行星式逆流层析法的初步研究

螺旋管行星式离心分离机(Coil Planet Centrifuge)及其逆流分配层析法(Countercurrent Partition Chromatography),现称行星式逆流层析法,系由Ito首创,后经Lightbown,Suther-land等改进,近几年Ito又有新的发展。这是一种不用担体的液-液分配层析法,其分离原理与Craig的逆流分配法相似,即根据物质在两相溶剂系统中的分配系数的差异,经过样品在此系统中反复分配传递而达到组分的分离;但其技术构成已大大区别于Craig法,样品在内径为几毫米至零点几毫米的螺旋状聚四氟乙烯毛细软管内通过特定的旋转运动在两相溶剂系统中进行分配层析。用此法分离分析几毫克至几克的混合物时理论板数可达数千至一万,类似组分可在数小时内完成     

水平螺旋管行星式离心分离机逆流分配层析法

一、引言液相色谱法是进行物理化学分析研究的基本手段之一。近代色谱法系以不同物质在两相中具有分配系数的差异作为特征,在实验过程     

应用新型立式逆流色谱制备分离南蛇藤中的南蛇藤素

A versatile countercurrent chromatography with upright multilayer coil planet centrifuge, named upright countercurrent chromatography (UCCC), was applied to the isolation and purification of celastrol from the roots of Celastrus orbiculatus Thunb. The crude celastrol was obtained by elution with petroleum ether from ethanol extracts using a 15 cm length and 5 cm I.D. of silica gel flash chromatography. Preparative UCCC (Fig. 1) with a two-phase system composed of petroleum ether (b. p. 60 ～ 90 ℃ )-ethyl acetate-tetrachloromethanemethanol-water ( 1:1:8:6: 1, v/v) was successfully performed, yielding 705 mg celastrol at 99.5 % purity from 1020 rng of the crude extract in one step separation.     

应用立式逆流色谱和反相液相色谱制备性分离荜菝中的酰胺类化合物

A versatile countercurrent chromatography with upright type-J multilayer coil planet centrifuge,named upright countercurrent chromatography (UCCC),was applied to the isolation and purification of amides from Piper Longum L., which is widely used as an anodyne and a treatment for stomach disease in China. After saponification by KOH of the ethanol extracts solution of crud drug "Piper Longi Fructus", the fruits of Piper Longum L., the remaining solution was extracted with petroleum ether and red crude oil was obtained. Using 2.5 g red crude oil as sample, the preparative UCCC with a two-phase system composed of petroleum ether (b. p. 60 ～ 90 ℃)-ethyl acetate-tetrachloromethanemethanol-water ( 1: 1: 8: 6: 1, v/v) was successfully performed, yielding ten fractions. Then these fractions were further purified by use of reversed-phase liquid chromatography with a glass column of 500 × 10 mm I. D.packed with reversed - phase silica gel. As a result,nine amides with more than 98% purity, i.e., (2E,4E)-N-isobutyl-eicosa-2,4-dienamide, (2E, 4E, 8Z)-N-isobutyl-eicosa-2,4,8-trienamide, (2E, 4E, 8Z)-N-isobutyl-ocatadeca-2,4,8-trienamide, guineensine, pipemonaline, ( 2E, 4E )-N-isobutyl-2, 4-didecadienamide, piperine, piperanine, and piperlonguminine were isolated preparatively.     

半制备型高速逆流色谱分离制备铁棒锤根中的一种咪唑生物碱

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。     

一种用于色谱分离的凝胶型苯乙烯系阳离子树脂的制备方法

<正>公开号:CN103433010A公开日:2013.12.11申请人:凯瑞化工股份有限公司摘要本发明提供一种用于色谱分离的凝胶型苯乙烯系阳离子树脂的制备方法,选用凝胶型苯乙烯系阳离子交换树脂生产标准工艺配方中的升温方法及反应时间,生产出苯乙烯系聚合白球,然后在磺化时于硫酸中加入螯合剂及氟硼酸,经标准工艺磺化反应可得色谱分离树脂。相对于磺化时未加入螯合剂和氟硼酸的凝胶型苯乙烯系阳离子交换树脂,本发明制备色谱分离树脂吸附能力可提高50%;机械强度可提高20%。本发明制备的色谱分离树脂可提高果糖的含量,     

一种凝血酶亲和色谱介质的制备方法

对凝血酶-琼脂糖亲和色谱介质的制备方法进行了研究。首先使用凝血酶和溴化氰活化的琼脂糖制备凝血酶-琼脂糖亲和色谱介质,然后用生色底物法考察亲和色谱介质上凝血酶的活性,以凝血酶活性为指标对最佳偶联条件进行了优化。结果表明最佳条件为使用pH 8.3的Na₂CO₃-NaHCO₃溶液(含0.5 mol/L NaCl)为缓冲溶液,凝血酶用量为每1 g色谱介质加入凝血酶200 U,室温反应10 h。在最佳条件下所制备的色谱介质有较好的稳定性,在4℃条件下存放40天,亲和介质上的凝血酶活性仍有70.6%保留。该亲和色谱介质可广泛用于含凝血酶抑制剂的天然药物筛选和分离纯化。     

