Peris-Díaz Manuel David, Alcoriza-Balaguer Maria Isabel, García-Cañaveras Juan Carlos, Santonja Francisco, Sentandreu Enrique, Lahoz Agustín
Biomarkers and Precision Medicine Unit, Analytical Unit, Instituto de Investigación Sanitaria, Fundación Hospital La Fe, Valencia, Spain.
Departamento de Estadística e Investigación Operativa, Facultat de Ciencias Matematicas, Universidad de Valencia, Burjassot, Spain.
Electrophoresis. 2017 Nov;38(22-23):2985-2995. doi: 10.1002/elps.201700180. Epub 2017 Jul 26.
Characterization of chromatographic columns using the traditional van Deemter method is limited by the necessity of calculating extra-column variance, issue particularly relevant when modeling asymmetrical peaks eluted from monolithic columns. A novel R package that implements Parabolic Variance Modified Gaussian approach for accurate peak modeling, van Deemter equation and two alternatives approaches, based on van Deemter, has been developed to calculate the height equivalent to a theoretical plate (HETP). To assess package capabilities conventional packed reverse-phase and monolithic HPLC columns were characterized. Peaks eluted from the monolithic column showed a high value of factor asymmetry due, in part, to the contribution of extra-column factors. Such deviation can be circumvented by the two alternatives approaches implemented in the R-package. Furthermore, increased values of eddy diffusion and mass transfer kinetics terms in HETP were observed for the packed column, while accuracy was below 9% in all cases. These results showed the usefulness of the R-package for both modeling chromatographic peaks and assessing column efficiency. The RpeakChrom package could become a helpful tool for testing new stationary phases during column development and to evaluate column during its lifetime. This R tool is freely available from CRAN (https://CRAN.R-project.org/package=RpeakChrom).
使用传统范德姆特方法对色谱柱进行表征受到计算柱外方差必要性的限制，在对整体柱上洗脱的不对称峰进行建模时，这个问题尤为突出。已开发出一种新颖的R包，它基于范德姆特方法实现了抛物线方差修正高斯方法用于精确峰建模、范德姆特方程以及两种替代方法，以计算理论塔板高度（HETP）。为了评估该包的性能，对传统填充反相和整体式高效液相色谱柱进行了表征。从整体柱上洗脱的峰显示出较高的不对称因子值，部分原因是柱外因素的影响。R包中实现的两种替代方法可以规避这种偏差。此外，观察到填充柱的HETP中涡流扩散和传质动力学项的值增加，而在所有情况下精度均低于9%。这些结果表明该R包对于色谱峰建模和评估柱效均有用。RpeakChrom包可能成为在柱开发过程中测试新固定相以及在柱的使用期内评估柱的有用工具。这个R工具可从CRAN（https://CRAN.R-project.org/package=RpeakChrom）免费获取。
