(R)-α-phenylethylammonium (-)-(1R, 2S)-(1,2-epoxypropyl)phosphonate monohydrate

Endpoint summary

Description of key information

Acute toxicity: oral. Key study: Test method according to EU Method B.1 and OECD Guideline 401. GLP study. The oral LD50 of the test article in Sprague-Dawley rats was > 2000 mg/kg bw.

Acute toxicity: dermal. Key study: Test method according to EU Method B.3 and OECD Guideline 402. GLP study. The LD50 value of test article in Sprague-Dawley rats by dermal route at 24 h exposure period is > 2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 23, 1994 - July 29, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A.
- Age at study initiation: 7/9 weeks
- Weight at study initiation: Males: 225-250 g. Females: 200-225 g
- Fasting period before study: 16 hours
- Housing: 5 animals/sex/cage in T11C air-conditioned room in grill cages 40.5x38.5x18h cm with stainless feeder.
- Diet (e.g. ad libitum): Ad libitum (GLP 4RF21 top certificate pellet diet)
- Water (e.g. ad libitum): Ad libitum (from the municipal water main system, filtered).
- Acclimation period: Five days before the start of the test

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ºC
- Humidity (%): 55 ± 10 %
- Air changes (per hr): 20 per hour (filtered on HEPA 99.97 %)
- Photoperiod (hrs dark / hrs light): 12 h dark/12 h light (7 a.m - 7 p.m)

Route of administration:

oral: gavage

Vehicle:

other: Methocel (methylcellulosae aqueous solution)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 100 mg/ml

MAXIMUM DOSE VOLUME APPLIED: 20 ml/kg

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical signs and mortality: at 30 minutes, 2, 4 and 6 hours on the first day after the administration (day 1) and then twice a day up to termination of the observation period.
Weighing: twice pre-trial (at randomization and on day 1 just before administration) and on days 3, 8 and 14.
- Necropsy of survivors performed: Yes. All surviving animals (fasted overnight) were killed by excision of the femoral arteries, after i.p. overdosage anesthesia with 5% sodium pentobarbital.
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Clinical signs and behaviour, body weight and gross pathology examinations.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No animals died during the observation period.

Clinical signs:

Signs involving the CNS (hypoactivity, muscular tremors, palpebral closure and hypothermia), the respiratory apparatus (shallow breathing) and piloerection, hunched posture and salivation were clinically observed in the treated rats starting from 30 minutes - 4 hours and lasting up to 6-24 hours after the test article administration. Sporadic cases of ataxia, reddish nasal discharge and chromodacryorrhea were also observed in some animals during the first days of the study. Recovery of all treated rats was achieved within 24 (females) - 48 hours (males) of treatment.

Body weight:

Low body weight gain was observed in some animals only at the day 3 weighing. Body weight gain returned to normal at the subsequent weighings.

Gross pathology:

No appreciable macroscopic findings were evident in any treated rat.

Table 1. Clinical signs frequency (cumulative)

Clinical signs	No. of rats affected	Signs observed (time)
		From*	To**
Hypoactivity	10	30 m	6 h
Ataxia	2	2 h	6 h
Reddish nasal discharge	2	2 d	-
Shallow breathing	3	2 h	2 d
Piloerection	10	30 m	2 d
Hunched posture	10	30 m	2 d
Muscular tremors	6	30 m	6 h
Salivation	6	30 m	6 h
Chromodacryorrhea	1	2 d	-
Palpebral closure	4	4 h	6 h
Hypothermia	3	2 h	2 d
Recovery	10	2 d	3 d

* first observation in one or more animals; ** last observation in one or more animals.

(day of treatment = day 1)

 

Table 2. Body weight (g). Dose: 2000 mg/kg.

Day	1 M	2 M	3 M	4 M	5 M	6F	7F	8F	9F	10F
Pre-trial	269	260	269	274	267	242	228	224	228	236
1*	247	235	248	250	238	220	209	209	210	214
3	258	251	269	260	249	226	209	214	215	224
8	298	287	305	285	301	248	229	236	261	248
14	362	344	352	353	344	263	252	253	281	270

 

Table 3. Gross pathology examination.

Dose (mg/kg)	Animals	Macroscopic findings
2000	Males No. 1-5	No appreciable macroscopic findings
	Females No. 6-10	No appreciable macroscopic findings

Interpretation of results:

GHS criteria not met

Remarks:

EU criteria.

Conclusions:

The oral LD50 of the test article in rats was higher than 2000 mg/kg bw.

Executive summary:

A toxicity study in Sprague Dawley Crl:CD (SD) rats (5 males and 5 females) with a single oral administration of the test article Fosfomycin PEA salt at the dosage of 2000 mg/kg bw (limit dose) was performed in accordance with EU Method B.1 and OECD Guideline 401. The test article was administered as a suspension in 0.4% methylcelllulosae aqueous solution at the constant volume of 20 ml/kg. All rats were treated after a 16 hour fasting period. The day of treatment was considered day 1 of the study. The animals were weighed twice before treatment (at randomization and on day 1 just before treatment) and on days 3, 8 and 14. They were clinically observed for 14 days following the treatment. On day 15 all treated rats were killed by excision of the femoral arteries, after i.p. overdosage anesthesia with 5% sodium pentobarbital and they were submitted to a thorough autopsy.

No animals died during the observation period.

The main clinical signs observed were: hypoactivity, muscular tremors, palpebral closure and hypothermia), the respiratory apparatus (shallow breathing) and piloerection, hunched posture starting from 30 minutes - 4 hours and lasting up to 6-24 hours after the test article administration. Sporadic cases of ataxia, reddish nasal discharge and chromodacryorrhea were also observed in some animals during the first days of the study. Recovery of all treated rats was achieved within 24 (females) - 48 hours (males) of treatment.

