离野地瓜茎中的绿原酸

建立了基于大孔吸附树脂快速富集,制备高效液相色谱高效分离野地瓜茎中绿原酸的制备方法:AB-8,D101,HPD600,CN206和NKA-Ⅱ5种树脂中,经静态吸附-解吸附试验发现NKA-Ⅱ型大孔树脂对目标化合物具有较好的吸附率和解析率;采用NKA-Ⅱ型大孔树脂,经4倍柱体积(bed volume,BV)5%(V/V)乙醇除杂后,用7 BV 10%(V/V)乙醇洗脱得到目标化合物组分,HPLC分析目标化合物的峰面积比达到80.8%;由制备高效液相色谱对目标化合物做进一步纯化并开发了重复进样分离模式,提高了分离效率,经纯化后目标化合物纯度达到98.6%;1H NM R和13C NM R鉴定目标化合物为绿原酸。该方法适合于野地瓜中绿原酸化合物的大规模制备。     

高速逆流色谱法快速分离制备枸杞中莨菪亭

建立了用高速逆流色谱(HSCCC)从枸杞中快速分离莨菪亭的方法。将枸杞的乙醇提取物经D-101大孔树脂初步纯化后直接进行高速逆流色谱分离,用薄层色谱-荧光法考察了莨菪亭在不同溶剂体系中的分配情况。结果表明,最佳的溶剂体系为氯仿-甲醇-水(10:7:3, v/v/v),取上相为固定相,下相为流动相,在主机转速为850 r/min、流速为1.5 mL/min、检测波长为365 nm的条件下,从200 mg样品中一次性分离制备可得到10.2 mg纯度达到98.3%的莨菪亭。制备所得的莨菪亭与对照品的高效液相色谱(HPLC)保留时间一致,且经核磁共振氢谱、碳谱鉴定结构;纯度经HPLC法测定。研究发现,氯仿-甲醇-水(10:7:3, v/v/v)体系可连续二次进样而样品的峰形未受明显的影响。实验结果表明用薄层色谱-荧光法可快速选定HSCCC溶剂体系,进而可快速、简便地制备高纯度的莨菪亭。     

Separation and purification of neohesperidin from the albedo of Citrus reticulata cv. Suavissima by combination of macroporous resin and high-speed counter-current chromatography

In this article, a simple and efficient protocol for rapid preparation and separation of neohesperidin from the albedo of Citrus reticulata cv. Suavissima was established by the combination of macroporous resin column chromatography and high-speed counter-current chromatography (HSCCC). Six types of resin were investigated by adsorption and desorption tests, and D101 macroporous resin was selected for the first cleaning-up procedure, in which 55% aqueous ethanol was used to elute neohesperidin. After treatment with D101 resin, the neohesperidin purity increased 11.83-fold from 4.92% in the crude extract to 58.22% in the resin-refined sample, with a recovery of 68.97%. The resin-refined sample was directly subjected to HSCCC purification with a two-phase solvent system composed of ethyl acetate-n-butanol-water (4:1:5, v/v), and 23.6 mg neohesperidin with 97.47% purity was obtained from 60 mg sample in only one run. The recovery of neohesperidin in HSCCC separation procedure was 65.85%. The chemical structure of the purified neohesperidin was identified by both HPLC and LC-MS. The established purification process will be helpful for further characterization and utilization of Citrus neohesperidin.     

Separation of injectable salidroside by column chromatography of macroporous resins for treating myocardial ischemia

The objective of the present study is to develop a method for large-scale separating and purifying salidroside from rhodiola kirilowii roots and for preparing injectable medicinal ingredient.Crude extract of salidroside was prepared by water-ethanol system,and purified by column chromatography of macroporous resins.Static adsorption and desorption studies were performed on six kinds of macroporous resins,and SP825 resin was chosen,followed by optimizing process parameters.The optimum sample volume,feed concentration,ratio of diameter to height,and feeding flow rate were 1.5 bed volumes(BV),15 mg/mL,1:10 and 1 BV/h,respectively.Dynamic desorption was performed consecutively with 8 BV of distilled water,3 BV of 5% ethanol and 8 BV of 10% ethanol at a flow rate of 2 BV/h.After three cycles in separating 3.5 tons of rhodiola kirilowii roots,salidroside purity was increased from 3.4% in the crude extract to 93.6% in purified salidroside product.This study provides a novel method to separate salidroside for injectable use.     

