Ethyl Glucuronide (EtG) and Ethyl Sulfate (EtS) in Oral Fluid on Raptor EtG/EtS

	Peaks	t_R (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Ethyl-β-D-glucuronide-d5	0.77	100	225.9	84.7	-
2.	Ethyl-β-D-glucuronide	0.78	500	220.9	74.9	85.0
3.	Ethyl sulfate-d5	1.75	100	129.8	97.7	-
4.	Ethyl sulfate	1.78	500	124.8	96.8	79.7

Column

Raptor EtG/EtS (cat.# 9325A12)

Dimensions:

100 mm x 2.1 mm ID

Particle Size:

2.7 µm

Pore Size:

90 Å

Guard Column:

UltraShield UHPLC precolumn filter (cat.# 25809)

Temp.:

30 °C

Standard/Sample

Ethyl-β-D-glucuronide (EtG) (cat.# 34101)

Ethyl-β-D-glucuronide-d5 (EtG-d5) (cat.# 34102)

Ethyl sulfate sodium salt (EtS) (cat.# 34103)

Ethyl sulfate-d5 sodium salt (EtS-d5) (cat.# 34104)

Diluent:

0.1% Formic acid in water

Conc.:

500 ng/mL

Inj. Vol.:

10 µL

Mobile Phase

0.1% Formic acid in water

0.1% Formic acid in acetonitrile

Time (min)	Flow (mL/min)	%A	%B
0.00	0.5	95	5
2.50	0.5	65	35
2.51	0.5	95	5
4.00	0.5	95	5

Detector	MS/MS
Ion Mode:	ESI-
Mode:	MRM
Instrument	HPLC
Sample Preparation	A 500 ng/mL standard was prepared in oral fluid. 50 µL of the standard was diluted with 950 µL of a working internal standard (100 ng/mL EtG-d5 and EtS-d5 in 0.1% formic acid in water). The sample was vortexed at 3000 rpm for 10 seconds to mix. The sample was then centrifuged at 3000 rpm for 10 minutes at 10 °C. The autosampler needle was adjusted to inject from the supernatant.

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