Vanillin

Endpoint summary

Description of key information

LD50 oral (rat): 3978 mg/kg bw. Vanilline is not harmful by ingestion according to CLP criteria.
LD50 dermal (rat): > 2000 mg/kg bw (no mortality observed at this dose). Vanilline is not harmful by dermal route according to CLP criteria.

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:

from 12-JUN-1991 to 04-MAR-1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according a recognised guideline and under GLP conditions. Howerer, some deviations occurred and the purity of the test substance is unknown.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Cited as Directive 84/449/EEC, B.1

Deviations:

yes

Remarks:

variation of temperature, variation of humidity beyond the norms

GLP compliance:

yes

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:
- Source: Iffa-Crédo (L'Arbresle, France)
- Age at study initiation: 5 - 7 weeks old
- Weight at study initiation: 132 - 224 g
- Fasting period before study: 16-17 hours
- Housing: by sex and in groups of 5, in polycarbonate cages (365 x 225 x 180 mm)
- Food consumption: complete pelleted rat-mouse maintenance, ad libitum
- Water consumption: softened and filtered mains water, ad libitum
- Acclimation period: at least 9 days before the start of the treatment

ENVIRONMENTAL CONDITIONS:
- Temperature: 19 - 26.5 °C
- Humidity: 30 - 74 %
- Air changes: at least 8 per hour
- Photoperiod: 12 hr light / 12 hr dark

In-life dates: from 06-NOV-1991 to 20-NOV-1991

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

* Vehicle:
- Concentration in vehicle: 20, 25.1, 31.6 and 39.8%
- Amount of vehicle: 10 mL/kg
- Justification for choice of vehicle: data not available
- Lot/batch no.: 85133
- Purity: data not available

* Maximum dose volume applied: 10 mL/kg bw

Doses:

0, 2000, 2510, 3160 and 3980 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- mortality: 15 minutes after administration of the treatment, then at 1, 2 and 4 hours, and daily thereafter
- clinical signs: daily
- weighing: on D-1, D1, D8 and D15, and on day of death
- Necropsy of survivors performed: yes

Statistics:

Bliss' Method (more reliable) and Litchfield & Wilcoxon's Method

Preliminary study:

3 groups each composed of 2 males and 2 females were treated under the same conditions as those employed in the main study, at the dose levels
of 500, 1000 and 2000 mg/kg. Animals were observed for 15 days
No death were seen at a dose level of 2000 mg/kg.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 3 978 mg/kg bw

Based on:

test mat.

95% CL:

> 2 484 - < 6 368

Remarks on result:

other: Litchfield & Wilcoxon's method

Mortality:

In main study for limit test and principal test, at all doses tested mortality was observed.
Mortality increased with dose level

Clinical signs:

other: Lethargy, prostration or subdued behaviour were observed in all groups, and sialorrhea was observed in some animals

Gross pathology:

Some animals that died during the study presented congestioned lungs at necropsy. No other abnormalities were recorded

 

Table 1: Number of animals dead and with evident toxicity, and range within which mortality occured for limit test

 

Dose (mg/kg bw)	Mortality (# dead/total)			Time range of deaths (hours)	Number with evident toxicity(#/total)
	Male	Female	Combined		Male	Female	Combined
2000	1/5	1/5	2/10	D1	5/10	5/10	10/10

 

 

Table 2: Number of animals dead and with evident toxicity, and time range within which mortality occurred for principal study

 

Dose (mg/kg bw)	Mortality (# dead/total)			Time range of deaths (hours)	Number with evident toxicity(#/total)
	Male	Female	Combined		Male	Female	Combined
Control	0/5	0/5	0/10		0/5	0/5	0/10
2000	1/5	0/5	1/10	D1	5/5	5/5	10/10
2510	2/5	2/5	4/10	D1-D2	5/5	5/5	10/10
3160	1/5	2/5	3/10	D1	5/5	5/5	10/10
3980	3/5	2/5	5/10	D1	5/5	5/5	10/10

 

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Based on CLP criteria the substance is not classified. Based on UN GHS criteria the substance is classified Acute Tox. Cat. 5, H303.

