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 L.
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 -