Glutaral

Endpoint summary

Description of key information

Valid data for the skin and eye irritation potential of glutaraldehyde 50% aqueous solution are available. The data showed that under the test conditions used, glutaraldehyde 50% was corrosive to the skin of rabbits; the application of glutaraldehyde 50% unchanged into the eyes of rabbits resulted in severe lesions which were not reversible within the observation period. Thus, the findings were indicative of a risk of serious damage to eyes; in the respiratory sensory test conducted with mice, peripheral sensory irritation of the respiratory tract by glutaraldehyde 25% vapor was evident, indicating that the substance is a moderately potent peripheral sensory irritant. These observations were confirmed by human data reporting irritation symptoms of the eye, skin, nose, throat and/or lower respiratory tract related to glutaraldehyde exposure.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Scientifically acceptable; the test conduct fulfilled the requirements of the OECD TG 404 as adopted 2015 without deviations

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

(as adopted 28 July 2015)

Deviations:

no

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): Glutaraldehyde
- Physical state: clear liquid
- Lot/batch No.: ZA1955S4D1
- Other: following dilutions were used:
4% Glutaraldehyde in water
15% Glutaraldehyde in water
24% Glutaraldehyde in water
45% Glutaraldehyde in water

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Robinson Services, Inc., Clemmons NC
- Age at study initiation: no data
- Weight at study initiation: 2126 to 2590 g
- Housing: single housed in suspended stainless steel caging with mesh floors
- Diet: Harlan Teklad Certified Global High Fiber Rabbit Diet®) #2031C, appr. 150 g/day
- Water: filtered tap water, ad libitum
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-22
- Humidity (%): 23-48 (Humidity was below the targeted lower limit for 3 days during the study. A portable humidifier was used to raise the humidity levels during this time.)
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): 4%, 15% and 24% Glutaraldehyde in water

Duration of treatment / exposure:

3 min, 1 hour and 4 hours (three test sites on each animal)

Observation period:

30-60 min, 24, 48 and 72 hours, 7, 10 and 14 days

Number of animals:

5 (all females; 1 for 24% Glutaraldehyde in water, 1 for 15% Glutaraldehyde in water and 3 for 4% Glutaraldehyde in water)

Details on study design:

TEST SITE
- Area of exposure: 6 cm²
- Type of wrap if used: test substance was covered with 1-inch x 1-inch, 4-ply gauze pad. The pads and entire trunk of the animal were then wrapped with semi-occlusive 3-inch Micropore™ tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with 3% soap solution followed by water and clean paper towels
- Time after start of exposure: 3 minute, 1 hour or 4 hours, respectively

SCORING SYSTEM: according to guideline

Irritation parameter:

erythema score

Remarks:

(4-hour exposure)

Basis:

mean

Remarks:

(3 animals)

Time point:

other: 24h - 48h - 72h

Score:

2.87

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: 4% Glutaraldehyde in water

Irritation parameter:

edema score

Remarks:

(4-hour exposure)

Basis:

mean

Remarks:

(3 animals)

Time point:

other: 24h- 48h - 72h

Score:

3

Max. score:

4

Reversibility:

not reversible

Remarks:

in 2 of 3 animals

Remarks on result:

other: 4% Glutaraldehyde in water

Irritation parameter:

erythema score

Remarks:

(3-minute exposure)

Basis:

animal #2

Time point:

other: 24h - 48h -72h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: 15% Glutaraldehyde in water

Irritation parameter:

edema score

Remarks:

(3-minute exposure)

Basis:

animal #2

Time point:

other: 24h - 48h - 72h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

other: 15% Glutaraldehyde in water

Irritation parameter:

erythema score

Remarks:

(1-hour exposure)

Basis:

animal #2

Time point:

other: 24h - 48h - 72h

Score:

3

Max. score:

4

Reversibility:

other: euthanized after 72 hours

Remarks on result:

other: 15% Glutaraldehyde in water

Irritation parameter:

edema score

Remarks:

