Reaction products of 2-(chloromethyl)oxirane and glycerol

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Key, one-generation study, oral, non TG, non-GLP, rat, ReA substance Epichlorohydrin: LOAEL 12.5 mg/kg/day, damage to male fertility.

Supp, two-generation reproduction study, inhalation, non-TG, non-GLP, rat, ReA substance phenylglycidyl-ether (PGE): no significant effects on reproductive parameters.

Link to relevant study records

Reference

Endpoint:

one-generation reproductive toxicity

Remarks:

similar to OECD Guideline 415 (One-Generation Reproduction Toxicity Study)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Deviations:

not specified

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Long-Evans

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories (Portage, MI).
- Age at study initiation: males (80-90 days old) and females (70-80 days old)
- Weight at study initiation: no data
- Fasting period before study: no
- Housing:Males were singly housed in stainless-steel hanging cages. Females were housed three to a cage until pregnant, when they were transferred to individual shoebox cages.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): Purina Lab Chow 5001 ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 2 to 3 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25
- Humidity (%): 55
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 10/14 with lights on at 20:00 h (males) and 12/12 with light commencing at 6:00 h (females)

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Dosing solutions were made fresh weekly and stored in the refrigerator. Chemical analyses showed that
these solutions were stable for this period.

VEHICLE
- Justification for use and choice of vehicle (if other than water): the test material is well soluble in corn oil.
- Concentration in vehicle: Concentrations were adjusted so that animals received 1 mL solution per kilogram of body weight.
- Amount of vehicle (if gavage): 1 mL

Details on mating procedure:

- M/F ratio per cage: 1/3
- Length of cohabitation: In the mating trials designed to evaluate pre-and postimplantation loss, treated males were cohabited overnight (0 and 50 mg/kg bw only); In the trials to evaluate female fertility, an untreated male was placed in each female cage to cohabit overnight.
- Proof of pregnancy: [vaginal plug or sperm in vaginal smear] referred to as [day 1] of pregnancy;
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: no.
During the third week of treatemnt, males were allowed 5 days to mate, with a different proestrous female introduced each day until each male had copulated successfully with one female.
- Further matings after two unsuccessful attempts: no data
- After successful mating each pregnant female was caged (how): individually in shoebox cages.
- Any other deviations from standard protocol: see below in "Any other information on materials and methods, incl. tables"

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Males: 21 days;
Females: 2 weeks prior to mating, 1 week during mating and during the gestation period until delivery.

Frequency of treatment:

once daily/5 days a week

Details on study schedule:

Not applicable

Dose / conc.:

100 mg/kg bw/day

Remarks:

females

Dose / conc.:

50 mg/kg bw/day

Remarks:

females

Dose / conc.:

25 mg/kg bw/day

Remarks:

females

Dose / conc.:

50 mg/kg bw/day

Remarks:

males

Dose / conc.:

25 mg/kg bw/day

Remarks:

males

Dose / conc.:

12.5 mg/kg bw/day

Remarks:

males

No. of animals per sex per dose:

Males: 20
Females: no data

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: preliminary studies.
Males:
Epichlorohydrin given for 15 days resulted in severe toxicity in male rats (weight loss, nephrosis, testicular edema, epididymal granulomas, decreased testicular and epididymal sperm counts). Treatment levels for the expanded study were, therefore, set at 0, 12.5, 25, and 50 mg/kg body wt (n = 20/group).
Females:
Epichlorohydrin was lethal, causing death in 10/10 animals within 48 hr of initiation of treatment. Administering epichlorohydrin for 15 days at levels of 50 and 100 mg/kg caused dose-dependent increases in liver, kidney, adrenal, and spleen organ-to-body weight ratios. In the 100 mg/kg group, 5/10 animals were so moribund as to warrant their sacrifice after 7-9 days treatment. The remaining 5 animals did not show weight loss or other overt toxic signs, but on sacrifice after 15 days treatment they displayed pale and enlarged kidneys. Since half of the females treated with 100 mg/kg/day survived, this was selected as the high dose, while 25 and 50 mg/kg/day were chosen as the low and middoses, respectively.

- Rationale for animal assignment (if not random): the animals were assigned to treatment groups such that mean body weights were the same across groups.

Positive control:

No

Parental animals: Observations and examinations:

The animals were weighed on alternate days (except pregnant females which were weighed daily).

Oestrous cyclicity (parental animals):

Estrous cyclicity and copulatory behaviour were examined.

Sperm parameters (parental animals):

Parameters examined in [P] male parental generations:
testis weight, epididymis weight, daily sperm production, sperm count in testes, sperm count in epididymides, enumeration of cauda epididymal sperm reserve, sperm motility and sperm morphology.

Litter observations:

Litters were sexed and weighed on Postnatal Day 1 (day of delivery). Pups were weighed weekly thereafter. At Day 4, litters were culled to 8 pups (approximately 4 male and 4 female). At postnatal Day 21, dams were sacrificed and litters reduced to 1 male and 1 female. Weanlings were weighed weekly until sacrifice at Postnatal Day 42. No further data.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals: on Day 21; On day 23 (an additional group of males which was used for the measurement of cauda epididymal spem motion.
- Maternal animals: All surviving animals: One group of females was sacrificed on Day 15 of pregnancy and corpora lutea, implantations,
and resorptions were counted. Other females were sacrificed on Postnatal Day 21.

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]: no data

HISTOPATHOLOGY / ORGAN WEIGHTS
Males:
At sacrifice, the following organs were excised and weighed: liver, kidney, adrenals, spleen. heart, testes, accessory organs (prostate and seminal vesicles), vas deferens, and epididymis.

Postmortem examinations (offspring):

No data

Statistics:

Body weights for all animals were analyzed using a repeated-measures analysis of variance (BMDP, Los Angeles. CA). Fertility and pregnancy rates were analyzed by X2 methods (Snedecor and Cochran, 1980). All other data were analyzed by one-way analysis of variance followed by Tukey’s multiple comparison procedure (SAS, Gary, NC), with the following parameters being transformed before analysis: mount latency period and ejaculation latency period (logarithmic): numbers of mounts and intromissions (square root); and percentage normal sperm morphology; and percentage motile sperm (arcsin). CellSoft-generated sperm motion parameters were analyzed using t tests (control vs treated groups) with a modified Bonferroni correction (Simes, 1986). Dose dependence was determined by linear regression for trend.

