CLINICAL/FORENSICS
26
Steroids
GC
Steroids
Anabolic steroidscanbeanalyzedasTMS-deriva-
tized or as underivatized compounds. In either
case, this analysis is usuallydone at high temper-
ature by GC. Analysis time can be reduced
through theuseof thin film GC columns. Lower
polarity GC columns will help to reduce the
effective elution temperature. Rtx
®
-5 columns
provide sufficient selectivity to resolvemanyana-
bolic steroids, and have the stability necessary to
withstand the high temperatures needed.
Allure™ Biphenyl columns are an excellent
choice for steroid analyses. Through
π
-
π
interac-
tionswithdoublebonds in the steroid ring struc-
ture, the biphenyl stationary phase greatly
improves selectivity, relative to alkyl phases (e.g.,
C18). For example analyses, request the free
ApplicationsNote listedbelow.
Steroids also can be analyzed by HPLC on an
octadecylsilyl (C18) stationary phase. Specially
developed for HPLC/MS. Allure
™
C18 columns
increase the sensitivityof the analysis by allowing
higher concentrations of organic content in the
mobile phase.
6
30m, 0.25mm ID, 0.10µmRtx
®
-5 (cat.# 10208)
0.5µL split injection of anabolic steroids
Conc.:
1000ng/µL
Oven temp.:
180°C to 340°C@ 10°C/min. (hold 3min.)
Inj. / det. temp.:
280°C / 340°C
Carrier gas:
helium
Linear velocity:
35cm/sec. set@ 180°C
FID sensitivity:
1.28 x 10
-10
AFS
Split ratio:
50:1
1. 5-androstene-3
β
,17
β
-diol
2. 17
α
-methyl-5-androstene-3
β
,17
β
-diol
3. 5
α
-androstan-17
β
-ol-3-one
4. 19-nortestosterone
5. 17
α
-methylandrostan-17
β
-ol-3-one
6. mesterolone
7. testosterone
8. 17
α
-methyltestosterone
9. 1-dehydrotestosterone
10. 1-dehydro-17
α
-methyltestosterone
11. bolasterone
12. oxymethalone
13. 19-nortestosterone-17-propionate
14. testosterone propionate
15. fluoxymesterone
16. 4-chlorotestosterone-17-acetate
17. testosterone-17
β
-cypionate
18. 1-dehydrotestosterone benzoate
19. 1-dehydrotestosterone undecylenate
Steroids,Anabolic (Underivatized)
Rtx®-5
1
19
18
17
2
3
5
7
8
9
10,11
12
13
14
15
16
4
min.
2
4
6
8
10
12
14
16
18
GC_PH00255
HPLC
LC_0111
Data Courtesy of Pfizer Inc
Peak List:
1. deoxycorticosterone (DCC) acetate
2. DCC glucoside
3. DCC
Column:
Cat.#:
9164552
Dimensions:
50 x 2.1mm
Particle size:
5µm
Pore size:
60Å
Conditions:
Scan range:
320-520amu
Flow rate:
0.4mL/min.
LC/MS Interface:positive ion ESI-TOF
Voltage:
3800V
Nozzle temp.:
160V
Nozzle volt.:
190V
Allure™C18
water:methanol (33:67, v/v)
4191.1 cps
ConventionalC18
water:methanol (40:60, v/v)
3324.0 cps
Corticosteroids
Allure™C18vs.ConventionalC18
(LC/MS)
12% increase inorganic=
26% increase inLC/MSsensitivity
1
2
3
1
2
3
0
1
2
3
4 min.
0
1
2
3
4 min.
free
literature
ImprovedHPLCAnalysis of Steroids, Using Restek's Unique Allure
™
Biphenyl Column
lit. cat.# 580020
Call Restek at 800-356-1688 or 814-353-1300, ext. 5, or contact
your Restek representative, to request your free copy!
