A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots
The role of therapeutic drug monitoring (TDM) of valaciclovir (VA)/aciclovir (A) and valganciclovir/ganciclovir (VG/G) in critically ill patients is still a matter of debate. More data on the dose–concentration relationship might therefore be useful, especially in pediatrics where clinical practice...
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MDPI AG
2021-10-01
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| Online Access: | https://www.mdpi.com/2227-9059/9/10/1379 |
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| author | Federica Pigliasco Alessia Cafaro Raffaele Simeoli Sebastiano Barco Alberto Magnasco Maura Faraci Gino Tripodi Bianca Maria Goffredo Giuliana Cangemi |
| author_facet | Federica Pigliasco Alessia Cafaro Raffaele Simeoli Sebastiano Barco Alberto Magnasco Maura Faraci Gino Tripodi Bianca Maria Goffredo Giuliana Cangemi |
| author_sort | Federica Pigliasco |
| collection | DOAJ |
| description | The role of therapeutic drug monitoring (TDM) of valaciclovir (VA)/aciclovir (A) and valganciclovir/ganciclovir (VG/G) in critically ill patients is still a matter of debate. More data on the dose–concentration relationship might therefore be useful, especially in pediatrics where clinical practice is not adequately supported by robust PK studies. We developed and validated a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) micro-method to simultaneously quantify A and G from plasma and dried plasma spots (DPS). The method was based on rapid organic extraction from DPS and separation on a reversed-phase C-18 UHPLC column after addition of deuterated internal standards. Accurate analyte quantification using SRM detection was then obtained using a Thermo Fisher Quantiva triple-quadrupole MS coupled to an Ultimate 3000 UHPLC. It was validated following international (EMA) guidelines for bioanalytical method validation and was tested on samples from pediatric patients treated with A, VG, or G for cytomegalovirus infection following solid organ or hematopoietic stem cell transplantation. Concentrations obtained from plasma and DPS were compared using Passing–Bablok and Bland–Altman statistical tests. The assay was linear over wide concentration ranges (0.01–20 mg/L) in both plasma and DPS for A and G, suitable for the expected therapeutic ranges for both Cmin and Cmax, accurate, and reproducible in the absence of matrix effects. The results obtained from plasma and DPS were comparable. Using an LC-MS/MS method allowed us to obtain a very specific, sensitive, and rapid quantification of these antiviral drugs starting from very low volumes (50 μL) of plasma samples and DPS. The stability of analytes for at least 30 days allows for cost-effective shipment and storage at room temperature. Our method is suitable for TDM and could be helpful for improving knowledge on PK/PD targets of antivirals in critically ill pediatric patients. |
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| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-10T06:42:05Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
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| series | Biomedicines |
| spelling | doaj.art-0ff16d830e7e4b1eb1dc97051444d7872023-11-22T17:30:58ZengMDPI AGBiomedicines2227-90592021-10-01910137910.3390/biomedicines9101379A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma SpotsFederica Pigliasco0Alessia Cafaro1Raffaele Simeoli2Sebastiano Barco3Alberto Magnasco4Maura Faraci5Gino Tripodi6Bianca Maria Goffredo7Giuliana Cangemi8Chromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyChromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyDepartment of Pediatric Specialties, Division of Metabolic Biochemistry, Children’s Hospital Bambino Gesù, IRCCS, 00165 Rome, ItalyChromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyPediatric Nephrology, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyHematopoietic Stem Cell Unit, Department of Pediatric Hematology and Oncology, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyChromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyDepartment of Pediatric Specialties, Division of Metabolic Biochemistry, Children’s Hospital Bambino Gesù, IRCCS, 00165 Rome, ItalyChromatography and Mass Spectrometry Section, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyThe role of therapeutic drug monitoring (TDM) of valaciclovir (VA)/aciclovir (A) and valganciclovir/ganciclovir (VG/G) in critically ill patients is still a matter of debate. More data on the dose–concentration relationship might therefore be useful, especially in pediatrics where clinical practice is not adequately supported by robust PK studies. We developed and validated a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) micro-method to simultaneously quantify A and G from plasma and dried plasma spots (DPS). The method was based on rapid organic extraction from DPS and separation on a reversed-phase C-18 UHPLC column after addition of deuterated internal standards. Accurate analyte quantification using SRM detection was then obtained using a Thermo Fisher Quantiva triple-quadrupole MS coupled to an Ultimate 3000 UHPLC. It was validated following international (EMA) guidelines for bioanalytical method validation and was tested on samples from pediatric patients treated with A, VG, or G for cytomegalovirus infection following solid organ or hematopoietic stem cell transplantation. Concentrations obtained from plasma and DPS were compared using Passing–Bablok and Bland–Altman statistical tests. The assay was linear over wide concentration ranges (0.01–20 mg/L) in both plasma and DPS for A and G, suitable for the expected therapeutic ranges for both Cmin and Cmax, accurate, and reproducible in the absence of matrix effects. The results obtained from plasma and DPS were comparable. Using an LC-MS/MS method allowed us to obtain a very specific, sensitive, and rapid quantification of these antiviral drugs starting from very low volumes (50 μL) of plasma samples and DPS. The stability of analytes for at least 30 days allows for cost-effective shipment and storage at room temperature. Our method is suitable for TDM and could be helpful for improving knowledge on PK/PD targets of antivirals in critically ill pediatric patients.https://www.mdpi.com/2227-9059/9/10/1379aciclovirganciclovirvalganciclovirvalaciclovirtherapeutic drug monitoringLC-MS/MS |
| spellingShingle | Federica Pigliasco Alessia Cafaro Raffaele Simeoli Sebastiano Barco Alberto Magnasco Maura Faraci Gino Tripodi Bianca Maria Goffredo Giuliana Cangemi A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots Biomedicines aciclovir ganciclovir valganciclovir valaciclovir therapeutic drug monitoring LC-MS/MS |
| title | A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots |
| title_full | A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots |
| title_fullStr | A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots |
| title_full_unstemmed | A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots |
| title_short | A UHPLC–MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots |
| title_sort | uhplc ms ms method for therapeutic drug monitoring of aciclovir and ganciclovir in plasma and dried plasma spots |
| topic | aciclovir ganciclovir valganciclovir valaciclovir therapeutic drug monitoring LC-MS/MS |
| url | https://www.mdpi.com/2227-9059/9/10/1379 |
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