HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF THE STABILIZED CYCLOPHOSPHAMIDE METABOLITE 4-HYDROXYCYCLOPHOSPHAMIDE IN PLASMA AND RED BLOOD CELLS

Autor: Thomas Kerbusch, Matthijs M. Tibben, A. D. R. Huitema, Sjoerd Rodenhuis, Jos H. Beijnen, J. J. Kettenes van den Bosch
Rok vydání: 2000
Předmět:
Zdroj: Journal of Liquid Chromatography & Related Technologies. 23:1725-1744
ISSN: 1520-572X
1082-6076
DOI: 10.1081/jlc-100100448
Popis: A reversed-phase high performance liquid chromatographic (HPLC) method for the determination of the activated cyclophosphamide (CP) metabolite 4-hydroxycyclophosphamide (4-OHCP) in human plasma and red blood cells is described. 4-OHCP is very unstable in biological matrices. Therefore, it was treated, immediately after sampling, with semicarbazide to form a stable semicarbazone derivative, which was identified with electron spray mass spectrometry. The derivative was analysed with HPLC with ultraviolet (UV) detection at 230 nm. Sample pre-treatment consisted of a liquid-liquid extraction with ethyl acetate, the chromatography system was a 25 cm C8 column (particle size 5 μm) with a mobile phase of acetonitrile-0.025 M potassium phosphate buffer (15:85 v/v) pH 6.0. After assay optimisation, the method was validated and stability studies were carried out. The method proved linear in clinically relevant concentrations (50-5000 ng/mL). The lower limit of quantitation was 50 ng/mL. Accuracy and precision were ...
Databáze: OpenAIRE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje