Methyl anthranilate

Endpoint summary

Description of key information

Acute oral toxicity:

An acute oral toxicity study was carried out on rats to determine the toxic nature of test chemical.50% mortality was observed at a dose level of 2800 mg/kg bw with 95% confidence limit of 2300 to 3300 mg/kg bw. Thus, the LD50 value was >2000 mg/kg bw indicating that the test chemical is not toxic to rats when administered orally and can be classified under the category ‘Not classified’.

Acute inhalation toxicity:

An acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 1.599 Pa (0.012 mmHg). Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver.

Acute dermal toxicity:

An acute dermal toxicity dose (LD50) was considered based on different studies conducted on rats and rabbits for the test chemical. The studies concluded that LD50 value is >5000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

Data is from a NTRL report

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

40 CFR Part 163.81-1.
(ENVIRONMENTAL PROTECTION AGENCY. PESTICIDE PROGRAMS. Proposed Guidelines for Registering Pesticides in the U.S.; Hazard Evaluation:
Humans and Domestic Animals. Acute Oral Toxicity Study.)

Principles of method if other than guideline:

To determine to Acute oral toxicity of test chemical in rats.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable) : No
- Source: Charles River Breeding Laboratories. Inc;Wilmington. Mass
- Females (if applicable) nulliparous and non-pregnant: Not specified
- Rationale for use of males: Not specified
- Age at study initiation: Not specified
- Weight at study initiation:
Males: approx 230-230gm
Females: 180-250 gm
- Fasting period before study: 18 hours
- Housing: Individual in suspended stainless steel
- Historical data: Not specified
- Diet (e.g. ad libitum): Purina Laboratory Chow. ad libitum
- Water (e.g. ad libitum): Elizabethtown water Co. ad libitum
- Acclimation period: Not specified
- Microbiological status when known : no
- Method of randomisation in assigning animals to test and control groups : The animals are randomly selected for each study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not specified
- Humidity (%): Not specified
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): Not specified

IN-LIFE DATES: From: To:Not specified

Route of administration:

oral: gavage

Vehicle:

not specified

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Not specified
- Amount of vehicle (if gavage):Not specified
- Justification for choice of vehicle:Not specified
- Lot/batch no. (if required): Not specified
- Purity: Not specified

MAXIMUM DOSE VOLUME APPLIED: 5000 mg/kg

DOSAGE PREPARATION (if unusual): Liquids are administered as received.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Not specified

Doses:

Single dose: 5.0 g/kg (5000 mg/kg bw)
Range-finding: 0.05, 0.10, 0.50, 1.0 and 2.0 g/kg (50, 100, 500, 1000 and 2000 mg/kg bw)
LD50: 1.2. 1.7. 2.5 and 3.5 g/kg (1200, 1700, 2500 and 3500 mg/kg bw)

No. of animals per sex per dose:

Total 80 animals (males and females)
Single dose level (5 g/kg): Ten (five/sex)
Range-finding Screen: Ten (one/sex/dose level)
LD50 Determination: Fifty (five/sex/dose level)
Control group: 10 (5 males, 5 females)

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The test animals were observed approximately 1, 2 and 4 hours after dosing, and daily thereafter for fourteen days.
- Necropsy of survivors performed: yes; Gross post mortem examinations were performed on all surviving animals on Day 14 and on all animals which die or were found dead during the study. All abnormalities were recorded but no tissues were saved.
- Clinical signs including body weight: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Body weight: Body weights were checked before fasting, post fasting (prior to dosing) and Terminal: Any animals which do not survive for 14 days were weighed at the time of death or at the time they were found dead.

Preliminary study:

A dose of 5 g/kg was administered initially to 10 animals (5/sex), unless the test substance was suspected to have an LD50 value of less than 5 g/kg.
If no animals die during the fourteen day post-dose observation period, no additional testing is required.
If deaths occur in animals dosed with 5 g/kg, or if the test substance is suspected to have an LD50 which is less than 5 g/kg, the following is done:
A preliminary range-finding screen is performed by administering five different dose levels (usually 0.05. 0.1. 0.5. 1.0 and 2.0 g/kg) to ten rats (one male; one female per dose level). If results do not provide adequate information for selecting doses for the LD50 determination, additional screening studies are performed. Animals were observed for viability for seven days after dosing. Based on the results of this screen, five geometrically spaced dose levels were selected and each dose was administered to ten animals (5/sex) for a determination of the LD50.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 800 mg/kg bw

Based on:

test mat.

