双柱吸附法对莲子心总碱的分离纯化研究

研究了大孔吸附树脂和阳离子交换树脂双柱吸附法对莲心碱、异莲心碱和甲基莲心碱的吸附行为。利用阳离子交换树脂D72制备出莲心碱、异莲心碱和甲基莲心碱,含量分别为10.6%、10.6%和28.0%,总含量为49.2%的莲子心总碱提取物,其中杂质主要是水溶性较大的生物碱成分。利用AKS-W大孔吸附树脂能去除极性较大的生物碱,而D72阳离子交换树脂能去除非生物碱的特点,采用双柱联合吸附法,制备出3种吸附质含量分别为33.1%、15.0%和34.5%,总含量为82.6%的莲子心总碱提取物。     

Fragmentation study on the phenolic alkaloid neferine and its analogues with anti-HIV activities by electrospray ionization tandem mass spectrometry with hydrogen/deuterium exchange and its application for rapid identification of in vitro microsomal metabolites of neferine

The application of mass spectrometry in drug discovery, especially in drug metabolites, is very important. This present paper is at first focused on the elucidation of fragmentation patterns of the phenolic bisbenzyltetrahydroisoquinoline alkaloid, neferine, together with its analogues isoliensinine and liensinine with anti-HIV activities using electrospray ionization tandem mass spectrometry (ESI-MS/MS) and hydrogen/deuterium (H/D) exchange. All title compounds displayed major diagnostic fragments that formed by the cleavage of the C1'--C9' bond resulting in positive group CD, and the loss of 4-ethyl-1-phenol or 4-ethyl-1-methoxybenzene following rearrangements. Their ESI-MS/MS spectra also showed the relatively stable fragment ions formed by the elimination of H2O, CH3NH2, CH3OH, and CH3-N==CH2. Secondly, the metabolites of neferine from dog hepatic microsomal incubations were analyzed and characterized by high-performance liquid chromatography (HPLC) and data-dependent ESI-MS/MS. Based on fragmentation patterns and compared with their retention times in LC, molecular weights and ultraviolet (UV) absorbances with standard compounds, six metabolites were identified as isoliensinine, liensinine and four novel bisbenzyltetrahydroisoquinoline alkaloids named as 6-O-desmethylneferine, 2'-N-desmethylneferine, 2'-N-6-O-didesmethylneferine, and 6,13-O-didesmethylneferine. All metabolites were desmethyl or didesmethyl products of neferine. The possible metabolic pathways for neferine have been proposed. The results suggest that N-demethylation and O-demethylation are two important metabolic pathways of neferine in dog hepatic microsomal incubations. This is critical for screening and development of phenolic bisbenzyltetrahydroisoquinoline alkaloids with anti-HIV activities such as neferine and its analogues isoliensinine and liensinine.     

Preparative separation and purification of liensinine,isoliensinine and neferine from seed embryo of Nelumbo nucifera GAERTN using high‐speed counter‐current chromatography

A preparative high‐speed counter‐current chromatography method for separation and purification of liensinine, isoliensinine and neferine from seed embryo of Nelumbo nucifera GAERTN was successfully established by using n‐hexane‐ethyl acetate‐methanol‐water (5:8:4:5, v/v, containing 0.5% NH₄OH) as the two‐phase solvent system. From 200 mg of crude extract, 18.4 mg of liensinine, 19.6 mg of isoliensinine and 58.4 mg of neferine were obtained with the purity of 96.8, 95.9, and 98.6%, respectively. The identification of the three alkaloids was performed with ¹H NMR and ¹³C NMR.     