A simple and efficient protocol for large‐scale preparation of three flavonoids from the flower of Daphne genkwa by combination of macroporous resin and counter‐current chromatography

In this paper, macroporous resin column chromatography and counter‐current chromatography (CCC) were applied for large‐scale preparative separation of three flavonoids from the flower of Daphne genkwa, a famous Chinese medicinal herb. Nine kinds of resins were investigated by adsorption and desorption tests and D101 macroporous resin was selected for the first cleaning‐up, in which 40% aqueous ethanol was used to remove the undesired constituents and 90% aqueous ethanol was used to elute the targets. The crude extract after the first step was directly subjected to the preparative CCC purification using the solvent system composed of n‐hexane–ethyl acetate–methanol–water (4:5:4:5, v/v). The compounds apigemin (823 mg), 3‐hydroxyl‐genkwanin (842 mg) and genkwanin (998 mg) with the purities of 98.79, 97.71 and 93.53%, respectively, determined by HPLC were produced from 3‐g crude extract only in one CCC run. Their chemical structures were identified by MS, UV and the standards.     

Separation and purification of isorhamnetin 3-sulphate from Flaveria bidentis (L.) Kuntze by counter-current chromatography comparing two kinds of solvent systems

The first preparative separation of a flavonoid sulphate isorhamnetin 3-sulphate from Flaveria bidentis (L.) Kuntze by counter-current chromatography (CCC) was presented. Two kinds of solvent systems were used. A conventional organic/aqueous solvent system n-butanol-ethyl acetate-water (4:1:5, v/v) was used, yielding isorhamnetin 3-sulphate 2.0 mg with a purity of 93.4% from 83 mg of pre-enriched crude extract obtained from 553 mg ethanol extract by macroporous resin. A one-component organic/salt-containing system composed of n-butanol-0.25% sodium chloride aqueous solution (1:1, v/v) was also used, and the LC column packed with macroporous resin has been employed for desalination of the target compound purified from CCC. As a result, 2.1 mg of isorhamnetin 3-sulphate with a purity of over 97% has been isolated from 402 mg of crude extract without pre-enrichment. Compared with the conventional organic/aqueous system, the one-component organic/salt-containing aqueous system was more suitable for the separation of isorhamnetin 3-sulphate, and purer target compound was obtained from the crude extract without pre-enrichment using the new solvent system. The chemical structure was confirmed by ESI-MS and (1)H, (13)C NMR. In summary, our results indicated that CCC using one-component organic/salt-containing aqueous solution is very promising and powerful for high-throughput purification of isorhamnetin 3-sulphate from Flaveria bidentis (L.) Kuntze.     

Separation of capsaicin from capsaicinoids by macroporous resin adsorption chromatography

The aim of present study is to develop an efficient and low‐cost method for capsaicin production isolated from capsaicinoids by macroporous resin adsorption chromatography. HZ816 resin has shown the best adsorption and desorption capacities for capsaicin among other resins. To optimize the operating parameters for separation, initial concentration, diameter‐to‐height ratio, mobile phase ratio, and crystallization method were investigated. When capsaicinoids solution (5 g/L) was loaded onto the column (diameter‐to‐height ratio = 1:12) with ethanol/1% w/w NaOH (4:6, v/v) as the mobile phase, capsaicin was purified most effectively. By using acid neutralization as the crystallization method, the purity of capsaicin improved from 90.3 to 99.5% with 82.3% yield. In conclusion, this study provides a simple and low‐cost method for the industrial‐scale production of high‐purity capsaicin.     

An effective high-speed countercurrent chromatographic method for preparative isolation and purification of mollugin directly from the ethanol extract of the Chinese medicinal plant Rubia cordifolia

The medicinal plant Rubia cordifolia has been used widely in traditional Chinese medicine (TCM) for its antibacterial, antioxidant and anti-inflammatory activities. In this study, a preparative high-speed countercurrent chromatography (HSCCC) method for isolation and purification of the bioactive component mollugin directly from the ethanol extract of R. cordifolia was successfully established by using light petroleum (bp 60-90 degrees C)/ethanol/diethyl ether/water as the two-phase solvent system. The upper phase of light petroleum/ethanol/diethyl ether/water (5:4:3:1 v/v) was used as the stationary phase of HSCCC. Under the optimum conditions, 46 mg of mollugin at 98.5% purity, as determined by HPLC, could be yielded from 500 mg of the crude extract in a single HSCCC separation. The peak fraction of HSCCC was identified by 1H NMR and 13C NMR.     

