Simultaneous liquid-chromatographic determination of prednisone and prednisolone in plasma.
This high-performance liquid-chromatographic (HPLC) method for simultaneous determination of prednisone and its metabolite, prednisolone, in plasma is a modification of the method of Frey et al. (Clin Chem 1979;25:1944-7). Heparinized plasma (1.0 mL) with 0.1 mL of internal standard solution (11-deoxy-17-hydroxycorticosterone, 2 mg/L) is extracted with 7.0 mL of dichloromethane, then washed sequentially with 0.1 mol/L HCl, 0.1 mol/L NaOH, and deionized water, 2.0 mL each. The extract is evaporated and the residue reconstituted with 75 microL of mobile phase, methanol/H2O (40/60 by vol). Thirty microliters of this is injected onto a reversed-phase C6 column, which is eluted at 1.4 mL/min. Analytical recoveries of prednisone and prednisolone were 94-98% and 102-106%, respectively. Day-to-day precision (CV) was 3.8% for prednisone, 6.1% for prednisolone. We encountered no interference from the 21 other steroids and 25 drugs tested. This method is simple, accurate, and precise.
Clinical Laboratory, Childrens Hospital, Los Angeles, CA 90027.
M H
MH
Cheng
W Y
WY
Huang
A I
AI
Lipsey
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D010672:Phenytoin / Q000097:blood; D011239:Prednisolone / Q000097:blood*; D011241:Prednisone / Q000097:blood*; D011786:Quality Control
D010672:Phenytoin; D011239:Prednisolone; D011241:Prednisone