Preparative counter-current chromatography isolation of liensinine and its analogues from embryo of the seed of Nelumbo nucifera GAERTN. using upright coil planet centrifuge with four multilayer coils connected in series

Preparative counter-current chromatography (CCC) isolation of liensinine and its analogues, isoliensinine and neferine from embryo of the seed of Nelumbo nucifera GAERTN. has been successfully performed for the first time using upright coil planet centrifuge with four multilayer coils connected in series with 1600 mL capacity. Two kinds of two-phase solvent systems were applied to preparative CCC isolation. The first was the system composed of light petroleum (b.p. 60-90 degrees C)-ethyl acetate-tetrachloromethane-chloroform-methanol-water (1:1:4:4:6:2, v/v) which was very suitable for fast and small-scale CCC isolation. The second was the system composed of ethyl acetate-tetrachloromethane-methanol-water (1:6:4:1, v/v), which was the optimum for large-scale CCC isolation. Using the first system, 1102 mg of the crude alkaloid was purified in one-step separation of 150 min, yielding 350 mg neferine, 100 mg isoliensinine and 95 mg liensinine with over 95% purity. While using the second solvent system, 5850 mg of the crude alkaloid was purified in one-step separation of 9 h, yielding 2545 mg neferine, 698 mg isoliensinine and 650 mg liensinine with over 97% purity. Structures of the compounds were identified by electrospray ionization multiple mass spectrometry, one- and two-dimensional NMR.     

Bisbenzylisoquinoline Alkaloids from Nelumbo nucifera

From the embryos of the seeds of Nelumbo nucifera, three bisbenzylisoquinoline alkaloids, nelumboferine and nelumborines A and B, were isolated along with four known compounds, neferine, liensinine, isoliensinine and anisic acid. The structures of the new alkaloids were determined mainly by spectroscopic methods.     

Zn2＋型磺酸树脂对维生素B12的配位吸附及洗脱性能

原位吸附;分离;Zn2＋型磺酸树脂对维生素B12的配位吸附及洗脱性能     

A new bisbenzylisoquinoline alkaloid from Plumulanelumbinis

One new bisbenzylisoquinoline alkaloid, neferine N-oxide(1), along with three known compounds,neferine(2), liensinine(3), and isoliensinine(4), were isolated from Plumulanelumbinis – the embryo of the seed of Nelumbonucifera. Their structures were elucidated by extensive spectroscopic analysis(MS,UV, IR, NMR), and the absolute configurations were determined by experimental circulardichroism(CD)spectra. Compound 1 is a new naturally occurring bisbenzylisoquinoline alkaloid with an N-oxide functionality, and the CD spectrum of(1R, 1'R) neferine is first reported. The antioxidant activities of compounds 1–4 were evaluated using the ORAC assay, which illustrated that all these compounds showed the different levels of oxygen radical absorbance capacity.     

大孔树脂分离纯化丹酚酸的研究

比较了D301R、D392、D380大孔阴离子交换树脂和X-5.AB-8、NKA-9、SP825大孔吸附树脂对丹参水溶性成分的吸附和解吸能力,筛选出效果较好的SP825进行分离纯化丹酚酸的研究.实验表明,大孔吸附树脂SP825能分离出纯度为95.32%的丹参素,在梯度洗脱条件下可得到以丹参素(水洗脱)和丹酚酸B(乙醇洗脱)为主的产品.在最佳吸附与解吸工艺参数下,丹参素、紫草酸、迷迭香酸、丹酚酸A和丹酚酸B的收率分别为:36.92%、80.39%、82.45%、43.07%和41.03%.     

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     

SYNTESIS OF THE COMPLEXES OF MACROPOROUS SULFONATED RESINS WITH FERRIC CHLORIDE AND THEIR CATALYTIC BEHAVIOR FOR SETERIFICATION OF ACETIC ACID WITH BUTANOL

The complex resins prepared from macroporous sulfonated resin D72(H^+ form) with ferric chloride or ferric chloride hexahydrate have both sites of Bronsted acid and Lewis acid.In the catalysis of exterification of acetic acid with butanol the complex resins show to have much higher catalytic activity than that of its matrix.a conventional sulfonated cation exchange resin D72.     

