Quantification of vincristine and its major metabolite in human plasma by high-performance liquid chromatography/tandem mass spectrometry

Jennifer B. Dennison, Jamie Renbarger, David O. Walterhouse, David R. Jones, Stephen D. Hall

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

An analytical method using electrospray ionization and high-performance liquid chromatography/tandem mass spectrometry (LC/ESI-MS/MS) was developed to quantify vincristine and M1, the CYP3A-mediated metabolite of vincristine, in human plasma. Vinblastine (internal standard), vincristine, and M1 in plasma were extracted in methylene chloride after acidification with TCAA. The analytes were separated on an Inertsil ODS-3 C18 column (2.1 × 150 mm) with a 5-μm particle size using a gradient elution with a run time of 20 min. The initial mobile phase composition was 0.2% formic acid/water (80:20, v/v) with a final composition of 0.2% formic acid/water (20:80, v/v). Detection was accomplished with multiple reaction monitoring for vinblastine (m/z 406.3→ 271.7), vincristine (m/z 413.2→ 362.2), and M1 (m/z 397.3 → 376.2). At three concentrations of vincristine and M1, the inter-day and intra-day accuracy and precision were within the acceptable limits for validation (106.8 ± 9.6% for intra-day, n = 5 each concentration; 90.9 ± 10.9% for inter-day, n = 4 each concentration). For both vincristine and M1, the concentration limits of quantification and detection were 12 pg/mL and 6 pg/mL, respectively. Stability studies indicated that 80% of M1 degraded in plasma after 15 hours at room temperature (n = 3, high and low QC concentrations). Therefore, short plasma processing times (<30 min) are recommended. The assay was used successfully to quantify vincristine and M1 in pediatric plasma samples up to 24 hours after vincristine administration. Vincristine and M1 concentrations were within the limits of quantification for all patient plasma samples.

Original languageEnglish
Pages (from-to)357-364
Number of pages8
JournalTherapeutic Drug Monitoring
Volume30
Issue number3
DOIs
StatePublished - Jun 2008

Fingerprint

Plasma (human)
High performance liquid chromatography
Vincristine
Metabolites
Tandem Mass Spectrometry
Mass spectrometry
High Pressure Liquid Chromatography
formic acid
Plasmas
Vinblastine
Cytochrome P-450 CYP3A
Plasma applications
Electrospray ionization
Pediatrics
Water
Acidification
Methylene Chloride
Particle Size
Phase composition
Limit of Detection

Keywords

  • Tandem mass spectrometry
  • Vincristine
  • Vincristine metabolism

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Pharmacology
  • Toxicology
  • Health, Toxicology and Mutagenesis
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Public Health, Environmental and Occupational Health

Cite this

Quantification of vincristine and its major metabolite in human plasma by high-performance liquid chromatography/tandem mass spectrometry. / Dennison, Jennifer B.; Renbarger, Jamie; Walterhouse, David O.; Jones, David R.; Hall, Stephen D.

In: Therapeutic Drug Monitoring, Vol. 30, No. 3, 06.2008, p. 357-364.

Research output: Contribution to journalArticle

Dennison, Jennifer B. ; Renbarger, Jamie ; Walterhouse, David O. ; Jones, David R. ; Hall, Stephen D. / Quantification of vincristine and its major metabolite in human plasma by high-performance liquid chromatography/tandem mass spectrometry. In: Therapeutic Drug Monitoring. 2008 ; Vol. 30, No. 3. pp. 357-364.
@article{884446b25dbc46a6a3302b52ffc4fe57,
title = "Quantification of vincristine and its major metabolite in human plasma by high-performance liquid chromatography/tandem mass spectrometry",
abstract = "An analytical method using electrospray ionization and high-performance liquid chromatography/tandem mass spectrometry (LC/ESI-MS/MS) was developed to quantify vincristine and M1, the CYP3A-mediated metabolite of vincristine, in human plasma. Vinblastine (internal standard), vincristine, and M1 in plasma were extracted in methylene chloride after acidification with TCAA. The analytes were separated on an Inertsil ODS-3 C18 column (2.1 × 150 mm) with a 5-μm particle size using a gradient elution with a run time of 20 min. The initial mobile phase composition was 0.2{\%} formic acid/water (80:20, v/v) with a final composition of 0.2{\%} formic acid/water (20:80, v/v). Detection was accomplished with multiple reaction monitoring for vinblastine (m/z 406.3→ 271.7), vincristine (m/z 413.2→ 362.2), and M1 (m/z 397.3 → 376.2). At three concentrations of vincristine and M1, the inter-day and intra-day accuracy and precision were within the acceptable limits for validation (106.8 ± 9.6{\%} for intra-day, n = 5 each concentration; 90.9 ± 10.9{\%} for inter-day, n = 4 each concentration). For both vincristine and M1, the concentration limits of quantification and detection were 12 pg/mL and 6 pg/mL, respectively. Stability studies indicated that 80{\%} of M1 degraded in plasma after 15 hours at room temperature (n = 3, high and low QC concentrations). Therefore, short plasma processing times (<30 min) are recommended. The assay was used successfully to quantify vincristine and M1 in pediatric plasma samples up to 24 hours after vincristine administration. Vincristine and M1 concentrations were within the limits of quantification for all patient plasma samples.",
keywords = "Tandem mass spectrometry, Vincristine, Vincristine metabolism",
author = "Dennison, {Jennifer B.} and Jamie Renbarger and Walterhouse, {David O.} and Jones, {David R.} and Hall, {Stephen D.}",
year = "2008",
month = "6",
doi = "10.1097/FTD.0b013e31816b92c9",
language = "English",
volume = "30",
pages = "357--364",
journal = "Therapeutic Drug Monitoring",
issn = "0163-4356",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - Quantification of vincristine and its major metabolite in human plasma by high-performance liquid chromatography/tandem mass spectrometry

