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Column	Ascentis^® Express OH5, 10 cm x 2.1 mm I.D., 2.7 µm particles (53757-U)
Column Temperature	30 °C
Mobile Phase	[A] 5 mM ammonium formate and 0.1% formic acid in 95:5, acetonitrile:water; [B] 5 mM ammonium formate and 0.1% formic acid in 80:20, acetonitrile:water
Gradient	0 to 100% B in 1 min, held at 100% B for 4 min, to 0% B, held for 5 min
Flow Rate	0.4 mL/min
Pressure	1450 psi (100 bar)
Sample	Urine spiked with ethyl sulfate and ethyl-ÃŸ-D-glucuronide, both at 50 ng/mL, diluted 20:1 in acetonitrile containing deuterated internal standards at 100 ng/mL
Injection	5 µL
Detector	MS, ESI(-), 125.0 m/z, 126.1 m/z, 221.1 m/z, 226.2 m/z

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Categories	Analytical Chromatography, Alcohols, Drugs of Abuse
Featured Industry	Clinical Forensics and Toxicology
General description	Ethyl sulphate (EtS) and ethyl glucuronide (EtG) are direct ethanol metabolites and may indicate recent alcohol consumption. The two compounds differ in their pathways for formation and degradation. Being polar compounds, they are poorly retained by C18 phases and elute early in the chromatogram along with matrix compounds. This leads to poor or unreliable quantification. The method shown here uses HILIC mode on an Ascentis Express OH5 column to retain both analytes well, making it very likely to be more robust and reliable, as well as highly MS-friendly. Cerilliant CRMs provided reliable quantification. The internal standard was needed for accurate quantification. External calibration resulted in a large excess of sulfate.
Suitability	Application for LC-MS

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