95% CL:

>= 2 300 - <= 3 300

Remarks on result:

other: 50% mortality observed

Mortality:

Single dose: All ten animals died after receiving 5 grams/kg of the test material.
Dose range finding: Only one animal died at 2000 mg/kg bw dose level.

Clinical signs:

other: Some animals in all groups exhibited motor activity decrease, (from the 1 hour interval to Day 7), piloerection (from the 2 hour interval to Day 7). respiratory rate decrease (from the 1 hour interval to Day 2), and ataxia (at the 4 hour interval and on D

Gross pathology:

Necropsy observations are presented in table:

Other findings:

From Day 8 to termination no abnormalities were recorded for any animals.

Table 1: Dose levels and mortality for the preliminary range-finding study

Dose level (g/kg)	Mortality
0.05	0/2
0.10	0/2
0.50	0/2
1.0	0/2
2.0	1/2

 

Table 2: Dose levels, mortality data and LD50 calculations for the LD50 study

Dose level Gm/kg	Mortality
	Male	Female	Total
1.2	0/5	0/5	0/10
1.7	0/5	1/5	1/10
2.5	0/5	3/5	3/10
3.5	4/5	5/5	9/10
LD50 (gm/kg)	3.10	2.15	2.8
95% Confidence limits (gm/kg)	2.60 to 3.50	1.55 to 2.75	2.3 to 3.3

 

Table 3: Body weights (grams) and day of death; Dose level 1.2 gm/kg

Dose level (g/kg)	Animal no. and sex	Pretest weights		Interim deaths			Survivors
		Pre fast	Post fast	Terminal weight	Day of death	Change^a	Day 7	Change^a	Day 14	Change^a
1.2	2504 M	257	228				320	63	372	115
	2510 M	242	216				309	67	355	113
	2514 M	241	217				283	42	312	71
	2516 M	250	220				287	37	322	72
	2525 M	241	219				273	32	308	67
	2593 F	240	218				266	26	267	27
	2599 F	227	204				228	1	243	16
	2604 F	239	216				246	7	256	17
	2609 F	237	215				249	12	255	18
	2613 F	236	215				241	5	249	13

^aChange from prefasted weight (grams).

 

Table 4: Body weights (grams) and day of death; Dose level 1.7 gm/kg

Dose level (g/kg)	Animal no. and sex	Pretest weights		Interim deaths			Survivors
		Pre fast	Post fast	Terminal weight	Day of death	Change^a	Day 7	Change^a	Day 14	Change^a
1.7	2526M	232	210				266	34	305	73
	2534M	234	210				268	34	310	76
	2540M	230	205				271	41	304	74
	2541M	247	220				306	59	360	113
	2546M	251	220				296	45	340	89
	2616F	238	220				247	9	258	20
	2619F	145	221				266	21	295	50
	2628F	224	205				226	2	259	35
	2630F	231	215	NR	Day 2	-
	2595F	221	204				239	18	246	25

^aChange from prefasted weight (grams).

NR: Not recorded

Table 5: Body weights (grams) and day of death; Dose level 2.5 gm/kg

 

Dose level (g/kg)	Animal no. and sex	Pretest weights		Interim deaths			Survivors
		Pre fast	Post fast	Terminal weight	Day of death	Change^a	Day 7	Change^a	Day 14	Change^a
2.5	2512M	253	226				303	50	370	117
	2520M	236	211				288	52	330	94
	2523M	247	230				287	40	323	76
	2533M	231	210				266	35	305	74
	2539M	249	226				249	0	296	47
	2600F	235	212				229	-6	245	10
	2607F	230	208	200	Day 1	-30
	2610F	250	229	NR	Day 2	-
	2615F	249	225				254	5	266	17
	2621F	240	224	217	7 hrs	-23

^aChange from prefasted weight (grams).

