Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MS
A simple liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI/MS) method has been developed to analyze oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (ox-PAPC) products on the lipoproteins of uremic patients. The native PAPC standard was in vitro oxidized by the F...
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MDPI AG
2022-07-01
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| author | Chiz-Tzung Chang I-Ting Chang Min-Hui Hsia Chun-Cheng Wang Chao-Jung Chen Hsin-Bai Zou Yu-Qing Chen Wen-Chien Chiu Zhi-Ru Lin Mine-Yine Liu |
| author_facet | Chiz-Tzung Chang I-Ting Chang Min-Hui Hsia Chun-Cheng Wang Chao-Jung Chen Hsin-Bai Zou Yu-Qing Chen Wen-Chien Chiu Zhi-Ru Lin Mine-Yine Liu |
| author_sort | Chiz-Tzung Chang |
| collection | DOAJ |
| description | A simple liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI/MS) method has been developed to analyze oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (ox-PAPC) products on the lipoproteins of uremic patients. The native PAPC standard was in vitro oxidized by the Fenton reaction, and the ox-PAPC products were analyzed by LC- ESI/MS. For LC, a C<sub>8</sub> column and a mobile phase (acetonitrile-isopropanol containing 0.1% formic acid (70:30, <i>v</i>/<i>v</i>)) were selected. For ESI/MS, the optimal conditions included sheath gas pressure (10 psi), capillary temperature (270 °C), and injection time (1000 ms). The identification of ox-PAPC products on human lipoproteins was based on the extracted ion chromatograms (EICs) and the ESI-MS spectra of the in vitro oxidation products of PAPC standard. The EICs and ESI-MS spectra showed good repeatability and sensitivity. A total of 21 ox-PAPC products was determined. Linear analysis has been performed for the phospholipid standard, 1, 2-Di-O-hexadecyl-sn-glycero-3-phosphocholine (PC(O-16:0/O-16:0)). The linear range was 5.0–100.0 µg/mL, and the coefficient of determination (R<sup>2</sup>) was 0.989. The concentration limit of detection (LOD) was 1.50 µg/mL, and the concentration limit of quantitation (LOQ) was 4.54 µg/mL. The selected 21 ox-PAPC products have been identified and quantified in very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) of uremic and healthy subjects. Interestingly, the results showed that the levels of 18 products in VLDL, one product in LDL, and 19 products in HDL were significantly higher for uremic patients than healthy controls. This simple LC-ESI/MS method might accelerate the searching for biomarkers of uremia in the future. |
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| spelling | doaj.art-8c65a55581444afb81ce6b208406cc502023-12-03T14:27:57ZengMDPI AGSeparations2297-87392022-07-019819210.3390/separations9080192Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MSChiz-Tzung Chang0I-Ting Chang1Min-Hui Hsia2Chun-Cheng Wang3Chao-Jung Chen4Hsin-Bai Zou5Yu-Qing Chen6Wen-Chien Chiu7Zhi-Ru Lin8Mine-Yine Liu9Department of Medicine, China Medical University Hospital, Taichung 40447, TaiwanDepartment of Chemistry, National Changhua University of Education, Changhua 50058, TaiwanDepartment of Chemistry, National Changhua University of Education, Changhua 50058, TaiwanDepartment of Medicine, China Medical University Hospital, Taichung 40447, TaiwanGraduate Institute of Integrated Medicine, China Medical University, Taichung 40447, TaiwanDepartment of Chemistry, National Changhua University of Education, Changhua 50058, TaiwanDepartment of Chemistry, National Changhua University of Education, Changhua 50058, TaiwanDepartment of Chemistry, National Changhua University of Education, Changhua 50058, TaiwanDepartment of Chemistry, National Changhua University of Education, Changhua 50058, TaiwanDepartment of Chemistry, National Changhua University of Education, Changhua 50058, TaiwanA simple liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI/MS) method has been developed to analyze oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (ox-PAPC) products on the lipoproteins of uremic patients. The native PAPC standard was in vitro oxidized by the Fenton reaction, and the ox-PAPC products were analyzed by LC- ESI/MS. For LC, a C<sub>8</sub> column and a mobile phase (acetonitrile-isopropanol containing 0.1% formic acid (70:30, <i>v</i>/<i>v</i>)) were selected. For ESI/MS, the optimal conditions included sheath gas pressure (10 psi), capillary temperature (270 °C), and injection time (1000 ms). The identification of ox-PAPC products on human lipoproteins was based on the extracted ion chromatograms (EICs) and the ESI-MS spectra of the in vitro oxidation products of PAPC standard. The EICs and ESI-MS spectra showed good repeatability and sensitivity. A total of 21 ox-PAPC products was determined. Linear analysis has been performed for the phospholipid standard, 1, 2-Di-O-hexadecyl-sn-glycero-3-phosphocholine (PC(O-16:0/O-16:0)). The linear range was 5.0–100.0 µg/mL, and the coefficient of determination (R<sup>2</sup>) was 0.989. The concentration limit of detection (LOD) was 1.50 µg/mL, and the concentration limit of quantitation (LOQ) was 4.54 µg/mL. The selected 21 ox-PAPC products have been identified and quantified in very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) of uremic and healthy subjects. Interestingly, the results showed that the levels of 18 products in VLDL, one product in LDL, and 19 products in HDL were significantly higher for uremic patients than healthy controls. This simple LC-ESI/MS method might accelerate the searching for biomarkers of uremia in the future.https://www.mdpi.com/2297-8739/9/8/192oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholinelipoproteinsuremiaLC-ESI/MS |
| spellingShingle | Chiz-Tzung Chang I-Ting Chang Min-Hui Hsia Chun-Cheng Wang Chao-Jung Chen Hsin-Bai Zou Yu-Qing Chen Wen-Chien Chiu Zhi-Ru Lin Mine-Yine Liu Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MS Separations oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine lipoproteins uremia LC-ESI/MS |
| title | Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MS |
| title_full | Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MS |
| title_fullStr | Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MS |
| title_full_unstemmed | Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MS |
| title_short | Analysis of Oxidized 1-Palmitoyl-2-Arachidonoyl-Sn-Glycero-3 Phosphocholine Products in Uremic Patients by LC-ESI/MS |
| title_sort | analysis of oxidized 1 palmitoyl 2 arachidonoyl sn glycero 3 phosphocholine products in uremic patients by lc esi ms |
| topic | oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine lipoproteins uremia LC-ESI/MS |
| url | https://www.mdpi.com/2297-8739/9/8/192 |
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