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AN63738_E 10/13S
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0
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0 5000 10000 15000 20000 25000 30000 35000 40000 45000 50000
Y = 8.337e-3X + 3.048e-1; R^2: 0.9946; Origin: Ignore; W: 1/X; Area
0
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0 10000 20000 30000 40000 50000 60000 70000 80000 90000 100000
Y = 7.43e-4X + 1.18e-2; R^2: 0.9973; Origin: Ignore; W: 1/X; Area
EtG
Calibrator %Recovery
25 ng/mL -3.45
50 ng/mL 9.81
100 ng/mL -3.28
250 ng/mL 3.96
500 ng/mL 4.39
1000 ng/mL -12.4
5000 ng/mL -10.1
10,000 ng/mL 12.4
50,000 ng/mL -1.28
EtS
Calibrator %Recovery
100 ng/mL 0.93
250 ng/mL 8.64
500 ng/mL -11.8
1000 ng/mL 0.14
5000 ng/mL 2.41
10,000 ng/mL 9.65
50,000 ng/mL -7.26
100,000 ng/mL 2.58
Application Note 589
Figure 2. Representative chromatograms showing quantifier and confirming ions for
EtG and EtS at 100 and 25 ng/mL, respectively
Figure 1. Representative calibration curves for EtG and EtS in urine
1.6 1.8 2.0 2.2 2.4
Time (min)
0
20
40
60
80
100
RelativeAbundance
0
20
40
60
80
100
RelativeAbundance
RT: 2.10
AA: 1742
SN: 329
RT: 2.10
AA: 889
SN: 131
0
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60
80
100
RelativeAbundance
0
20
40
60
80
100
RelativeAbundance
RT: 2.75
AA: 8229
SN: 169
RT: 2.74
AA: 2444
SN: 249
2.5 2.6 2.7 2.8 2.9 3.0 3.1 3.2
Time (min)
EtG quanƟfier ion at 100 ng/mL
EtS quanƟfier ion at 25 ng/mL
EtG confirming ion at 100 ng/mL EtS confirming ion at 25 ng/mL
Conclusion
• This method provides limits of detection in urine of
100 ng/mL for EtG and 25 ng/mL for EtS.
• SPE extraction helps remove interferences in urine.
• Addition of an ion-pairing reagent helps
chromatographically separate the compounds from
interferences on the solvent front, thereby improving
limits of detection.
• This method is suitable for forensic toxicology.
References
1. LC/MS Method For Extracting Ethyl Glucuronides
From Urine Using: 200 mg Clean Screen® Extraction
Column; Part #: CSETG203; UTC, Inc. Bristol, PA;
2008
2. Thermo Scientific Application Note 488b,
Quantitation of Urinary Ethyl Glucuronide and Ethyl
Sulfate Using Ultrahigh Resolution LC-MS, http://
planetorbitrap.com/data/uploads/ZFS1327868383151_
AN488b.pdf
For Forensic Toxicology Use Only.