Simultaneous quantification of seven prostanoids using liquid chromatography/tandem mass spectrometry: the effects of arachidonic acid on prostanoid production in mouse bone marrow-derived mast cells.
We have developed a method for the simultaneous estimation of the levels of the prostanoids 6-keto prostaglandin (PG) Flalpha, PGB2, PGD2, PGE2, PGF2(alpha), PGJ2, and thromboxane (TX) B2 in blood- or serum-containing medium using liquid chromatography-tandem mass spectrometry. These prostanoids and their deuterium derivatives, which were used as internal standards, were subjected to solid-phase extraction using Empore C18 HD disk cartridges and analyzed in the selected reaction-monitoring mode. A linear response curve starting at 10 pg of prostanoid/tube was observed for each prostanoid. The accuracy of the method was demonstrated with samples containing known amounts of the prostanoids. Furthermore, we used this method to analyze the prostanoids produced in mouse bone marrow-derived mast cells stimulated with arachidonic acid, which resulted in the production of PGD2, PGE2, PGF2alpha, and TXB2. The results suggest that this simultaneous quantification method is useful for the analysis of the production of biomedically important prostanoids.
Hishinuma
Takanori
T
Graduate School of Pharmaceutical Sciences, Tohoku University, and Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai, Japan.
Suzuki
Kaori
K
Saito
Masayoshi
M
Yamaguchi
Hiroaki
H
Suzuki
Naoto
N
Tomioka
Yoshihisa
Y
Kaneko
Izumi
I
Ono
Masao
M
Goto
Junichi
J
D000818:Animals; D016718:Arachidonic Acid / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000166:cytology; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D008297:Male; D013058:Mass Spectrometry; D008407:Mast Cells / Q000737:chemistry* / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D011453:Prostaglandins / Q000032:analysis* / Q000145:classification / Q000378:metabolism; D012680:Sensitivity and Specificity
D011453:Prostaglandins; D016718:Arachidonic Acid