NR: Not recorded

   

Table 6: Body weights (grams) and day of death; Dose level 3.5 gm/kg

Dose level (g/kg)	Animal no. and sex	Pretest weights		Interim deaths			Survivors
		Pre fast	Post fast	Terminal weight	Day of death	Change^a	Day 7	Change^a	Day 14	Change^a
3.5	2545M	230	214	200	Day 1	-30
	2559M	246	220	201	Day 1	-45
	2519M	230	205				254	24	298	68
	2537M	246	218	200	Day 1	-46
	2543M	248	223	206	Day 1	-42
	2625F	241	220	212	Day 1	-29
	2629F	235	315	208	Day 1	-27
	2697F	234	210	195	Day 1	-39
	2602F	219	200	187	Day 1	-32
	2603F	247	225	212	Day 1	-35

^aChange from prefasted weight (grams).

 

Table 7: Summary of in-life observations

Dose level (g/kg)	Observations	Time interval
		Hours			Days
		1	2	4	1	2	3	4	5-7	8-14
1.2	Ataxia	0	0	1	0	0	0	0	0	0
	Red oral discharge	1	0	0	0	0	0	0	0	0
	Respiratory Rate Increase	0	1	0	0	0	0	0	0	0
	Respiratory Rate Decrease	1	1	1	0	0	0	0	0	0
	Soft Stool	0	1	0	0	0	0	0	0	0
	Piloerection	0	1	0	0	0	0	0	1	0
	Abdominal Griping	0	0	0	1	0	0	0	0	0
	Motor Activity Decrease	7	5	1	2	0	0	0	1	0
1.7	Dead	0	0	0	0	1	1	1	1	1
	Ataxia	0	0	0	1	0	0	0	0	0
	Fine Tremors	0	0	0	1	0	0	0	0	0
	Clear oral discharge	1	0	0	0	0	0	0	0	0
	Respiratory Rate Decrease	0	2	1	4	0	0	0	0	0
	Piloerection	0	1	0	4	1	0	1	1	0
	Motor Activity Increase	0	0	0	0	0	0	1	0	0
	Motor Activity Decrease	4	4	5	5	4	0	0	0	0
2.5	Dead	0	0	0	2	3	3	3	3	3
	Ataxia	0	0	0	1	0	0	0	0	0
	Fine Tremors	0	0	0	1	0	0	0	0	0
	Clear nasal discharge	0	1	0	0	0	0	0	0	0
	Respiratory Rate Decrease	0	3	2	4	0	0	0	0	0
	Clear ocular discharge	0	0	1	0	0	0	0	0	0
	Fecal staining	0	0	0	1	1	0	0	0	0
	Soft stool	0	0	0	1	0	0	0	0	0
	Piloerection	0	4	1	3	4	4	0	2	0
	Motor Activity Increase	0	0	0	0	0	1	0	1	0
	Motor Activity Decrease	5	8	3	6	7	1	1	2	0
	Prostration	0	0	2	0	0	0	0	0	0
	Aggressive	0	0	0	0	0	1	0	0	0
3.5	Dead	0	0	0	7	9	9	9	9	9
	Ataxia	0	0	0	2	1	0	0	0	0
	Fine Tremors	0	0	0	1	0	0	0	0	0
	Clear oral discharge	2	1	0	1	0	0	0	0	0
	Respiratory Rate Decrease	2	5	1	2	1	1	0	0	0
	Clear ocular discharge	0	0	0	1	0	0	0	0	0
	Urinary staining	0	0	0	1	1	0	0	0	0
	Piloerection	0	7	4	2	0	1	0	0	0
	Abdominal griping	0	1	0	0	0	0	0	0	0
	Motor activity decrease	6	9	8	2	1	1	0	0	0
	Prostration	0	0	0	1	0	0	0	0	0
	Rales	1	1	0	0	0	0	0	0	0

 

Table 8: Day of death and necropsy observations (1.2 gm/kg)

