A simple assay for determination of levetiracetam in rat dried blood spots by LC–MS/MS

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A simple LC–MS/MS method was developed and validated for the quantification of levetiracetam (LEV, Keppra®), a broad-spectrum antiepileptic drug (AED) in rat dried blood spots (DBS). LEV was simply extracted with methanol spiked with adenosine (ADE) as IS before LC–MS/MS analysis. The correlation between the DBS and plasma concentrations of LEV was also determined.


Linearity was from 0.067–60 µg/ml for LEV in DBS samples. The intra- and inter-day accuracy and precision of the assay met validation acceptance criteria. The developed assay was applied to monitor levetiracetam DBS levels in Sprague–Dawley rats after intravenous administration. DBS concentrations were well correlated to the plasma concentrations (R2 = 0.9399), as fraction of LEV bound to blood cells remains very constant (0.466 ± 0.041) over a wide concentration range.


The study illustrated that DBS could be used as alternative matrix for monitoring LEV in preclinical studies.

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