Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies
MPI8, a peptidyl aldehyde, is a potent antiviral agent against coronavirus. Due to unique tri-peptide bonds and the formyl functional group, the bioassay of MPI8 in plasma was challenged by a strong interference from water MPI8. Using QTOF LC-MS/MS, we identified MPI8•H<sub>2</sub>O as t...
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MDPI AG
2022-05-01
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| Online Access: | https://www.mdpi.com/1424-8247/15/6/676 |
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| author | Yang Wang Huan Xie Yugendar R. Alugubelli Yuying Ma Shiqing Xu Jing Ma Wenshe R. Liu Dong Liang |
| author_facet | Yang Wang Huan Xie Yugendar R. Alugubelli Yuying Ma Shiqing Xu Jing Ma Wenshe R. Liu Dong Liang |
| author_sort | Yang Wang |
| collection | DOAJ |
| description | MPI8, a peptidyl aldehyde, is a potent antiviral agent against coronavirus. Due to unique tri-peptide bonds and the formyl functional group, the bioassay of MPI8 in plasma was challenged by a strong interference from water MPI8. Using QTOF LC-MS/MS, we identified MPI8•H<sub>2</sub>O as the major interference form that co-existed with MPI8 in aqueous and biological media. To avoid the resolution of MPI8 and MPI8•H<sub>2</sub>O observed on reverse phase columns, we found that a Kinetex hydrophilic interaction liquid chromatography (HILIC) column provided co-elution of both MPI8 and MPI8•H<sub>2</sub>O with a good single chromatographic peak and column retention of MPI8 which is suitable for quantification. Thus, a sensitive, specific, and reproducible LC-MS/MS method for the quantification of MPI8 in rat plasma was developed and validated using a triple QUAD LC-MS/MS. The chromatographic separation was achieved on a Kinetex HILIC column with a flow rate of 0.4 mL/min under gradient elution. The calibration curves were linear (r<sup>2</sup> > 0.99) over MPI8 concentrations from 0.5–500 ng/mL. The accuracy and precision are within acceptable guidance levels. The mean matrix effect and recovery were 139% and 73%, respectively. No significant degradation of MPI8 occurred under the experimental conditions. The method was successfully applied to a pharmacokinetic study of MPI8 after administration of MPI8 sulfonate in rats. |
| first_indexed | 2024-03-09T22:47:35Z |
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| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-09T22:47:35Z |
| publishDate | 2022-05-01 |
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| series | Pharmaceuticals |
| spelling | doaj.art-9cafdd3cf5144515b12e0359ebad12f72023-11-23T18:26:50ZengMDPI AGPharmaceuticals1424-82472022-05-0115667610.3390/ph15060676Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic StudiesYang Wang0Huan Xie1Yugendar R. Alugubelli2Yuying Ma3Shiqing Xu4Jing Ma5Wenshe R. Liu6Dong Liang7Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004, USADepartment of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004, USADepartment of Chemistry, Texas A&M University, College Station, TX 77843, USADepartment of Chemistry, Texas A&M University, College Station, TX 77843, USADepartment of Chemistry, Texas A&M University, College Station, TX 77843, USADepartment of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004, USADepartment of Chemistry, Texas A&M University, College Station, TX 77843, USADepartment of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004, USAMPI8, a peptidyl aldehyde, is a potent antiviral agent against coronavirus. Due to unique tri-peptide bonds and the formyl functional group, the bioassay of MPI8 in plasma was challenged by a strong interference from water MPI8. Using QTOF LC-MS/MS, we identified MPI8•H<sub>2</sub>O as the major interference form that co-existed with MPI8 in aqueous and biological media. To avoid the resolution of MPI8 and MPI8•H<sub>2</sub>O observed on reverse phase columns, we found that a Kinetex hydrophilic interaction liquid chromatography (HILIC) column provided co-elution of both MPI8 and MPI8•H<sub>2</sub>O with a good single chromatographic peak and column retention of MPI8 which is suitable for quantification. Thus, a sensitive, specific, and reproducible LC-MS/MS method for the quantification of MPI8 in rat plasma was developed and validated using a triple QUAD LC-MS/MS. The chromatographic separation was achieved on a Kinetex HILIC column with a flow rate of 0.4 mL/min under gradient elution. The calibration curves were linear (r<sup>2</sup> > 0.99) over MPI8 concentrations from 0.5–500 ng/mL. The accuracy and precision are within acceptable guidance levels. The mean matrix effect and recovery were 139% and 73%, respectively. No significant degradation of MPI8 occurred under the experimental conditions. The method was successfully applied to a pharmacokinetic study of MPI8 after administration of MPI8 sulfonate in rats.https://www.mdpi.com/1424-8247/15/6/676MPI8MPI8•H<sub>2</sub>O adductSARS-CoV-2LC-MS/MSmethod development and validationpharmacokinetics |
| spellingShingle | Yang Wang Huan Xie Yugendar R. Alugubelli Yuying Ma Shiqing Xu Jing Ma Wenshe R. Liu Dong Liang Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies Pharmaceuticals MPI8 MPI8•H<sub>2</sub>O adduct SARS-CoV-2 LC-MS/MS method development and validation pharmacokinetics |
| title | Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies |
| title_full | Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies |
| title_fullStr | Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies |
| title_full_unstemmed | Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies |
| title_short | Accurate Mass Identification of an Interfering Water Adduct and Strategies in Development and Validation of an LC-MS/MS Method for Quantification of MPI8, a Potent SARS-CoV-2 Main Protease Inhibitor, in Rat Plasma in Pharmacokinetic Studies |
| title_sort | accurate mass identification of an interfering water adduct and strategies in development and validation of an lc ms ms method for quantification of mpi8 a potent sars cov 2 main protease inhibitor in rat plasma in pharmacokinetic studies |
| topic | MPI8 MPI8•H<sub>2</sub>O adduct SARS-CoV-2 LC-MS/MS method development and validation pharmacokinetics |
| url | https://www.mdpi.com/1424-8247/15/6/676 |
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