Executive summary:

In an acute oral toxicity study (Lheritier, 1992), groups of fasted, 5-7 weeks old Sprague-Dawley rats (5/sex) were given a single oral dose of crystallised Vanilline (purity unknown) in CMC at doses of 0, 2000, 2510, 3160 and 3980 mg/kg bw and observed for 15 days. Lethargy, prostration or subdued behaviour were observed in all groups, and sialorrhea was observed in some animals. Death occurred in animals tested of main experiment: for limit test 2/10 at 2000 mg/kg bw for principal study : 0/10 at 0 mg/kg bw 1/10 at 2000 mg/kg bw 2/10 at 2510 mg/kg bw 3/10 at 3160 mg/kg bw 5/10 at 3980 mg/kg bw Oral LD50 Combined Males/Females = 3978 mg/kg bw (95% C.I. 2484 - 6368 mg/kg bw) according to Bliss' method

In the preliminary study, 3 groups of 2 males and 2 females/group were treated under the same conditions as those employed in the main study, at the dose levels of 500, 1000 and 2000 mg/kg. Animals were observed for 15 days. No clinical signs nor death were seen at all doses tested up to 2000 mg/kg in the preliminary study.

Endpoint conclusion

Dose descriptor:

LD50

Value:

3 978 mg/kg bw

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:

14-JUN-1991 to 13-NOV-1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according a recognised guideline and under GLP conditions. Howerer, some deviations occurred and the purity of the test substance is unknown.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

variation of humidity beyond the norms

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:

- Source: Iffa-Crédo (L'Arbresle, France)
- Age at study initiation: 5 - 8 weeks old
- Weight at study initiation: 204 - 271 g
- Fasting period before study: data not available
- Housing: housed individually in polycarbonate cages (305 x 180 x 184 mm)
- Diet: complete pelleted rat-mouse maintenance diet, ad libitum
- Water: softened and filered mains drinking water, ad libitum
- Acclimation period: 7 days before the start of treatment

ENVIRONMENTAL CONDITIONS:
- Temperature: 20 - 24 °C
- Humidity: 43 - 85 %
- Air changes: at least 8 per hr
- Photoperiod: 12 hrs dark / 12 hrs light (artificial)

In-life dates: from 26-AUG-1991 to 09-SEP-1991

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

* Test site:
- Area of exposure: data not available
- % coverage: apprimately 10 %
- Type of wrap if used: perforated adhesive band 10 cm wide, applied onto an elastic crepe bandage covering the entire shaved area

* Removal of test substance:
- Washing: yes, with lukewarm water
- Time after start of exposure: 24 hours

* Test material:
- Amount(s) applied: 2000 mg/kg bw
- Concentration: 69.56 % (w/v)
- For solids, paste formed: yes
- pH: 4.7

* Vehicle:
- Amount applied: data not available
- Purity: purified water

Duration of exposure:

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:
- mortality and clinical signs: 15 minutes after administration, then at 1, 2 and 4 hours, and then daily for 14 days
- cutaneous lesions: daily from days 2 to 15
- weighing: immediately before application and at days 8 and 15
- Necropsy of survivors performed: yes

Statistics:

Not applicable

Preliminary study:

2 groups each composed of 2 males and 2 females were treated under the same conditions as those employed in the main study, at the dose levels of 1000 and 2000 mg/kg. No deaths were seen at the dose level of 2000 mg/kg.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

no mortality occurred

Clinical signs:

other: none

Gross pathology:

no effects

Other findings:

no cutaneous lesions were observed. Only a yellowish coloration was noted.

Table 1: Number of animals dead during limit test

 

Dose (mg/kg bw)	Mortality (# dead/total)			Time range of deaths (hours)
	Male	Female	Combined
1000	0/2	0/2	0/4	/
2000	0/2	0/2	0/4	/

 

 

Table 2: Number of animals dead and with evident toxicity for principal study = limit test

 

Dose (mg/kg bw)	Mortality (# dead/total)			Time range of deaths (hours)	Number with evident toxicity(#/total)
	Male	Female	Combined		Male	Female	Combined
2000	0/5	0/5	0/10	/	0/5	0/5	0/10

 

Interpretation of results:

GHS criteria not met

Conclusions:

The substance is not classified since no mortality occured at the dose level of 2000 mg/kg/day.