Low body weight gain was observed in some animals only at the day 3 weighing. Body weight gain returned to normal at the subsequent weighings. No appreciable macroscopic findings were evident in any treated rat.

In conclusion, the oral LD50 of the test article in rats was higher than 2000 mg/kg bw and transient clinical signs involving the CNS and the respiratory apparatus were the main findings.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

The study is GLP compliant and has a Klimisch score 1.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 13, 1994 - July 28, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A.
- Age at study initiation: 7/9 weeks
- Weight at study initiation: Males: 225-250 g. Females: 200-225 g
- Housing: Individual caging in air-conditioned rooms. Grill cages 40.5x38.5x18h cm with stainless steel feeder.
- Diet (e.g. ad libitum): Ad libitum (GLP 4 RF21 top certificate pelleted diet).
- Water (e.g. ad libitum): Ad libitum (from the municipal water main system, filtered).
- Acclimation period: 5 days before the start of the test.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ºC
- Humidity (%): 55 ± 10 %
- Air changes (per hr): 20 per hour (filtered on HEPA 99.97 %)
- Photoperiod (hrs dark / hrs light): 12 h dark/12 h light (7 a.m. - 7 p.m.)

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 6x5 cm of the body dorsal surface.
- % coverage: 10 %
- Type of wrap if used: the treated area was covered with a porous gauze dressing fixed to the skin with hypoallergenic non-irritation tape. The test side was further covered in a suitable manner in order to ensure that the animals could not ingest the test substance.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period the residual test substance was wiped off.
- Time after start of exposure: about 24 h.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000mg/kg bw. Individual dosages were based on body weight taken just before treatment.
- For solids, paste formed: No, the test article was administrated by uniform application onto the cleared area.

Duration of exposure:

About 24 hours.

Doses:

2000 mg/kg bw.

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical signs and mortality: at 30 minutes, 2, 4 and 6 hours on the first day after the administration on exposure day and then twice a day up to termination of the observation period.
Weighing: twice pre-trial (at randomization and on day 1 just before treatment), on days 8 and 15.
- Necropsy of survivors performed: Yes, animals (fasted overnight) were killed by excision of the femoral arteries, after i.p. overdosage anesthesia with 5 % sodium pentobarbital, at the end of the observation period.
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: clinical signs and behaviour, body weight and gross pathology.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No animals died during the study.

Clinical signs:

No clinical signs nor behavioural alterations were observed in any treated rat. Slight erythema was observed at the application site during the first days of the study in two male and one female rats.

Body weight:

Body weight gain of all treated animals was considered within normal limits.

Gross pathology:

No changes were observed in the animals killed at the end of the study.

Table 1. Clinical signs frequency (cumulative)

Clinical signs	No. of rats affected	Signs observed (time)
		From*	Toº
Treatment site: erythema	3	2d	3d
Recovery	3	3d	4d

*) First observation in one or more animals

º) Last observation in one or more animals

d) Days

Table 2. Body weight (g). Dose: 2000 mg/kg.

Day	1 M	2 M	3 M	4 M	5 M	6F	7F	8F	9F	10F
Pre-trial	270	273	271	268	255	237	229	220	233	214
1*	280	285	282	288	279	241	243	235	250	225
8	333	342	341	324	323	253	248	250	256	227
15	379	400	388	366	378	277	264	284	278	238

*pre-dosing.

Table 3. Gross pathology examination.

Dose (mg/kg)	Animals	Macroscopic findings
2000	Males No. 1-5	No appreciable macroscopic findings
	Females No. 6-10	No appreciable macroscopic findings

 

Interpretation of results:

GHS criteria not met

Remarks:

EU criteria.

Conclusions:

LD50 value of test article in rats by dermal route at 24 h exposure period is higher than 2000 mg/kg bw.

Executive summary:

A toxicity study in Sprague Dawley Crl:CD (SD) rats (5 males and 5 females) with a single dermal administration of the test article Fosfomycin PEA salt at the dosage of 2000 mg/kg bw (24 hours exposure) was performed in accordance with EU Method B.3 and OECD Guideline 402. The test article was administered by uniform application on the cleared area and covered for the exposure period of 24 hours. The day of treatment was considered day 1 of the study. The animals were weighed twice before treatment (at randomization and on day 1 just before treatment) and on days 8 and 15. They were clinically observed for 24 days after patch removal. On day 15 all treated rats were killed by excision of the femoral arteries, after i.p. overdosage anesthesia with 5% sodium pentobarbital and they were submitted to a thorough autopsy.

No deaths occurred as a result of treatment.

No clinical signs or behaviour alterations were observed during the study.

Slight transient erythema was occasionally observed at the application site in some animals during the first day of the study.

The body weight gain appeared normal.

No changes were found at the gross pathology examination performed at the end of the observation period.

In conclusion, the test article Fosfomycin PEA salt, when administrated by dermal route to rat, under the conditions adopted in this experiment, did not cause mortality nor show systemic toxic effects at the limit dose of 2000 mg/kg bw. The LD50 by dermal route is higher than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

The study is a GLP compliant and has a Klimisch score 1.

Additional information

Acute toxicity: oral. Key study: Test method according to EU Method B.1 and OECD Guideline 401. GLP study. The oral LD50 of the test article in Sprague-Dawley rats was > 2000 mg/kg bw.

Acute toxicity: dermal. Key study: Test method according to EU Method B.3 and OECD Guideline 402. GLP study. The LD50 value of test article in Sprague-Dawley rats by dermal route at 24 h exposure period is > 2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint

Only one study available.

Justification for selection of acute toxicity – dermal endpoint

Only one study available.

Justification for classification or non-classification

Based on the available data oral and dermal acute toxicity, the substance is not classified for acute toxicity.