Sensitive fluorometric detection of prostaglandins by high performance liquid chromatography after precolumn labelling with 4-(N,N-dimethylaminosulphonyl)-7-(1-piperazinyl)-2,1,3-benzoxadiazole (DBD-PZ).
A highly sensitive and simple method for the determination of prostaglandins (PGs) by HPLC with fluorescence detection is described. PGs are converted to the corresponding fluorescence derivatives by the reaction with 4-(N,N-dimethylaminosulphonyl)-7-(1-piperazinyl)-2,1,3-benzoxadiaz ole (DBD-PZ) in the presence of 2,2'-dipyridyl disulphide and triphenyl phosphine in acetonitrile. The reaction is completed at room temperature after 30 min. The DBD derivatives of nine PGs are separated within a single 45 min chromatographic run on a reversed phase ODS column with a linear gradient elution using water and acetonitrile. The detection limits (signal-to-noise ratio of 3) calculated from the standard mixture of PGs (6-keto-F1 alpha, F1 alpha, F2 alpha, E1, E2, D2, limaprost, A1 and B1) are in the range 1.7-5.0 fmol. The applicability of the proposed procedure is evaluated to the detection of PGs added to rat plasma.
Toyo'oka
T
T
Division of Drugs, National Institute of Hygienic Sciences, Tokyo, Japan.
Ishibashi
M
M
Terao
T
T
Imai
K
K
D000818:Animals; D001583:Benzoxazoles; D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D010069:Oxadiazoles; D010879:Piperazines; D011453:Prostaglandins / Q000097:blood*; D051381:Rats; D013050:Spectrometry, Fluorescence; D013449:Sulfonamides
D001583:Benzoxazoles; D007202:Indicators and Reagents; D010069:Oxadiazoles; D010879:Piperazines; D011453:Prostaglandins; D013449:Sulfonamides; C076762:4-(N,N-dimethylaminosulphonyl)-7-(1-piperazinyl)-2,1,3-benzoxadiazole