Analysis of the thromboxane/prostacyclin balance in human urine by gas chromatography/selected ion monitoring: abnormalities in diabetics.
We microanalyzed 2,3-dinor-6-keto-prostaglandin F1 alpha (2,3-dinor-6-keto-PGF1 alpha 1) and 11-dehydrothromboxane B2 (11-dehydro-TXB2, 2) in human urine. Samples containing a [2H4]-analogue as an internal standard were extracted by chromatography using Sep Pak tC18 and silica gel. The compounds were then analysed by means of the lactone ring opening reaction and dimethylisopropylsilylation. The conversion of 1 to 1-methyl ester (ME)-propylamide (PA)-9, 12, 15-dimethylisopropylsilyl (DMIPS) ether derivative and of 2 to 1-ME-6-methoxime (MO)-9, 12, 15-tris-DMIPS ether derivative was followed by gas chromatography/selected ion monitoring (GC/SIM). Interfering substances from the urine matrix were eliminated during GC/SIM analysis using a DB-5 column. We were able to detect 1 (222-1031 pg/mg creatinine) and 2 (18-155 pg/mg creatinine) in human urine. Furthermore, the thromboxane/prostacyclin (IX/PGI) ratio in the urine of diabetics was higher than that of healthy volunteers. This method can be used to determine the TX/PGI balance in human urine.
Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai, Japan.
T
T
Hishinuma
G S
GS
Yu
M
M
Takabatake
Y
Y
Nakagawa
K
K
Ito
M
M
Nishikawa
M
M
Ishibashi
K
K
Suzuki
M
M
Matsumoto
T
T
Toyoda
M
M
Mizugaki
D015121:6-Ketoprostaglandin F1 alpha / Q000031:analogs & derivatives / Q000652:urine; D000328:Adult; D002849:Chromatography, Gas / Q000379:methods*; D003903:Deuterium; D003924:Diabetes Mellitus, Type 2 / Q000652:urine*; D011464:Epoprostenol / Q000652:urine*; D005260:Female; D006801:Humans; D008297:Male; D012016:Reference Values; D013929:Thromboxane B2 / Q000031:analogs & derivatives* / Q000652:urine
D013929:Thromboxane B2; D015121:6-Ketoprostaglandin F1 alpha; C028290:2,3-dinor-6-ketoprostaglandin F1alpha; C049235:11-dehydro-thromboxane B2; D003903:Deuterium; D011464:Epoprostenol