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LC-MS Quantitative Screening Method for 18 Anabolic Steroids in Oral Fluid Using MS2 Spectra Data Collected with Q Exactive Orbitrap Mass Spectrometer
For Research Use Only. Not for use in diagnostic procedures.
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This information is not intended to encourage use of these products in any manners that might infringe the
intellectual property rights of others.
tion range (2 ng/mL, 15 ng/mL, 90 ng/mL,
C samples were analyzed in 5 replicates in 3
ay precision (Table 3).
iking blank oral fluid with all analytes at
/mL and analyzing these samples with
/mL
10 ng/mL
100 ng/mL
21
131
107
01
123
101
D
97.7
93.5
.0
104
103
02
92.4
94.3
01
103
99.6
00
127
115
.5
124
109
.5
105
96.8
.5
99.8
102
.5
92.6
95.3
.0
81.5
92.8
.0
100
95.9
.0
109
113
.7
110
118
.5
101
104
.5
92.0
105
.5*
102
104
Q
Donor Samples
Testosterone and Epitestosterone in negative tested oral fluid processed with LLE.
Testosterone- 0.206 ng/mL (extrapolated)
Epitestosterone- 0.016 ng/mL (extrapolated)
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Compounds detected in selected positive tested samples prepared in collaborator
lab with protein precipitation method.
Conclusion
We developed sensitive and robust quantitative screening method to analyze
anabolic steroids in human oral fluid.
• Implementation of the ultra high resolution Q Exactive mass spectrometer to
collect MS2 spectra and ion ratio confirmation results in high confidence in
compound identification.
• Method was validated using LLE for sample preparation, but we also detected all
analytes in positive tested samples processed with protein precipitation and
provided by collaborator laboratory.
Testosterone- 2270 ng/mL (extrapolated)
Epitestosterone 886 ng/mL
Boldenone- 7500 ng/mL (extrapolated)
Methandrosterone- 6600 ng/mL (extrapolated)
Testosterone 1.6 ng/mL
Epitestosterone- not detected
Oxandrolone 23.7 ng/mL
Clenbuterol 11 ng/mL
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s
oral fluid
Acknowledgement
We would like to thank Erica Guice, Research Director, Western Slope
Laboratory, for scientific advice and for providing samples for method testing.
References
1. Draft SANCO 1805/2000 Rev.1 [Revised Commission Decision 93/256 of 14 April 1993]
laying down performance criteria for analytical methods to be used for certain substances and
residues there of in live animals and animals products according to Council Directive 96/23/EC
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Inter assay
L 450 ng/mL 2 ng/mL 15 ng/mL 90 ng/mL 450 ng/mL
<15.1
12.6
8.4
5.8
11.0
5
17.9
16.3
9.4
12.0
14.1
<13.0
NA
12.7
10.0
10.4
9
<18.3
11.5
12.7
13.9
17.3
5
<20.0
14.5
10.8
9.5
13.6
1
<12.9
12.6
11.3
16.1
18.2
5
<9.7
14.2
10.2
10.2
8.9
9
<19.9
10.6
10.4
10.2
13.7
9
<19.1
11.5
7.6
9.5
16.7
8
<13.2
14.3
9.8
7.7
8.3
0
<18.1
18.7
13.5
14.3
19.9
5
<15.2
19.9
10.9
8.2
13.1
0
<16.9
16.3
11.1
7.5
13.4
<15.0
24.5
9.0
4.9
12.6
3
<12.8
14.1
11.0
6.6
9.7
6
<19.2
24.0
9.0
7.5
14.0
<12.5
24.8
8.0
5.8
11.0
<13.7
NA
9.1
9.4
14.2