Development and Validation of a Liquid Chromatography Method with Electrochemical Detection for Hydroxyurea Quantification in Human Plasma and Aqueous Solutions

Abstract

Hydroxyurea is the unique drug having demonstrated a significant efficacy in sickle cell disease treatment. We developed a liquid chromatography method with electrochemical detection for hydroxyurea analysis in plasma and aqueous solutions. Analytical goals included an analytical range from 2 to 50 mg/L, a total imprecision lower than 15% and a total error lower than 30%. After protein precipitation with acetonitrile, the separation was performed on a C18 Atlantis T3 column and eluted with sodium acetate 25 mM, acetonitrile 2.5%, pH 6.5. Thioacetamide was used as internal standard. The method was linear for drug concentrations ranging from 0.5 to 50 mg/L and recovery was comprised between 100 and 120%. The intra-day precision was lower than 6.0% and between-day precision was lower than 11%. The detection limit was 0.18 and 0.63 mg/L for aqueous solution and plasma, respectively and the quantification limit was 1.0 and 1.2 mg/L for aqueous solution and plasma, respectively. No interference from urea was observed. The liquid chromatography method developed can be used for pharmacokinetic studies in plasma and other biological samples such as saliva or urine. It requires low sample volumes and a simple pre-treatment and it allows a direct measure of non-derivatized hydroxyurea.

Hydroxyurea (hydroxycarbamide) genotoxicity in pediatric patients with sickle cell disease

“…Blood samples were collected in EDTA tubes 30 minutes, 60 minutes, and 180 minutes post drug administration. Quantification was performed by liquid chromatography, as reported by Rodriguez et al . Maximal concentration (C max ) of HU on plasma was registered for each patient.…”

Hydroxyurea-Lactose Interaction Study: In Silico and In Vitro Evaluation

“…The stationary phase was a Phenomenex Luna C 18 column (250 mm × 4.6 mm) with a particle size of 5 μm. For analysis of hydroxyurea, the mobile phase was 25 mM sodium acetate and 2.5% acetonitrile at pH 6.5 with a flow rate of 1.0 mL/min (20). For analysis of the adduct, the mobile phase was water/methanol (95:5, v/v) with a flow rate of 1.0 mL/min (21).…”

“…3.) with solvent system sodium acetate 25 mM, acetonitrile 2.5%, pH 6.5 (20) with detection at 214 nm. The adduct resolution was not obtained.…”

Spectrophotometric Determination of Hydroxyurea in Pharmaceutical Preparations

“…Different techniques have been reported for the determination of hydroxyurea. These include high performance liquid chromatography with electrochemical detection (Jong et al, 2003;Manouilov et al, 1998;Rodriguez et al, 2014) and gas chromatography-mass spectrometry (Kettani, 2009) for the determination of hydroxyurea in human plasma (Legrand et al, 2017) as well as the determination of hydroxyurea in capsules and biological fluids by ion-selective potentiometry and fluorimetry (El-Kosasy, 2003). Another method is the analysis of hydroxyurea by proton nuclear magnetic resonance (NMR) spectroscopy has also been proposed (Main et al, 1987).…”