HPLC Analysis of Pancuronium Bromide on Reverese-Phase and Mixed-Mode Columns
反相柱和混模柱上潘库溴铵的HPLC分析
Mixed-mode chromatography usually adds secondary and tertiary interactions which help to achieve controlled retention, unique selectivity and allow to adjust retention time of various compounds by changing the amount of organic, buffer concentration, buffer pH and buffer nature(混合模式色谱法通常增添二重和三重相互作用,有助于实现可控的保留,独特的选择性,并通过改变有机溶剂的含量,缓冲浓度,缓冲pH值和缓冲盐的性质来调整各种化合物的保留时间。 ). Presence of ionizable groups on the column also helps improve peak shape for various analytes, which sometimes is impossible to achieve with single mode columns.(色谱柱上存在可电离基团也有助于改善各种分析物的峰形,一些时候,这在单一模式色谱柱上是不可能实现的) Pancuronium bromide is a hydrophobic compound with two strong basic groups. It shows a poor peak shape when RP columns are used(泮库溴铵是一种具有两个强碱性基团的疏水化合物。当采用RP柱时,峰形较差。). The poor peak shape is attributed to secondary interaction between analyte and silanols on the surface of RP columns (分析物与在RP柱表面的硅烷醇二次相互作用导致峰形较差) (Fig. 1). Sometimes you need to understand what secondary interaction to add to improve your chromatography(有时,你需要了解增加什么样的二次相互作用来改善你的色谱方法). In case of pancuronium bromide, addition of cation-exchange mechanism did not work and target compound never eluted, (Fig. 2) even with very strong mobile phase from the column due to three strong interactions (reversed-phase and two cation-exchange interactions)(在泮库溴铵的例子下,添加阳离子交换机制不起作用,目标化合物也没有洗脱(图2)。即使柱内流动相洗脱能力很强,存在三种强烈的相互作用(反相作用和两种阳离子交换作用),化合物也没有得到保留.)Problem of elution and peak shape was addressed by changing cation-exchange interaction to cation-exclusion interaction. This allowed to retain and analyze both the drug and counterion with simple isocratic conditions (Fig. 3)(通过将阳离子交换相互作用转变为阳离子排斥相互作用,解决了洗脱和峰形问题。这样就可以在简单的等度条件下保留和分析药物和反离子).
以上应用所用到的产品可以咨询菲罗门,菲罗门提供色谱柱品牌、型号、价格。