Dose level g/kg	Animal no. and sex	Day of death	Type of death^a	Observations
1.2	2504 M	Day 1	T	Lungs: dark red foci, mottled dark red 20%; liver: mottled pale 30%; kidneys: pale; adrenals: pale red.
	2510 M	Day 14	T	Lungs: dark red foci, mottled dark red 20%; adrenals: dark red.
	2514 M	Day 14	T	Lungs: mottled dark red 80%; kidneys: pale; adrena1s: pale red.
	2516 M	Day 14	T	Lungs: dark red foci; liver: mottled pale 30%; kidneys: mottled pale 60%; adrenals: pale red.
	2525 M	Day 14	T	Lungs: dark red foci; liver: mottled pale 50%; kidneys: mottled pale 40%; adrenals: pale red.
	2593 F	Day 14	T	Lungs: dark red foci; adrenals: dark red foci.
	2599 F	Day 14	T	Lungs: dark red foci, mottled dark red 30%; liver: mottled pale 40%; adrenals: pale red.
	2604 F	Day 14	T	Lungs: dark red foci, mottled dark red 30%; liver: mottled pale 30%; adrenals: pale red.
	2609 F	Day 14	T	Lungs: mottled dark red 60%; liver: mottled pale 30%; adrenals: pale red.
	2613 F	Day 14	T	Lungs: dark red foci; adrenals: dark red foci.

T= Terminal (Killed on day 14)

Table 9: Day of death and necropsy observations (1.7 gm/kg)

Dose level g/kg	Animal no. and sex	Day of death	Type of death^a	Observations
1.7	2526M	Day 1	T	Lungs: dark red foci; adrenals: dark red; mesentary lymph nodes: bright red.
	2534M	Day 14	T	Lungs: dark red foci; adrenals: pale red.
	2540M	Day 14	T	Lungs: dark red foci, mottled dark red 20%; adrenals: pale red.
	2541M	Day 14	T	Adrenals: dark red foci.
	2546M	Day 14	T	Lungs: dark red foci; mottled pale 10%; kidneys: mottled pale 50%; adrenals: pale red.
	2616F	Day 14	T	Lungs: dark red foci, mottled dark red 20% adrenals: dark red foci.
	2619F	Day 14	T	Lungs: dark red foci; adrenals: pale red.
	2628F	Day 14	T	Lungs: dark red foci, mottled dark red 60%; adrenals: pale red.
	2630F	Day 2	F	Clear oral discharge; lungs: pale, tan patches 50%; liver: mottled tan; stomach: contains dark brown fluid; intestines: contain brown fluid; kidneys: tan; adrenals: dark red; urinary bladder: contains brown fluid.
	2595F	Day 14	T	Lungs: dark red foci.

T= Terminal (Killed on day 14)

F=Found dead (Day given is day found dead).

 

Table 10: Day of death and necropsy observations (2.5 gm/kg)

Dose level g/kg	Animal no. and sex	Day of death	Type of death^a	Observations
2.5	2512M	Day 14	T	Lungs: dark red foci; liver: mottled pale 40%; kidneys: mottled pale 30%; adrenals: pale red.
	2520M	Day 14	T	Lungs: dark red foci; liver: mottled pale 40%; kidneys: mottled pale 20%; adrenals: pale red.
	2523M	Day 14	T	Lungs: dark red foci, mottled dark red 30%; liver: anterior edges clear; kidneys: mottled pale 60%; adrenals: pale red.
	2533M	Day 14	T	Lungs: dark red foci; liver: mottled pale 40%; kidneys: mottled pale 50%; adrenals: pale red.
	2539M	Day 14	T	Lungs: dark red foci; kidneys: mottled pale 70%; adrenals: pale red.
	2600F	Day 14	T	Lungs: dark red foci; liver: mottled pale 20%; adrenals: pale red.
	2607F	Day 1	F	Brain: blood vessels injected; lungs: mottled dark red and pale 70%; stomach: contains thick tan material; intestines: contain reddish yellow viscous fluid; kidneys: pale; adrenals: dark red.
	2610F	Day 2	F	Clear oral discharge; clear nasal discharge; yellow urinary staining; lungs: pale red; liver: mottled tan, dark red foci; intestines: contain dark red-black fluid; spleen: diminished, black wrinkled; kidneys: tan; adrenals: dark red.
	2615F	Day 14	T	Lungs: dark red foci; adrenals: pale red.
	2621F	7 hours	F	Stomach: contains tan fluid; jujunum: contains reddish yellow fluid; duodenum: contains tan fluid; adrenals: dark red.

T= Terminal (Killed on day 14)

F=Found dead (Day given is day found dead).

