Determination and confirmation of selective estrogen receptor modulators (SERMs), anti-estrogens and aromatase inhibitors in bovine and porcine urine using UHPLC-MS/MS

Figures & data

Table 1. Compounds, structures, chemical names, compound group (SERM, AE or AI) and MS parameters.

Compound	Chemical structure	Chemical name	Compound group	Precursor ion (m/z)	Product ion (m/z)	Cone voltage (V)	Collision energy (eV)
Raloxifene		[6-Hydroxy-2-(4-hydroxyphenyl)-1-benzothiophen-3-yl]{4-[2-(1-piperidinyl)ethoxy]phenyl}methanone	SERM	474.1	112.1	30	25
					84.2		40
Tamoxifen		2-{4-[(1Z)-1,2-diphenyl-1-buten-1-yl]phenoxy}-N,N-dimethylethanamine	SERM	372.3	72.2	40	13
					129.1		30
Toremifene		2-{4-[(1Z)-4-chloro-1,2-diphenyl-1-buten-1-yl]phenoxy}-N,N-dimethylethanamine	SERM	406.1	72.2	40	12
					205.2		22
4-Hydroxytamoxifen		4-(1-{4-[2-(Dimethylamino)ethoxy]phenyl}-2-phenyl-1-buten-1-yl)phenol	SERM	388.2	72.2	40	12
					129.2		25
Fulvestrant		(7α,17β)-7-{9-[(4,4,5,5,5-Pentafluoropentyl) sulfinyl]nonyl}estra-1(10),2,4-triene-3,17-diol	AE	607.3	467.1	40	25
					493.2		25
Clomiphene		2-{4-[(E)-2-chloro-1,2-diphenylvinyl]phenoxy}-N,N-diethylethanamine	AE	406.2	86.2	40	25
					100.2		10
Mesterolone		(1α,5α,17β)-17-Hydroxy-1-methylandrostan-3-one	AE	305.3	269.3	40	15
					95.2		30
Dromostanolone		(2α,5α,17β)-17-Hydroxy-2-methylandrostan-3-one	AE	305.4	269.2	40	15
					95.2		30
4-Hydroxycyclofenil		4,4ʹ-[(4-Hydroxycyclohexylidene)methylene]diphenol	AE	297.2	133.1	30	20
Anastrozole		2,2ʹ-[5-(1H-1,2,4-triazol-1-ylmethyl)-1,3-phenylene]bis(2-methylpropanenitrile)	AI	294.2	225.2	40	35
					209.0		35
Exemestane		6-Methyleneandrosta-1,4-diene-3,17-dione	AI	297.2	121.1	35	18
					149.1		15
Letrozole		4,4ʹ-(1H-1,2,4-triazol-1-ylmethylene)dibenzonitrile	AI	286.2	217.2	25	15
					190.1		35
DL-aminoglutethimide		3-(4-Aminophenyl)-3-ethyl-2,6-piperidinedione	AI	233.2	188.1	40	15
					146.1		25
Formestane		4-Hydroxyandrost-4-ene-3,17-dione	AI	303.2	113.1	45	25
					173.1		18
Androsta-1,4,6-triene-3,17-dione		Androsta-1,4,6-triene-3,17-dione	AI	283.2	97.2	40	15
					265.1		10
Testolactone		(4aS,4bR,10aR,10bS,12aS)-10a,12a-dimethyl-3,4,4a,5,6,10a,10b,11,12,12a-decahydro-2H-naphtho[2,1-f]chromene-2,8(4bH)-dione	AI	301.0	121.1	30	10
					147.1		10
1Alpha-methyl-5alpha-androstan-3alpha-ol-17 one^a		1Alpha-methyl-5alpha-androstan-3alpha-ol-17 one	AAS	287.3	269.2	30	10
					161.1		15
2Alpha-methyl-5alpha-androstan-3alpha-ol-17 one^b		2Alpha-methyl-5alpha-androstan-3alpha-ol-17 one	AAS	287.3	269.2	30	10
					161.1		15

Notes: ^aMetabolite of mesterolone.

^bMetabolite of dromostanolone.

Table 2. CCα and CCβ (ng ml^–1) in bovine and porcine urine.

Compound	CCα (ng ml^–1)		CCß (ng ml^–1)
	Bovine	Porcine	Bovine	Porcine
Raloxifene	< 0.05	< 0.05	< 0.05	< 0.05
Tamoxifen	0.45	0.44	0.90	0.89
Toremifene	2.37	1.49	4.74	2.98
4-Hydroxytamoxifen (peak 1^a)	< 0.625	< 0.625	< 0.625	< 0.625
4-Hydroxytamoxifen (peak 2^a)	< 0.625	< 0.625	< 0.625	< 0.625
Fulvestrant	< 5.0	< 5.0	< 5.0	< 5.0
Clomiphene (total)^b	0.82	< 1.25	1.63	< 1.25
Mesterolone	< 12.5	< 12.5	< 12.5	< 12.5
Dromostanolone	< 12.5	< 12.5	< 12.5	< 12.5
4-Hydroxycyclofenil (1 ion)	< 50		< 50
Anastrozole	0.85	0.72	1.71	1.43
Exemestane	7.0	3.47	14.0	7.47
Letrozole	< 1.25	0.92	< 1.25	1.83
DL-aminoglutethimide	< 2.5	< 2.5	< 2.5	< 2.5
Formestane	< 12.5	< 12.5	< 12.5	< 12.5
Androsta-1,4,6-triene-3,17 dione	11.0	< 12.5	23.8	< 12.5
Testolactone	10.9	< 12.5	21.8	< 12.5
1Alpha-Methyl-5alpha-androstan-3alpha-ol-17 one	< 25	< 12.5	< 25	< 12.5
2Alpha-Methyl-5alpha-androstan-3alpha-ol-17 one	< 25	< 12.5	< 25	< 12.5

Notes: ^aNot known which peak is the E or Z isomer. The distribution of the E and Z isomers in the standard is 50/50.

^bE and Z isomers are not separated.

Figure 1. (colour online) Chromatograms of a blank porcine urine (bottom), a blank porcine urine with addition of 25 ng ml⁻¹ formestane (middle) and a porcine urine from the survey samples that contain formestane. The areas on the y-axes of the chromatograms are not normalised.