ArticleEinstein (Sao Paulo, Brazil)2025
Validation of an ultra-high performance liquid chromatography/UV method to quantify busulfan in plasma: application to therapeutic drug monitoring.
Article in Einstein (Sao Paulo, Brazil), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundOlivo et al. validated an in-house precise UHPLC/UV method for quantifying busulfan in human plasma for therapeutic monitoring. The method shows linearity (0.5-10 μg/mL) with a lower limit of quantification of 0.5 μg/mL, demonstrating accuracy and precision. It effectively supported therapeutic drug monitoring in a Brazilian public hospital by providing rapid and reliable results. ■ We validated the UHPLC/UV method for accurate busulfan quantification in plasma. ■ Inaccuracy and imprecision were below 15%, ensuring reliable therapeutic drug monitoring results. ■ This enables effective pharmacokinetic studies with rapid turnaround times in patient samples.
objectiveThis study aimed to validate a sensitive, accurate, and precise bioanalytical ultra-high-performance liquid chromatography coupled with ultraviolet (UHPLC/UV) method for the determination of busulfan in human plasma using 1,6-bis-(methanesulfonyloxy) hexane as an internal standard for therapeutic drug monitoring.
methodsPlasma samples were deproteinized with acetonitrile (1:2, v/v) and, after derivatization with sodium diethyl dithiocarbamate, submitted to liquid-liquid extraction with ethyl acetate and evaporation at 50ºC under a nitrogen stream. Analyses were performed on a Shimadzu® system using a C18 column and isocratic elution with methanol/water (70:30, v/v) at a flow rate of 0.4mL min-1 and detection at 277nm.
resultsThe retention times of busulfan and the IS were approximately 2.87 and 6.35 min, respectively. The plasma calibration curves were linear in the concentration range of 0.5-10 µg mL-1 with a coefficient of determination greater than 0.99. The lower limit of quantification was 0.5 µg mL-1. The inaccuracies and imprecisions of this method are less than 15%. The applicability of this method to pharmacokinetic studies was confirmed using patient samples obtained after 4 h of 3.2-5.4 mg kg-1 busulfan intermittent infusion.
conclusionThis method demonstrated the feasibility of quantifying samples within the target concentration range and quickly releasing results to allow for busulfan therapeutic monitoring.
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