Abstract:
:A reversed-phase HPLC method is reported for simultaneous quantitation of methylprednisolone (MP), MP succinate (MPS), and endogenous corticosterone (CST) in plasma of rats. Additionally, the 11-keto metabolite of MP (methylprednisone, MPN) is resolved from the other analytes. After addition of internal standard (triamcinolone acetonide: IS) and an initial clean up step, the analytes of interest are extracted into methylene chloride. The steroids are then resolved on a reversed-phase polymer column using a mobile phase of 0.1 M acetate buffer (pH 5.7): acetonitrile (77:23) which is pumped at a flow rate of 1.5 ml min-1. Sample detection was accomplished using an UV detector at a wavelength of 250 nm. All the five components (MPS, MP, MPN, CST and IS) were baseline resolved from each other and other components of plasma. Linear relationships were found between the steroids: IS peak area ratios and plasma concentrations in the range of 0.1-4 mircog ml-1 for MP and MPS and 0.1-1.0 microg ml-1 for MPN and CST. The assay is accurate as intra- and inter-run error values were < +/- 8% for all the components. Further, the intra- and inter-run CVs of the assay were < 16% at all the concentrations and for all the components. The application of the assay was demonstrated after the injection of a single 5 mg kg-1 (MP equivalent) dose of MPS or a macromolecular prodrug of MP to rats.
journal_name
J Pharm Biomed Analjournal_title
Journal of pharmaceutical and biomedical analysisauthors
Mehvar R,Dann RO,Hoganson DAdoi
10.1016/s0731-7085(00)00253-3subject
Has Abstractpub_date
2000-07-01 00:00:00pages
1015-22issue
6eissn
0731-7085issn
1873-264Xpii
S0731708500002533journal_volume
22pub_type
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