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; Eun Hee Lee

doi:10.15263/jlmqa.2022.44.4.226

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자료유형: 학술저널

저자정보: Ahram Yi (Green Cross Laboratories (GC Labs)) Jun Hyung Lee (Green Cross Laboratories (GC Labs)) Hyeon Gyeong Nam (Green Cross Laboratories (GC Labs)) Jungsun Han (Green Cross Laboratories (GC Labs)) Sung-Eun Cho (Green Cross Laboratories (GC Labs)) Sang Gon Lee (Green Cross Laboratories (GC Labs)) Eun Hee Lee (Green Cross Laboratories (GC Labs))

저널정보: 대한임상검사정도관리협회 Journal of Laboratory Medicine And Quality Assurance Laboratory Medicine and Quality Assurance 제44권 제4호

발행연도: 2022.12

수록면: 226 - 229 (4page)

DOI: 10.15263/jlmqa.2022.44.4.226

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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 UHPLCMS/ 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 (R 2=0.9997) and from 0.17 to 99.98 μg/mL for LAM (R 2=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.

#Levetiracetam #Lamotrigine #Liquid chromatography-tandem mass spectrometry #Method validation

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