High-performance liquid chromatography analysis, preliminary pharmacokinetics, metabolism and renal excretion of methylprednisolone with its C6 and C20 hydroxy metabolites in multiple sclerosis patients receiving high-dose pulse therapy

• Published in: Journal of chromatography. B, Biomedical sciences and applications Biomedical sciences and applications, 1999-09, Vol.732 (2), p.337-348
• Main Authors: Vree, Tom B., Lagerwerf, Aart J., Verwey-van Wissen, Corrien P.W.G.M., Jongen, P.Jozef H.
• Format: Article
• Language: English
• Subjects: Anti-Inflammatory Agents - metabolism; Anti-Inflammatory Agents - pharmacokinetics; Anti-Inflammatory Agents - therapeutic use; Biological and medical sciences; Bones, joints and connective tissue. Antiinflammatory agents; Chromatography, High Pressure Liquid - methods; Female; Humans; Kidney - metabolism; Medical sciences; Metabolic Clearance Rate; Methylprednisolone; Methylprednisolone - metabolism; Methylprednisolone - pharmacokinetics; Methylprednisolone - therapeutic use; Multiple Sclerosis - blood; Multiple Sclerosis - drug therapy; Multiple Sclerosis - metabolism; Multiple Sclerosis - urine; Pharmacology. Drug treatments
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(99)00292-3

A gradient eluent HPLC analysis in human plasma and urine was developed and validated for methylprednisolone (MP), its prodrug methylprednisolone-21-hemisuccinate (MPS) with the metabolites 6β-hydroxy-6α-methylprednisolone (MPA), 20-hydroxymethylprednisolone (MPC), 6β-hydroxy-20α-hydroxymethylprednisolone (MPB), 6β-hydroxy-20β-hydroxymethylprednisolone (MPE), 20-carboxymethylprednisolone (MPD), methylprednisolone-glucuronide (MPF) and 21-carboxymethylprednisolone (MPX). The column was Cp Spherisorb C8 5 μm, 250 mm×4.6 mm I.D. (Chrompack, Bergen op Zoom, The Netherlands) with a guard column 75 mm×2.1 mm, packed with pellicular reversed-phase. The eluent was a mixture of acetonitrile and 0.067 M KH 2PO 4 buffer, pH 4.5. At t=0, the eluent consisted of 2% acetonitrile and 98% buffer (v/v). Over the following 35 min the eluent changed linearly until it attained a composition of 50% acetonitrile and 50% buffer (v/v). At 37 min ( t=37) the eluent was changed over 5 min to the initial composition, followed by equilibration over 3 min. The flow-rate was 1.5 ml/min and UV detection was achieved at 248 nm. Preliminary pharmacokinetic data were obtained from one patient who showed illustrative plasma concentration–time curves and renal excretion-time profiles after a short-lasting infusion (0.5 h) of 1 g of methylprednisolone hemisuccinate. The half-life of prodrug methylprednisolone-21-hemisuccinate (MPS) was 0.3 h, that of metabolite MPX (21-carboxy MP) was 0.4 h and that of the parent drug methylprednisolone (MP) was 1.4 h. The half-lives of the metabolites are almost similar (4 h). The main compounds in the urine are methylprednisolone hemisuccinate (prodrug, 15.0%), methylprednisolone (parent drug, 14.6%), metabolite MPD (20-carboxy, 11.7%), and metabolite MPB (13.2%). The renal clearance values of metabolites MPB, MPC and MPD are approximately 500 ml/min, that of MP is 100 ml/min.