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Urinary profile of methylprednisolone acetate metabolites in patients following intra-articular and intramuscular administration

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Abstract

A study on urinary metabolites of methylprednisolone acetate (MPA) has been performed by liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS) in precursor ion scanning (PIS) and neutral loss (NL) modes. Patients suffering from joint inflammation have been treated with Depo-Medrol® (MPA marketed suspension, 40 mg) intra-articularly (IA) and after a wash-out period, intramuscularly (IM) at the same dose. Urine samples have been collected after both the administration routes. Metabolites were identified in PIS mode by setting the fragment ion at m/z 161 which is specific for MPA, methylprednisolone (MP), methylprednisolone hemisuccinate, and in NL mode by selecting the losses of 54, 72, 176 and 194 Da. The MP-related structure of each target ion detected in both the MS modes was then confirmed by MS/MS acquisitions, and by accurate mass experiments. By using this approach, 13 MPA metabolites (M1–M13) have been identified, nine already reported in the literature and four unknown and for which the chemical structures have been proposed. No differences in the metabolic pattern of MPA when administered IM or IA were observed. The relative abundances of metabolites compared with the internal standard (MP-D2) were monitored by multiple reaction monitoring analysis for 19 days after both the administration routes.

Urine of patients before (dotted lines) and after (1st day, thick lines) the IM administration of Depo-Medrol®. Extracted ion chromatograms of glucuronic metabolites of MP with [M+H]+ at m/z 551. PIS m/z 161 and NL 194 Da traces (25 and 20 eV collision energies).

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Abbreviations

MP:

Methylprednisolone

MPA:

Methylprednisolone acetate

LC–ESI–MS/MS:

Liquid chromatography–electrospray ionization–tandem mass spectrometry

LC–APCI–MS/MS:

Liquid chromatography–atmospheric pressure chemical ionization–tandem mass spectrometry

PIS:

Precursor ion scanning

NL:

Neutral loss

MRM:

Multiple reaction monitoring

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Acknowledgements

The Italian National Institute of Health (ISS) is thanked for the financial support to A.P. during her Doctorate leave at the University of Milan. This work was supported by the Italian Ministry of Health (Progetto di Ricerca 2006 sui farmaci, sulle sostanze e sulle pratiche mediche utilizzabili a fini doping nelle attività sportive) and was carried out at the University of Milan.

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Authors and Affiliations

  1. Dipartimento del Farmaco, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy

    Alessia Panusa

  2. Dipartimento di Scienze Farmaceutiche “P. Pratesi”, Universita’ degli Studi di Milano, 20133, Milan, Italy

    Luca Regazzoni, Giancarlo Aldini, Marica Orioli, Paola Minghetti & Marina Carini

  3. Istituto di Medicina e Scienza dello Sport del CONI, Largo Piero Gabrielli 1, 00197, Rome, Italy

    Arrigo Giombini & Carlo Tranquilli

Authors
  1. Alessia Panusa
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  2. Luca Regazzoni
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  3. Giancarlo Aldini
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  4. Marica Orioli
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  5. Arrigo Giombini
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  6. Paola Minghetti
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  7. Carlo Tranquilli
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  8. Marina Carini
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Correspondence to Alessia Panusa.

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Panusa, A., Regazzoni, L., Aldini, G. et al. Urinary profile of methylprednisolone acetate metabolites in patients following intra-articular and intramuscular administration. Anal Bioanal Chem 400, 255–267 (2011). https://doi.org/10.1007/s00216-011-4744-6

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  • DOI: https://doi.org/10.1007/s00216-011-4744-6

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