Simultaneous quantification of prostaglandins in human synovial cell-cultured medium using liquid chromatography/tandem mass spectrometry.
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).
Takabatake
M
M
Department of Pharmaceutical Sciences, Tohoku University Hospital, Aoba-ku, Sendai, Japan.
Hishinuma
T
T
Suzuki
N
N
Chiba
S
S
Tsukamoto
H
H
Nakamura
H
H
Saga
T
T
Tomioka
Y
Y
Kurose
A
A
Sawai
T
T
Mizugaki
M
M
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*
D017077:Culture Media, Conditioned; D011453:Prostaglandins