Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-Lactams
<b>Background/Introduction:</b> Beta-lactam antibiotics are among the most frequently prescribed drugs in clinical practice, yet their therapeutic drug monitoring remains underutilized despite high interindividual pharmacokinetic variability, especially in critically ill patients. <b&...
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MDPI AG
2025-06-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/14/6/613 |
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| author | Iria Varela-Rey Marta Martínez-Guitián Gonzalo Hermelo-Vidal Enrique Bandín-Vilar Ignacio Novo-Veleiro Pablo Manuel Varela-García Irene Zarra-Ferro Miguel González-Barcia Cristina Mondelo-García Anxo Fernández-Ferreiro |
| author_facet | Iria Varela-Rey Marta Martínez-Guitián Gonzalo Hermelo-Vidal Enrique Bandín-Vilar Ignacio Novo-Veleiro Pablo Manuel Varela-García Irene Zarra-Ferro Miguel González-Barcia Cristina Mondelo-García Anxo Fernández-Ferreiro |
| author_sort | Iria Varela-Rey |
| collection | DOAJ |
| description | <b>Background/Introduction:</b> Beta-lactam antibiotics are among the most frequently prescribed drugs in clinical practice, yet their therapeutic drug monitoring remains underutilized despite high interindividual pharmacokinetic variability, especially in critically ill patients. <b>Methods:</b> To address this, we developed and validated an ultra-high-performance liquid chromatography (UHPLC-UV/Vis) method for the simultaneous quantification of six beta-lactams (cefepime, ceftolozane, ceftazidime, meropenem, ampicillin, and ertapenem) in plasma. <b>Results:</b> This method uses a single gradient mobile phase and a photodiode array detector, ensuring accurate separation, minimal interference, and robust analyte identification. Validation followed EMA bioanalytical guidelines, demonstrating selectivity, precision, accuracy, and linearity within clinically relevant ranges (1.0–50.0 mg/L). Stability tests showed that the analytes were stable in plasma for up to seven days at 4 °C and one month at −20 °C. Pilot clinical implementation in 35 patients revealed significant interindividual variability, supporting the need for routine beta-lactam monitoring. Approximately 26% of trough concentrations were below the minimal inhibitory concentration, while others exceeded thresholds associated with potential toxicity. <b>Discussion</b>: This study represents the first UHPLC-UV/Vis method for the simultaneous determination of these six beta-lactams, overcoming limitations of prior methods that required different mobile phases or excluded clinically relevant antibiotics. The method is universally applicable and easily transferable to routine clinical practice. <b>Conclusions:</b> These findings underline the importance of beta-lactam monitoring in optimizing treatment outcomes and combating antibiotic resistance in vulnerable populations. Further studies to assess free drug concentrations are warranted to enhance clinical applicability. |
| format | Article |
| id | doaj-art-e577d56fbab24104ab495596073a1f96 |
| institution | Kabale University |
| issn | 2079-6382 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj-art-e577d56fbab24104ab495596073a1f962025-08-20T03:26:57ZengMDPI AGAntibiotics2079-63822025-06-0114661310.3390/antibiotics14060613Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-LactamsIria Varela-Rey0Marta Martínez-Guitián1Gonzalo Hermelo-Vidal2Enrique Bandín-Vilar3Ignacio Novo-Veleiro4Pablo Manuel Varela-García5Irene Zarra-Ferro6Miguel González-Barcia7Cristina Mondelo-García8Anxo Fernández-Ferreiro9FarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, SpainFarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, SpainFarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, SpainFarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, SpainHome Hospitalization Unit, University Clinical Hospital of Santiago de Compostela (SERGAS), 15706 Santiago de Compostela, SpainInternal Medicine Department, University Clinical Hospital of Santiago de Compostela (SERGAS), 15706 Santiago de Compostela, SpainFarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, SpainFarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, SpainFarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, SpainFarmaCHUSLab, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, Spain<b>Background/Introduction:</b> Beta-lactam antibiotics are among the most frequently prescribed drugs in clinical practice, yet their therapeutic drug monitoring remains underutilized despite high interindividual pharmacokinetic variability, especially in critically ill patients. <b>Methods:</b> To address this, we developed and validated an ultra-high-performance liquid chromatography (UHPLC-UV/Vis) method for the simultaneous quantification of six beta-lactams (cefepime, ceftolozane, ceftazidime, meropenem, ampicillin, and ertapenem) in plasma. <b>Results:</b> This method uses a single gradient mobile phase and a photodiode array detector, ensuring accurate separation, minimal interference, and robust analyte identification. Validation followed EMA bioanalytical guidelines, demonstrating selectivity, precision, accuracy, and linearity within clinically relevant ranges (1.0–50.0 mg/L). Stability tests showed that the analytes were stable in plasma for up to seven days at 4 °C and one month at −20 °C. Pilot clinical implementation in 35 patients revealed significant interindividual variability, supporting the need for routine beta-lactam monitoring. Approximately 26% of trough concentrations were below the minimal inhibitory concentration, while others exceeded thresholds associated with potential toxicity. <b>Discussion</b>: This study represents the first UHPLC-UV/Vis method for the simultaneous determination of these six beta-lactams, overcoming limitations of prior methods that required different mobile phases or excluded clinically relevant antibiotics. The method is universally applicable and easily transferable to routine clinical practice. <b>Conclusions:</b> These findings underline the importance of beta-lactam monitoring in optimizing treatment outcomes and combating antibiotic resistance in vulnerable populations. Further studies to assess free drug concentrations are warranted to enhance clinical applicability.https://www.mdpi.com/2079-6382/14/6/613beta-lactamUHPLC-UV/Vistherapeutic monitoringsimultaneous quantification |
| spellingShingle | Iria Varela-Rey Marta Martínez-Guitián Gonzalo Hermelo-Vidal Enrique Bandín-Vilar Ignacio Novo-Veleiro Pablo Manuel Varela-García Irene Zarra-Ferro Miguel González-Barcia Cristina Mondelo-García Anxo Fernández-Ferreiro Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-Lactams Antibiotics beta-lactam UHPLC-UV/Vis therapeutic monitoring simultaneous quantification |
| title | Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-Lactams |
| title_full | Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-Lactams |
| title_fullStr | Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-Lactams |
| title_full_unstemmed | Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-Lactams |
| title_short | Development of a UHPLC-UV/Vis Method for Simultaneously Determining Six Beta-Lactam Antibiotics in Plasma: A Tool for the Clinical Implementation of Therapeutic Monitoring of Beta-Lactams |
| title_sort | development of a uhplc uv vis method for simultaneously determining six beta lactam antibiotics in plasma a tool for the clinical implementation of therapeutic monitoring of beta lactams |
| topic | beta-lactam UHPLC-UV/Vis therapeutic monitoring simultaneous quantification |
| url | https://www.mdpi.com/2079-6382/14/6/613 |
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