Determination of procarbazine in human plasma by liquid chromatography with electrospray ionization mass spectrometry

X. He, T. T. Batchelor, Stuart A Grossman, J. G. Supko

Research output: Contribution to journalArticle

Abstract

Procarbazine is a cytotoxic chemotherapeutic agent used in the treatment of lymphomas and brain tumors. Its pharmacokinetic behavior remains poorly understood even though more than 30 years have elapsed since the drug was approved for clinical use. To characterize the pharmacokinetics of procarbazine in brain cancer patients during a phase I trial, a method for determining the drug in human plasma by reversed-phase high-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry (ESI-MS) was developed and thoroughly validated. Plasma samples were prepared for analysis by precipitating proteins with trichloroacetic acid and washing the protein-free supernatant with methyl tert-butyl ether to remove excess acid. The solution was separated on a Luna C-18 analytical column using methanol-25mM ammonium acetate buffer, pH 5.1 (22:78, v/v) as the mobile phase at 1.0ml/min. A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M+H]+ ions at m/z 222.2 for procarbazine and at m/z 192.1 for the internal standard (3-dimethylamino-2-methylpropiophenone). Procarbazine and the internal standard eluted as sharp, symmetrical peaks with retention times (mean±S.D.) of 6.3±0.1 and 9.9±0.3min, respectively. Calibration curves of procarbazine hydrochloride in human plasma at concentrations ranging from 0.5 to 50ng/ml exhibited excellent linearity. The mean absolute recovery of the drug from plasma was 102.9±1.0%. Using a sample volume of 150μl, procarbazine was determined at the 0.5ng/ml (1.9nM) lower limit of quantitation with a mean accuracy of 105.2% and an interday precision of 3.60% R.S.D. on 11 different days over 5 weeks. During this same time interval, the between-day accuracy for determining quality control solutions of the drug in plasma at concentrations of 2.0, 15 and 40ng/ml ranged from 97.5 to 98.2% (mean±S.D., 97.9±0.4%) and the precision was 3.8-6.2% (mean±S.D., 5.1±1.2%). Stability characteristics of the drug were thoroughly evaluated to establish appropriate conditions to process, store and prepare clinical specimens for chromatographic analysis without inducing significant chemical degradation. The sensitivity achieved with this assay permitted the plasma concentration-time profile of the parent drug to be accurately defined following oral administration of standard doses to brain cancer patients.

Original languageEnglish (US)
Pages (from-to)281-291
Number of pages11
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume799
Issue number2
DOIs
StatePublished - Jan 25 2004

Fingerprint

Procarbazine
Plasma (human)
Electrospray ionization
Electrospray Ionization Mass Spectrometry
Liquid chromatography
Liquid Chromatography
Mass spectrometry
Brain Neoplasms
Pharmaceutical Preparations
Plasmas
Brain
Pharmacokinetics
Chromatographic analysis
Ions
Drug Stability
Trichloroacetic Acid
Cytotoxins
High performance liquid chromatography
Mass spectrometers
Reverse-Phase Chromatography

Keywords

  • Procarbazine

ASJC Scopus subject areas

  • Biochemistry

Cite this

@article{d838d21436d946b38c7ed6e759fa307f,
title = "Determination of procarbazine in human plasma by liquid chromatography with electrospray ionization mass spectrometry",
abstract = "Procarbazine is a cytotoxic chemotherapeutic agent used in the treatment of lymphomas and brain tumors. Its pharmacokinetic behavior remains poorly understood even though more than 30 years have elapsed since the drug was approved for clinical use. To characterize the pharmacokinetics of procarbazine in brain cancer patients during a phase I trial, a method for determining the drug in human plasma by reversed-phase high-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry (ESI-MS) was developed and thoroughly validated. Plasma samples were prepared for analysis by precipitating proteins with trichloroacetic acid and washing the protein-free supernatant with methyl tert-butyl ether to remove excess acid. The solution was separated on a Luna C-18 analytical column using methanol-25mM ammonium acetate buffer, pH 5.1 (22:78, v/v) as the mobile phase at 1.0ml/min. A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M+H]+ ions at m/z 222.2 for procarbazine and at m/z 192.1 for the internal standard (3-dimethylamino-2-methylpropiophenone). Procarbazine and the internal standard eluted as sharp, symmetrical peaks with retention times (mean±S.D.) of 6.3±0.1 and 9.9±0.3min, respectively. Calibration curves of procarbazine hydrochloride in human plasma at concentrations ranging from 0.5 to 50ng/ml exhibited excellent linearity. The mean absolute recovery of the drug from plasma was 102.9±1.0{\%}. Using a sample volume of 150μl, procarbazine was determined at the 0.5ng/ml (1.9nM) lower limit of quantitation with a mean accuracy of 105.2{\%} and an interday precision of 3.60{\%} R.S.D. on 11 different days over 5 weeks. During this same time interval, the between-day accuracy for determining quality control solutions of the drug in plasma at concentrations of 2.0, 15 and 40ng/ml ranged from 97.5 to 98.2{\%} (mean±S.D., 97.9±0.4{\%}) and the precision was 3.8-6.2{\%} (mean±S.D., 5.1±1.2{\%}). Stability characteristics of the drug were thoroughly evaluated to establish appropriate conditions to process, store and prepare clinical specimens for chromatographic analysis without inducing significant chemical degradation. The sensitivity achieved with this assay permitted the plasma concentration-time profile of the parent drug to be accurately defined following oral administration of standard doses to brain cancer patients.",
keywords = "Procarbazine",
author = "X. He and Batchelor, {T. T.} and Grossman, {Stuart A} and Supko, {J. G.}",
year = "2004",
month = "1",
day = "25",
doi = "10.1016/j.jchromb.2003.10.061",
language = "English (US)",
volume = "799",
pages = "281--291",
journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",
issn = "1570-0232",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Determination of procarbazine in human plasma by liquid chromatography with electrospray ionization mass spectrometry

AU - He, X.