(1-hour exposure)

Basis:

animal #2

Time point:

other: 24h - 48h - 72h

Score:

3

Max. score:

3

Reversibility:

other: euthanized after 72 hours

Remarks on result:

other: 15% Glutaraldehyde in water

Irritation parameter:

erythema score

Remarks:

(4-hour exposure)

Basis:

animal #2

Time point:

other: 24h - 48h - 72h

Score:

3.7

Max. score:

4

Reversibility:

other: euthanized after 72 hours

Remarks on result:

other: 15% Glutaraldehyde in water

Irritation parameter:

edema score

Remarks:

(4-hour exposure)

Basis:

animal #2

Time point:

other: 24h - 48h - 72h

Score:

4

Max. score:

4

Reversibility:

other: euthanized after 72 hours

Remarks on result:

other: 15% Glutaraldehyde in water

Irritation parameter:

erythema score

Remarks:

(3-minute exposure)

Basis:

animal #1

Time point:

other: 24h

Score:

2

Max. score:

2

Reversibility:

other: euthanized after 24 hours

Remarks on result:

other: 24% Glutaraldehyde in water

Irritation parameter:

edema score

Remarks:

(3-minute exposure)

Basis:

animal #1

Time point:

other: 24h

Score:

2

Max. score:

2

Reversibility:

other: euthanized after 24 hours

Remarks on result:

other: 24% Glutaraldehyde in water

Irritation parameter:

erythema score

Remarks:

(1-hour and 4-hours exposure)

Basis:

animal #1

Time point:

other: 24h

Score:

4

Max. score:

4

Reversibility:

other: euthanized after 24 hours

Remarks on result:

other: 24% Glutaraldehyde in water

Irritation parameter:

edema score

Remarks:

(1-hour and 4-hours exposure)

Basis:

animal #1

Time point:

other: 24h

Score:

4

Max. score:

4

Reversibility:

other: euthanized after 24 hours

Remarks on result:

other: 24% Glutaraldehyde in water

Irritant / corrosive response data:

24% concentration (one animal tested = animal #1):
3-minute exposure and 24-hour observation period no corrosion was observed
1-hour exposure and 24-hour observation period corrosion was observed
4-hour exposure and 24-hour observation period corrosion was observed

15% concentration (one animal tested = animal #2):
3-minute exposure and 72-hour observation period no corrosion was observed
1-hour exposure and 72-hour observation period no corrosion was observed
4-hour exposure and 72-hour observation period corrosion was observed

4% concentration (three animals tested):
4-hour exposure and 14-day observation period no corrosion was observed in all three animals

Details on findings and scores:

24% Glutaraldehyde in water - Erythema/Edema
Animal No.	Exposure time	30-60 min after patch removalErythema/Edema	24 hours after removal of 4-hours patchErythema/Edema	Findings
1	3 min	1/1*	2/2
1	1 hour	2/2*	4/4**	* Dark yellow discolouration in and around the dose site. **Green discolouration in and/or around the dose site. Blanching in the dose site.
1	4 hours	2/3*	4/4**	* Dark yellow discolouration in and around the dose site. **Green discolouration in and/or around the dose site. Blanching in the dose site.

15% Glutaraldehyde in water - Erythema/Edema
Animal No.	Exposure time	30-60 min after patch removalErythema/Edema	After removal of 4-hour patchErythema/Edema			Findings
			24h	48h	72h
2	3 min	1/0	1/0	1/0	1/0
2	1 hour	2/2^1	2/3^2	3/3^2	4/3^3,4	1 Yellow discoloration in the dose site.2 Green discoloration in the dose site.3 Hyperkeratosis present at the dose site.4 Dark brown discoloration in the dose site.
2	4 hours	2/4^2	3/4^2	4/4^2,3	4/4^3,4