Reproductive indices:

In females which were sacrificed on Day 15 of pregnancy, corpora lutea, implantations, and resorptions were counted.

Offspring viability indices:

No data

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

all females in the 100 mg/kg group died or were moribund.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Males in the 50 mg/kg bw group exhibited a 7% decline in weight gain compared to controls. Females: no effects.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Males in the 50 mg/kg bw group exhibited a 7% decline in weight gain compared to controls. Females: no effects.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

not examined

Other effects:

not examined

Description (incidence and severity):

Test substance intake: not applicable

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

effects observed, treatment-related

Description (incidence and severity):

In the group of males, used for sperm motion analysis, cauda epididymal sperm counts were slightly but significantly decreased (50 mg/kg bw)

Reproductive performance:

no effects observed

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
Within the first week of treatment, all females in the 100 mg/kg group died or were so moribund as to warrant sacrifice. Animals in the remaining groups appeared healthy and in good condition.

Males: All animals appeared healthy and active throughout the experiment.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Males in the 50 mg/kg bw group exhibited a 7% decline in weight gain compared to controls. This difference was not significant (Table 1).
Females: Mean body weights across groups showed no differences during pregestational exposure nor during pregnancy (data not shown).


REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
The fertility rate and litter outcome data are shown in Table 6. Although mating success was somewhat low across all groups (since proestrous females were not chosen for mating trials), no differences were seen in the number of females mating or in number of successful pregnancies. Litter size and mean pup birth weight were not significantly different. One dam in the 25 mg/kg group neglected her pups, which all died by Postnatal Day 4. This accounts for the slightly decreased percentage survival seen in the 25 mg/kg group. Male and female pup weights, measured weekly, did not significantly differ during lactation or in the 3 weeks postweaning.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
There were no significant differences in cauda epididymal sperm count, testicular spermatid count, or epididymal sperm morphology in treated animals (Table 3). In the second group of males used for sperm motion analyses, testis and epididymis organ-to-body weight ratios were unchanged. However, cauda epididymal spem counts were slightly, but significantly decreased in the 50 mg/kg bw group (p<0.05) (Table 4). Table 5 shows shows the effects of ECH treatment on cauda epididymal sperm motion parameters as measured with the Cell-Soft computer-assisted sperm motion analysis
system. Consistent with the analyses of ejaculated sperm, there was no change in the percentage of motile cells in the cauda epididymides of animals from any dose group. Mean curvilinear velocity and straight-line velocity of cauda epididymal sperm were reduced across all groups in a dose-dependent fashion (p = 0.0001). Table 5 also shows theeffect of ECH treatment of male rats on specific motion endpoints related to sperm swimming pattern: linearity, amplitude of lateral head displacement, flagellar beat frequency, and number of cells displaying circular motion. Sperm track linearity was reduced only at the 50 mg/kg dose level. However, a dose-dependent trend was evident (p = 0.000 1). The amplitude of lateral head displacement
(ALH) was decreased and the beat/cross frequency was increased in all dose groups in a dose-dependent manner (p < 0.0006). No effect was seen on the percentage of sperm swimming in a circular motion.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Mating performance was unimpaired by ECH treatment, as were copulatory plug weight, ejaculated sperm count, percentage motil sperm, and sperm morphology (Table 2). In the mating trials designed to evaluate pre- and postimplantation loss, all control and 50 mg/kg - males mated, as verified by a copulatory plug or vaginal sperm. However, none of the females that mated with ECH males was pregnant, compared to a 90% pregnancy rate in controls (data not shown).

ORGAN WEIGHTS (PARENTAL ANIMALS)
Males: liver, kidney, and epididymal organ-to-body weight ratios were significantly higher (p<0.05) in 50 mg/kg bw animals relative to controls, although all differences were marginal. In the second group of males used for sperm motion analyses, testis and epididymis organ-to-body weight ratios were unchanged. However, cauda epididymal spem counts were slightly, but significantly decreased in the 50 mg/kg bw group (p<0.05) (Table 4).

GROSS PATHOLOGY (PARENTAL ANIMALS)
Females: Autopsy of females from the 100 mg/kg bw dose group that died showed hemorrhagic stomachs and kidneys with a very pale cortex and hemorrhagic medulla.

Dose descriptor:

NOAEL

Effect level:

50 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Impaired fertility (none of females mated with the treated males from 50 mg/kg bw dose group were pregnant)

Remarks on result:

not determinable

Remarks:

no NOAEL identified

Dose descriptor:

NOAEL

Effect level:

50 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No effects in any parameter examined.

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not specified

Histopathological findings:

not specified

VIABILITY (OFFSPRING)
Litter size and mean pup birth weight were not significantly different. One dam in the 25 mg/kg group neglected her pups, which all died by Postnatal Day 4. This accounts for the slightly decreased percentage survival seen in the 25 mg/kg group.

CLINICAL SIGNS (OFFSPRING)
No detailed description.

BODY WEIGHT (OFFSPRING)

Mean pup birth weight were not significantly different. Male and female pup weights, measured weekly, did not significantly differ during lactation or in the 3 weeks postweaning (Table 7).

SEXUAL MATURATION (OFFSPRING) Not reported in details

Remarks on result:

other: no information available

Reproductive effects observed:

not specified

TABLE 1
Male Body Weight and Organ/ 100 g Final Body Weight Ratios following Treatment with Epichlorohydrin (Means ± SD)
		Dose (mg/kg po)
	0 ^a	12.5 ^a	25 ^a	50 ^a
	(« = 20)	(n= 20)	(n = 20)	(n= 20)
Initial body weight (g)	356 ±37.7	353 ±37.1	352 ± 28.6	355 ±40.3
Final body weight (g)	403 ±38.5	390 + 34.6	386 + 24.2	377 ± 42.0
Liver	3.580 ±0.282	3.654 + 0.362^b	3.722 ±0.377 ^b	3.876 ±0.224 ^b
Kidney	0.413 ±0.025	0.425 ±0.049 ^b	0.412 ±0.026	0.445 ±0.037 ^b
Adrenal	0.0134 ±0.0028	0.0148 ±0.0033	0.0148 ±0.0031	0.0143 ±0.0034
Spleen	0.174 ±0.032	0.165 ±0.021	0.167 ±0.022	0.175 ±0.029
Heart	0.298 ± 0.023	0.303 ± 0.024	0.303 ±0.017	0.314 ±0.020
Testis	0.409 ±0.043	0.406 ±0.042	0.420 ±0.042	0.423 ±0.045
Vas deferens	0.0263 ±0.0025	0.0266 ± 0.0039	0.0283 ±0.0031 ^b	0.0291 ±0.0044 ^b
Accessory organs	0.735 ±0.107	0.771 ±0.094	0.791 ±0.092	0.782 ±0.144
Epididymis	0.134±0.015	0.138 + 0.01^b	0.143 + 0.012 ^b	0.148 ±0.020 ^b
Note.Organ weights expressed as g/100 g body wt.   ^a,b Groups with different superscripts are significantly different(p <0.05).

