Quantitation of 4-hydroxycyclophosphamide/aldophosphamide in whole blood

L. W. Anderson, S. M. Ludeman, O. M. Colvin, L. B. Grochow, J. M. Strong

Research output: Contribution to journalArticle

Abstract

There is considerable interest in determining 4-hydroxycylcophosphamide/aldophosphamide (4-HO-CP/AP) blood levels in patients receiving the prodrug, cyclophosphamide (CP). Phosphoramide mustard (PM), the alkylating metabolite of CP, is relatively impermeable to cell membranes and it is generally believed that circulating intermediary metabolites, including aldophosphamide, the immediate precursor of PM, is transported by circulating blood to tumor tissue. Therefore, circulating 4-HO-CP/AP blood levels should more closely reflect the oncostatic and cytotoxic effects of CP than the parent drug. We have developed a gas chromatographic electron-impact mass spectrometric (GC-EIMS) method suitable for routine monitoring of 4-HO-CP/AP levels in whole blood over the range 0.085 μM (25 ng/ml) to 34 μM (10 μ/ml). The unstable metabolites were derivatized with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine-HCl to form a stable aldophosphamide oxime derivative (PBOX). [2H4]PBOX was used as an internal standard. For clinical samples, tubes were prepared prior to blood drawing, which contained the derivatizing reagent solution and the internal standard. These solutions were stable for up to 3 months when stored at room temperature. Following addition of blood to the reaction tubes, PBOX formation was rapid and the resulting derivative was stable under these conditions for up to 8 days at room temperature. Application of the method was demonstrated by quantitating 4-HO-CP/AP blood levels in patients receiving 4 g/m2 intravenous infusion of CP over a period of 90 min.

Original languageEnglish (US)
Pages (from-to)247-257
Number of pages11
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume667
Issue number2
DOIs
StatePublished - May 19 1995

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4-hydroxycyclophosphamide
Cyclophosphamide
Blood
Metabolites
Derivatives
Oximes
aldophosphamide
Prodrugs
Cell membranes

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Quantitation of 4-hydroxycyclophosphamide/aldophosphamide in whole blood. / Anderson, L. W.; Ludeman, S. M.; Colvin, O. M.; Grochow, L. B.; Strong, J. M.

In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 667, No. 2, 19.05.1995, p. 247-257.

Research output: Contribution to journalArticle

Anderson, L. W. ; Ludeman, S. M. ; Colvin, O. M. ; Grochow, L. B. ; Strong, J. M. / Quantitation of 4-hydroxycyclophosphamide/aldophosphamide in whole blood. In: Journal of Chromatography B: Biomedical Sciences and Applications. 1995 ; Vol. 667, No. 2. pp. 247-257.
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