大孔树脂精制6种含黄连复方水提液体系的吸附动力学比较研究

研究含黄连的不同复方水提液体系经大孔吸附树脂精制后,黄连中主要指标性成分生物碱的变化,阐明不同复方组合-配伍对药方中组成药味指标性成分的影响,为大孔树脂精制技术在中药复方中的应用提供实验依据.选用黄连用量和复方总用量均相同,组方药味不同的复方1～6.通过考察5种树脂AB-8、S-8、X-5、NKA-II、NKA-9对黄连中盐酸小檗碱的静态吸附率,确定AB-8树脂作为复方1～6吸附动力学比较研究的树脂;通过比较复方1～6水提液体系在相同树脂环境下的静态吸附过程,考察6种不同复方组成药味对复方中指标性成分分离纯化的影响.实验结果表明,由于体系本身溶液环境的差异,复方1～6各指标性成分达到吸附平衡的时问均有差异.AB～8树脂对复方1～6水提液体系的静态吸附在开始阶段吸附量的增加比较明显,在40min后,由于吸附竞争点饱和的原因,吸附量的增加均比较平稳;黄连与不同药材配伍时,虽然配伍药材所含的化合物类型差异较大,但黄连中生物碱类成分均可以在树脂上被选择性吸附,可知在复方水提液复杂体系中可以选择性地分离纯化生物碱类有效成分.     

两种大孔吸附树脂结合分离纯化京尼平甙

比较了H103、NKA-II、HPD100A、HPD400A及D141等5种大孔吸附树脂对栀子提取液中栀子黄色素和京尼平甙的吸附性能。在通过静态吸附实验研究其吸附量、吸附动力学特征的基础上,确定了用H103和HPD100A两种非极性大孔树脂进行京尼平甙的分离纯化,并确定了工艺参数。首先用H103树脂吸附京尼平甙,用蒸馏水洗脱杂质,再用一定浓度的乙醇洗脱;所得的京尼平甙洗脱液再用HPD100A树脂吸附,进一步除去栀子黄色素等杂质,得到的京尼平甙纯度达到81.3%,回收率为88.5%。     

Preparative separation of isoquinoline alkaloids from Corydalis impatiens using a middle‐pressure chromatogram isolated gel column coupled with two‐dimensional liquid chromatography

We established a two‐dimensional strong cation exchange/reversed‐phase liquid chromatography protocol to isolate and purify isoquinoline alkaloids from Corydalis impatiens. Isoquinoline alkaloids were first enriched from a C. impatiens extract in which liposoluble components were removed using a medium‐pressure chromatographic tower containing middle chromatogram isolated gel. A strong cation exchange column was employed to separate and obtain 30 fractions. We chose fractions 22–29 for reversed‐phase liquid chromatography purification using characteristic isoquinoline alkaloid ultraviolet absorption spectra. Several isoquinoline alkaloid fractions (22–29) were further separated, and those of low resolution were isolated via two‐dimensional liquid chromatography in the orthogonal plane. A total of eight novel isoquinoline alkaloids with characteristic ultraviolet spectra were obtained from C. impatiens. We thus demonstrate the benefits of off‐line two‐dimensional strong cation exchange/reversed‐phase liquid chromatography to isolate isoquinoline alkaloids from C. impatiens.     

Adsorption and Desorption Characteristics of Total Flavonoids from Acanthopanax senticosus on Macroporous Adsorption Resins

We examined the application of six different resins with the aim of selecting a macroporous resin suitable for purifying Acanthopanax senticosus total flavonoids (ASTFs) from Acanthopanax senticosus crude extract (EAS) by comparing their adsorption/desorption capacities, which led to the selection of HPD-600. Research on the adsorption mechanism showed that the adsorption process had pseudo-second-order kinetics and fit the Freundlich adsorption model. Moreover, the analysis of thermodynamic parameters indicated that the adsorption process is spontaneous and endothermic. The optimal conditions for purification of ASTFs were determined as sample pH of 3, 60% ethanol concentration, and 3 BV·h⁻¹ flow rate, for both adsorption and desorption, using volumes of 2.5 and 4 BV, respectively. The application of macroporous resin HPD-600 to enrich ASTFs resulted in an increase in the purity of total flavonoids, from 28.79% to 50.57%. Additionally, the antioxidant capacity of ASTFs was higher than that of EAS, but both were lower than that of L-ascorbic acid. The changes in ASTFs compositions were determined using ultra-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS), with the results illustrating that the levels of seven major flavonoids of ASTFs were increased compared to that in the crude extract.     