TABLE 2 Male Mating Behavior and Ejaculated Semen Parameters after 21 Days of Epichlorohydrin Treatment (Means ± SD)
	Dose (mg/kg po)
	0(n= 20)^a	12.5(n =20)^a	25(n =20)^a	50(n= 20)^a
Number of mounts	2.6 ±2.33	1.6 ± 1.54	2.2+1.56	2.9 + 2.02
Number of intromissions	15.8 ± 4.10 ^b	12.6 ±3.69	15.0 ± 3.49 ^b	14.8±3.90 ^b
Mount latency (sec)	7.8 ± 4.22	15.0 ±30.52	12.6 ± 12.36	10.3 ± 8.6
Ejaculation latency (sec)	280 ±81.2	267 ± 196.5	249 ± 140.4	247 ± 73.4
Plug weight (g)	0.12 ±0.022	0.12 ±0.022	0.12 ±0.020	0.12 ±0.017
Sperm count X 10^6	48 ± 13.3	53 ± 12.1	54 ±9.2	45± 15.1
% Normal morphology	98 + 2.0	98 ± 1.8	98 ± 1.6	98±0.1
% Motile	45± 1.1	37 ± 1.4	42 ±0.8	41 ± 1.0
^a,bGroups with different superscripts are significantly different(p <0.05).

TABLE 3 Sperm Parameters Obtained at Sacrifice (Means ± SD)"
	Dose (mg/kg po)
	0	12.5	25	50
Cauda epididymal sperm count (X10^6/g cauda)	645 ± 188.7  (n = 10)	698 ± 95.1 (n=19)	736 ± 113.4 (n = 20)	612±199.4 (n = 20)
Spermatid count (X10^6/g testis)	119 ± 21.8 (n = 20)	110 ± 15.9 (n = 19)	120 + 21.7 (n = 20)	120+ 12.7 (n = 20)
% Normal morphology	96.7 ± 0.02 (n = 10)	94.6 ± 0.07 (n = 16)	96.0 ± 0.05 (n = 18)	97.3 + 0.33 (n = 18)
'' No significant differences observed between groups on any of the sperm parameters.

TABLE 4. Male Body Weight, Organ-to-Body Weight Ratios, and Cauda Epididymal Sperm Counts  for Second Group of Males Used in Motion Analyses (Means ± SD)
		Dose (mg/kg po)
		0'' (n = 18)	12.5'' (n = 16)	25''(n = 18)	50 (n= 15)
Initial body weight (g)		434 + 16.8	428 ± 18.8	429 ± 14.9	432 ± 25.1''
Final body weight (g)		445 ± 17.4	444 ± 26.2	438 ± 21.1	426 ± 36.1''
Testis		0.38 ± 0.044"	0.38 ± 0.020	0.38 ± 0.023	0.40 ± 0.030''
Epididymis		0.13±0.013	0.13 ± 0.011	0.13 ± 0.010	0.14 + 0.015''
Sperm count (X10^6/g cauda)		574 ± 93.5	568 ± 93.8	648 ± 117.9	446 ± 115.8*
'',* Groups with different superscripts are significantly different(p <0.05).

TABLE 6 Fertility Rates and Litter Outcomes in Females Treated with Epichlorohydrin (Means ± SD)''
	Dose (mg/kg po)
	0		25	50
Fertility rate*	60		70	57
Litter size	14.4 ± 1.4 (n = 12)		13.5 ± 3.3 (n = 12)	13.6 ± 1.8  (n = 10)
% Survival at Day 4	97  (n = 12)		90 (n = 12)	99 (n = 10)
Birth weight	5.8 ± 0.48 (n = 12)		5.7 ± 0.46 (n = 12)	5.9 ±0.52 (n = 10)
Weaning weight (Day 21)	44.4 ± 3.28 (n=12)		46.1 ± 4.18 (n= 11)	44.6 ± 2.00 (n = 10)
" Groups did not differ significantly on parameters. * Fertility rate = (No. pregnant/No. mated) x 100.

TABLE 7
Body Weights of Male and Female Pups Postweaning (Means ± SD)''
	Dose (mg/kg po)
	0	25	50
Day 28
Male	78 ±5.12	80 ± 6.32	78 ± 5.66
	(n = 11)	(n = 11)	(n = 10)
Female	69 ± 3.59	72 ±6.41	71 ±3.72
	(n = 12)	(n = 11)	(n = 10)
Day 35
Male	129 ±8.85	132 ±9.75	124 ± 12.84
	(n = 11)	(n = 11)	(n =10)
Female	109 ±3.88	111 ± 8.29	110 ±7.48
	(n = 12)	(n = 11)	(n = 10)
Day 42
Male	185 + 10.84	188 ± 14.39	178 ± 12.74
	(n = 11)	(n = 11)	(n = 10)
Female	139 ± 5.18	141 ± 12.60	138 ± 10.40
	(n = 12)	(n = 11)	(n = 10)
" Groups did not differ significantly on parameters.

Conclusions:

Epichlorohydrin reduced sperm motion parameters at a dose as low as 12.5 mg/kg/day and total infertility at 50 mg/kg/day fertility in exposed male rats for 21 days. No reproductive or developmental parameters were affected in femal rats.

