. Author manuscript; available in PMC: 2019 Oct 1.

Published in final edited form as: Biochem Pharmacol. 2018 Aug 4;156:32–42. doi: 10.1016/j.bcp.2018.08.003

Table 2.

Optimized multiple reaction monitoring (MRM) parameters used for quantitative analysis of testosterone metabolites.

Metabolite/internal standard	Precursor ion (m/z )	Daughter Ion(s) (m/z)	DP (V)	CE (eV)	LLOQ^a (ng/mL)
6β-Hydroxytestosterone	305.3	269.1, 121, 109	90	30, 21, 30	1 [0]; 0.5 [0]

6β-Hydroxytestosterone-d3	308.3	272.2, 290.2	80	30

Androstenedione	287.2	109.1, 97.1	80	30	50 [0]; 50 [0]

Androsterone	291.2	255.1	60	25	10 [2]; 5 [8]
Androsterone	273.1	255.1, 147	60	15, 20	10 [2]; 5 [8]

Androsterone glucuronide	449.1	255.2	70	25	5 [1]; 5 [11]
Androsterone glucuronide	484.3	255.2, 273.2	70	35	5 [1]; 5 [11]

Androsterone-d2	275.2	275.1, 147.1	80	5, 30

Androsterone glucuronide-	453.2	259.2, 277.4	80	30, 22
d4	488.3	259.3	80	30

5α-Dihydrotestosterone	291.1	255.4, 159.1	70	22, 28	5 [0]; 1 [1]
5α-Dihydrotestosterone	273.1	255.4	70	22	5 [0]; 1 [1]

5α-Dihydrotestosterone glucuronide	467.3	255.2	70	35	5 [0]; 1 [7]
5α-Dihydrotestosterone glucuronide	484.3	273.2	70	35	5 [0]; 1 [7]

5α-Dihydrotestosterone-d3	294.3	91, 258.2	70	69, 25

5α-Dihydrotestosterone glucuronide-d3	470.2	294.2, 276.2, 159.2	80	30

Etiocholanolone	273.2	255.1, 147	80	25	5 [5]; 1 [16]
Etiocholanolone	291.1	255.1	80	30	5 [5]; 1 [16]

Etiocholanolone glucuronide	484.3	273.2, 255.2	70	35	0.5 [2]; 5 [16]
Etiocholanolone glucuronide	467.3	255.2	70	35	0.5 [2]; 5 [16]

Testosterone	289.1	109.1, 97.1	80	30

Testosterone glucuronide	465.2	289.2, 271.2, 109.1	70	25, 30, 35	0.5 [0]; 0.5 [0]

Testosterone-d3	292.2	109.1, 97.1	80	30

Testosterone glucuronide-d3	468.2	292.2, 274.2, 256.2	70	25, 30, 35

Lower limit of quantification (LLOQ) in hepatocytes; enterocytes. Value in brackets indicates number of samples below LLOQ, which were assigned limit of detection (LOD) as one-third of LLOQ values.