Simultaneous determination of Levetiracetam and its acid metabolite (ucb L057) in serum/plasma by liquid chromatography tandem mass spectrometry

Damodara Rao Mendu, Steven J. Soldin

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Objective: Levetiracetam and its acid metabolite have almost identical MRMs. They therefore need to be separated chromatographically prior to quantitation. Research design and methods: The sample is deproteinized with acetonitrile containing Ritonavir as internal standard, centrifuged and the supernatant diluted with water (1:2 v/v). Sixty microliters of the supernatant is injected into the LC-MS/MS and Levetiracetam (LEV) and LEV metabolite separated chromatographically at room temperature employing a Supelco C18 column and a 0.1% formic acid methanol gradient at pH of 2.5. Results: The retention times for LEV metabolite, LEV and Ritonavir were 4.50, 5.38 and 9.18 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 0-50 μg/mL for LEV and 0.0-5.0 μg/mL for LEV metabolite. Intra- and inter-run imprecision (n = 10) gave CVs of 2.3-4.7%, 3.4-8.9% for LEV and 2.9-3.9%, 3.3-7.4% for LEV metabolite. Recoveries of both LEV and LEV metabolite were close to 100%. Results for LEV were compared with those obtained by a commercial reference laboratory (r = 0.974). Conclusion: The procedure is reliable, quick, and inexpensive. LEV and LEV metabolite co-elute using C-18 columns at pHs > 3.0 and previously published methods employing these conditions could therefore be subject to metabolite interference. In this method LEV and LEV metabolite are separated at pH 2.5. The total run time including the washing step is 10 min/sample, making this method suitable when moderate throughput is needed such as in clinical or commercial reference laboratories.

Original languageEnglish (US)
Pages (from-to)485-489
Number of pages5
JournalClinical Biochemistry
Volume43
Issue number4-5
DOIs
StatePublished - Mar 2010
Externally publishedYes

Fingerprint

etiracetam
Liquid chromatography
Metabolites
Tandem Mass Spectrometry
Liquid Chromatography
Mass spectrometry
Plasmas
Acids
Serum
Ritonavir
formic acid

Keywords

  • Levetiracetam
  • Levetiracetam metabolite antiepileptic drug
  • Tandem mass spectrometry

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Simultaneous determination of Levetiracetam and its acid metabolite (ucb L057) in serum/plasma by liquid chromatography tandem mass spectrometry. / Mendu, Damodara Rao; Soldin, Steven J.

In: Clinical Biochemistry, Vol. 43, No. 4-5, 03.2010, p. 485-489.

Research output: Contribution to journalArticle

@article{4e191423fdbc482c804a0ba6edfadc9e,
title = "Simultaneous determination of Levetiracetam and its acid metabolite (ucb L057) in serum/plasma by liquid chromatography tandem mass spectrometry",
abstract = "Objective: Levetiracetam and its acid metabolite have almost identical MRMs. They therefore need to be separated chromatographically prior to quantitation. Research design and methods: The sample is deproteinized with acetonitrile containing Ritonavir as internal standard, centrifuged and the supernatant diluted with water (1:2 v/v). Sixty microliters of the supernatant is injected into the LC-MS/MS and Levetiracetam (LEV) and LEV metabolite separated chromatographically at room temperature employing a Supelco C18 column and a 0.1{\%} formic acid methanol gradient at pH of 2.5. Results: The retention times for LEV metabolite, LEV and Ritonavir were 4.50, 5.38 and 9.18 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 0-50 μg/mL for LEV and 0.0-5.0 μg/mL for LEV metabolite. Intra- and inter-run imprecision (n = 10) gave CVs of 2.3-4.7{\%}, 3.4-8.9{\%} for LEV and 2.9-3.9{\%}, 3.3-7.4{\%} for LEV metabolite. Recoveries of both LEV and LEV metabolite were close to 100{\%}. Results for LEV were compared with those obtained by a commercial reference laboratory (r = 0.974). Conclusion: The procedure is reliable, quick, and inexpensive. LEV and LEV metabolite co-elute using C-18 columns at pHs > 3.0 and previously published methods employing these conditions could therefore be subject to metabolite interference. In this method LEV and LEV metabolite are separated at pH 2.5. The total run time including the washing step is 10 min/sample, making this method suitable when moderate throughput is needed such as in clinical or commercial reference laboratories.",
keywords = "Levetiracetam, Levetiracetam metabolite antiepileptic drug, Tandem mass spectrometry",
author = "Mendu, {Damodara Rao} and Soldin, {Steven J.}",
year = "2010",
month = "3",
doi = "10.1016/j.clinbiochem.2009.11.008",
language = "English (US)",
volume = "43",
pages = "485--489",
journal = "Clinical Biochemistry",
issn = "0009-9120",
publisher = "Elsevier Inc.",
number = "4-5",

