Abstract
In the present study a selective determination of Thromboxane B2 (TXB2) and 6-Keto-Prostag, landin F1α (6KF) in plasma was performed using the improved resolving power of highperformance thin layer chromatography (HPTLC) and sensitive radioimmunoassay.
TXB2 and 6KF were extracted with ethyl acetate pH3.0, applied on a HPTLC plate and developped with ethyl acetate/acetic acid. The Rf values of TXB2 and 6KF were 0.37 and 0.14 respectively and they could be separated from prostaglandin A2 (PGA2), PGB2, PGE2 and PGF2α. The recoveries at each step of extraction by ethyl acetate and HPTLC fractionation were about 95% and 80%, and total recovery was about 72%. Normal ranges of TXB2 and 6KF by this method were 154.6±34.5 (SD, n=24)pg/ml and 123.3±46.9 (SD, n=15)pg/ml respectively.
Plasma TXB2 concentration in cerebral ischemia measured by this method rose appreciably in relation with its attack.
The features of the present method are the following: 1) The purification procedure by ion exchange column chromatography prior to HPTLC can be eliminated. 2) Simultaneous separation of TXB2, 6KF, other prostaglandins and contaminants can be achieved on a HPTLC plate by ethyl acetate/acetic acid. 3) The simplified procedure brought a considerable reduction in time.