 

 

Table 11: Day of death and necropsy observations (3.5 gm/kg)

Dose level g/kg	Animal no. and sex	Day of death	Type of death^a	Observations
3.5	2545M	Day 1	F	Brain: blood vessels injected; stomach: pyloric region - reddish tint, pronounced vascularization, contains thick tan material; intestines: appeared reddened, contain reddish yellow fluid; kidneys: pale; adrenals: dark red.
	2559M	Day 1	F	Brain: blood vessels injected; lungs: wrinkled; stomach: contains thick grey green material; intestines: vascu1arized, contains thick brown fluid; spleen: extremely diminished; kidneys: mottled pale 30%; adrenals: dark red.
	2519M	Day 14	T	Adrenals: pale red
	2537M	Day 1	F	Clear oral discharge; red nasal discharge; brain: blood vessels injected; lungs: dark red foci; mottled dark red 20%; stomach: pyloric region - vascularized, contains thick tan substance; intestines: vascularized, contain red fluid; adrenals: pale red; testes: undescended.
	2543M	Day 1	F	Fecal staining; brain: blood vessels injected; left lung; mottled black; liver: mottled pale, clear edges; stomach: distended with gas, contains grey green fluid; intestines: pronounced vascularization, contain thin brown fluid; spleen: diminished; adrenals: extremely dark red.
	2625F	Day 1	F	Brain: blood vessels injected; lungs: dark red foci; liver: clear edges; stomach: pyloric region - reddish tint, contains thick tan material; intestines: contain reddish yellow viscous fluid; kidneys: pale; adrenals: pale red.
	2629F	Day 1	F	Lungs: mottled tan 50%; liver: clear edges; stomach: distended with gas, contains viscous tan substance; kidneys: mottled pale 50%; adrenals: dark red.
	2697F	Day 1	F	Clear oral discharge; lungs: mottled dark red 30%; liver: mottled pale 30%; stomach: distended with yellow tan fluid; intestines: contain yellow tan fluid; spleen: roughened; kidneys: pale; adrenals: pale red.
	2602F	Day 1	F	Brain: blood vessels injected; lungs: mottled dark red 30%, dark red foci; liver: clear edges; stomach: distended with gas, contains thick tan material and black material; intestines: black material in duodenum; kidneys: pale; adrenals: dark red.
	2603F	Day 1	F	Lungs: dark red foci 100%; stomach: contains viscous tan material; intestines: contains reddish yellow fluid; kidneys: mottled pale 40%; adrenals: pale red.

 

T= Terminal (Killed on day 14)

F=Found dead (Day given is day found dead).

Table 12: Body weights (grams) of Control group

Animal no.	Sex	Pre fast	Post fast	Day 7	Change^a	Day 14	Change^a
2509	M	259	230	318	59	350	91
2527	M	250	221	290	40	317	67
2549	M	251	227	310	59	350	99
2554	M	252	222	315	63	352	100
2556	M	251	225	297	46	322	71
2571	F	235	212	253	18	264	29
2577	F	238	217	253	15	278	40
2581	F	254	228	262	8	280	26
2584	F	233	210	249	16	242	9
2588	F	210	201	230	20	255	45

^aChange from initial (prefasted) weight.

Table 13: NECROPSY OBSERVATIONS - CONTROL ANIMALS (TERMINAL - DAY 14)

Animal no.	Sex	Observations
2509	M	Kidneys: mottled pale 30%; adrenals: dark red foci
2527	M	Lungs: dark red foci throughout; liver: mottled pale 20%; kidneys: mottled pale 50%; adrenals: dark red.
2549	M	Lungs: dark red foci; adrenals: pale red
2554	M	Lungs: dark red foci; kidneys: mottled pale 70%; adrenals: dark red foci.
2556	M	Lungs: dark red foci throughout; liver: mottled pale 60%; kidneys: pale; adrena1s: pale red.
2571	F	Lungs: dark red foci throughout; adrenals: dark red foci.
2577	F	Adrenals: pale red.
2581	F	Kidneys: mottled pale 50%; adrenals: pale red
2584	F	Lungs: mottled dark red 40%; adrenals: pale red
2588	F	Lungs: mottled bright red 30%; adrenals: pale red.

Interpretation of results:

other: Not classified

Conclusions:

Acute oral toxicity study was carried out on rats to determine the toxic nature of test chemical.50% mortality was observed at a dose level of 2800 mg/kg bw with 95% confidence limit of 2300 to 3300 mg/kg bw. Thus, the LD50 value was >2000 mg/kg bw indicating that the test chemical is not toxic to rats when administered orally and can be classified under the category ‘Not classified’.

