Triazolines. XXIII. High-performance liquid chromatographic assay in rat blood for a novel triazoline anticonvulsant (ADD17014).
A sensitive and specific high-performance liquid chromatographic (HPLC) method for the analysis of 1-(4-chlorophenyl)-5-(4-pyridyl)-delta 2-1,2,3-triazoline (ADD17014, I), a novel anticonvulsant agent, in rat blood is described. Compound I and the internal standard (dipyridamole) were extracted into diethyl ether (5 ml) from alkalinised blood (0.25 ml of blood plus 0.75 ml of pH 10.7 buffer), with extractability nearing 100% under these conditions. The assay is based on reversed-phase HPLC (25 cm x 0.46 cm I.D. Spherisorb 5-ODS) using a mobile phase of methanol-acetonitrile-McIlvaine's citric acid-phosphate buffer (pH 8.0, 0.005 M) (30:30:40, v/v) and ultraviolet detection at 290 nm. Calibration curves were linear and reproducible (correlation coefficient greater than 0.999). Measurement of I in rat blood (250 microliters sample size) was linear in the range 0-40 microgram/ml and the coefficient of variation was less than 5%. The minimum detectable level was about 0.1 microgram/ml; however, a larger blood sample size (1-2 ml) allowed measurement of levels as low as 10 ng/ml, especially for estimation of drug levels in samples withdrawn at later time points (24 h).
Stevenson
P J
PJ
Chelsea Department of Pharmacy, King's College London, U.K.
Hamelijnck
M A
MA
Kadaba
P K
PK
Damani
L A
LA
D000818:Animals; D000927:Anticonvulsants / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D014230:Triazoles / Q000097:blood*; D014466:Ultraviolet Rays
D000927:Anticonvulsants; D014230:Triazoles; C055990:ADD 17014