AU - Dennison, Jennifer B.

AU - Renbarger, Jamie

AU - Walterhouse, David O.

AU - Jones, David R.

AU - Hall, Stephen D.

PY - 2008/6

Y1 - 2008/6

N2 - An analytical method using electrospray ionization and high-performance liquid chromatography/tandem mass spectrometry (LC/ESI-MS/MS) was developed to quantify vincristine and M1, the CYP3A-mediated metabolite of vincristine, in human plasma. Vinblastine (internal standard), vincristine, and M1 in plasma were extracted in methylene chloride after acidification with TCAA. The analytes were separated on an Inertsil ODS-3 C18 column (2.1 × 150 mm) with a 5-μm particle size using a gradient elution with a run time of 20 min. The initial mobile phase composition was 0.2% formic acid/water (80:20, v/v) with a final composition of 0.2% formic acid/water (20:80, v/v). Detection was accomplished with multiple reaction monitoring for vinblastine (m/z 406.3→ 271.7), vincristine (m/z 413.2→ 362.2), and M1 (m/z 397.3 → 376.2). At three concentrations of vincristine and M1, the inter-day and intra-day accuracy and precision were within the acceptable limits for validation (106.8 ± 9.6% for intra-day, n = 5 each concentration; 90.9 ± 10.9% for inter-day, n = 4 each concentration). For both vincristine and M1, the concentration limits of quantification and detection were 12 pg/mL and 6 pg/mL, respectively. Stability studies indicated that 80% of M1 degraded in plasma after 15 hours at room temperature (n = 3, high and low QC concentrations). Therefore, short plasma processing times (<30 min) are recommended. The assay was used successfully to quantify vincristine and M1 in pediatric plasma samples up to 24 hours after vincristine administration. Vincristine and M1 concentrations were within the limits of quantification for all patient plasma samples.

AB - An analytical method using electrospray ionization and high-performance liquid chromatography/tandem mass spectrometry (LC/ESI-MS/MS) was developed to quantify vincristine and M1, the CYP3A-mediated metabolite of vincristine, in human plasma. Vinblastine (internal standard), vincristine, and M1 in plasma were extracted in methylene chloride after acidification with TCAA. The analytes were separated on an Inertsil ODS-3 C18 column (2.1 × 150 mm) with a 5-μm particle size using a gradient elution with a run time of 20 min. The initial mobile phase composition was 0.2% formic acid/water (80:20, v/v) with a final composition of 0.2% formic acid/water (20:80, v/v). Detection was accomplished with multiple reaction monitoring for vinblastine (m/z 406.3→ 271.7), vincristine (m/z 413.2→ 362.2), and M1 (m/z 397.3 → 376.2). At three concentrations of vincristine and M1, the inter-day and intra-day accuracy and precision were within the acceptable limits for validation (106.8 ± 9.6% for intra-day, n = 5 each concentration; 90.9 ± 10.9% for inter-day, n = 4 each concentration). For both vincristine and M1, the concentration limits of quantification and detection were 12 pg/mL and 6 pg/mL, respectively. Stability studies indicated that 80% of M1 degraded in plasma after 15 hours at room temperature (n = 3, high and low QC concentrations). Therefore, short plasma processing times (<30 min) are recommended. The assay was used successfully to quantify vincristine and M1 in pediatric plasma samples up to 24 hours after vincristine administration. Vincristine and M1 concentrations were within the limits of quantification for all patient plasma samples.

KW - Tandem mass spectrometry

KW - Vincristine

KW - Vincristine metabolism

UR - http://www.scopus.com/inward/record.url?scp=47249104198&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=47249104198&partnerID=8YFLogxK

U2 - 10.1097/FTD.0b013e31816b92c9

DO - 10.1097/FTD.0b013e31816b92c9

M3 - Article

C2 - 18520608

AN - SCOPUS:47249104198

VL - 30

SP - 357

EP - 364

JO - Therapeutic Drug Monitoring

JF - Therapeutic Drug Monitoring

SN - 0163-4356

IS - 3

ER -