Semi‐preparative high‐speed counter‐current chromatography separation of alkaloids from embryo of the seed of Nelumbo nucifera Gaertn by pH‐gradient elution

A new high‐speed counter‐current chromatography method for semi‐preparative separation and purification of alkaloids from embryo of the seed of Nelumbo nucifera Gaertn was developed by using pH‐gradient elution mode. Diethyl ether was used as the stationary phase of the two‐phase solvent system and Na₂HPO₄/NaH₂PO₄ buffer solution with pH values of 7.5 and 7.2 in gradient mode as the mobile phase. Consequently, 33 mg of liensinine, 42 mg of isoliensinine, and 67 mg of neferine were obtained from 200 mg of crude extracts. The purities of them were all over 98% as determined by HPLC area normalization method, and the structures were identified by ¹H‐NMR and ¹³C‐NMR.     

大孔吸附树脂分离提取多杀菌素

采用大孔吸附树脂法分离提取多杀菌素.从11种大孔吸附树脂中筛选出DM11进行了静态、动态吸附性能实验,并考察了不同吸附、解吸条件的影响.结果表明,DM11的静态吸附容量为25.63mg/g(wet resin),其吸附等温线符合Langmuir吸附等温式.采用丙酮做洗脱剂,洗脱率为97.5%,动态吸附最佳吸附pH为9.5,吸附流速为6BV/h,穿透吸附容量为21.2mg/ml(wet resin),洗脱流速1.5BV/h.     

大孔吸附树脂富集板蓝根中(R,S)-告依春工艺研究

采用静态吸附法考察了D101、AB-8、NKA-2、NKA-9、HPD 100、HPD600等6种大孔吸附树脂对(R,S)-告依春的吸附及解吸性能,筛选出效果最佳的AB-8树脂,并对其进行动态考察.最佳富集条件为:上样液pH 6,生药质量-体积浓度为0.200g/mL,解吸液为2BV量70％乙醇,在优化条件下(R,S)-告依春在浸膏中含量可从0.76％提高到12.48％.结果表明,AB-8型大孔吸附树脂可用来从板蓝根水提取液中富集(R,S)-告依春.     

阴离子交换树脂回收水相中有机磷酸萃取剂

研究了阴离子交换树脂对水相中有机磷酸萃取剂的吸附。 通过比较不同的离子交换树脂对水相中2-乙基己基膦酸-单-2-乙基己基酯(P507)的去除率,发现大孔强碱性阴离子交换树脂(D201-OH)从水溶液中去除P507的能力最强,去除率可达99.24%。 而且当溶液在pH=1.0时,D201-OH对P507的吸附主要是分子吸附,其吸附等温线更适用于Langmuir模型;当溶液在pH=5.0时,阴离子交换反应占主导地位,其吸附等温线更适用于Freundlich模型。 研究还表明,D201-OH对P507的吸附在20 min内即达到吸附平衡时99.8%的吸附量。 通过动力学研究表明,拟一级动力学模型(R²>0.99)更适用于描述实验数据,并且吸附速率主要受膜扩散控制。 此外,吸附-解吸附循环8次后,D201-OH的吸附能力仍然保持在93%以上。 综上所述,D201-OH是有机磷酸类萃取剂的良好吸附剂,其吸附性能高效,循环过程稳定,因此可用于实际生产过程中回收有机磷酸萃取剂。     

一种基于R语言的大孔吸附树脂预测模型的构建

针对精准筛选高选择性分离中药多糖的大孔吸附树脂时间冗长、操作繁琐的现状,该文构建了一种快速筛选大孔吸附树脂的预测模型.采用R语言建立大孔吸附树脂参数、葡聚糖分子量与吸附量之间的多源信息模型,利用多源信息模型快速预测和筛选对中药多糖具有最佳吸附效果的大孔吸附树脂类型.所建多源信息模型的相关系数(r2)为0.901 2,P...     

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.