4% Glutaraldehyde in water - Erythema/Edema
Animal No.	30-60 min after patch removal Erythema/Edema	After removal of 4-hour patch Erythema/Edema
		24h	48h	72h	Day 7	Day 10	Day 14
3	2/3^1,2	2/4^3,4	4/4^3,4	4/4^3,4,5	3/2^3,5	4/2^3,5	4/1^3,5,6
4	2/2	2/2^7	3/2^7	3/2^8	2/1^5,8	1/0^5,7	1/0^5,7
5	2/2	2/3^7	3/3^7	3/3^8	2/2^5,8	1/2^5,7	1/2^5,7
Total	6/7	6/9	10/9	10/9	7/5	6/4	6/3
Mean	2.0/2.3	2.0/3.0	3.3/3.0	3.0/3.0	2.3/1.7	2.0/1.3	2.0/1.0
1 Greenish yellow discolouration in the dose site.2 Small light brown areas in the dose site.3 Area of brown discolouration in the dose site.4 Area of superficial eschar in the dose site.5 Hyperkeratosis present at the dose site.6 Shiny skin evident at the dose site.7 Light brown discolouration in the dose site.8 Dark brown discolouration in the dose site.

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

Under the test conditions used, Glutaraldehyde 24% and 15% were corrosive and Glutaraldehyde 4% was irritating to the skin of rabbit.
Thus, Glutaraldehyde 24% and 15% are assigned R 34 according to EU classification and labelling, and are classified in Categories 1B and 1C "Corrosive" according to GHS (2005), respectively. Glutaraldehyde 4% is assigned R38 according to EU classification and labelling, and is classified in Category 2 "Irritant" according to GHS (2005).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

original German report dated 22 Dec 1981

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study conducted according to the Fed. Reg. 38, No. 187, § 1500.42, p. 27019 of Sept. 27, 1973.

Qualifier:

according to guideline

Guideline:

other: Fed. Reg. 38, No. 187, § 1500.42, p. 27019 of Sept. 27, 1973

Principles of method if other than guideline:

Draize Test

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted

Specific details on test material used for the study:

- Name of test material (as cited in study report): Glutaraldehyde approx. 50% sol.
- Physical state: liquid
- Analytical purity: 50.0% glutaraldehyde
- Lot/batch No.: the est substance No. was 80/265
- Stability under test conditions: about 12 months

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Gaukler, Offenbach/Main, Germany (animal breeder)
- Weight at study initiation: 2.62 - 2.99 kg
- Housing: individually, in cages of stainless steel with wire mesh walk floor (floor area was 40 cm x 51 cm), no bedding, sawdust in the waste trays
- Diet (e.g. ad libitum): Ovator Solikanin 4 mm; Muskator-Werke, Düsseldorf, FRG; ca. 130 g per animal per day
- Water (e.g. ad libitum): about 250 ml per animal per day of fully demineralized water on working days, tap water on week-ends
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26 °C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12 h/ 12 h

Vehicle:

unchanged (no vehicle)

Controls:

other: The untreated eye of each test animal served for control.

Amount / concentration applied:

0.1 ml of the test substance was applied into the conjunctival sac of the right eye.

Duration of treatment / exposure:

The test substance was not washed out.

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

6 (3 per sex)

Details on study design:

A volume of 0.1 ml of the unchanged test substance was instilled into the right eye of each of the 6 white Vienna rabbits;.
The test substance was not washed out and the animals were scored for corneal changes, iris effects and conjunctival reaction, after 1, 24, 48 and 72 hours, and after 8 days following test substance application.
The assessment of the findings was based on the scoring system of Draize JH ((Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. In Dermal Toxicity, 46-59, 1959).