Executive summary:

In an acute dermal toxicity study (Hazleton, 1991), groups of 5 -7 weeks old Sprague-Dawley rats (male / female) were dermally exposed to Vanilline for 24 hours to approximately 10% area of the body at doses of 2000  mg/kg bw.  Animals then were observed for 14 days.

 In the preliminary study, 2 males and 2 females per group were treated under the same conditions as those employed in the main study, at the dose levels of 1000 and 2000 mg/kg. No deaths were seen at all dose level tested of 2000 mg/kg.

Dermal LD50 Combined > 2000mg/kg bw. No clinical sign, except yellowish coloration of the skin and no mortality occurred at this dose.

     

According to classification criteria of EC regulation 1272/2008 Vanillin should not be classified for acute dermal toxicity based on the LD50 in male and female rats.

Endpoint conclusion

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Additional information

Acute oral toxicity

Seven studies were available which 3 of them were of validity 2.

All data gave LD50 results above 2000 mg/kg, except results in Jenner (1964) publication (1580 mg/kg in rat and 1400 mg/kg in guinea pig) which was not taken into consideration, because it was validity 3 according to Klimish scale.

The three studies with validity 2 (Hazleton 1992, Monsanto 1955 and 1976,) were selected as key studies. The results were similar, the study of Hazleton(1992), was the most detailed.

In this study groups of fasted, 5-7 weeks old Sprague-Dawley rats (5/sex) were given a single oral dose of crystallised Vanilline (purity unknown) in CMC at doses of 0, 2000, 2510, 3160 and 3980 mg/kg bw and observed for 15 days. Lethargy, prostration or subdued behaviour were observed in all groups, and sialorrhea was observed in some animals. Death occurred in animals tested of main experiment: for limit test 2/10 at 2000 mg/kg bw for principal study: 0/10 at 0 mg/kg bw 1/10 at 2000 mg/kg bw 2/10 at 2510 mg/kg bw 3/10 at 3160 mg/kg bw 5/10 at 3980 mg/kg bw. In the preliminary study, 3 groups of 2 males and 2 females per group were treated under the same conditions as those employed in the main study, at the dose levels of 500, 1000 and 2000 mg/kg. Animals were observed for 15 days. Neither clinical signs nor death were seen at all dose level tested.

Oral LD50 Combined Males/Females = 3978 mg/kg bw (95% C.I. 2484 - 6368 mg/kg bw) according to Bliss' method

 

The key studies provided a LD50 range from 3300 to 3978 mg/kg.

According to classification criteria of EC regulation 1272/2008 Vanillin should not be classified for acute oral toxicity. However based on the LD50 the substance is classified Acute Tox Cat. 5 for oral route according to UN GHS criteria.

 

Inhalation route:

Only one study on rat (Makaruk, 1980) was available. In this study, only one dose was tested which is the saturated vapour concentration. It gave a result of LC50 above 41.7 mg/m³after 4 hours exposure for rats and 2 hours for mouse with no other details. It was in validity 3 according to Klimish scale, and no purpose flag was selected.

 

Dermal route:

In an acute dermal toxicity study (Hazleton, 1991) of validity 2, groups of 5 -7 weeks old Sprague-Dawley rats (male / female) were dermaly exposed to Vanillin for 24 hours to approximately 10% area of the body at doses of 2000  mg/kg bw.  Animals then were observed for 14 days.

 In the preliminary study, 2 males and 2 females per group were treated under the same conditions as those employed in the main study, at the dose levels of 1000 and 2000 mg/kg. No deaths were seen at all dose level tested of 2000 mg/kg.

 

Dermal LD50 Combined > 2000mg/kg bw. No clinical sign, except yellowish coloration of the skin and no mortality occurred at this dose.

 

According to the classification criteria of EC regulation 1272/2008 Vanillin should not be classified for acute dermal toxicity based on the LD50 in male and female rats. No classification is required according to UN GHS criteria since no mortality occured at the dose level of 2000 mg/kg.

Justification for classification or non-classification

Regarding data available, vanillin was not classified for acute toxicity by oral and dermal routes according to CLP criteria.