Quantitation of Humalog Insulin by Reversed-Phase High-Performance Liquid Chromatography
| Autor: | Tom Dziubla, Robert J. Murray, Marc C. Torjman, Anthony M. Lowman, Jeffrey I. Joseph, Carey Munsick |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
medicine.medical_specialty
Serial dilution Endocrinology Diabetes and Metabolism Coefficient of variation medicine.medical_treatment Biomedical Engineering 030209 endocrinology & metabolism Bioengineering High-performance liquid chromatography 03 medical and health sciences 0302 clinical medicine Internal medicine Technology Reports Internal Medicine medicine Insulin lispro 030212 general & internal medicine Detection limit Chromatography business.industry Insulin Endocrinology business Nonlinear regression Stock solution medicine.drug |
| Zdroj: | Journal of Diabetes Science and Technology. 1:603-607 |
| ISSN: | 1932-2968 |
| DOI: | 10.1177/193229680700100420 |
| Popis: | INTRODUCTION The methodology for in vitro testing of insulin delivery systems for long-term infusion of lispro insulin requires insulin flow studies over time, with the measurement of lispro concentrations in wells or other laboratory fluid collection systems. We postulated that the efficiency of the insulin assay could be improved if the insulin collected in the wells could be measured without having to perform dilutions of insulin samples. The manufacturer's method for the identification of Humalog(R) insulin by high-performance liquid chromatography (HPLC) does not provide a detailed method for the quantitation of the clinical formulation. No reports could be found in the literature describing a method to quantitate lispro insulin's entire concentration range, nor could we find detailed information on pH effects. The purpose of this study was to investigate the use of a reversed-phase HPLC method to quantitate Humalog insulin at three levels of pH. METHODS Serial dilutions of Humalog insulin stock solution were prepared at pH levels of 2.6 and 7.4 (concentrations 0.20-100 IU/ml) to construct calibration curves. Samples were also prepared at lower concentrations (0.23-15.0 IU/ml) at pH levels of 4.0 and 7.4. Areas under curve data were plotted against known concentrations, and the limit of quantitation and precision of the assay were determined. RESULTS Calibration curves for the 0.20-100 IU/ml concentration (pH 7.4 and 2.6) revealed no significant differences (p < 0.01) between those curves. pH and concentration did not have an effect on the overall precision of the method. Linearity was not conserved past the 15-IU/ml concentration. Coefficient of variation (CV) values were generally |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|