Executive summary:

Effects of Epichlorohydrin on Male and Female Reproduction in Long-Evans Rats was studied (Toth et al., 1989). The animals were treated with epichlorohydrin (ECH) by oral gavage (males: 12.5, 25, and 50 mg/kg/day; females: 25, 50, and 100 mg/kg/day) for 21 and 14 days, respectively, prior to mating trials with untreated animals. Treated females were further dosed until delivery. Fertility was assayed in the high-dose males only and was found to be totally impaired. None of the females that mated with ECH males was pregnant, compared to a 90% pregnancy rate in controls. However, treated males showed normal copulatory behavior. Sperm morphology and percentage motile sperm were not statistically different from control values in both ejaculated and cauda epididymal samples from ECH-treated animals. The number of sperm in ejaculates was normal while cauda epididymal sperm count was slightly decreased in males at the 50 mg ECH/kg dose level. The decreased cauda epididymal sperm count is unlikely to be responsible for the observed infertility. However, it may in part be a result of the effects of ECH on the vigor and swimming pattern of cauda epididymal sperm. Mean curvilinear velocity, straight-line velocity, and amplitude of lateral head displacement of cauda epididymal sperm were significantly reduced by ECH at 12.5 mg/kg/day and above. Sperm track linearity was also reduced, but only at 50 mg/kg/day. Beat/cross frequency of sperm was significantly increased at 12.5 mg/kg/day and above. All of the above sperm motion parameters showed dose-dependent trends. At necropsy, liver, kidney, and epididymal organ-to-body weight ratios were significantly higher in 50 mg/kg males (p<0.05) relative to controls, although all differences were marginal. Liver and kidneys are known targets for epichlorohydrin toxicity. Increased epididymis-to-body weight ratios are possibly a result of inflammation and swelling from spermatocoeles and sperm granulomas.In the second group of males used for sperm motion analyses, testis and epididymis organ-to-body weight ratios were unchanged.

Within the first week of treatment, all females in the 100 mg/kg group died or were so moribund as to warrant sacrifice. Autopsy showed hemorrhagic stomachs and kidneys with a very pale cortex and haemorrhagic medulla. Animals in the remaining groups appeared healthy and in good condition. Mean body weights across groups showed no differences during pregestational exposure nor during pregnancy. No measured parameters of female reproduction were changed relative to controls. No differences were seen in the number of females mating or in number of successful pregnancies. Litter size and mean pup birth weight were not significantly different. One dam in the 25 mg/kg group neglected her pups, which all died by Postnatal Day 4. This accounts for the slightly decreased percentage survival seen in the 25 mg/kg group. Male and female pup weights, measured weekly, did not significantly differ during lactation or in the 3 weeks postweaning.

In conclusion, no evidence for ECH-induced female reproductive or developmental toxicity was observed at doses up to 100 mg/kg/day. However, male rats showed reductions in sperm motion parameters at a dose as low as 12.5 mg/kg/day and total infertility at 50 mg/kg/day.

Effect on fertility: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEL

12.5 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

There are a lot studies available for structurally similar epoxides. Therefore the overall quality of the database is high.

Effect on fertility: via inhalation route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

11 ppm

Study duration:

subchronic

Species:

rat

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Several non-guideline-compliant studies on "read-across" substances are available. There is a one-generation study on epichlorohydrin (CAS 106 -89 -8) (Toth, 1989) considered as key information. Beside the one-generation study on on epichlorohydrin (CAS 106 -89 -8) , there is a two-generation inhalation study on phenylglycidyl ether (CAS 122 -60 -1) (Terril, 1982) and a developmental toxicity study with epichlorohydrin (Marks, 1982) available. Also, there are QSAR predictions that provide further supporting information. Taken all available data into account, this endpoint is expected to be adequately address.

There is no study on reproductive toxicity potential of the target substance available. Therefore, read-across from the related substances (containing glycidyloxy moiety in their chemical structures) was performed to evaluate this endpoint. Data on epichlorohydrin (CAS 106 -89 -8) and phenyl glycidyl ether (CAS 122 -60 -1) are available. 

The epoxy compounds are known to be potential toxicants to male fertility (please refer to read-across statement). They reduce sperm motility and sperm count and produce testicular atrophy in treated animals (NTP TR 374, 1990; NTP TR 376, 1990; BAuA, 2012; Screening Assessment, 2010; NIOSH, 1988a,b; HPV, 2002). Low molecular weight epoxides produce adverse effects in male fertility in animal studies while high molecular weight compound (containing a hydrophobic hydrocarbon chain) do not.

 

Data on epichlorohydrin (CAS 106 -89 -8)

 

Effects of epichlorohydrin on male and female reproduction in Long-Evans rats was studied (Toth et al., 1989). The animals were treated with epichlorohydrin (ECH) by oral gavage (males: 12.5, 25, and 50 mg/kg/day; females: 25, 50, and 100 mg/kg/day) for 21 and 14 days, respectively, prior to mating trials with untreated animals. Treated females were further dosed until delivery. Fertility was assayed in the high-dose males only and was found to be totally impaired. None of the females that mated with ECH males was pregnant, compared to a 90% pregnancy rate in controls. However, treated males showed normal copulatory behaviour. Sperm morphology and percentage motile sperm were not statistically different from control values in both ejaculated and cauda epididymal samples from ECH-treated animals. The number of sperm in ejaculates was normal while cauda epididymal sperm count was slightly decreased in males at the 50 mg ECH/kg dose level. The decreased cauda epididymal sperm count is unlikely to be responsible for the observed infertility. However, it may in part be a result of the effects of ECH on the vigour and swimming pattern of cauda epididymal sperm. Mean curvilinear velocity, straight-line velocity, and amplitude of lateral head displacement of cauda epididymal sperm were significantly reduced by ECH at 12.5 mg/kg/day and above. Sperm track linearity was also reduced, but only at 50 mg/kg/day. Beat/cross frequency of sperm was significantly increased at 12.5 mg/kg/day and above. All of the above sperm motion parameters showed dose-dependent trends. At necropsy, liver, kidney, and epididymal organ-to-body weight ratios were significantly higher in 50 mg/kg males (p<0.05) relative to controls, although all differences were marginal. Liver and kidneys are known targets for epichlorohydrin toxicity. Increased epididymis-to-body weight ratios are possibly a result of inflammation and swelling from spermatocoeles and sperm granulomas.In the second group of males used for sperm motion analyses, testis and epididymis organ-to-body weight ratios were unchanged.

Within the first week of treatment, all females in the 100 mg/kg group died or were so moribund as to warrant sacrifice. Autopsy showed haemorrhagic stomachs and kidneys with a very pale cortex and hemorrhagic medulla. Animals in the remaining groups appeared healthy and in good condition. Mean body weights across groups showed no differences neither during pregestational exposure nor during pregnancy. No measured parameters of female reproduction were changed relative to controls. No differences were seen in the number of females mating or in number of successful pregnancies. Litter size and mean pup birth weight were not significantly different. One dam in the 25 mg/kg group neglected her pups, which all died by Postnatal Day 4. This accounts for the slightly decreased percentage survival seen in the 25 mg/kg group. Male and female pup weights, measured weekly, did not significantly differ during lactation or in the 3 weeks postweaning.

