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

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.

Applications and Challenges in Using LC–MS/MS Assays for Quantitative Doping Analysis

“…HILIC may help solve this problem because compared with RPLC, polar and hydrophilic analytes are more greatly retained on an HILIC column but few studies exist describing quantification of hydrophilic target analytes like morphine and codeine and their glucuronide [41], cathinone-designer drugs (11 chemicals) [77], etilefrine and oxilofrine [78], ephedrines [79,51] and narcotics [80]. The few studies reported suggest that HILIC requires simple sample preparation, offers good sensitivity and robustness and only requires adequate column equilibration and precondition time.…”

Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis

“…Furthermore, reliable HILIC method was used for the detection of therapeutic agent and/or their metabolites in more than one matrix . For example, antihypertensive drug etilefrine and oxilofrine in equine blood plasma and urine were determined with a HALO HILIC column (50 × 2.1 mm id, particle size 2.7 μm) coupled with ESI‐MS/MS detector, the method showed relatively low LOD (≤ 0.1 ng/mL) and LOQ (≤ 1.0 ng/mL) in both matrices .…”

Facile Preparation of Ionic Liquid‐coated Copper Nanowire‐modified Carbon Paste Electrode for Electrochemical Detection of Etilefrine Drug

“…Growing the ratio of these deaths were correlated with a prescription drug overdose. The various analytical techniques are used to detect the ET‐HCl include spectrophotometry, flow injection chemiluminescence, proton magnetic resonance spectroscopy, HPLC, and GC–MS . The numerous analytical methods have been used to detect PC in different matrices including chromatography, spectrophotometry, titrimetry, chemiluminescent .…”