Executive summary:

Acute oral toxicity study was carried out on rats to determine the toxic nature of test chemical. A total of 80 Male and female Sprague-Dawley rats were used during the study. The test chemical was administered orally by gavage at various dose ranges for 14 days. The study was carried out in three different stages; Single dose study, a dose of 5 g/kg is administered initially to 10 animals (5/sex), unless the test substance is suspected to have an LD50 value of less than 5 g/kg. If no animals die during the fourteen day post-dose observation period, no additional testing is required; Range finding study, If deaths occur in animals dosed with 5 g/kg, or if the test substance is suspected to have an LD50 which is less than 5 g/kg, the following is done: A preliminary range-finding screen is performed by administering five different dose levels (usually 50, 100, 500, 1000 and 2000 mg/kg) to ten rats (one male; one female per dose level). If results do not provide adequate information for selecting doses for the LD50 determination, additional screening studies are performed; and LD50 determination study, animals were observed for viability for seven days after dosing. Based on the results of this screen, four geometrically spaced dose levels ie. 1200, 1700, 2500 and 3500 mg/kg bw were administered and each dose was administered to ten animals (5/sex) for a determination of the LD50. The test chemical was administered by oral intubation, using a ball-tipped intubation needle. Doses were calculated using pre-fasted body weights. Whenever possible, a constant concentration of test substance was administered at all doses (i.e., varying doses were achieved by varying the volume of the dosing mixture) 60 ml of test substance per kg of body weight was considered the maximum volume which can be administered to a rat; doses in excess of this amount were not administered. In order to provide control data, ten animals (5/sex) were dosed with methylcellulose in a volume of 45.0 ml/kg [Because maximum dose specified in protocol (60 ml/kg) was found to be excessive, dose was reduced to 45 ml/kg]. Mortality data, in-life observations (14 days) and necropsy observations for these animals are tabulated. The test animals were observed approximately 1, 2 and 4 hours after dosing, and daily thereafter for fourteen days. Gross post mortem examinations were performed on all surviving animals on Day 14 and on all animals which die or were found dead during the study. All abnormalities were recorded but no tissues were saved. Body weights were checked before fasting, post fasting (prior to dosing) and Terminal; any animals which do not survive for 14 days were weighed at the time of death or at the time they were found dead. Mortality was observed in all animals at dose level of 5000 mg/kg bw and only one animal died at 2000 mg/kg bw dose in range finding study. Some animals in all groups exhibited motor activity decrease, (from the 1 hour interval to Day 7), piloerection (from the 2 hour interval to Day 7), respiratory rate decrease (from the 1 hour interval to Day 2), and ataxia (at the 4 hour interval and on Days 1 and 2). All surviving animals exhibited body weight gains at Day 14, the majority of animals exhibited body weight gains at Day 7, except for one animal which exhibited a slight weight loss and one animal which showed no change. Animals dying prior to termination exhibited weight losses. From Day 8 to termination no abnormalities were recorded for any animals. 50% mortality was observed at a dose level of 2800 mg/kg bw with 95% confidence limit of 2300 to 3300 mg/kg bw. Thus, the LD50 value was >2000 mg/kg bw indicating that the test chemical is not toxic to rats when administered orally and can be classified under the category ‘Not classified’.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimish rating 2 and from publication

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Waiver

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from publication

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

To determine the acute dermal toxicity of test chemical in rabbits.

GLP compliance:

not specified

Test type:

other: not specified

Species:

rabbit

Strain:

not specified

Sex:

not specified

Details on test animals or test system and environmental conditions:

not specified

Type of coverage:

not specified

Vehicle:

not specified

Details on dermal exposure:

not specified

Duration of exposure:

not specified

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

not specified

Control animals:

not specified

Details on study design:

not specified

Statistics:

not specified

Preliminary study:

not specified

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

not specified

Clinical signs:

other: not specified

Gross pathology:

not specified

Other findings:

not specified

Interpretation of results:

other: Not classified

Conclusions:

Acute dermal toxicity study of the test chemical was performed on rabbits. An LD50 of >5000 mg/kg bw was observed indicating that the test chemical is not classified.

