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Simultaneous determination of cyclophosphamide and 4-hydroxycyclophosphamide in human plasma by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry – application to Chinese systemic lupus erythematosus patients
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Wenying Shu
Veröffentlicht/Copyright:
9. September 2011
Abstract
Background: A pharmacogenomics study of cyclophosphamide in systemic lupus erythematosus patients is being conducted in our laboratory in which the plasma concentrations of cyclophosphamide and its active metabolite 4-hydroxycyclophosphamide should be assayed rapidly and sensitively.
Methods: A rapid, stable and sensitive liquid chromato-graphy/electrospray ionization tandem mass spectrometry method was developed to simultaneously determine cyclophosphamide and 4-hydroxycyclophosphamide in human plasma with ifosfomide as an internal standard. After a protein precipitation with cold acetonitrile and stabilization of 4-hydroxycyclophosphamide by ansyldrazine and extraction with ethyl acetate, separation was performed on a C18 3.5 μm 2.1×50 mm column with mobile phase of acetonitrile and water (50:50, v/v) with 0.1% formic acid at 200 μL/min. The chromatographic run time was 3 min.
Results: The linear calibration curves ranged from 5 to 5000 ng/mL for cyclophosphamide and 5–500 ng/mL for 4-hydroxycyclophosphamide. The recoveries of the liquid extraction were 54.5%–58.5% for cyclophosphamide and 103.5%–105.5% for 4-hydroxycyclophosphamide. The lower limit of quantification was 5 ng/mL for both analytes. The intra- and inter-day precision was <15% for quality control samples at 4000, 500, 50 ng/mL for cyclophosphamide and 4-hydroxycyclophosphamide at 400, 100, 20 ng/mL. The method was applied in this pharmacogenomics study in Chinese systemic lupus erythematosus patients treated with low-dose cyclophosphamide.
Conclusions: The method was efficient with shorter running time and lower limit of quantification compared to previous reports and has been successfully applied in this pharmacogenomics study.
Keywords: cyclophosphamide; 4-hydroxycyclophosphamide; liquid chromatography/tandem mass spectrometry; pharmacokinetics; systemic lupus erythematosus
Received: 2011-5-18
Accepted: 2011-8-17
Published Online: 2011-9-9
Published in Print: 2011-12-1
©2011 by Walter de Gruyter Berlin Boston
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Schlagwörter für diesen Artikel
cyclophosphamide;
4-hydroxycyclophosphamide;
liquid chromatography/tandem mass spectrometry;
pharmacokinetics;
systemic lupus erythematosus
Artikel in diesem Heft
- Editorial
- Appropriate labelling of blood collection tubes: a step ahead towards patient’s safety
- Reviews
- New insights into adipose tissue dysfunction in insulin resistance
- Role of biomarkers in cardioncology
- Minireviews
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- High-sensitivity troponin assays in the evaluation of patients with acute chest pain in the emergency department
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- Guidelines and Recommendations
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