Determination of 4-methylpyrazole in plasma using solid phase extraction and HPLC.
A rapid method for the determination of 4-methylpyrazole in plasma is described. Internal standard (3-methylpyrazole) is added to the plasma which is subsequently applied to a BondElut SCX column. After washing the column the 3- and 4-methylpyrazoles are eluted with a phosphate buffer and analyzed by isocratic reversed-phase high performance liquid chromatography with UV detection. The concentration of 4-methylpyrazole is calculated from the 4-methylpyrazole/3-methylpyrazole peak height ratio. The method is linear between 2.5 and 100 mumol/L with a within-day precision (CV) of 2.2% (n = 10) and a day-to-day precision of 2.8% (n = 30). The sensitivity is sufficient for analysis of plasma levels in the low micromolar range.
D002851:Chromatography, High Pressure Liquid; D000077604:Fomepizole; D006801:Humans; D011720:Pyrazoles / Q000097:blood*
D011720:Pyrazoles; D000077604:Fomepizole