Identification and quantification of thymol metabolites in plasma, liver and duodenal wall of broiler chickens using UHPLC-ESI-QTOF-MS.
In the present study, we aimed to develop a method for thymol sulfate and thymol glucuronide determination in plasma, liver and duodenal wall of broiler chickens after feeding with a Thymus vulgaris essential oil at the different concentrations (0.01, 0.05 and 0.1% w/w). UHPLC coupled with accurate-mass QTOF-MS was used for identification and quantification of thymol metabolites. Novel Waters Oasis Prime HLB solid-phase extraction cartridges were applied to sample clean-up with extraction recoveries ranged from 85 to 92%. The presence of thymol glucuronide was confirmed by MS software according to molecular formula, score, mass error and double bond equivalent. In terms of validation, calibration curves of thymol sulfate were constructed in matrix samples with linearity from 3.91 to 250.0 ng/mL and correlation coefficients were within the range of 0.9979-0.9995. Limits of detection were 0.97, 0.29 and 0.63 ng/mL and limits of quantification were 3.23, 0.97 and 2.09 ng/mL for plasma, liver and duodenal wall, respectively. Intra-day and inter-day precision expressed as relative standard deviation were <4.35%. To highlight, thymol metabolites were directly detected for the first time in liver and duodenal wall and this method was shown to be successfully applicable for investigation of thymol metabolism in chickens after thyme essential oil ingestion.
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 04001, Košice, Slovakia.|Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University, 4306 Walker Building, Auburn, AL, 36849, USA.
Jana
J
Pisarčíková
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 04001, Košice, Slovakia.
Vladimíra
V
Oceľová
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 04001, Košice, Slovakia.
Štefan
Š
Faix
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 04001, Košice, Slovakia.
Iveta
I
Plachá
Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University, 4306 Walker Building, Auburn, AL, 36849, USA.
Angela I
AI
Calderón
D000818:Animals; D002138:Calibration; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D004355:Drug Stability; D004386:Duodenum / Q000737:chemistry*; D020719:Glucuronides / Q000032:analysis; D008099:Liver / Q000737:chemistry; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards; D013943:Thymol / Q000031:analogs & derivatives / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics*; D014018:Tissue Distribution
D020719:Glucuronides; C000620477:thymol glucuronide; C000620476:thymol sulfate; D013943:Thymol
UHPLC-ESI-QTOF-MS; biological samples; quantitation; thymol metabolites; validation