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Steroids in Serum on Raptor Biphenyl by LC-MS/MS

LC_CF0599
PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct IonProduct Ion
1.Estriol1.44100289.1107.0253.2
2.Cortisol2.1150363.2121.0327.2
3.17β-Estradiol2.4550255.3159.1133.1
4.17α-Hydroxypregnenolone2.60100315.2297.2159.2
5.Aldosterone2.6850361.4315.2157.2
6.11-Deoxycortisol2.9550347.397.1109.1
7.Corticosterone3.0750347.4329.2121.1
8.Estrone3.0750271.2253.2133.1
9.Boldenone3.1125287.2121.0135.0
10.Epitestosterone3.1625289.2109.297.1
11.Dehydroepiandrosterone (DHEA)3.1650289.1253.1271.1
12.17α-hydroxyprogesterone3.3050331.297.0109.0
13.Testosterone3.3525289.297.0109.0
14.17β-trenbolone3.3650271.2253.1199.0
15.17α-Methyltestosterone3.4125303.297.0109.0
16.5α-Dihydrotestosterone (DHT)3.4150291.2255.2159.2
17.Nandrolone3.4250275.1109.0257.1
18.25-Hydroxy vitamin D33.71100383.3365.3257.2
19.Androstenedione3.7925287.197.0109.0
20.11-Deoxycorticosterone3.8125331.297.1109.1
21.Progesterone4.2125315.297.0109.0
Conc. listed is the final concentration following sample preparation.
ColumnRaptor Biphenyl (cat.# 9309A12)
Dimensions:100 mm x 2.1 mm ID
Particle Size:2.7 µm
Guard Column:Raptor Biphenyl EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9309A0252)
Temp.:35 °C
Sample
Conc.: 500 µL of mobile phase B was added to 250 µL of fortified beagle serum (100-400 ng/mL). The sample was vortexed for 5 minutes and centrifuged at 4,000 rpm for 10 minutes (10 °C). 100 µL of supernatant was diluted in 100 µL of water. Final dilution factor was 6x.
Inj. Vol.:2 µL
Mobile Phase
A:Water + 0.1% formic acid
B:Methanol + 0.1% formic acid
Time (min)Flow (mL/min)%A%B
0.000.65050
4.000.60100
4.500.60100
4.510.65050
6.500.65050
DetectorMS/MS
Ion Mode:ESI+
InstrumentUHPLC