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pISSN 2950-9114 eISSN 2950-9122
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Evaluation Brief

Lab Med Qual Assur 2022; 44(4): 226-229

Published online December 31, 2022

https://doi.org/10.15263/jlmqa.2022.44.4.226

Copyright © Korean Association of External Quality Assessment Service.

Implementation and Validation of an Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Quantifying Levetiracetam and Lamotrigine in Serum Specimens

Ahram Yi* , Jun Hyung Lee* , Hyeon Gyeong Nam , Jungsun Han , Sung-Eun Cho , Sang Gon Lee , and Eun Hee Lee

Department of Laboratory Medicine, Green Cross Laboratories (GC Labs), Yongin, Korea

Correspondence to:Sung-Eun Cho
Department of Laboratory Medicine, Green Cross Laboratories (GC Labs), 107 Ihyeon-ro 30beon-gil, Giheung-gu, Yongin 16924, Korea
Tel +82-31-260-0947
E-mail secho1206!!@gclabs.co.kr
*These two authors contributed equally to this work.

Received: July 20, 2022; Revised: August 3, 2022; Accepted: August 9, 2022

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

We validated an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for quantifying levetiracetam (LEV) and lamotrigine (LAM) in serum specimens. We developed an UHPLC-MS/MS method using Triple Quad 4500MD (SCIEX, Singapore) with 10 μM LEV-2H3 (LEV-internal standard [IS]) and 10 μM LAM 13C3, d3 (LAM-IS) as internal standards. The mass spectrometer detected the transitions from the precursor to product ions (mass-to-charge ratio [m/z] 171.0→126.0 for LEV, m/z 177.1→132.1 for LEV-IS, m/z 256.0→210.9 for LAM, and m/z 262.1→216.8 for LAM-IS, respectively). We used a Kinetex (2.6 μm C18 100Å, 100×3.0 mm; Phenomenex, USA) column on an ExionLC AD UHPLC. The UHPLC-MS/MS method yielded a linear response from 0.60 to 340.13 μg/mL for LEV (R2=0.9997) and from 0.17 to 99.98 μg/mL for LAM (R2=0.9995). The lower limits of quantification using this method were 0.60 and 0.17 μg/mL for LEV and LAM, respectively. The recovery of LEV and LAM UHPLC-MS/MS method measurements were within ±6% of the targeted values. The intra-day and inter-day coefficients of variation were all acceptable with values of less than 7% in both cases. Carry-over was not observed in any of the results. Ion suppression or enhancement was not observed in the blank and six samples for both LEV and LAM test results. The UHPLC-MS/MS LEV and LAM assay showed adequate recovery, precision, and sensitivity, and an adequate AMR, and thus is suitable for routine clinical work.

Keywords: Levetiracetam, Lamotrigine, Liquid chromatography-tandem mass spectrometry, Method validation

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