Merck
  • Home
  • Search Results
  • Development and validation of a hydrophilic interaction liquid chromatography with tandem mass spectrometry method for the simultaneous detection and quantification of etilefrine and oxilofrine in equine blood plasma and urine.

Development and validation of a hydrophilic interaction liquid chromatography with tandem mass spectrometry method for the simultaneous detection and quantification of etilefrine and oxilofrine in equine blood plasma and urine.

Journal of separation science (2014-08-22)
Noel Kong, Rong Yi, Sarah Zhao, Jasmeet Sandhu, Geoffrey Lam, Devan Loganathan, Barbara Morrissey
ABSTRACT

A sensitive hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry method was developed and validated for the simultaneous detection and quantification of etilefrine and oxilofrine in equine blood plasma and urine. The method is highly sensitive and specific with good precision and accuracy. In plasma the limit of detection and limit of quantification are 0.03 and 0.1 ng/mL, respectively, for both analytes. In urine the limit of detection and limit of quantification are 0.3 and 1 ng/mL, respectively, for both analytes. The suitability of the method for doping control analysis in equine species is demonstrated by analyzing postadministration samples collected after a single intravenous administration of 50 mg etilefrine to a standardbred mare. Etilefrine was detected up to 120 h in urine and up to 48 h in plasma. Etilefrine is highly conjugated in equine urine whereas it exists in the free form in equine plasma. Therefore, enzyme hydrolysis prior to sample preparation is recommended for the detection and quantification of etilefrine and oxilofrine in equine urine.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methanol, anhydrous, 99.8%
Sigma-Aldrich
Ammonium formate, BioUltra, ≥99.0% (calc. based on dry substance, NT)
Supelco
Ammonium formate, eluent additive for LC-MS, LiChropur, ≥99.0%
Supelco
tert-Butyl methyl ether, analytical standard
Supelco
Methanol, analytical standard
Sigma-Aldrich
Ammonia, puriss., anhydrous, ≥99.95%
Sigma-Aldrich
Ammonia, puriss., anhydrous, ≥99.9%
Supelco
(1S,2R)-(+)-Ephedrine hydrochloride, 1.0 mg/mL in methanol (as free base), ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
Methanol, HPLC Plus, ≥99.9%, poly-coated bottles
Sigma-Aldrich
Ammonia-14N, 99.99 atom % 14N
Supelco
Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Ammonium formate, Vetec, reagent grade, ≥96%
Supelco
Residual Solvent - Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid
Sigma-Aldrich
Ammonia, anhydrous, ≥99.98%
Sigma-Aldrich
tert-Butyl methyl ether, anhydrous, 99.8%
Sigma-Aldrich
Ammonium formate, ≥99.995% trace metals basis
Sigma-Aldrich
Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC
Millipore
Acetonitrile solution, suitable for HPLC, acetonitrile:water 5:95% (v/v), 10 mM Ammoniumbicarbonate, pH 10,0
Sigma-Aldrich
Ammonia solution, 4 M in methanol
Sigma-Aldrich
Ammonia solution, 0.4 M in THF
Sigma-Aldrich
tert-Butyl methyl ether, ACS reagent, ≥99.0%
Sigma-Aldrich
Methanol, NMR reference standard
Ephedrine hydrochloride (racemic), European Pharmacopoeia (EP) Reference Standard
Supelco
tert-Butyl Methyl Ether, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Methanol, Laboratory Reagent, ≥99.6%
Sigma-Aldrich
Methanol, Absolute - Acetone free
Sigma-Aldrich
Ammonia solution, 0.4 M in dioxane
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Methanol, BioReagent, ≥99.93%