title: Evaluation of a monolithic silica column operated in the hydrophilic interaction chromatography mode with evaporative light scattering detection for the separation and detection of counter-ions.
abstract: In this work a monolithic silica column operated in the hydrophilic interaction chromatography (HILIC) mode in conjunction with an evaporative light scattering detector (ELSD) was investigated. Lithium, sodium and potassium were used as the test counter-ions for this evaluation. Chromatographic properties of this column operated in the HILIC mode were determined by varying key mobile phase parameters, such as pH, flow rate, buffer strength, acid and organic modifier. As organic content was increased from 60 to 90% acetonitrile, retention time increased on average by a factor of seven for the test cations listed above. Buffer concentration and pH were also observed to have an effect, although not as significant as the HILIC effect that was observed by changing organic content. Flow rates up to 5 mL/min were utilized to perform counter-ion separations in less than 3 min. After examining the changes in retention, resolution, and peak shape an optimized method was established and then further evaluated for linearity, reproducibility, and limit of detection (LOD) for sodium. Linearity was acceptable with an R2 value of 0.999 across the working-standard range and a LOD of 0.1 microg/mL was calculated. The reproducibility on the counter-ion determination from pharmaceutical sodium salts was 1.6% R.S.D. on average, and the accuracy of the counter-ion prediction was approximately 3% from theory when salt content was corrected for potency.
authors.affiliation: Eli Lilly and Company, Pharmaceutical Product Research and Development, Lilly Research Laboratories, Indianapolis, IN 46285, USA. packbw@lilly.com
authors.forename: Brian W
authors.initials: BW
authors.lastname: Pack
authors.forename: Donald S
authors.initials: DS
authors.lastname: Risley
mesh_terms: D002021:Buffers; D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D006863:Hydrogen-Ion Concentration; D008027:Light; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D002021:Buffers; D012822:Silicon Dioxide