Spiral tube assembly for countercurrent chromatography

用于逆流色谱的螺旋管组件

基本信息

批准号：
8557952
负责人：
Yoichiro Ito
金额：
$ 3.64万
依托单位：
NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

项目摘要

Existing two-phase solvent systems for high-speed countercurrent chromatography cover the separation of hydrophobic to moderately polar solvent systems, but often fails to provides a suitable partition coefficient values for highly polar compounds such as sulfonic dyes, catecholamines and twitter ions. The present paper introduces a new solvent series which can be applied for separation of these polar compounds by spiral tube countercurrent chromatography. It is composed of 1-butanol, ethanol, saturated ammonium sulfate and water at various volume ratios. The system consists of 10 steps which are arranged according to the polarity of the solvent system so that the two-phase solvent system with suitable K values for the target compound(s)can be found in few steps of search. Each solvent system gives proper volume ratio and high density difference between the two phases, and provides a satisfactory level of retention of the stationary phase in the spiral column assembly. The method is validated by partition coefficient measurement of four typical polar compounds including methyl green (basic dye), tartazine (sulfonic dye), tyrosine (twitter ion) and epinephrine (catecholamine), all of which show low partition coefficient values in the polar 1-butanol-water system. Recently, the above ultra polar solvent systems (Organic/high ionic strength aqueous solvent system) has been improved by increasing the solvent composition in 20 steps (1 to 10 and 11 to 20). Using this new system, a graphic determination of two-phase systems can be done in two steps, i.e., first the sample is partitioned in system 10. If the K value is over 1, the second K determination is performed at system 1 whereas if the K value in system 1 is below unity, the second K determination is performed at system 20. By drawing the line between these two points, the optimum K value of near unity can be easily obtained from the crossing point with the K = 1 line in each case. The method was successfully applied to separation of tartrazine (K=0.77), tryptophan (K=1.00), methyl green (K= 0.93), tyrosine (0.81), metanephrine (K=0.89), tyramine (K=0.98), and normetanephrine (K=0.96). Three sulfonic acid components in D&C Green No. 8 were separated by HSCCC using the graphic selection of the two-phase solvent system This new method was also applied for separation of ultra polar samples polar compounds including nucleobases, nucleosides and nucleotides.

现有的用于高速逆流色谱的两相溶剂系统涵盖了疏水性向中度极性溶剂系统的分离，但通常无法为高极性化合物（如硫磺染料，儿茶酚胺和Twitter离子）提供合适的分区系数值。本文引入了一个新的溶剂系列，可以通过螺旋管逆电色谱法将这些极性化合物分离。它由1-丁醇，乙醇，硫酸铵和水组成，以各种体积比。该系统由10个步骤组成，这些步骤根据溶剂系统的极性排列，因此可以在几个搜索步骤中找到具有合适k值的两相溶剂系统。每个溶剂系统都给出了两个阶段之间适当的体积比和高密度差，并在螺旋柱组件中提供了令人满意的固定相的保留水平。该方法通过分区系数测量的四种典型极性化合物进行验证，包括甲基绿色（碱性染料），塔尔塔嗪（硫磺染料），酪氨酸（Twitter离子）和肾上腺素（Catecholamine），所有这些（catecholamine）在极性1-二硫代水系统中均显示出低分配系数。最近，通过以20个步骤（1至10和11至20）增加溶剂成分，可以改善上述超极性溶剂系统（有机/高离子强度水性系统）。 Using this new system, a graphic determination of two-phase systems can be done in two steps, i.e., first the sample is partitioned in system 10. If the K value is over 1, the second K determination is performed at system 1 whereas if the K value in system 1 is below unity, the second K determination is performed at system 20. By drawing the line between these two points, the optimum K value of near unity can be easily obtained from the crossing point with the K = 1 line在每种情况下。该方法成功地应用于tarrazine（K = 0.77），色氨酸（K = 1.00），甲基绿色（K = 0.93），酪氨酸（0.81），Metanephrine（K = 0.89），酪氨酰胺（K = 0.98）和Normetanephrine（K = 0.96）。 D＆C绿色8号中的三个磺酸成分通过HSCCC分离，使用两相溶剂系统的图形选择该新方法也用于分离超极性样品极性化合物，包括核苷酸酶，核苷和核苷酸。