Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and Sweeping
In this study, we established a novel capillary electrophoresis method for monitoring the concentration of doripenem in human plasma. As a time-dependent antibiotic, doripenem maximizes its antibacterial effects and minimizes the potential for antibiotic resistance through careful therapeutic drug m...
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MDPI AG
2023-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/18/13751 |
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| author | Hsin-Hua Liang Yu-Chao Lin Chin-Chuan Hung Yu-Chi Hou Yi-Hui Lin |
| author_facet | Hsin-Hua Liang Yu-Chao Lin Chin-Chuan Hung Yu-Chi Hou Yi-Hui Lin |
| author_sort | Hsin-Hua Liang |
| collection | DOAJ |
| description | In this study, we established a novel capillary electrophoresis method for monitoring the concentration of doripenem in human plasma. As a time-dependent antibiotic, doripenem maximizes its antibacterial effects and minimizes the potential for antibiotic resistance through careful therapeutic drug monitoring. Two online preconcentration techniques, field-enhanced sample stacking (FESS) and sweeping, were coupled to enhance the detection sensitivity. Briefly, an uncoated fused silica capillary (40 cm × 50 μm i.d) was rinsed with a high conductivity buffer (HCB) composed of 150 mM phosphate buffer (NaH<sub>2</sub>PO<sub>4</sub>, pH 2.5) and 20% methanol. A large sample plug prepared in a low-conductivity phosphate buffer (50 mM NaH<sub>2</sub>PO<sub>4</sub>, pH 2.5) was then hydrodynamically injected (5 psi, 80 s) into the capillary. Under an applied voltage of −30 kV, the analyte was accumulated at the FESS boundary and swept by the negatively charged micelles toward the UV detector. Plasma samples were pretreated by solid-phase extraction (SPE) to eliminate endogenous interferences. The validation results demonstrated a high coefficient of determination (r<sup>2</sup> > 0.9995) for the regression curve with impressive precision and accuracy: relative standard deviation (RSD) <5.86% and relative error <4.63%. The limit of detection (LOD, S/N = 3) for doripenem was determined to be 0.4 μg/mL. Compared to the conventional micellar electrokinetic chromatography method, our developed method achieved a sensitivity enhancement of up to 488-fold for doripenem. Furthermore, the newly developed method successfully quantified doripenem concentrations in plasma samples obtained from patients accepting doripenem regimens, proving its application potential in the clinical realm. |
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| language | English |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-939d06491f334db5a82ff38b7652c12c2023-11-19T11:02:38ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-09-0124181375110.3390/ijms241813751Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and SweepingHsin-Hua Liang0Yu-Chao Lin1Chin-Chuan Hung2Yu-Chi Hou3Yi-Hui Lin4School of Pharmacy, China Medical University, Taichung 406040, TaiwanDivision of Pulmonary and Critical Care Medicine, Department of Internal Medicine, China Medical University Hospital, Taichung 404332, TaiwanSchool of Pharmacy, China Medical University, Taichung 406040, TaiwanSchool of Pharmacy, China Medical University, Taichung 406040, TaiwanSchool of Pharmacy, China Medical University, Taichung 406040, TaiwanIn this study, we established a novel capillary electrophoresis method for monitoring the concentration of doripenem in human plasma. As a time-dependent antibiotic, doripenem maximizes its antibacterial effects and minimizes the potential for antibiotic resistance through careful therapeutic drug monitoring. Two online preconcentration techniques, field-enhanced sample stacking (FESS) and sweeping, were coupled to enhance the detection sensitivity. Briefly, an uncoated fused silica capillary (40 cm × 50 μm i.d) was rinsed with a high conductivity buffer (HCB) composed of 150 mM phosphate buffer (NaH<sub>2</sub>PO<sub>4</sub>, pH 2.5) and 20% methanol. A large sample plug prepared in a low-conductivity phosphate buffer (50 mM NaH<sub>2</sub>PO<sub>4</sub>, pH 2.5) was then hydrodynamically injected (5 psi, 80 s) into the capillary. Under an applied voltage of −30 kV, the analyte was accumulated at the FESS boundary and swept by the negatively charged micelles toward the UV detector. Plasma samples were pretreated by solid-phase extraction (SPE) to eliminate endogenous interferences. The validation results demonstrated a high coefficient of determination (r<sup>2</sup> > 0.9995) for the regression curve with impressive precision and accuracy: relative standard deviation (RSD) <5.86% and relative error <4.63%. The limit of detection (LOD, S/N = 3) for doripenem was determined to be 0.4 μg/mL. Compared to the conventional micellar electrokinetic chromatography method, our developed method achieved a sensitivity enhancement of up to 488-fold for doripenem. Furthermore, the newly developed method successfully quantified doripenem concentrations in plasma samples obtained from patients accepting doripenem regimens, proving its application potential in the clinical realm.https://www.mdpi.com/1422-0067/24/18/13751carbapenemsdoripenemFESS-sweeping MEKChuman plasmaonline preconcentration |
| spellingShingle | Hsin-Hua Liang Yu-Chao Lin Chin-Chuan Hung Yu-Chi Hou Yi-Hui Lin Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and Sweeping International Journal of Molecular Sciences carbapenems doripenem FESS-sweeping MEKC human plasma online preconcentration |
| title | Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and Sweeping |
| title_full | Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and Sweeping |
| title_fullStr | Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and Sweeping |
| title_full_unstemmed | Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and Sweeping |
| title_short | Method Development for Determination of Doripenem in Human Plasma via Capillary Electrophoresis Coupled with Field-Enhanced Sample Stacking and Sweeping |
| title_sort | method development for determination of doripenem in human plasma via capillary electrophoresis coupled with field enhanced sample stacking and sweeping |
| topic | carbapenems doripenem FESS-sweeping MEKC human plasma online preconcentration |
| url | https://www.mdpi.com/1422-0067/24/18/13751 |
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