Desenvolvimento e validação de método para a quantificação de vancomicina em soro por CLAE-DAD e elaboração de protocolo de monitorização terapêutica em UTI neonatal
Therapeutic monitoring of vancomycin concentration can improve the safety and efficiency of the treatment with this antibiotic. High-performance liquid chromatography (HPLC) is considered gold standard for quantification for this drug. However the analysis requires multiple procedures for purificati...
| Autor principal: | Vilas Boas, Rogério Rodrigues |
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| Formato: | Dissertação |
| Idioma: | Português |
| Publicado em: |
Universidade Tecnológica Federal do Paraná
2016
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| Assuntos: | |
| Acesso em linha: |
http://repositorio.utfpr.edu.br/jspui/handle/1/1839 |
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| Resumo: |
Therapeutic monitoring of vancomycin concentration can improve the safety and efficiency of the treatment with this antibiotic. High-performance liquid chromatography (HPLC) is considered gold standard for quantification for this drug. However the analysis requires multiple procedures for purification and analysis of the sample, which reduces their applicability for clinical use. There`s currently no consensus on vancomycin serum target levels and dose adjustment among newborns. This paper proposes a vancomycin therapeutic monitoring protocol and develops and validates a low cost, simplified method, with smaller sample volume requirement for serum concentration of vancomycin determination by using HPLC with diode array. Samples were extracted by protein precipitation with trifluoroacetic acid (TFA), supernatant was injected directly into the chromatographic system. Mobile phase consisted of TFA 0.014% (pH 2.800) in ultrapure water and acetonitrile (92:8, v /v) under flow rate of 2 mL / min eluting in isocratic method in 40oC. Response were monitored at a wavelength of 230.9 nm. Validation was conducted in accordance with the resolution of the RDC No. 27 of May 17, 2012 ANVISA. Vancomycin and internal standard (7-hydroxycoumarin) were eluted at approximately 5.1 min. and 10.3 min respectively. Linearity was assessed between 2 and 70 mg /L with linear correlation greater than 0.999. The method proved to be suitable for the proposed application, with the advantages of less sample volume required, simplified extraction and achieved satisfactory results of precision, accuracy, stability and selectivity. This method was tested in 17 real samples from 10 patients with satisfactory results. The drug monitoring protocol was successfully developed. |
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