Executive summary:

Acute dermal toxicity test was performed to determine the toxic nature of test chemical on rabbits. The test chemical was applied on skin of rabbits. An LD50 of >5000 mg/kg bw was observed indicating that the test chemical is not toxic in nature and can be categorized as 'Not classified' according to CLP regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

Data is Klimish rating 2 and from publication

Additional information

Acute oral toxicity:

In different studies, the given test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats, mice and Guinea pig for test chemical. The studies are summarized as below –

1. Acute oral toxicity study was carried out on rats to determine the toxic nature of test chemical. A total of 80 Male and female Sprague-Dawley rats were used during the study. The test chemical was administered orally by gavage at various dose ranges for 14 days. The study was carried out in three different stages; Single dose study, a dose of 5 g/kg is administered initially to 10 animals (5/sex), unless the test substance is suspected to have an LD50 value of less than 5 g/kg. If no animals die during the fourteen day post-dose observation period, no additional testing is required; Range finding study, If deaths occur in animals dosed with 5 g/kg, or if the test substance is suspected to have an LD50 which is less than 5 g/kg, the following is done: A preliminary range-finding screen is performed by administering five different dose levels (usually 50, 100, 500, 1000 and 2000 mg/kg) to ten rats (one male; one female per dose level). If results do not provide adequate information for selecting doses for the LD50 determination, additional screening studies are performed; and LD50 determination study, animals were observed for viability for seven days after dosing. Based on the results of this screen, four geometrically spaced dose levels ie. 1200, 1700, 2500 and 3500 mg/kg bw were administered and each dose was administered to ten animals (5/sex) for a determination of the LD50. The test chemical was administered by oral intubation, using a ball-tipped intubation needle. Doses were calculated using pre-fasted body weights. Whenever possible, a constant concentration of test substance was administered at all doses (i.e., varying doses were achieved by varying the volume of the dosing mixture) 60 ml of test substance per kg of body weight was considered the maximum volume which can be administered to a rat; doses in excess of this amount were not administered. In order to provide control data, ten animals (5/sex) were dosed with methylcellulose in a volume of 45.0 ml/kg [Because maximum dose specified in protocol (60 ml/kg) was found to be excessive, dose was reduced to 45 ml/kg]. Mortality data, in-life observations (14 days) and necropsy observations for these animals are tabulated. The test animals were observed approximately 1, 2 and 4 hours after dosing, and daily thereafter for fourteen days. Gross post mortem examinations were performed on all surviving animals on Day 14 and on all animals which die or were found dead during the study. All abnormalities were recorded but no tissues were saved. Body weights were checked before fasting, post fasting (prior to dosing) and Terminal; any animals which do not survive for 14 days were weighed at the time of death or at the time they were found dead. Mortality was observed in all animals at dose level of 5000 mg/kg bw and only one animal died at 2000 mg/kg bw dose in range finding study. Some animals in all groups exhibited motor activity decrease, (from the 1 hour interval to Day 7), piloerection (from the 2 hour interval to Day 7), respiratory rate decrease (from the 1 hour interval to Day 2), and ataxia (at the 4 hour interval and on Days 1 and 2). All surviving animals exhibited body weight gains at Day 14, the majority of animals exhibited body weight gains at Day 7, except for one animal which exhibited a slight weight loss and one animal which showed no change. Animals dying prior to termination exhibited weight losses. From Day 8 to termination no abnormalities were recorded for any animals. 50% mortality was observed at a dose level of 2800 mg/kg bw with 95% confidence limit of 2300 to 3300 mg/kg bw. Thus, the LD50 value was >2000 mg/kg bw indicating that the test chemical is not toxic to rats when administered orally and can be classified under the category ‘Not classified’.

2. Acute oral toxicity study was performed in rats to determine the toxic nature of test chemical. Groups of 10 young adult Osborne-Mendel rats of both sexes were used in the study. The rats were fasted 18 hours prior to treatment and had access to water ad. libitum, and food was replaced in cages as soon as animals received their respective doses. All doses were given by oral intubation. All animals were maintained under close observation for 14 days for recording toxic signs and time of death, observation was continued until animals appeared normal and showed weight gain. LD50'S (slope function) were computed by the method of Litchfield & Wilcoxon (1949). Mortality was observed within 1-2 days and Clinical signs like depression and coma were observed. According to the graph, slope function was determined to be 1.4 with 95% confidence limit of 0.9 -2.0. In 14 days observation period, an LD50 of 2910 mg/kg bw with 95% confidence limit of 2500 to 3400 mg/kg bw was observed. Thus the test chemical can be categorised as 'Not classified' according to CLP regulation.