芦荟色酮的高速逆流色谱分离制备方法研究

芦荟色酮是在芦荟叶中特有的一类具有抗炎和抑制酪氨酸酶等作用的活性物质。以芦荟全叶为原料,经过一系列的预处理手段,从脱色剂活性炭中获得芦荟色酮粗提物,再经过溶剂分配和富集后采用高速逆流色谱（HSCCC）对其中的色酮成分进行分离纯化。研究结果表明,采用氯仿-甲醇-水(体积比为4∶3∶2)混合溶液和二氯甲烷-甲醇-水(体积比为5∶4∶2)混合溶液作为溶剂分离系统,经过两步HSCCC可以分离纯化出色谱纯度在95％以上的芦荟色酮单体。经过紫外检测、快原子轰击质谱及核磁共振等方法的结构分析鉴定,证实分离所得物质为肉桂酰基-C-葡萄糖甙芦荟色酮。     

高良姜中二苯基庚烷类化合物的高速逆流色谱分离制备

应用高速逆流色谱法(HSCCC)分离纯化了高良姜中3种二苯基庚烷类化合物。以正己烷-乙酸乙酯-甲醇-水(2:3:1.75:1, v/v/v/v)为两相溶剂系统,下相为固定相,上相为流动相,在主机转速为858 r/min、流速1.5 mL/min的条件下,从122.20 mg高良姜石油醚萃取物中经一步HSCCC分离可制备得到5R-羟基-7-(4-羟基-3-甲氧基苯基)-1-苯基-3-庚酮(7.37 mg)、7-(4-羟基-3-甲氧基苯基)-1-苯基-4E-烯-3-庚酮(9.11 mg)和1,7-二苯基-4E-烯-3-庚酮(15.44 mg),经高效液相色谱分析,纯度均大于93%,各化合物的结构由质谱和核磁共振氢谱、碳谱鉴定确证。该方法简便、快速、高效,可用于高良姜中二苯基庚烷类化合物的快速分离制备。     

高速逆流色谱对芦荟有效成分的制备性分离研究

1　引　　言由于高速逆流色谱具有两大突出优点 :(1 )聚四氟乙烯管中的固定相不需要载体 ,因而消除了液相色谱中由于使用载体而带来的吸附现象 ,特别适用于分离极性物质和具有生物活性的物质 ;(2 )由于其与一般色谱的分离方式不同 ,使其特别适用于制备性分离。芦荟的叶汁干燥品为一传统的常用中药。它除用作药物外 ,在化妆品行业己将芦荟系列作为我国今后的主要化妆品系列之一。另外 ,在饮料及食品等工业中 ,它得到了广泛应用。其化学有效成分主要是芦荟甙 ,以及一些少量的芦荟大黄素、异芦荟甙、β 芦荟甙等蒽醌类活性成分。2　实验部分2 .…     

Improved g-level calculations for coil planet centrifuges

Calculation of the g-level is often used to compare CCC centrifuges, either against each other or to allow for comparison with other centrifugal techniques. This study shows the limitations of calculating the g-level in the traditional way. Traditional g-level calculations produce a constant value which does not accurately reflect the dynamics of the coil planet centrifuge. This work has led to a new equation which can be used to determine the improved non-dimensional values. The new equations describe the fluctuating radial and tangential g-level associated with CCC centrifuges and the mean radial g-level value. The latter has been found to be significantly different than that determined by the traditional equation. This new equation will give a better understanding of forces experienced by sample components and allows for more accurate comparison between centrifuges. Although the new equation is far better than the traditional equation for comparing different types of centrifuges, other factors such as the mixing regime may need to be considered to improve the comparison further.     

Compact type-I coil planet centrifuge for counter-current chromatography

A compact type-I coil planet centrifuge has been developed for performing counter-current chromatography. It has a revolution radius of 10 cm and a column holder height of 5 cm compared with 37 and 50 cm in the original prototype, respectively. The reduction in the revolution radius and column length permits application of higher revolution speed and more stable balancing of the rotor which leads us to learn more about its performance and the future potential of type-I coil planet centrifuge. The chromatographic performance of this apparatus was evaluated in terms of retention of the stationary phase (S_f), peak resolution (R_s), theoretical plate (N) and peak retention time (t_R). The results of the experiment indicated that increasing the revolution speed slightly improved both the retention of the stationary phase and the peak resolution while the separation time is remarkably shortened to yield an excellent peak resolution at a revolution speed of 800 rpm. With a 12 ml capacity coiled column, DNP-DL-glu, DNP-β-ala and DNP-l-ala were resolved at R_s of 2.75 and 2.16 within 90 min at a flow rate of 0.4 ml/min. We believe that the compact type-I coil planet centrifuge has a high analytical potential.     

