| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1212125 | Journal of Chromatography B | 2015 | 9 Pages |
•Automated on-line SPE-POPLC method for determining IOM was developed.•More than 400 plasma samples could be analyzed with one on-line SPE column.•The validated method is likely to be a standard for assessing kidney function impairment.
Phase optimized liquid chromatography (POPLC) allows for the optimized combination of column segments of any length and stationary phases with different functionalities. In this study, a simple and rapid method using POPLC coupled with on-line solid-phase extraction (SPE) for the analysis of X-ray contrast media agent iomeprol (IOM) in human plasma was developed. Because the phenyl (PH) stationary phase has strong hydrophobic and π–π interactions with IOM and iopromide (IOP, internal standard), the best separation efficiency was achieved with a 250 mm × 3 mm homogenous PH POPLC-column. Different kinds of on-line SPE sorbents were studied, including restricted access material-alkyl diol silica (ADS), LiChrolut EN with excellent absorption capacity and hydrophilic–lipophilic-balanced Oasis HLB. The most efficient on-line sample clean-up was carried out using a fast-flow on-line purification approach with an Oasis HLB pre-column ((20 mm × 2 mm, 30 μm). This pre-column showed excellent durability and reproducibility. At least 400 samples could be analyzed with one pre-column. Each plasma sample was directly injected and analyzed within 15 min. The calibration curves were linear in the range of 10–1000 μg/mL. The limit of quantitation was 2.26 μg/mL. The inter-day precision of this method was excellent and less than 1.44%, and the intra-day precision was less than 4.44%. The inter-day and intra-day accuracy ranged from 94.33% to 104.36% and 94.60% to 101.71%, respectively. This validated method is expected to be useful in the analysis of human plasma samples for glomerular filtration rate (GFR) measurements and assessment of kidney function.
