title: Simultaneous quantification of prostaglandins in human synovial cell-cultured medium using liquid chromatography/tandem mass spectrometry.
abstract: A liquid chromatographic-tandem mass spectrometric (LC/MS-MS) method was developed for the simultaneous quantification of prostaglandin (PG) E(2), PGF(2alpha), 6-keto-PGF(lalpha) and thromboxane (TX) B(2). These eicosanoids and their deuterium derivatives, using as internal standards, were extracted by solid-phase extraction and analyzed using LC/MS-MS in the selected reaction-monitoring (SRM) mode. A good linear response over the range of 10 pg to 10 ng for each eicosanoid was demonstrated. The accuracy of added eicosanoids ranged from 94.1 to 106.6% and coefficients of variation ranged from 0.62 to 7.8%. Furthermore, we applied this method for the determination of eicosanoids in the human synovial cell-cultured medium, stimulated by lipopolysaccharide (LPS). LPS produced each eicosanoid and they increased in a time-dependent manner. The production levels after 24 h stimulation were 6-keto-PGF(1alpha) > PGE(2) > TXB(2) >> PGF(2alpha). This simultaneous quantification method is so useful to clarify the function of synovial cells in rheumatoid arthritis (RA).
authors.affiliation: Department of Pharmaceutical Sciences, Tohoku University Hospital, Aoba-ku, Sendai, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Takabatake
authors.forename: T
authors.initials: T
authors.lastname: Hishinuma
authors.forename: N
authors.initials: N
authors.lastname: Suzuki
authors.forename: S
authors.initials: S
authors.lastname: Chiba
authors.forename: H
authors.initials: H
authors.lastname: Tsukamoto
authors.forename: H
authors.initials: H
authors.lastname: Nakamura
authors.forename: T
authors.initials: T
authors.lastname: Saga
authors.forename: Y
authors.initials: Y
authors.lastname: Tomioka
authors.forename: A
authors.initials: A
authors.lastname: Kurose
authors.forename: T
authors.initials: T
authors.lastname: Sawai
authors.forename: M
authors.initials: M
authors.lastname: Mizugaki
mesh_terms: D002138:Calibration; D002460:Cell Line; D002853:Chromatography, Liquid; D017077:Culture Media, Conditioned / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry; D011453:Prostaglandins / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013582:Synovial Fluid / Q000166:cytology* / Q000378:metabolism*
chemical_list: D017077:Culture Media, Conditioned; D011453:Prostaglandins