TY - JOUR
T1 - Determination of levetiracetam in human plasma/serum/saliva by liquid chromatography-electrospray tandem mass spectrometry
AU - Guo, Tiedong
AU - Oswald, Lisa M.
AU - Mendu, Damodara Rao
AU - Soldin, Steven J.
N1 - Funding Information:
This study has been supported in part by NIH/NINDS RO1NS045656. It was also supported in part by a grant M01-RR13297 from the General Clinical Research Center Program of the National Center for Research Resources, National Institutes of Health, Department of Health and Human Services, Bethesda, MD. It was supported in part by grant 1 U10HD45993-02 of the National Institute of Child Health and Development, Bethesda, MD.
PY - 2007/1
Y1 - 2007/1
N2 - Background: Levetiracetam (Keppra) is a novel antiepileptic drug recently approved by the U.S. Food and Drug Administration as an add-on therapy in the treatment of partial onset seizures in patients. We developed and describe a simple and rapid high performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) assay for the determination of levetiracetam in human matrix (plasma, serum, or saliva). Methods: An API-3000 or API-4000 triple-quadrupole mass spectrometer (Sciex, Concord, Canada) coupled with the IonSpray source and Shimadzu HPLC system (Shimadzu Scientific Instruments, Columbia, MD) was used employing ritonavir as internal standard (IS) for levetiracetam. One hundred microliters of serum (or plasma, saliva) was deproteinized by adding 150 μl of acetonitrile containing internal standard. After centrifugation, 100 μl of supernatant was diluted with 300 μl of water and 10 μl aliquot was injected onto a C-18 column. After a 2.5 min wash the valve was activated to initiate the isocratic elution program which eluted the levetiracetam and internal standard into the MS/MS system. Quantitation by MRM analysis was performed in the positive ion mode. Within-day and between-day imprecision were evaluated for levetiracetam using three levels of in-house controls. Reliability and accuracy of this method were assessed by comparison of targets with external QC material (ChromSystems), between laboratory comparisons and by recovery studies. Results: Within-day coefficients of variation (CVs) were < 6.1% and between-day CVs were < 8.2%. The average spiked recoveries of levetiracetam added to the drug-free human plasma samples were 108% at low concentration level and 103% at high concentration level. Conclusions: The method was found both specific and sensitive for the rapid and accurate measurement of levetiracetam in human matrices and correlated well with the Quest/Chantilly tandem mass spectrometric method (r = 0.983).
AB - Background: Levetiracetam (Keppra) is a novel antiepileptic drug recently approved by the U.S. Food and Drug Administration as an add-on therapy in the treatment of partial onset seizures in patients. We developed and describe a simple and rapid high performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) assay for the determination of levetiracetam in human matrix (plasma, serum, or saliva). Methods: An API-3000 or API-4000 triple-quadrupole mass spectrometer (Sciex, Concord, Canada) coupled with the IonSpray source and Shimadzu HPLC system (Shimadzu Scientific Instruments, Columbia, MD) was used employing ritonavir as internal standard (IS) for levetiracetam. One hundred microliters of serum (or plasma, saliva) was deproteinized by adding 150 μl of acetonitrile containing internal standard. After centrifugation, 100 μl of supernatant was diluted with 300 μl of water and 10 μl aliquot was injected onto a C-18 column. After a 2.5 min wash the valve was activated to initiate the isocratic elution program which eluted the levetiracetam and internal standard into the MS/MS system. Quantitation by MRM analysis was performed in the positive ion mode. Within-day and between-day imprecision were evaluated for levetiracetam using three levels of in-house controls. Reliability and accuracy of this method were assessed by comparison of targets with external QC material (ChromSystems), between laboratory comparisons and by recovery studies. Results: Within-day coefficients of variation (CVs) were < 6.1% and between-day CVs were < 8.2%. The average spiked recoveries of levetiracetam added to the drug-free human plasma samples were 108% at low concentration level and 103% at high concentration level. Conclusions: The method was found both specific and sensitive for the rapid and accurate measurement of levetiracetam in human matrices and correlated well with the Quest/Chantilly tandem mass spectrometric method (r = 0.983).
KW - HPLC
KW - Levetiracetam
KW - Tandem mass spectrometry
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U2 - 10.1016/j.cca.2006.06.022
DO - 10.1016/j.cca.2006.06.022
M3 - Article
C2 - 16914128
AN - SCOPUS:33750066632
SN - 0009-8981
VL - 375
SP - 115
EP - 118
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
IS - 1-2
ER -