Skip to Content
Merck
  • 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

Supelco
Methanol, analytical standard
Sigma-Aldrich
Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid
Sigma-Aldrich
Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC
Sigma-Aldrich
Ammonium formate, ≥99.995% trace metals basis
Sigma-Aldrich
tert-Butyl methyl ether, anhydrous, 99.8%
Sigma-Aldrich
Ammonia, anhydrous, ≥99.98%
Sigma-Aldrich
Acetonitrile solution, contains 0.1 % (v/v) formic acid, suitable for HPLC
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Supelco
tert-Butyl methyl ether, HPLC grade, suitable for HPLC, 99.8%
Supelco
Ammonium formate, eluent additive for LC-MS, LiChropur, ≥99.0%
Sigma-Aldrich
Ammonium formate, BioUltra, ≥99.0% (calc. based on dry substance, NT)
Sigma-Aldrich
DL-Ephedrine hydrochloride, 99%
Sigma-Aldrich
(1S,2R)-(+)-Ephedrine hydrochloride, 99%
Sigma-Aldrich
Methanol, suitable for HPLC, gradient grade, 99.93%
Sigma-Aldrich
β-Glucuronidase from Helix pomatia, Type H-2, aqueous solution, ≥85,000 units/mL
Sigma-Aldrich
tert-Butyl methyl ether, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%
Sigma-Aldrich
Ammonia-14N, 99.99 atom % 14N
Sigma-Aldrich
Methanol, HPLC Plus, ≥99.9%, poly-coated bottles
Supelco
tert-Butyl methyl ether, analytical standard
Sigma-Aldrich
Ammonia solution, 0.4 M in THF
Sigma-Aldrich
Methanol, JIS 300, ≥99.8%, for residue analysis
Sigma-Aldrich
Acetonitrile solution, contains 0.05 % (w/v) ammonium formate, 5 % (v/v) water, 0.1 % (v/v) formic acid, suitable for HPLC
Sigma-Aldrich
Acetonitrile solution, contains 10.0% acetone, 0.05% formic acid, 40.0% 2-propanol
Sigma-Aldrich
Ammonium formate, SAJ first grade, ≥95.0%
Sigma-Aldrich
tert-Butyl methyl ether, SAJ special grade, ≥99.0%
Sigma-Aldrich
Methanol, SAJ special grade
Sigma-Aldrich
Ammonia solution, 4 M in methanol
Sigma-Aldrich
Methanol, suitable for HPLC
Sigma-Aldrich
Methanol, SAJ first grade, ≥99.5%
Sigma-Aldrich
Methanol, JIS special grade, ≥99.8%