In conclusion, no evidence for ECH-induced female reproductive or developmental toxicity was observed at doses up to 100 mg/kg/day. However, male rats showed reductions in sperm motion parameters at a dose as low as 12.5 mg/kg/day and total infertility at 50 mg/kg/day.

 

Data on phenyl glycidyl ether (CAS 122 -60 -1)

 

The effects of phenylglycidyl-ether (PGE) in rats following inhalation of atmospheres containing either 0, 2, 6, or 11 ppm were measured in a two-generation reproduction study (Terril et al., 1982). The potential teratogenicity effects of PGE were also evaluated in rats exposed to the same concentrations during the period of rapid organogenesis. No significant changes in reproduction parameters (fertility, progeny numbers and survival, lactational performance) were produced by PGE inhalation, and there was no evidence of dominant lethality. Exposure of pregnant rats to the compound, 6 hr/day from Gestation Days 4 through 15 resulted in no embryotoxicity or teratogenicity effect.

So, as normal fertility was observed, and there was no evidence of increased early embryonic death (dominant lethals) in the reproduction study, the test substance is not considered to bear a significant potential to change reproductive parameters. Test progenies were delivered in normal numbers; these animals survived and grew like the controls. A decrease in the number of pregnancies resulting from mating to males exposed at the highest level (11 ppm) the first week post exposure suggested a treatment relationship, but all other reproductive data appeared normal and it is concluded that this isolated finding was not related to exposure to PGE. A single pup (2 ppm) displayed an abnormal hair pattern (curly hair) which again was an isolated occurrence of doubtful biologic significance. Teratogenicity and cytogenetic (bone marrow cells) tests failed to reveal any effects which could be related to PGE exposure. It is recognized that the evaluation of the mutagenicity, teratogenicity, and reproductive effects can be measured in a wide variety of test systems. It is concluded that rats exposed to atmospheres of up to 11 ppm PGE showed no significant abnormalities in cytogenetic, teratogenicity, or reproductive evaluations.

 

Prediction of the reproductive toxicity by the OECD QSAR Toolbox (v3.1)

 

The target chemical was profiled as "Epoxides" by the "US EPA New Chemical Categories". According to the US EPA definition of glycidyls, the target substance represents triglycidyl ether of glycerine and therewith belongs also to the epoxide group of chemicals due to the presence of epoxide moiety in glycilydoxy group of the molecule. Common properties of glycidyls are high reactivity, cytotoxicity, and high probability of mutagenic potential and/or carcinogenicity. Chemicals with the same profiling result have been retrieved from the database. The chemicals containing other chemical elements in their structure and/or other organic functional groups were considered as dissimilar to the target chemical and have been removed from the domain. The predictions were based on the experimental data of chemicals with different units (mg/kg bw, mg/L air, and ppm analytical). Therefore the substances have been grouped in three different domains (three different predictions).

The target chemical is obtained by the reaction of epichlorohydrin with glycerol. Therefore, epichlorohydrin is considered to be a suitable candidate for read-across. Further substances used in the read-across were ethyleneoxide as the simplest representative of epoxy compounds, allyl,- and phenyl glycidyl ethers which are members of glycidyl ether category according to the Epoxy Resin Systems Task Group (ERSTG) (HPV, 2001). Moreover, their profiling results regarding binding to proteins and to DNA (properties which are likely responsible for reproductive toxicity) are similar to those of the target.

Epichlorohydrin caused reduced male fertility and an increase in the incidence of histopathological changes of the testis and epididymidis. Allyl glycidyl ether induced multiple adverse effects on the reproductive system of treated animals and is officially classified for reproductive toxicity Cat. 2 H361 "Suspected of damaging fertility or the unborn child". Phenyl glycidyl ether caused testicular atrophy as well as reduced reproductive capacity in treated animals if administered orally. Based on these data reproductive adverse effects of the target chemical cannot be ruled out.

Effects on developmental toxicity

Description of key information

Developmental toxicity, ReA epichlorohydrin and glycidol, albino mice and rats: no developmental effects

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: well documented publication, which meets basic scientific principles

Principles of method if other than guideline:

The present study was undertaken to determine whether epichlorohydrin and/or glycidol is teratogenic. Pregnant outbred albino rats (CD) and mice (CD-1) were given epichlorohydrin by gastric intubation on day 6-15 of gestation. The rats were killed on day 21 (day 18 for mice) and the offspring checked for gross, visceral, and skeletal malformations.

GLP compliance:

not specified

Limit test:

no

Species:

other: rat and mouse

Strain:

other: CD-rats and CD-1-mice

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- male and nulliparous female rats and mice
- Source: Charles River Breeding Laboratories (Wilmington, Mass.)
- Age at study initiation: 60 -90 d (mice)
- Weight at study initiation: 176-200 g (rats)
- Housing: Females werer housed in groups of 10 mice or 4 rats per polypropylene cage. The males were housed individually in polypropylene cages in seperated rooms by species.
- Diet (e.g. ad libitum): The animals were fed Purina Lab Chow ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature: 72-75 °F
- Humidity (%): 45-50 % relative humidity
- Photoperiod (hrs dark / hrs light): 12-h daily light cycle

Route of administration:

oral: gavage

Vehicle:

cotton seed oil

Details on exposure:

On day 6-15 of gestation, epichlorohydrin or glycidol was administered by gastric intubation at 40, 80, and 160 mg/kg/d in the rat study and at 80, 120, and 160 mg/kg/d in the mouse study. Glycidol was administered to mice at 100, 150, or 200 mg/kg/d. Each epichlorohydrin dosage except 120 mg/kg/d was studied independently of the other dosages in four or five replicates; 120 mg/kg/d was studied in only one replicate. The glycidol study was carried out in five replicates. Since no significant differences between replicates occurred, the results were combined in one table for each study. The dose that produced approximately 10% mortality was used as the maximum dose for each study; half this dose was used as a second dose. Later replicates included a third dose halfway between the high and low doses.
Epichlorohydrin was dissolved in cottonseed oil and glycidol was dissolved in sterile, distilled water (Gibco, Grand Island, N.Y.). In the rat study, the vehicle containing the epichlorohydrin was administered on a 0.1% (v/w) body weight basis. Epichlorohydrin and glycidol were administered to mice on a 1% (v/w) body weight basis. Since the animals and test (or control) groups were individually identified only by number, laboratory personnel knew neither the test agent nor the test group.

