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

	Peaks	t_R (min)	Conc. (ng/mL)	Precursor Ion	Product Ion	Product Ion
1.	Estriol	1.44	100	289.1	107.0	253.2
2.	Cortisol	2.11	50	363.2	121.0	327.2
3.	17β-Estradiol	2.45	50	255.3	159.1	133.1
4.	17α-Hydroxypregnenolone	2.60	100	315.2	297.2	159.2
5.	Aldosterone	2.68	50	361.4	315.2	157.2
6.	11-Deoxycortisol	2.95	50	347.3	97.1	109.1
7.	Corticosterone	3.07	50	347.4	329.2	121.1
8.	Estrone	3.07	50	271.2	253.2	133.1
9.	Boldenone	3.11	25	287.2	121.0	135.0
10.	Epitestosterone	3.16	25	289.2	109.2	97.1
11.	Dehydroepiandrosterone (DHEA)	3.16	50	289.1	253.1	271.1
12.	17α-hydroxyprogesterone	3.30	50	331.2	97.0	109.0
13.	Testosterone	3.35	25	289.2	97.0	109.0
14.	17β-trenbolone	3.36	50	271.2	253.1	199.0
15.	17α-Methyltestosterone	3.41	25	303.2	97.0	109.0
16.	5α-Dihydrotestosterone (DHT)	3.41	50	291.2	255.2	159.2
17.	Nandrolone	3.42	50	275.1	109.0	257.1
18.	25-Hydroxy vitamin D3	3.71	100	383.3	365.3	257.2
19.	Androstenedione	3.79	25	287.1	97.0	109.0
20.	11-Deoxycorticosterone	3.81	25	331.2	97.1	109.1
21.	Progesterone	4.21	25	315.2	97.0	109.0

Conc. listed is the final concentration following sample preparation.

Column

Raptor 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

Standard/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

Water + 0.1% formic acid

Methanol + 0.1% formic acid

Time (min)	Flow (mL/min)	%A	%B
0.00	0.6	50	50
4.00	0.6	0	100
4.50	0.6	0	100
4.51	0.6	50	50
6.50	0.6	50	50

Detector	MS/MS
Ion Mode:	ESI+
Instrument	UHPLC

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