Skip to Content
Merck
  • Detection and characterization of triamcinolone acetonide metabolites in human urine by liquid chromatography/tandem mass spectrometry after intramuscular administration.

Detection and characterization of triamcinolone acetonide metabolites in human urine by liquid chromatography/tandem mass spectrometry after intramuscular administration.

Rapid communications in mass spectrometry : RCM (2015-01-07)
Xavier Matabosch, Oscar J Pozo, Esther Papaseit, Magí Farré, Josep Marcos, Jordi Segura, Rosa Ventura
ABSTRACT

Glucocorticosteroids are prohibited in sports when used by systemic administrations (e.g. intramuscular, IM), whereas they are allowed using other ways of administration. Strategies to discriminate between administrations routes have to be developed by doping control laboratories. For this reason, the metabolism of triamcinolone acetonide (TA), one of the most used glucocorticosteroids, was studied using liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). Urine samples obtained after IM administration of TA were analyzed using two sample treatments: (a) hydrolysis with β-glucuronidase enzymes and liquid-liquid extraction under alkaline conditions, and (b) liquid-liquid extraction under acidic conditions. The extracts were analyzed by LC/MS/MS. TA, commercially available metabolites (6β-hydroxytriamcinolone acetonide, 6β-OH-TA, and triamcinolone), and their C20-reduced derivatives showed characteristic fragmentation behavior. Besides common product ions and neutral losses for corticosteroids containing fluorine, additional characteristic neutral losses (58 Da, loss of acetone; 44 Da, loss of acetaldehyde) were observed in positive electrospray ionization. Based on that behavior, two complementary approaches were applied to detect TA metabolites: (a) open detection by precursor ion and neutral loss scan methods and (b) targeted detection by selected reaction monitoring methods (SRM) containing theoretical ion transitions of the potential metabolites. Two main compounds, TA and 6β-OH-TA, and nine minor potential metabolites, were detected by open screening methods. Using SRM, two additional metabolites were detected. Some of the metabolites were characterized using reference standards and, for the rest of metabolites, feasible structures were proposed based on mass spectrometric data. Metabolites resulting from hydroxylation in C-6, oxidation of the 11-hydroxyl group, reduction of the Δ(4) double bond and oxidation of the side chain were detected. Some of them have not been previously described. Excretion profiles of the detected metabolites after IM administration are presented.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methanol, NMR reference standard
Pricing and availability is not currently available.
Triamcinolone acetonide, European Pharmacopoeia (EP) Reference Standard
Pricing and availability is not currently available.
Sigma-Aldrich
Sodium phosphate dibasic solution, BioUltra, 0.5 M in H2O
Pricing and availability is not currently available.
Supelco
Phosphate Standard for IC, TraceCERT®, 1000 mg/L phosphate in water (nominal concentration)
SKU
Pack Size
Availability
Price
Quantity
USP
Triamcinolone acetonide, United States Pharmacopeia (USP) Reference Standard
Pricing and availability is not currently available.
Sigma-Aldrich
Ammonium chloride, for molecular biology, suitable for cell culture, ≥99.5%
SKU
Pack Size
Availability
Price
Quantity
Supelco
Triamcinolone acetonide, analytical standard
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
Triamcinolone
Pricing and availability is not currently available.
Sigma-Aldrich
Methanol, anhydrous, 99.8%
SKU
Pack Size
Availability
Price
Quantity
Supelco
Methanol, analytical standard
Pricing and availability is not currently available.
Sigma-Aldrich
Ammonium chloride, tested according to Ph. Eur.
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
Ammonium chloride, 99.99% trace metals basis
Pricing and availability is not currently available.
Sigma-Aldrich
Ammonium chloride, 99.998% trace metals basis
Pricing and availability is not currently available.
Supelco
Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
Pricing and availability is not currently available.
USP
Triamcinolone, United States Pharmacopeia (USP) Reference Standard
SKU
Pack Size
Availability
Price
Quantity
Supelco
Residual Solvent - Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material
Pricing and availability is not currently available.
Triamcinolone acetonide for system suitability, European Pharmacopoeia (EP) Reference Standard
Pricing and availability is not currently available.
Dexamethasone for peak identification, European Pharmacopoeia (EP) Reference Standard
Pricing and availability is not currently available.
Supelco
Ammonium chloride, Pharmaceutical Secondary Standard; Certified Reference Material
Pricing and availability is not currently available.
Sigma-Aldrich
Ammonium chloride, BioUltra, for molecular biology, ≥99.5% (AT)
Pricing and availability is not currently available.
Sigma-Aldrich
Methanol, Absolute - Acetone free
Pricing and availability is not currently available.
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Pricing and availability is not currently available.
Sigma-Aldrich
Methanol, ACS spectrophotometric grade, ≥99.9%
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
Ammonium chloride, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, 99.5-100.5% (calc. to the dried substance)
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
Pricing and availability is not currently available.
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Pricing and availability is not currently available.
Sigma-Aldrich
Methanol, puriss., meets analytical specification of Ph Eur, ≥99.7% (GC)
Pricing and availability is not currently available.
Sigma-Aldrich
Methanol, BioReagent, ≥99.93%
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Pricing and availability is not currently available.
Triamcinolone, European Pharmacopoeia (EP) Reference Standard
Pricing and availability is not currently available.