3. Acute oral toxicity study was performed in rats to determine the toxic nature of test chemical. An LD50 of 3000 mg/kg bw indicated that the test chemical is not toxic in nature and can be categorised as 'Not classified'.

4. Acute oral toxicity study was performed in mice to determine the toxic nature of test chemical. The mice were treated on full stomach and had access to water ad. libitum, and food was replaced in cages as soon as animals received their respective doses. All doses were given by oral intubation. All animals were maintained under close observation for 14 days for recording toxic signs and time of death, observation was continued until animals appeared normal and showed weight gain. LD50'S (slope function) were computed by the method of Litchfield & Wilcoxon (1949). Mortality was observed within 4 -18 hours and Clinical signs like depression was observed. According to the graph, slope function was determined to be 1.5 with 95% confidence limit of 1.3 -1.7. In 14 days observation period, an LD50 of 3900 mg/kg bw with 95% confidence limit of 3260 to 4680 mg/kg bw was observed. Thus the test chemical can be categorized as 'Not classified' according to CLP regulation.

5. Acute oral toxicity study was performed in guinea pig to determine the toxic nature of test chemical. Guinea pig of both sexes were used in the study and were fasted 18 hours prior to treatment and had access to water ad. libitum, and food was replaced in cages as soon as animals received their respective doses. All doses were given by oral intubation. All animals were maintained under close observation for 14 days for recording toxic signs and time of death, observation was continued until animals appeared normal and showed weight gain. LD50'S (slope function) were computed by the method of Litchfield & Wilcoxon (1949). Mortality was observed within 4 hours to 4 days and Clinical signs like Depression, gasping, rapid respiration and irritated gastro-intestinal tract were observed.. According to the graph, slope function was determined to be 1.8 with 95% confidence limit of 1.4 -2.3. In 14 days observation period, an LD50 of 2780 mg/kg bw with 95% confidence limit of 2210 to 3500 mg/kg bw was observed. Thus the test chemical can be categorised as 'Not classified' according to CLP regulation.

6. Acute oral toxicity study was performed in guinea pig to determine the toxic nature of test chemical. An LD50 of 4000 mg/kg bw indicated that the test chemical is not toxic in nature and can be categorised as 'Not classified'.

Thus, based on the above summarized studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Acute inhalation toxicity: The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 1.599 Pa (0.012 mmHg). Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver.

Acute Dermal Toxicity:

In different studies, the given test chemical has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats and rabbits for test chemical. The studies are summarized as below –

Acute dermal toxicity test was performed to determine the toxic nature of test chemical on rabbits. The test chemical was applied on skin of rabbits. An LD50 of >5000 mg/kg bw was observed indicating that the test chemical is not toxic in nature and can be categorized as 'Not classified' according to CLP regulation.

Another study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Under the condition of this, it was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to male and female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Similarly, Acute dermal toxicity study was conducted by using the given test chemical as per OECD No. 402 in Wistar rats. Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Approximately, twenty four hours prior to dermal application of test item, greater than 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight based on the test item density (1.05513) and latest body weight was applied by single dermal application and observed for 14 days after treatment. On test day 0, calculated volume of test item was applied directly on the intact skin of clipped area of rats; the porous gauze dressing was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating tape. After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water. The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period and post dosing till the termination. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were re­corded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically. No mortality was observed in any animal till the end of the experimental period. At 2000 mg/kg, all the animals were observed normal throughout the experimental period. Mean body weight was observed with gain on day 7 and 14 of male and female animals, as compared to day 0. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Hence, the LD50 value was considered to be >2000 mg/kg bw, when male and female wistar rats were occlusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

Thus, based on the above summarized studies on test chemical, it can be concluded that LD50 value is >5000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw, for acute oral and acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral and acute dermal toxicity. For acute inhalation toxicity, exposure via inhalation route is not possible. Therefore a wavier for acute inhalation toxicity endpoint is added.