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Quantification of new antiepileptic drugs by liquid chromatography/electrospray ionization tandem mass spectrometry and its application to cellular uptake experiment using human placental choriocarcinoma BeWo cells.

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Title: Quantification of new antiepileptic drugs by liquid chromatography/electrospray ionization tandem mass spectrometry and its application to cellular uptake experiment using human placental choriocarcinoma BeWo cells.
Authors: Furugen, Ayako Browse this author →KAKEN DB
Kobayashi, Masaki Browse this author →KAKEN DB
Nishimura, Ayako Browse this author
Takamura, Shigeo Browse this author
Narumi, Katsuya Browse this author →KAKEN DB
Yamada, Takehiro Browse this author
Iseki, Ken Browse this author →KAKEN DB
Keywords: Cellular uptake experiment
Gabapentin
Lamotrigine
Levetiracetam
Liquid chromatography/tandem mass spectrometry
Topiramate
Issue Date: 1-Oct-2015
Journal Title: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
Volume: 1002
Start Page: 228
End Page: 233
Publisher DOI: 10.1016/j.jchromb.2015.08.031
PMID: 26343016
Abstract: A method for quantification of new antiepileptic drugs, including lamotrigine (LTG), levetiracetam (LEV), gabapentin (GBP), and topiramate (TPM), in cellular samples, using liquid chromatography/electrospray ionization tandem mass spectrometry was developed to better understand the membrane transport mechanisms of these drugs. Cell lysate was deproteinized by methanol containing LEV-d3 as an internal standard (IS). Chromatographic separation was performed on a C18 column using gradient elution with methanol-water-formic acid (10:90:0.1, v/v/v) and methanol-formic acid (100:0.1, v/v). Analytes were detected in positive ion electrospray mode with selected reaction monitoring (SRM). This method was applicable for a linear range of 5 to 500pmol for LTG; 5 to 1000pmol for LEV; 10 to 10,000pmol for GBP; and 5 to 5000pmol for TPM. The intra-day precision, inter-day precision, and accuracy data were assessed and found to be acceptable. This developed and validated method was then successfully applied to the investigation of uptake of the new antiepileptic drugs in placental choriocarcinoma BeWo cells. The intracellular concentration of these drugs in BeWo cells, accumulating over 30min at 37°C was in the order of GBP>LTG>LEV≈TPM. Furthermore, the uptake of GBP at 4°C was much lower than that at 37°C. The uptake of GBP was saturated at high concentrations. The kinetic parameters calculated for GBP uptake in BeWo cells were determined as Km of 105.4±6.4μM and Vmax at 8153±348pmol/mg protein/min. The novel method described here should enable investigators to elucidate the transport mechanisms of these antiepileptic drugs in BeWo cells.
Type: article (author version)
URI: http://hdl.handle.net/2115/75611
Appears in Collections:薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 古堅 彩子

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