AU - Batchelor, T. T.

AU - Grossman, Stuart A

AU - Supko, J. G.

PY - 2004/1/25

Y1 - 2004/1/25

N2 - Procarbazine is a cytotoxic chemotherapeutic agent used in the treatment of lymphomas and brain tumors. Its pharmacokinetic behavior remains poorly understood even though more than 30 years have elapsed since the drug was approved for clinical use. To characterize the pharmacokinetics of procarbazine in brain cancer patients during a phase I trial, a method for determining the drug in human plasma by reversed-phase high-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry (ESI-MS) was developed and thoroughly validated. Plasma samples were prepared for analysis by precipitating proteins with trichloroacetic acid and washing the protein-free supernatant with methyl tert-butyl ether to remove excess acid. The solution was separated on a Luna C-18 analytical column using methanol-25mM ammonium acetate buffer, pH 5.1 (22:78, v/v) as the mobile phase at 1.0ml/min. A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M+H]+ ions at m/z 222.2 for procarbazine and at m/z 192.1 for the internal standard (3-dimethylamino-2-methylpropiophenone). Procarbazine and the internal standard eluted as sharp, symmetrical peaks with retention times (mean±S.D.) of 6.3±0.1 and 9.9±0.3min, respectively. Calibration curves of procarbazine hydrochloride in human plasma at concentrations ranging from 0.5 to 50ng/ml exhibited excellent linearity. The mean absolute recovery of the drug from plasma was 102.9±1.0%. Using a sample volume of 150μl, procarbazine was determined at the 0.5ng/ml (1.9nM) lower limit of quantitation with a mean accuracy of 105.2% and an interday precision of 3.60% R.S.D. on 11 different days over 5 weeks. During this same time interval, the between-day accuracy for determining quality control solutions of the drug in plasma at concentrations of 2.0, 15 and 40ng/ml ranged from 97.5 to 98.2% (mean±S.D., 97.9±0.4%) and the precision was 3.8-6.2% (mean±S.D., 5.1±1.2%). Stability characteristics of the drug were thoroughly evaluated to establish appropriate conditions to process, store and prepare clinical specimens for chromatographic analysis without inducing significant chemical degradation. The sensitivity achieved with this assay permitted the plasma concentration-time profile of the parent drug to be accurately defined following oral administration of standard doses to brain cancer patients.

AB - Procarbazine is a cytotoxic chemotherapeutic agent used in the treatment of lymphomas and brain tumors. Its pharmacokinetic behavior remains poorly understood even though more than 30 years have elapsed since the drug was approved for clinical use. To characterize the pharmacokinetics of procarbazine in brain cancer patients during a phase I trial, a method for determining the drug in human plasma by reversed-phase high-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry (ESI-MS) was developed and thoroughly validated. Plasma samples were prepared for analysis by precipitating proteins with trichloroacetic acid and washing the protein-free supernatant with methyl tert-butyl ether to remove excess acid. The solution was separated on a Luna C-18 analytical column using methanol-25mM ammonium acetate buffer, pH 5.1 (22:78, v/v) as the mobile phase at 1.0ml/min. A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M+H]+ ions at m/z 222.2 for procarbazine and at m/z 192.1 for the internal standard (3-dimethylamino-2-methylpropiophenone). Procarbazine and the internal standard eluted as sharp, symmetrical peaks with retention times (mean±S.D.) of 6.3±0.1 and 9.9±0.3min, respectively. Calibration curves of procarbazine hydrochloride in human plasma at concentrations ranging from 0.5 to 50ng/ml exhibited excellent linearity. The mean absolute recovery of the drug from plasma was 102.9±1.0%. Using a sample volume of 150μl, procarbazine was determined at the 0.5ng/ml (1.9nM) lower limit of quantitation with a mean accuracy of 105.2% and an interday precision of 3.60% R.S.D. on 11 different days over 5 weeks. During this same time interval, the between-day accuracy for determining quality control solutions of the drug in plasma at concentrations of 2.0, 15 and 40ng/ml ranged from 97.5 to 98.2% (mean±S.D., 97.9±0.4%) and the precision was 3.8-6.2% (mean±S.D., 5.1±1.2%). Stability characteristics of the drug were thoroughly evaluated to establish appropriate conditions to process, store and prepare clinical specimens for chromatographic analysis without inducing significant chemical degradation. The sensitivity achieved with this assay permitted the plasma concentration-time profile of the parent drug to be accurately defined following oral administration of standard doses to brain cancer patients.

KW - Procarbazine

UR - http://www.scopus.com/inward/record.url?scp=0344665700&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0344665700&partnerID=8YFLogxK

U2 - 10.1016/j.jchromb.2003.10.061

DO - 10.1016/j.jchromb.2003.10.061

M3 - Article

C2 - 14670747

AN - SCOPUS:0344665700

VL - 799

SP - 281

EP - 291

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

SN - 1570-0232

IS - 2

ER -