Mansour Fotouh R, Kirkpatrick Christine L, Danielson Neil D
Department of Chemistry and Biochemistry, Miami University, Oxford, OH, 45056.
Chromatographia. 2013 Jun;76(11-12). doi: 10.1007/s10337-013-2461-3.
An ion exclusion chromatography (IELC) comparison between a conventional ion exchange column and an ultra-high performance liquid chromatography (UHPLC) dynamically surfactant modified C18 column for the separation of an aliphatic carboxylic acid and two aromatic carboxylic acids is presented. Professional software is used to optimize the conventional IELC separation conditions for acetylsalicylic acid and the hydrolysis products: salicylic acid and acetic acid. Four different variables are simultaneously optimized including HSO concentration, pH, flow rate, and sample injection volume. Thirty different runs are suggested by the software. The resolutions and the time of each run are calculated and feed back to the software to predict the optimum conditions. Derringer's desirability functions are used to evaluate the test conditions and those with the highest desirability value are utilized to separate acetylsalicylic acid, salicylic acid, and acetic acid. These conditions include using a 0.35 m HSO (pH 3.93) eluent at a flow rate of 1 mL min and an injection volume of 72 μL. To decrease the run time and improve the performance, a UHPLC C18 column is used after dynamic modification with sodium dodecyl sulfate. Using pure water as a mobile phase, a shorter analysis time and better resolution are achieved. In addition, the elution order is different from the IELC method which indicates the contribution of the reversed-phase mode to the separation mechanism.
本文介绍了使用常规离子交换柱和超高效液相色谱(UHPLC)动态表面活性剂改性C18柱进行离子排斥色谱(IELC)比较,以分离脂肪族羧酸和两种芳香族羧酸。使用专业软件优化乙酰水杨酸及其水解产物水杨酸和乙酸的常规IELC分离条件。同时优化四个不同变量,包括硫酸氢根浓度、pH值、流速和进样体积。软件建议进行30次不同的运行。计算每次运行的分离度和时间,并反馈给软件以预测最佳条件。使用Derringer合意函数评估测试条件,并使用合意值最高的条件分离乙酰水杨酸、水杨酸和乙酸。这些条件包括使用0.35 m硫酸氢根(pH 3.93)洗脱液,流速为1 mL/min,进样体积为72 μL。为了缩短运行时间并提高性能,在使用十二烷基硫酸钠进行动态改性后,使用UHPLC C18柱。以纯水作为流动相,实现了更短的分析时间和更好的分离度。此外,洗脱顺序与IELC方法不同,这表明反相模式对分离机制有贡献。