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Sampling and sample treatment At the end of 3-week treatment, mice from Groups 1 to 5 were sacrificed after overnight fasting followed by collecting their liver, gallbladder and bile

Hepatic lipids were extracted by adding 150 l ice-cold ethanol containing 1% 10 mM butylated hydroxytoluene and 2% Avanti SPLASH LIPIDOMIX MS Standards to each liver sample

Mice were euthanized and blood was collected by cardiac puncture and processed immediately for simultaneous determination of EtOH and acetaldehyde by headspace gas chromatography/mass spectrometry (HS-GC/MS) analysis