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Ultra high performance liquid chromatography-tandem mass spectrometry vs. commercial immunoassay for determination of vancomycin plasma concentration in children. Possible implications for everyday clinical practice
Authors:Sebastiano Barco  Iulian Gennai  Laura Barbagallo  Anna Loy  Gino Tripodi
Institution:1. Clinical Pathology Laboratory Unit, Istituto Giannina Gaslini, Genoa, Italy;2. Infectious Disease Unit, Istituto Giannina Gaslini, Genoa, Italy
Abstract:Background: Vancomycin therapeutic drug monitoring (TDM) is necessary for effective and safetherapy. The aim of the this paper was to develop a specific and robust ultra high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method for vancomycin quantification starting from low plasma volumes to be applied for the routine TDM in children.

Methods: Samples from children receiving intravenous vancomycin were analysed using a TSQ Quantum Access MAX Triple Quadrupole system coupled with an Accela 1250 UHPLC system after a rapid protein precipitation. Gradient separation chromatography was carried out using a Hypersil GOLD aQ C18 column (50 × 2.1 mm, particle size 1.9 μm). Method performance was validated following international guidelines.

Results: UHPLC–MS/MS allowed a rapid and specific quantification of vancomycin over the range 0.1–128 μg/mL from 50 μL of plasma with high reproducibility and accuracy in the absence of matrix effect. The comparison with the commercial immunoassay performed on 138 samples demonstrated the presence of a proportional bias. The concentrations of vancomycin measured with immunoassay were found to be 4.5% (95% CI: 1.3–7.7) higher than those determined with UHPLC–MS/MS. Importantly, a clinical discordance was found in about 10% of samples analysed.

Conclusions: This new UHPLC–MS/MS method is accurate and specific for the measurement of vancomycin starting from small (50 μL) plasma volumes. The use of UHPLC–MS/MS is recommended to prevent a misclassification of therapeutic or toxic vancomycin levels in paediatrics.

Keywords:Vancomycin  UHPLC–MS/MS  Method validation  Therapeutic drug monitoring
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