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

After one week of acclimatisation, the mating period was begun by placing two females into each male's cage. The following morning, mice with vaginal plugs and rats with spermatozoa in their vaginal lavage (day 1 of gestation) were caged with the other animals found pregnant on that day.
Before d 1 of treatment, the dams were divided into experimental and control groups such that body weight differences between groups were minimized. The animals were individually identified by crystal violet markings and a number.

On day 6 -15 the animals were dosed with epichlorohydrin or glycidol.

Duration of treatment / exposure:

gestation day 6 - 15

Frequency of treatment:

once daily

Duration of test:

gestation day 6 - 15

Remarks:

Doses / Concentrations:
40 mg/kg/day
Basis:
nominal conc.
Epichlorohydrin, rat study

Remarks:

Doses / Concentrations:
80 mg/kg/day
Basis:
nominal conc.
Epichlorohydrin, rat and mouse study

Remarks:

Doses / Concentrations:
120 mg/kg/day
Basis:
nominal conc.
Epichlorohydrin, mouse study

Remarks:

Doses / Concentrations:
160 mg/kg/day
Basis:
nominal conc.
Epichlorohydrin, rat and mouse study

Remarks:

Doses / Concentrations:
100 or 150 or 200 mg/kg/day
Basis:
nominal conc.
Glycidol, mouse study

Maternal examinations:

On day 18 (d 21 for rats) of gestation, the mice were killed by cervical dislocation (the rats were first rendered unconscious with C02) and their reproductive status was determined.

Ovaries and uterine content:

Implantation sites in each uterine horn were counted and the general condition of each conceptus was recorded.

Fetal examinations:

Live fetuses were weighed individually, sexed internally, and examined for external anomalies (variations and malformations). Live fetuses weighing <0.5 g ( < 1.0 g for rats) or weighing less than two-thirds the mean of their larger littermates were designated as "stunted." At least one-third of the fetuses of each litter, as well as all stunted fetuses and those having external malformations, were examined for visceral alterations. The bodies of all fetuses were then processed for skeletal examination. The heads of fetuses subjected to visceral examination (with the exception of any fetuses that had external head malformations) were cut off at the base and prepared for freehand sectioning.
The average percent of malformed fetuses was calculated for each test group and for the vehicle controls as 100 * Sum of ((no of malformed fetuses in litter)/(no. of fetuses in litter))/(total no. of litters)

Statistics:

Statistically significant differences between groups were determined by the Mann-Whitney Latest or the Student t-test. Jonckheere's test was employed to determine the significance of dose-response relations. Two-tailed tests were performed and p< 0.05 was selected as the level of significance.

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
In the rat study, epichlorohydrin at 160 mg/kg/d was lethal to 3 of 27 dams and also significantly (two-sided p < 0.05 versus the vehicle control group) increased the average liver weight of the dams. At 80 mg/kg/d, epichlorohydrin caused a significant decrease in the average weight gain during pregnancy.

Dose descriptor:

other: not specified

Basis for effect level:

other: effect type not specified

Remarks on result:

other: no further information available

Dose descriptor:

other: not specified

Basis for effect level:

other: effect type not specified

Remarks on result:

other: no further information available

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
At 80 mg/kg/d, no significant differences in the average percent of malformed fetuses occurred between the control (0.18%) and treated groups (<= 0.76 %). Only three malformations were observed in this study: hydronephrosis in a control fetus, agnathia in a fetus in the 40 mg/kg/d group, and general oedema in a fetus in the 160 mg/kg/d group.

Remarks on result:

other: not specified

Abnormalities:

not specified

Developmental effects observed:

not specified

In this study, the type and quantity of variations found were not out of the ordinary and therefore will not be presented or discussed in this report.

In the rat study, epichlorohydrin at 160 mg/kg/d was lethal to 3 of 27 dams and also significantly (two-sided p < 0.05 versus the vehicle control group) increased the average liver weight of the dams (Table 1). At 80 mg/kg/d, epichlorohydrin caused a significant decrease in the average weight gain during pregnancy. In spite of these indications of maternal toxicity, no significant differences in the average percent of malformed fetuses occurred between the control (0.18%) and treated groups (<= 0.76 %). Only three malformations were observed in this study: hydronephrosis in a control fetus, agnathia in a fetus in the 40 mg/kg*d group, and general edema in a fetus in the 160 mg/kg'd group.

TABLE 1. Effect of Epichlorohydrin on Reproduction in Rats^a
Measurement	Dose (mg/kg/d)^b
	Control	40	80	160
Number of dams receiving test agent^d	35	14	25	27
Number of dams alive on d 18	35	14	24	24
Number of dams pregnant^e	32	10	23	22
Average weight gain (g) during pregnancy^	95.0 ± 1.97	85.6 ± 4.48^g	84.0 ± 4.35^h	88.1 ± 2.84
Average liver weight (± SE)^e	15.0 + 0.35	14.9 ± 0.59	16.1 ± 0.33	16.4 ± 0.37^h
Number of implants (avg.)^j	436 (13.6)	135 (13.5)	316 (13.7)	288 (13.1)
Number of females with resorptions	16	4	11	12
Number of resorptions	24	4	17	16
Average percent resorptions per litter	5.5	3.0	5.4	5.6
Number of fetal deaths	0	0	0	0
Average percent fetal deaths per litter	0	0	0	0
Male/female live fetuses	230/182	66/65	166/132^j	128/144
Number of stunted fetuses	1	0	0	0
Average number of live fetuses per dam	12.9	13.1	13.0	12.3
Average fetal weight (g)^k	3.73 ± 0.051	3.82 ± 0.084	3.71 ± 0.080	3.70 ± 0.052
^aKiIled on d 21 of gestation after receiving epichlorohydrin (gavage) on d 6-15 of gestation.
^bFive of five dams died before d 4 of dosing at 240 mg/kg.
^cReceived vehicle (cottonseed oil) only.
^dThe criterion used to identify mated females was spermatozoa in the vaginal lavage.
^eIncludes dams with all resorptions.
^fDays 6-20 of gestation; excludes dams with all resorptions; mean ± SE.
^gFive dams were omitted because of failure to record their weights on d 6.
^hTwo-sidedp <0.05 versus vehicle control group.
^iPer pregnant rat.
^jThe sex of one fetus was not determined.
^kStunted but- not dead fetuses were included; average of mean fetus weights per litter ± SE.

