TY - JOUR
T1 - Quantification of intracellular and extracellular eicosapentaenoic acid-derived 3-series prostanoids by liquid chromatography/electrospray ionization tandem mass spectrometry
AU - Tanaka, Nobuaki
AU - Yamaguchi, Hiroaki
AU - Furugen, Ayako
AU - Ogura, Jiro
AU - Kobayashi, Masaki
AU - Yamada, Takehiro
AU - Mano, Nariyasu
AU - Iseki, Ken
N1 - Funding Information:
This work was in part supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant number 25460184 (Hiroaki Yamaguchi (HY)) and the Towa Foundation for Food Research (HY) .
PY - 2014/9
Y1 - 2014/9
N2 - 3-Series prostanoids are bioactive lipid mediators synthesized from eicosapentaenoic acid (EPA). Determination of intracellular and extracellular levels of prostanoids is needed to elucidate the mechanism of action, and we therefore developed a method for quantification of intracellular and extracellular levels of 3-series prostanoids (including prostaglandin E3 (PGE3), PGD3, PGF3α, thromboxane B3 (TXB3), and δ17-6-keto PGF1α) by using liquid chromatography/electrospray ionization tandem mass spectrometry. The separation of prostanoids was performed with a CAPCELL PAK C18 MG II column (2.0mm×150mm, 3μm) with an isocratic flow of acetonitrile/water/acetic acid (40:60:0.1, v/v/v). This method was validated for measurement of both extracellular and intracellular samples with high levels of precision and accuracy. We applied this method to human lung epithelial A549 cells stimulated with calcium ionophore A23187 under the condition of arachidonic acid or EPA treatment and we could measure PGE3 in both intracellular and extracellular samples.
AB - 3-Series prostanoids are bioactive lipid mediators synthesized from eicosapentaenoic acid (EPA). Determination of intracellular and extracellular levels of prostanoids is needed to elucidate the mechanism of action, and we therefore developed a method for quantification of intracellular and extracellular levels of 3-series prostanoids (including prostaglandin E3 (PGE3), PGD3, PGF3α, thromboxane B3 (TXB3), and δ17-6-keto PGF1α) by using liquid chromatography/electrospray ionization tandem mass spectrometry. The separation of prostanoids was performed with a CAPCELL PAK C18 MG II column (2.0mm×150mm, 3μm) with an isocratic flow of acetonitrile/water/acetic acid (40:60:0.1, v/v/v). This method was validated for measurement of both extracellular and intracellular samples with high levels of precision and accuracy. We applied this method to human lung epithelial A549 cells stimulated with calcium ionophore A23187 under the condition of arachidonic acid or EPA treatment and we could measure PGE3 in both intracellular and extracellular samples.
KW - 3-Series prostanoids
KW - Eicosapentaenoic acid
KW - Extracellular measurement
KW - Intracellular measurement
KW - LC/MS/MS
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U2 - 10.1016/j.plefa.2014.04.005
DO - 10.1016/j.plefa.2014.04.005
M3 - Article
C2 - 24996760
AN - SCOPUS:84905583293
SN - 0952-3278
VL - 91
SP - 61
EP - 71
JO - Prostaglandins Leukotrienes and Essential Fatty Acids
JF - Prostaglandins Leukotrienes and Essential Fatty Acids
IS - 3
ER -