TY - JOUR
T1 - Simultaneous quantification of cyclophosphamide, 4-hydroxycyclophosphamide, N,N′,N″-triethylenethio-phosphoramide (thiotepa) and N,N′,N″-triethylene-phosphoramide (tepa) in human plasma by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry
AU - de Jonge, Milly E.
AU - van Dam, Selma M.
AU - Hillebrand, Michael J.X.
AU - Rosing, Hilde
AU - Huitema, Alwin D.R.
AU - Rodenhuis, Sjoerd
AU - Beijnen, Josh H.
PY - 2004/3
Y1 - 2004/3
N2 - The alkylating agents cyclophosphamide (CP) and N, N′, N″,-triethylenethiophosphoramide (thiotepa) are often co-administered in high-dose chemotherapy regimens. Since these regimens can be complicated by the occurrence of severe and sometimes life-threatening toxicities, pharmacokinetically guided administration of these compounds, to reduce variability in exposure, may lead to improved tolerability. For rapid dose adaptations during a chemotherapy course, we have developed and validated an assay, using liquid chromatography coupled with electrospray tandem mass spectrometry (LC/MS/MS), for the routine quantification of CP, thiotepa and their respective active metabolites 4-hydroxycyclophosphamide (4OHCP) and N, N′, N″-triethylenephosphoramide (tepa) in plasma. Because of the instability of 4OHCP in plasma, the compound is derivatized with semicarbazide (SCZ) immediately after sample collection and quantified as 4OHCP-SCZ. Sample pretreatment consisted of protein precipitation with a mixture of methanol and acetronitrile using 100 μl of plasma. Chromatographic separation was performed on an Zorbax Extend C18 column (150 × 2.1 mm i.d., particle size 5 μm), with a quick gradient using 1 mM ammonia solution and acetonitrile, at a flow-rate of 0.4 ml min-1. The analytical run time was 10 min. The triple quadrupole mass spectrometer was operating in the positive ion mode and multiple reaction monitoring was used for drug quantification. The method was validated over the concentration ranges 200-40 000 ng ml-1 for CP, 50-5000 ng ml-1 for 4OHCP-SCZ and 5-2500 ng ml-1 for thiotepa and tepa, using 100 μl of human plasma. These dynamic concentration ranges proved to be relevant in daily practice. Hexamethylphosphoramide was used as an internal standard. The coefficients of variation were <12% for both intra-day and inter-day precisions for each compound. Mean accuracies were also between the designated limits (±15%). This robust and rapid LC/MS/MS assay is now successfully applied for routine therapeutic drug monitoring of CP, thiotepa and their metabolites in our hospital.
AB - The alkylating agents cyclophosphamide (CP) and N, N′, N″,-triethylenethiophosphoramide (thiotepa) are often co-administered in high-dose chemotherapy regimens. Since these regimens can be complicated by the occurrence of severe and sometimes life-threatening toxicities, pharmacokinetically guided administration of these compounds, to reduce variability in exposure, may lead to improved tolerability. For rapid dose adaptations during a chemotherapy course, we have developed and validated an assay, using liquid chromatography coupled with electrospray tandem mass spectrometry (LC/MS/MS), for the routine quantification of CP, thiotepa and their respective active metabolites 4-hydroxycyclophosphamide (4OHCP) and N, N′, N″-triethylenephosphoramide (tepa) in plasma. Because of the instability of 4OHCP in plasma, the compound is derivatized with semicarbazide (SCZ) immediately after sample collection and quantified as 4OHCP-SCZ. Sample pretreatment consisted of protein precipitation with a mixture of methanol and acetronitrile using 100 μl of plasma. Chromatographic separation was performed on an Zorbax Extend C18 column (150 × 2.1 mm i.d., particle size 5 μm), with a quick gradient using 1 mM ammonia solution and acetonitrile, at a flow-rate of 0.4 ml min-1. The analytical run time was 10 min. The triple quadrupole mass spectrometer was operating in the positive ion mode and multiple reaction monitoring was used for drug quantification. The method was validated over the concentration ranges 200-40 000 ng ml-1 for CP, 50-5000 ng ml-1 for 4OHCP-SCZ and 5-2500 ng ml-1 for thiotepa and tepa, using 100 μl of human plasma. These dynamic concentration ranges proved to be relevant in daily practice. Hexamethylphosphoramide was used as an internal standard. The coefficients of variation were <12% for both intra-day and inter-day precisions for each compound. Mean accuracies were also between the designated limits (±15%). This robust and rapid LC/MS/MS assay is now successfully applied for routine therapeutic drug monitoring of CP, thiotepa and their metabolites in our hospital.
KW - Bioanalysis
KW - Cyclophosphamide
KW - Liquid chromatography/tandem mass spectrometry
KW - Protein precipitation
KW - Thiotepa
UR - http://www.scopus.com/inward/record.url?scp=1642298855&partnerID=8YFLogxK
U2 - 10.1002/jms.570
DO - 10.1002/jms.570
M3 - Article
C2 - 15039933
AN - SCOPUS:1642298855
SN - 1076-5174
VL - 39
SP - 262
EP - 271
JO - Journal of Mass Spectrometry
JF - Journal of Mass Spectrometry
IS - 3
ER -