High-performance liquid chromatographic determination of a 21-aminosteroid antioxidant in plasma.
A novel 21-aminosteroid, 21-[4-(2,6-di-1-pyrrolidinyl-4-pyrimidinyl)-1- piperazinyl]-16 alpha-methylpregna-1,4,9(11)-triene-3,20-dione monomethanesulfonate (I), is under development for the treatment of central nervous system injury in humans. This report describes a reversed-phase high-performance liquid chromatographic (RP-HPLC) method using ultraviolet detection at 254 nm for the determination of I in plasma with low nanogram per milliliter sensitivity. Plasma was deproteinated by mixing with acetonitrile and centrifuging, and I was extracted from the supernatant with disposable bonded-phase columns. The extracts were chromatographed on an octylsilane bonded-phase HPLC column with an acetonitrile-water mobile phase containing triethylammonium acetate, pH5. Quantification was accomplished by peak-height ratio analysis using an analogue of I as the assay internal standard. The method was suitable for the determination of I following a 30 mg/kg intraperitoneal dose in the rat.
Cox
J W
JW
Drug Metabolism Research, Pharmaceutical Research and Development, Upjohn Company, Kalamazoo, MI 49001.
Pullen
R H
RH
D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid; D007274:Injections, Intraperitoneal; D008297:Male; D011281:Pregnatrienes / Q000008:administration & dosage / Q000097:blood*; D051381:Rats; D011919:Rats, Inbred Strains; D013056:Spectrophotometry, Ultraviolet
D000975:Antioxidants; D011281:Pregnatrienes; C053355:tirilazad