大孔吸附树脂对辣椒素类物质的富集

辣椒;辣椒素;辣椒素类物质;吸附树脂;分离     

Combined application of macroporous resin and high speed counter-current chromatography for preparative separation of three flavonoid triglycosides from the leaves of Actinidia valvata Dunn

In this paper, the combined techniques of macroporous resin column chromatography and high speed counter-current chromatography were applied for preparative separation of flavonoid triglycosides from the leaves of Actinidia valvata Dunn, a famous Chinese medicinal herb. Twelve kinds of macroporous resins were investigated by adsorption and desorption tests. HPD-300 resin showed the maximum effectiveness and thus was selected for the first cleaning-up, in which 20% ethanol was used to remove the undesired constituents and 60% ethanol to elute the targets. The crude extract was then purified by high speed counter-current chromatography with the solvent system composed of ethyl acetate-n-butanol-water (2:1:3 and 4:1:5, v/v). Three flavonoid triglycosides, namely, kaempferol 3-O-α-L-rhamnopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→6)-β-D-galactopyranoside, kaempferol 3-O-α-L-rhamnopyranosyl-(1→3)-(4-O-acetyl-α-L-rhamnopyranosyl)-(1→6)-β-D-galactopyranoside and kaempferol 3-O-α-L-rhamnopyranosyl-(1→3)-(2,4-di-O-acetyl-α-L-rhamnopyranosyl)-(1→6)-β-D-galactopyranoside, were obtained. The purities of the separated compounds were all over 95% as determined by HPLC area normalization method. Their chemical structures were confirmed by UV, MS, NMR, and the standards.     

Efficient Protocol for Large-Scale Purification of Naringin with High Recovery from Fructus aurantii by Macroporous Resin Column Chromatography and HSCCC

Several kinds of resins were investigated in the first step and D101 macroporous resin was selected for cleaning-up naringin (NAR), a major flavonoid glycoside from Fructus aurantii. In the subsequent column chromatography, 10% aqueous ethanol was first used to elute the column to remove the undesired constituents and 70% aqueous ethanol was used to elute the target. The content of NAR was 57.1% with 95.7% recovery in this process. In the second step, the obtained crude sample was directly isolated by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of ethyl acetate–n-butanol–water at a volume ratio of 2: 0.8: 3.2 (v/v/v), and 331 mg NAR with 98.3% purity was obtained from 600 mg crude extract in only one run. The recovery of the compound in this step was 95.0%. Thus, the total recovery of NAR was 90.9% after the two step purification. The established protocol for large-scale isolation and separation of NAR with high purity and recovery from F. aurantii was simple, efficient, and suitable for pharmace- utical and commercial use.     

Preparative isolation and purification of bergapten and imperatorin from the medicinal plant Cnidium monnieri using high-speed counter-current chromatography by stepwise increasing the flow-rate of the mobile phase

A high-speed counter-current chromatography (HSCCC) method was developed for the preparative separation and purification of bergapten and imperatorin from the Chinese medicinal plant Cnidium monnieri (L.) Cusson. The crude extract was obtained by extraction with ethanol from the dried fruits of Cnidium monnieri (L.) Cusson under sonication. Preparative HSCCC with a two-phase solvent system composed of n-hexane-ethyl acetate-ethanol-water (5:5:5:5, v/v/v/v) was successfully performed by increasing the flow-rate of the mobile phase stepwise from 1.0 to 2.0 ml min(-1) after 180 min. The components purified and collected were analyzed by high-performance liquid chromatography. The method yielded 45.8 mg of bergapten at 96.5% purity and 118.3 mg of imperatorin at 98.2% purity from 500 mg of the crude extract in a single run. The recoveries of bergapten and imperatorin were 92.1 and 93.7%, respectively.