In the mouse study, epichlorohydrin at 160 mg/kg/d was lethal to 3 of 32 dams Table 2). No significant adverse effects on the average weight gain during pregnancy occurred at any of the doses tested. The 120 mg/kg/d dose did produce a significant increase in the average liver weight of the dams. This dose, as well as 160 mg/kg/d, led to a significant decrease in average fetal weight. The effect of epichlorohydrin on this parameter, as well as on average dam liver weight, also showed a significant (p< 0.005) trend with increasing dose levels (Jonckheere's test). Nonetheless, this compound did not cause a significant increase in the average percent of malformed fetuses at any of the doses tested (<= 3.5 %) as compared with the vehicle control group (0.23%).

No malformations were observed in the group receiving 120 mg/kg/d. One fetus in the control group had gastroschisis, ectocardia, and no right carotid artery. Three fetuses in the 80 mg/kg/d group had exencephaly with one or both eyes open, and one of these also had gastroschisis and cleft face. Another fetus in this group had fused arches, fused centra, and fused ribs. These four fetuses were in four litters.

In the 160 mg/kg/d group, five fetuses (four in the same litter) had exencephaly and bilateral open eyes; two fetuses (separate litters) had cleft palate; and another fetus had one eye open. A total of eight malformed fetuses in four litters was observed at this dose.

TABLE 2.Effect of Epichlorohydrin on Reproduction in Mice^a
Measurement	Dose (mg/kg/d)
	Control^b	80	120	160
Number of dams receiving test agent	49	31	24	32
Number of dams alive on d 18	49	31	24	29
Number of dams pregnant^c	40	25	22^d	20
Average weight gain (g) during pregnancy^e	17.5 ± 0.47	19.9 ± 0.79^f	19.1 ± 0.84^f	15.2 ± 1.44
Average liver weight (±SE)^c,g	2.37 ± 0.32^h	2.47 ± 0.054	2.52 ± 0.089^f	2.50 ± 0.119
Total number of implants	509	325	288	239
Average number of implants^i	12.7	13.0	13.1	12.0
Number of resporptions	46	23.0	44	22
Percent resorptions of total number of implants	9.0	7.1	15.3	9.2
Number of fetal deaths	8	1	6	8
Percent fetal deaths of total number of implants	1.6	0.31	2.1	3.1
Male/female live fetuses	212/243	169/132	128/110	108/101
Number of stunted fetuses	2	4	0	6^j
Average number of live fetuses per dam^e	11.4	12.0	11.9	10.5
Average fetal weight (g)^g,k	0.991 ± 0.011	0.990 ± 0.021	0.925 ± 0.022^f	0.898 ± 0.034^f
^aKilled on d 18 of gestation after receiving epichlororhydrin (gavage) on d 6-15 of gestation.
^bReceived vehicle (cottonseed oil) only.
^cIncludes dams with all resorptions.
^dTwo dams had all resorptions (implant sites).
^eDays 6-15 of gestation; excludes dams with all resorptions; mean ± SE.
^fTwo-sidedp <0.05 versus vehicle control group.
^gSignificant trend (two-sidedp <0.005 versus vehicle control group) as dose increased (Jonckheere's test).
^hOne liver weight not determined.
^iPer pregnant mouse.
^jFive of these fetuses were in one litter.
^kStunted but not dead fetuses were included; average mean fetus weights per litter ± SE.

Glycidol produced a significant number of stunted mouse fetuses in the group receiving 200 mg/kg/d (Table 3), but all of the stunted fetuses were in the same litter. In addition, 5 of the 30 females given this dose died, or were moribund and had to be sacrificed by d 16 of gestation. Two other dams experienced ataxia on one or more days during the treatment period.

Although a statistically significant increase in the average percent of malformed fetuses was not obtained as the result of glycidol treatment, 6 of the 15 stunted littermates in the 200 mg/kg/d groups had cleft palate. Since these cleft palates were found in stunted fetuses, they were not scored as malformations. Thus, even at 200 mg/kg/d, a dose that killed 5 of the 30 dams treated, glycidol did not cause a significant increase in the average percent of malformed fetuses (<= 0.66 %) as compared with the vehicle control group (0.23%). In this study, one fetus in the control group had fused sternebrae; one fetus in the 100 mg/kg/d dose group had meningocele and a small hole in the heart, and another in a second litter had fused sternebrae; and one fetus in the 200 mg/kg/d group had cleft palate. Glycidol at 500 mg/kg/d was either lethal or resulted in morbidity in 9 of 9 pregnant mice by day 10 of gestation.

TABLE 3. Effect of Glycidol on Reproduction in Mice^a
Measurement	Dose (mg/kg/d)^b
	Control	100	150	200
Number of dams receiving test agent^d	32	37	31	30
Number of dams alive on d 18	32	37	31	25
Number of dams pregnant	29	34	30	21
Average weight gain (g) during pregnancy^e	17.1 ± 0.832	19.3 ± 0.522	19.8 ± 0.600	17.0 ± 1.25
Number of implants (avg.)^f	373 (12.9)	458 (13.5)	390 (13.0)	291 (13.9)
Number of females with resorptions	23	28	21	17
Number of resorptions	41	50	32	34
Average percent resorptions per litter	11.0	10.9	8.2	11.7
Number of fetal deaths	3	10	5	3
Average percent fetal deaths per litter	0.8	2.2	1.3	1.0
Male/female live fetuses	187/142	190/208	183/170	114/140
Number of stunted fetuses	1	1	1	15^g,h
Average number of live fetuses per dam	11.3	11.7	11.8	12.1
Average fetal weight (g)^i	1.01 ± 0.015	0.984 ± 0.017	0.980 ± 0.016	0.955 ± 0.027
^aKilled on d 18 of gestation, after receiving glycidol (gavage) on d 6-15 of gestation.
^bNine of nine dams receiving glycidol at 400 mg/kg died or were sacrificed (near death) by d 10.
^cReceived vehicle (sterile distilled water) only.
^dVaginal plug was the criterion used to identify mated females.
^eDays 6-20 of gestation; mean ± SE.
^fPer pregnant mouse.
^gAIl fetuses were in the same litter.
^hTwo-sidedp <0.05 versus vehicle control.
^iStunted but not dead fetuses were included; average of mean fetus weights per litter ± SE.

