Testosterone

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Additional information

All data presented are based in the result of a literature search (references are stated in the tables). For testosterone, only limited data on toxicity was available. Further data was found for testosterone propionate and other esters and used for classification. Although reproductive toxicity test data are not required to fulfil the Annex VII obligations for a 1 -10 tpa registration dossier, summary data from the open literature are included here to provide support for classification conclusions. Read-across justification summary is attached to this endpoint summary.

Effects on fertility:

Test system	Test Substance	Application	Dose/Duration	Effect	Literature
Rat	1) Testosterone  2) Testosterone propionate	Subcutaneous injection	day 10 - 20 of gestation for 4 - 8 d  1) 0.5 - 80 mg  2) 1.0 - 55 mg	Resorption, necrosis, lethality, post-partum mortality and masculinisation in female offspring. Correlation to dose and period of administration.	Greene et al., 1939 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Rat	Testosterone	Subcutaneous injection	Injection on day 5 - 11 of gestation (low dose) and on day 1, 5 and 9 (high dose)  4 mg/kg bw and 20 mg/kg bw	Prevention of implantation at early time points, foetal loss or delayed parturition. Foetal loss in all animals treated with the high dose.	Dreisbach, 1959 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International
Sprague-Dawley rats	testosterone	implanted subcutaneously into groups of five and seven female rats on day 10 of pregnancy	Ten mg	Implantation with testosterone resulted in complete resorption of embryos in all treated animals.	Sarkar, K., Kinson, G.A. & Rowsell, H.C. (1986) Embryo resorption following administration of steroidal compounds to rats in mid-pregnancy. Can. J. Vet. Res., 50, 433-437.In: Toxicological Evaluation Of Certain Veterinary Drug Residues. In Food Who Food Additives Series: 43 Prepared by the Fifty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA) World Health Organization, Geneva, 2000 IPCS - International Programme on Chemical Safety
Rats	testosterone	subcutaneous injections	.3, 3 or 30 mg testosterone 3 times weekly for 6 weeks	At the lowest dose but not the intermediate and high dose testosterone decreased testis weight, produced atrophy of the germinal epithelium and suppressed serum LH but not FSH.	Walsh PC and Swerdloff: (1973) Biphasic effect of testosterone on spermatogenesis in the rat, Invest Urol, 11: 190-193 In: Toxicological Evaluation Of Certain Veterinary Drug Residues. In Food Who Food Additives Series: 43 Prepared by the Fifty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA) World Health Organization, Geneva, 2000 IPCS - International Programme on Chemical Safety

Short description of key information:

All data presented are based in the result of a literature search (references are stated in the tables). Two reports of the effect of testosterone injection after gestation indicated a dose and time dependent effect on resorption, embryo lethality and caused masculinization of the female offspring.

Effects on developmental toxicity

Description of key information

All data presented is based in the result of a literature search (references are stated in the tables).   Treatment of rats with testosterone propionate (100 mg) on day 14 of gestation resulted insmall or absent mammary glands in both sexes and the absence of nipples in females. Analogous results were obtained by other experiments in rats, over all resulting in a time and dose dependent masculinization of the offspring.

Additional information

All data presented are based in the result of a literature search (references are stated in the tables). For testosterone, only limited data on toxicity was available. Further data was found for testosterone propionate and other esters and used for classification. Although reproductive toxicity test data are not required to fulfil the Annex VII obligations for a 1 -10 tpa registration dossier, summary data from the open literature are included here to provide support for classification conclusions. Read-across justification summary is attached to this endpoint summary.

Effects on developmental toxicity:

Test system	Test Substance	Application	Dose/Duration	Effect	Literature
Rat	Testosterone propionate	Injection	on day 14 of gestation  100 mg	Small or absent mammary glands in offspring of both sexes and absence of nipples in females	Jean & Jean, 1969 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Crossbreed ewe	Testosterone	Oral	single dose at day 14 of gestation  1.3 - 1.8 mg	No effect	Keeler & Binns, 1968 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Guinea pig, female	Testosterone with or without aromatase inhibitor ATD		from days 28-65 of gestation	Prenatal testosterone treatments altered the hormonal requirements for androgen-activated mounting in females such that they resembled normal males	Roy, 1952. Physiol Behav 51 (1): 105-9 as cited by Hazardous Substances Data Bank (HSDB, testosterone, 19, 2009
Rat	Testosterone propionate		Last week of pregnancy	Testosterone significantly increased anogenital distance and delayed vaginal opening of progeny	McCoy & Shirley, 1992. Life Sci 50 (9): 621-8 as cited by Hazardous Substances Data Bank (HSDB, testosterone, 19, 2009
Sprague Dawley Rat, female	Testosterone	Subcutaneous injection	Daily dosage for 20 d, start at day 21 after birth  0.05, 0.2 or 1.0 mg/kg bw/day	Rodent pubertal female assay Dose dependent delay of vaginal opening, irregular estrous cycle, reduction of ovarian (all doses) and uterine weight (only high dose),	Kim et al., 2002. Tox Sci 67: 52.62
Rats				Female offspring exposed to 0, 1.5, and 2.5 mg/kg/day testosterone propionate (TP) on GDs 14-18 displayed increased AGD at postnatal day (PND) 2 and decreased nipples at PND 13 and as adults. TP-induced changes in neonatal AGD and infant areola number were reliable indicators of permanently altered adult phenotype in female rats. Further, females in the two high-dose groups displayed increased incidences of external genital malformations and the presence of prostatic tissue, not normally found in female rats.	Hotchkiss AK; Lambright CS; Ostby JS; Parks-Saldutti L; Vandenbergh JG; and Gray LE Jr. 2007.  Prenatal testosterone exposure permanently masculinizes anogenital distance, nipple development, and reproductive tract morphology in female Sprague-Dawley rats. Toxicol Sci. Apr; 96(2):335-45.
Rats				Prenatal hyperandrogenization induced an anti-oxidant response that prevented a possible prooxidant status. The higher dose of testosterone induced a pro-inflammatory state in ovarian tissue mediated by increased levels of prostaglandin E (PG) and the protein expression of cyclooxygenase 2 (COX2, the limiting enzyme of PGs synthesis). In summary, our data show that the levels of testosterone prenatally injected modulate the uterine environment and that this, in turn, would be responsible for the endocrine and metabolic abnormalities and the phenotype of polycystic ovarian syndrome (PCOS) during the adult life.	Amalfi S; Velez LM; Heber MF; Vighi S; Ferreira SR; Orozco AV; Pignataro O; and Motta AB. 2012. Prenatal hyperandrogenization induces metabolic and endocrine alterations which depend on the levels of testosterone exposure. PLoS One. 2012; 7(5):e37658 (http://toxnet.nlm.nih.gov/cgi-bin/sis/search/f?./temp/~87RHfj:59:BODY)

Justification for classification or non-classification

In accordance with the androgenic effects of testosterone signs of masculinisation as well as disturbances to the cycle and fertility are to be expected in women following long-term exposure to doses which are pharmacologically effective. Impairment of the endogenous hormone production as well as disturbance of fertility (impairment of spermiogenesis) are also to be expected in men following long-term exposure. Exposure during pregnancy may lead to signs of masculinisation in the sex organs of a female child. If taken by nursing women testosterone may reach into the mother's milk and thus impair the development of the infant.

Classified as Category 1B; H361: Suspected of damaging fertility or the unborn child according to Regulation 1272/2008/EC (CLP).

According to the Directive 67/548/EEC testosterone is classified:

Category 1; R60 - May impair fertility.

Category 2; R61 - May cause harm to the unborn child.

R64 - May cause harm to breastfed babies