ABSTRACT
An ultra-performance liquid chromatography coupled with electrospray ionisation tandem mass spectrometry (UPLC-ESI-MS/MS) with pre-column derivatisation was developed and validated for the determination of semicarbazide in human urine. Urine samples were derivatised with 2-nitrobenzaldehyde and subsequently extracted with acetonitrile. Extracts were concentrated and then analysed by UPLC-MS/MS. The time per run was 7 min. Good results were observed for the linearity of matrix-matched calibration curves (R2 > 0.99) in the concentration range of 1-100 µg/L. The absolute recovery ranged from 98.7% to 108.6%, with the relative standard deviations (RSDs) of 2.2%-3.6%. The limit of detection and quantification for the semicarbazide was 0.5 µg/L and 1 µg/L, respectively. The method showed good extraction efficiency, high sensitivity, and good reproducibility. It was suitable for the detection of semicarbazide in human urine. Residues of semicarbazide were between 1.0 and 41.5 µg/L in children's 24-h urine. This work is the first report on the quantitative analysis of SEM in 24-h human urine samples.
Subject(s)
Semicarbazides/isolation & purification , Semicarbazides/urine , Acetonitriles/chemistry , Benzaldehydes/chemistry , Biosensing Techniques , Child , Child, Preschool , China , Chromatography, High Pressure Liquid , Female , Humans , Limit of Detection , Male , Reproducibility of Results , Sensitivity and Specificity , Tandem Mass SpectrometryABSTRACT
A simple and sensitive UPLC-MS/MS assay was developed and validated for rapid determination of thiosemicarbazide derivative of isoniazid (TSC-INH), a potent anti-candidal agent in rat plasma, tissues, urine and feces. All biological samples were prepared by protein precipitation method using celecoxib as an internal standard (IS). Chromatographic separation was achieved on Acquity BEH™ C18 (50×2.1 mm, 1.7 µm) column using gradient mobile phase of acetonitrile and water (containing 0.1% formic acid) at flow rate of 0.3 mL/min. The MRM transitions were monitored at m/z 305.00â135.89 for TSC-INH and m/z 380.08â316.03 for IS in ESI negative mode. All validation parameter results were within the acceptable range described in guideline for bioanalytical method validation. The pharmacokinetic study showed that the compound TSC-INH was orally active with 66% absolute bioavailability in rats. It was rapidly absorbed with peak plasma concentration of 1985.92 ng/mL achieved within 1 h after single oral dose (10 mg/kg) administration. TSC-INH exhibited rapid distribution across the body with highest levels in liver and lungs. Penetration in brain tissues suggests that TSC-INH crossed the blood brain barrier. Only 5.23% of the orally administered drug was excreted as unconverted form in urine and feces implying that TSC-INH was metabolized extensively before excretion. With the preliminary knowledge of in vivo pharmacokinetics and disposition properties, this study will be beneficial for further development of compound TSC-INH in future studies.