Conclusions:

The results of the present study indicate that epichlorohydrin is not teratogenic in albino mice and rats. The 160 mg/kg/d dose was lethal to some of the pregnant rats. The rat dams experienced significant reductions in weight gain with doses as low as 80 mg/kg/d. However, the average foetal weight, average number of live foetuses per day, average per cent of resorptions per litter, and number of stunted fetuses were not significantly affected by this compound. Thus epichlorohydrin apparently was not toxic to the rat embryo or foetus, even at doses that were toxic to the pregnant rat. Epichlorohydrin did cause a statistically significant decrease in average foetal (mouse) weight at 120 and 160 mg/kg/d. In addition, the overall dose-response effect on this parameter was also statistically significant (Jonckheere's test). The effects were confounded by the fact that both dose levels caused maternal toxicity. The 160 mg/kg/d dose killed 3 of 32 dams and the 120 mg/kg/d dose produced a statistically significant increase in the average liver weight in the treated mice. Thus signs of embryotoxicity were observed at doses that were toxic to the dam.

Executive summary:

Pregnant outbred albino rats (CD) and mice (CD-1) were given epichlorohydrin by gastric intubation on day 6-15 of gestation (Marks, 1982). The rats were killed on day 21 (day 18 for mice) and the offspring checked for gross, visceral, and skeletal malformations. Epichlorohydrin caused a significant reduction in the weight gain of pregnant rats at 80 mg/kg/d as compared with the control group treated only with the vehicle. However, there was no evidence of teratogenicity in the rat fetuses even at a dose level (160 mg/kg/d) that caused the death of some of the treated dams. Epichlorohydrin also did not produce a statistically significant increase in the average percent of malformed mouse foetuses, even at 160 mg/kg/d, a dose that killed 3 of 32 treated dams. The 120 and 160 mg/kg/d levels did cause a significant (p < 0.05) reduction in the average foetal weight as compared with controls. In addition, the 120 mg/kg/d dose produced a statistically significant increase in the liver weight of the pregnant mouse. These observations indicate that the 120 and 160 mg/kg/d dose levels were toxic toward the dams and their unborn offspring. In a similar mouse study, glycidol showed no evidence of teratogenicity. There was a significant increase in the number of stunted foetuses at 200 mg/kg/d, but all of these were present in a single litter. Further, the same dose killed 5 of 30 dams.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Study duration:

subchronic

Species:

other: rat, mouse

Quality of whole database:

There are a lot studies publically available for structurally similar epoxides. Therefore the overall quality of the database is high.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

There is no developmental study available for the target substance. Read-across substances epichlorohydrin and glycidol were not developmental toxicants in the animal studies. A variety of other related epoxides e related substances (containing glycidyloxy moiety in their chemical stuctures) was performed to evaluate this endpoint. The epoxy compounds are not developmental toxicants (please refer to read-across statemnent).

The target chemical was profiled as "Epoxides" by the "US EPA New Chemical Categories". Common properties of epoxides are high reactivity, cytotoxicity, and high probability of mutagenic potential and/or carcinogenicity. Therefore chemicals with the same profiling result have been retrieved from the database. The chemicals containing other chemical elements in their structure and/or other organic functional groups were considered as dissimilar to the target chemical and have been removed from the domain.

The target chemical is obtained by the reaction of epichlorohydrin with glycerol. Therefore, epichlorohydrin is considered to be a suitable candidate for read-across. Glycidol is the simplest representative of glycidyl ethers category (HPV, Epoxy Resin Systems Task Group (ERSTG), 2001). 7-oxabicyclo-hept-3-ylmethyl 7-oxabicyclo-heptane-3-carboxylate is more lipophilic representative of glycidyl ethers category. Moreover, the profiling results of the category members regarding their ability to bind to proteins and to DNA (properties which are likely responsible for developmental toxicity) are similar to those of the target chemical.

Epichlorohydrin if administered to pregnant rats caused no embryotoxic, fetotoxic or teratogenicity effects (Marks et al., 1982; John, 1983). No developmental malformations were noted in mice treated with glycidol in a developmental study (Marks et al., 1982; IARC Monographs, Volume 77). No influence on embryonic or pup development was observed in the 13- week repeated dose toxicity and fertility study conducted with triglycidyl isocyanurate (HPV, No. 201 -15759).

 

Pregnant outbred albino rats (CD) and mice (CD-1) were given epichlorohydrin by gastric intubation on day 6-15 of gestation (Marks et al., 1982). The rats were killed on day 21 (day 18 for mice) and the offspring checked for gross, visceral, and skeletal malformations. Epichlorohydrin caused a significant reduction in the weight gain of pregnant rats at 80 mg/kg/d as compared with the control group treated only with the vehicle. However, there was no evidence of teratogenicity in the rat foetuses even at a dose level (160 mg/kg/d) that caused the death of some of the treated dams. Epichlorohydrin also did not produce a statistically significant increase in the average per cent of malformed mouse foetuses, even at 160 mg/kg/d, a dose that killed 3 of 32 treated dams. The 120 and 160 mg/kg/d levels did cause a significant (p < 0.05) reduction in the average foetal weight as compared with controls. In addition, the 120 mg/kg/d dose produced a statistically significant increase in the liver weight of the pregnant mouse. These observations indicate that the 120 and 160 mg/kg/d dose levels were toxic toward the dams and their unborn offspring. In a similar mouse study, glycidol showed no evidence of teratogenicity. There was a significant increase in the number of stunted foetuses at 200 mg/kg/d, but all of these were present in a single litter. Further, the same dose killed 5 of 30 dams.

Justification for classification or non-classification

Toxicity to reproduction

Experimental data revealed that the related substance Epichlorohydrin induces adverse effects on male fertility at 12.5 mg/kg/day. Related substances that contain glycidyloxy moieties in their structures produced damage of male fertility and testicular atrophy. The pattern of toxicity was similar in numerous animal studies. Based on the significant body of evidence, reproductive toxicity of the target substance GE-100 can not be ruled out. Therefore, it does meet the criteria for classification and will require labelling for this endpoint, according to the European regulation (EC) No. 1272/2008.

Proposed C&L: Toxicity to reproduction, Cat 2, H361f (suspected of damaging fertility (males).

 

Developmental toxicity

 

No evidence of developmental toxicity was found in the numerous studies available for structurally similar chemicals. Therefore, in analogy to other epoxides, no developmebntal toxicity can be assigned for the target substance. Therefore, it does not meet the criteria for classification and will not require labelling for this endpoint, according to the European regulation (EC) No. 1272/2008.