Evaluation on the performance of four different column models mounted on the compact type-I coil planet centrifuge

Optimal positions of coiled separation columns on the type-I centrifuge were determined for four typical two-phase solvent systems to obtain the best separation efficiency (resolution and retention of stationary phase) for each with a suitable set of test samples. A set of short coiled columns is connected in series and mounted around the holder hub in four different ways: (model A) the tail of one unit with left-handedness was connected to the head of the next unit with right-handedness (TL-HR); (model B) the tail of one unit with left-handedness was connected to the tail of the next unit with right-handedness (TL-TR); (model C) the tail of one unit with left-handedness was connected to the tail of the next unit with left-handedness (TL-TL); (model D) the tail of one unit with left-handedness was connected to the head of the next unit with left-handedness (TL-HL). The results indicated that the performance of model D was the best among the four models. High revolution speed (800 rpm) is favorable to separation using the moderately hydrophobic solvent system of hexane-ethyl acetate-methanol-0.1M HCl (1:1:1:1, v/v) (HEMW), while lower revolution speed (600 rpm) is beneficial to the separation with polar solvent system of 1-butanol-acetic acid-water (19:1:20, v/v) (BAW).     

Counter-current chromatographic separation of nucleic acid constituents with a hydrophilic solvent system

Nucleic acid constituents such as nucleobases, nucleosides and nucleotides were separated by counter-current chromatography using type J coil planet centrifuge. The separation was performed with a hydrophilic solvent system composed of 1-propanol/800 mM potassium phosphate buffer (pH 7.4) (1:1, v/v) by eluting the lower aqueous phase at a flow-rate of 0.5 ml/min. Eight selected nucleic acid constituents (4.0 mg, 0.5 mg of each), uridine monophosphate (UMP), adenosine monophosphate (AMP), deoxyadenosine monophosphate (dAMP), uridine, urasile, deoxy uridine, adenosine and adenine were well resolved within 160 min.     

Critical β-values of all coil planet centrifuges

I-type, J-type and non-synchronous centrifuges are all coil planet centrifuges. Analysing the motion of I-type and J-type centrifuges has advanced the understanding of how to manufacture and use these centrifuges. This paper analyses the motion of non-synchronous centrifuges producing equations of motion that can be applied to all coil planet centrifuges. This has also produced simple equations to determine the critical β-values for any coil planet centrifuge. This paper also demonstrates that I-type centrifuges also have 2 critical β-values when it was thought that β-value did not influence the understanding of the processes within I-type centrifuges. For the I-type instrument both of these critical values are at bobbin radii approaching infinity. In practice this means all I-types function within one β-value range hence the unilateral distribution and type/effectiveness of the mixing is consistent. Finally the paper shows the influence that the tangential velocity has on the Archimedean screw effect and thus the unilateral distribution of the upper and lower phases in the columns of coil planet centrifuges. This explains why the maximum stationary phase retention in an I-type centrifuge is limited to 50%.     

Preparative isolation and purification of celastrol from Celastrus orbiculatus Thunb. by a new counter-current chromatography method with an upright coil planet centrifuge

A new counter-current chromatography (CCC) method with an upright coil planet centrifuge, which holds four identical multilayer coil columns in the symmetrical positions around the centrifuge axis, was applied to the isolation and purification of celastrol from the roots of Celastrus orbiculatus Thunb. The crude celastrol was obtained by elution with light petroleum from ethanol extracts using 15 cm x 5 cm i.d. silica gel flash chromatography. Preparative CCC with a two-phase system composed of light petroleum (bp 60-90 degrees C)-ethyl acetate-tetrachloromethane-methanol-water (1:1:8:6:1, v/v) was successfully performed, yielding 798 mg celastrol at 99.5% purity from 1020 mg of the crude sample in one step separation.     

Fiber-packed needle-type sample preparation device designed for gas chromatographic analysis

A miniaturized sample preparation technique that uses a fine-fiber-packed needle as the extraction medium is reviewed, especially in relation to its application to the analysis of volatile organic compounds by gas chromatography. When the needle was packed longitudinally with a bundle of fine filaments (12 μm o.d.) which were also surface-coated with polymeric materials, successful sample preconcentration was obtained. Improved sensitivity was also established by introducing simultaneous derivatization reactions into the extraction process in the fiber-packed needle. The storage performance of the needle clearly demonstrated the potential of the technique for typical on-site sampling during environmental analysis. In this short review, the fiber-packed extraction needle developed by the authors is summarized along with applications that use the fiber-packed needle as a miniaturized extraction device.     

样品前处理技术在气相色谱分析中的应用进展

气相色谱法是当前应用最广泛的分析技术之一。使用气相色谱对复杂基体进行分析时的样品前处理步骤往往繁琐耗时,易引起误差,已成为制约分析效率和准确度提升的关键环节。本文综述了2009-2013年几种主要的样品前处理技术,包括吹扫捕集、固相萃取、固相微萃取、液相微萃取技术以及微波辅助萃取、超声波辅助萃取等场辅助萃取技术在气相色谱分析中的应用研究进展。     

Sample preparation for gas chromatographic analysis of organic solvents in aerosol cans

Summary A method for the sampling of chemical products from aerosol cans is described. An aerosol can is frozen in liquid nitrogen, followed by puncturing the can and allowing the propellant to distill off. The conditions for the smaple preparation have been optimized. Solvent content in the products were analysed by headspace gas chromatography-mass spectrometry.