}

TY - JOUR

T1 - Simultaneous determination of Levetiracetam and its acid metabolite (ucb L057) in serum/plasma by liquid chromatography tandem mass spectrometry

AU - Mendu, Damodara Rao

AU - Soldin, Steven J.

PY - 2010/3

Y1 - 2010/3

N2 - Objective: Levetiracetam and its acid metabolite have almost identical MRMs. They therefore need to be separated chromatographically prior to quantitation. Research design and methods: The sample is deproteinized with acetonitrile containing Ritonavir as internal standard, centrifuged and the supernatant diluted with water (1:2 v/v). Sixty microliters of the supernatant is injected into the LC-MS/MS and Levetiracetam (LEV) and LEV metabolite separated chromatographically at room temperature employing a Supelco C18 column and a 0.1% formic acid methanol gradient at pH of 2.5. Results: The retention times for LEV metabolite, LEV and Ritonavir were 4.50, 5.38 and 9.18 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 0-50 μg/mL for LEV and 0.0-5.0 μg/mL for LEV metabolite. Intra- and inter-run imprecision (n = 10) gave CVs of 2.3-4.7%, 3.4-8.9% for LEV and 2.9-3.9%, 3.3-7.4% for LEV metabolite. Recoveries of both LEV and LEV metabolite were close to 100%. Results for LEV were compared with those obtained by a commercial reference laboratory (r = 0.974). Conclusion: The procedure is reliable, quick, and inexpensive. LEV and LEV metabolite co-elute using C-18 columns at pHs > 3.0 and previously published methods employing these conditions could therefore be subject to metabolite interference. In this method LEV and LEV metabolite are separated at pH 2.5. The total run time including the washing step is 10 min/sample, making this method suitable when moderate throughput is needed such as in clinical or commercial reference laboratories.

AB - Objective: Levetiracetam and its acid metabolite have almost identical MRMs. They therefore need to be separated chromatographically prior to quantitation. Research design and methods: The sample is deproteinized with acetonitrile containing Ritonavir as internal standard, centrifuged and the supernatant diluted with water (1:2 v/v). Sixty microliters of the supernatant is injected into the LC-MS/MS and Levetiracetam (LEV) and LEV metabolite separated chromatographically at room temperature employing a Supelco C18 column and a 0.1% formic acid methanol gradient at pH of 2.5. Results: The retention times for LEV metabolite, LEV and Ritonavir were 4.50, 5.38 and 9.18 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 0-50 μg/mL for LEV and 0.0-5.0 μg/mL for LEV metabolite. Intra- and inter-run imprecision (n = 10) gave CVs of 2.3-4.7%, 3.4-8.9% for LEV and 2.9-3.9%, 3.3-7.4% for LEV metabolite. Recoveries of both LEV and LEV metabolite were close to 100%. Results for LEV were compared with those obtained by a commercial reference laboratory (r = 0.974). Conclusion: The procedure is reliable, quick, and inexpensive. LEV and LEV metabolite co-elute using C-18 columns at pHs > 3.0 and previously published methods employing these conditions could therefore be subject to metabolite interference. In this method LEV and LEV metabolite are separated at pH 2.5. The total run time including the washing step is 10 min/sample, making this method suitable when moderate throughput is needed such as in clinical or commercial reference laboratories.

KW - Levetiracetam

KW - Levetiracetam metabolite antiepileptic drug

KW - Tandem mass spectrometry

UR - http://www.scopus.com/inward/record.url?scp=76749105200&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=76749105200&partnerID=8YFLogxK

U2 - 10.1016/j.clinbiochem.2009.11.008

DO - 10.1016/j.clinbiochem.2009.11.008

M3 - Article

C2 - 19941845

AN - SCOPUS:76749105200

VL - 43

SP - 485

EP - 489

JO - Clinical Biochemistry

JF - Clinical Biochemistry

SN - 0009-9120

IS - 4-5

ER -