TY - JOUR
T1 - A sensitive and rapid ultra high-performance liquid chromatography with tandem mass spectrometric assay for the simultaneous quantitation of cyclophosphamide and the 4-hydroxycyclophosphamide metabolite in human plasma
AU - Hall, O. Morgan
AU - Peer, Cody J.
AU - Fitzhugh, Courtney D.
AU - Figg, William D.
N1 - Funding Information:
This study was funded in part by the Intramural Research Program of the NIH , National Cancer Institute and the National Heart Lung and Blood Institute ( ZIC SC 006536 ). This is a US Government work and there are no restrictions on its use. The views expressed within this paper do not necessarily reflect those of the US Government. We thank the nursing and clinical support staff of the NCI and NHLBI for their care of the patients. Most importantly, we appreciate the patients who enroll in investigational trials to advance the knowledge of this disease.
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/6/1
Y1 - 2018/6/1
N2 - Analysis of cyclophosphamide (CP) and its metabolite, 4-hydroxycyclophosphamide (4OHCP), in a single assay has the ability to improve sampling techniques benefitting both the patients who are receiving the drug and the clinicians drawing samples. Due to instability in plasma (t1/2 = 4 min), immediate stabilization of 4OHCP with phenylhydrazine is necessary upon sample draw. After stabilization, 4OHCP and the stable CP prodrug concentrations can be analytically measured to elucidate the pharmacokinetics, including half-life and exposure parameters (Cmax and AUC). For this purpose, a sensitive analytical method was developed to measure both the prodrug and active metabolite. A liquid-liquid extraction recovered the analytes prior to analysis with an ultra HPLC-MS/MS. A Thermo Scientific™ Hypersil™ BDS C18, 2.1 × 100 mm, 3.0 μm column was used for compound separation. Mass transitions for CP (m/z 261.0 ➔ 140.0), the internal standard d4-CP (m/z 265.0 ➔ 140.0), 4OHCP (m/z 367.3 ➔ 147.1), and the internal standard AZD7451 (m/z 383.4 ➔ 341.1) were monitored over a calibration range of 34.24–34,240 ng/mL and 3.424–3424 ng/mL for CP and 4OHCP, respectively. Each calibration range proved accurate (<15% deviation) and precise (<15% RSD) for the desired compound. Using this method, CP and 4OHCP plasma levels can be measured in clinical samples from patients receiving this therapy.
AB - Analysis of cyclophosphamide (CP) and its metabolite, 4-hydroxycyclophosphamide (4OHCP), in a single assay has the ability to improve sampling techniques benefitting both the patients who are receiving the drug and the clinicians drawing samples. Due to instability in plasma (t1/2 = 4 min), immediate stabilization of 4OHCP with phenylhydrazine is necessary upon sample draw. After stabilization, 4OHCP and the stable CP prodrug concentrations can be analytically measured to elucidate the pharmacokinetics, including half-life and exposure parameters (Cmax and AUC). For this purpose, a sensitive analytical method was developed to measure both the prodrug and active metabolite. A liquid-liquid extraction recovered the analytes prior to analysis with an ultra HPLC-MS/MS. A Thermo Scientific™ Hypersil™ BDS C18, 2.1 × 100 mm, 3.0 μm column was used for compound separation. Mass transitions for CP (m/z 261.0 ➔ 140.0), the internal standard d4-CP (m/z 265.0 ➔ 140.0), 4OHCP (m/z 367.3 ➔ 147.1), and the internal standard AZD7451 (m/z 383.4 ➔ 341.1) were monitored over a calibration range of 34.24–34,240 ng/mL and 3.424–3424 ng/mL for CP and 4OHCP, respectively. Each calibration range proved accurate (<15% deviation) and precise (<15% RSD) for the desired compound. Using this method, CP and 4OHCP plasma levels can be measured in clinical samples from patients receiving this therapy.
KW - 4-hydroxycyclophosphamide
KW - Cyclophosphamide
KW - Sickle cell disease
KW - Tandem mass spectrometry
KW - Ultra-high performance liquid chromatography
UR - http://www.scopus.com/inward/record.url?scp=85045402417&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2018.04.016
DO - 10.1016/j.jchromb.2018.04.016
M3 - Article
C2 - 29656084
AN - SCOPUS:85045402417
SN - 1570-0232
VL - 1086
SP - 56
EP - 62
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
ER -