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

(for each animal)

Time point:

other: 24 h - 48 h - 72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: Animal 1: 2; Animal 2: 2; Animal 3: 2; Animal 4: 2; Animal 5: 2; Animal 6: 2.3

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

(for each animal)

Time point:

other: 24 h - 48 h - 72 h

Score:

2.15

Max. score:

3

Reversibility:

not reversible

Remarks on result:

other: Animal 1: 2; Animal 2: 2; Animal 3: 2.3; Animal 4: 2; Animal 5: 2.3; Animal 6: 2.3

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

(for each animal)

Time point:

other: 24 h - 48 h - 72 h

Score:

2.3

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: Animal 1: 2; Animal 2: 2.6; Animal 3: 2.3; Animal 4: 2.3; Animal 5: 2.3; Animal 6: 2.3

Irritation parameter:

iris score

Basis:

mean

Remarks:

(for each animal)

Time point:

other: 24 h - 48 h - 72 h

Score:

1

Max. score:

2

Reversibility:

not reversible

Remarks on result:

other: Animal 1: 1; Animal 2: 1; Animal 3: 1; Animal 4: 1; Animal 5: 1; Animal 6: 1

Irritant / corrosive response data:

- Reading of the cornea revealed increasing opacity, ranging from scattered or diffuse (24 h) to nearly complete (8 d); the involved area was > 0.75 - <= 1;
- Reading of the iris revealed folds above normal, swelling and circum-corneal injection over the whole observation period, the iris was still reacting to light;
- Reading of the conjunctiva revealed redness (diffuse, deeper crimson red, with individual vessels, which were not easily discernible); swelling (obvious swelling with partial eversion of the lids during the first 48 hours, and swelling with half-closed lids seen until day 8) and eye discharge, which was above normal during the first 48 hours and resulted in moistening of lids and adjacent hairs within the following days (until day 8).
- Additional effects included contraction of pupils, small retractions in the eyelid, suppuration and loss of hair in the margin of the eyelid.
- These effects increased with time and no reversibility could be evidenced.

Summary of the eye scores for each animal at different reading time points, following application of glutaraldehyde:

Reading time point (h: hour; d: day)	Animal	Treated eyes
		Conjunctiva			Cornea		Iris	Additional findings
		Erythema	Chemosis	Secretion	Opacity	Area
24 h	1	2	2	1	1	4	1	none
	2	2	2	1	1	4	1	none
	3	2	2	1	1	4	1	pupil contracted
	4	2	2	1	1	4	1	pupil contracted
	5	2	2	2	1	4	1	none
	6	2	2	1	2	4	1	none
48 h	1	2	2	1	2	4	1	none
	2	2	3	1	2	4	1	small retractions in eyelid
	3	2	2	1	2	4	1	pupil contracted
	4	2	2	1	2	4	1	pupil contracted
	5	2	2	2	2	4	1	pupil contracted
	6	2	2	1	2	4	1	none
72 h	1	2	2	2	3	4	1	suppuration
	2	2	3	2	3	4	1	small retractions in eyelid & suppuration
	3	3	3	2	3	4	1	suppuration
	4	2	3	2	3	4	1	suppuration
	5	3	3	2	3	4	1	small retractions in eyelid & suppuration
	6	3	3	2	3	4	1	suppuration & small retractions in eyelid
8 d	1	2	2	2	4	4	1*	suppuration & small retractions in eyelid & loss of hair at the margins of the eyelid
	2	2	3	2	3	4	1	small retractions in eyelid & suppuration & loss of hair at the margins of the eyelid
	3	2	3	3	4	4	1*	small retractions in eyelid & loss of hair at the margins of the eyelid & suppuration
	4	2	3	2	4	4	1*	small retractions in eyelid & loss of hair at the margins of the eyelid & suppuration
	5	3	3	2	3	4	1	small retractions in eyelid & suppuration & loss of hair at the margins of the eyelid
	6	3	3	2	3	4	1	suppuration & small retractions in eyelid & loss of hair at the margins of the eyelid

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

Skin irritation/corrosion:

The Dow Chemical Company (2012) performed a study with 24%, 15% and 4% glutaraldehyde in water. Equivalent to OECD guideline 404, New Zealand White rabbits were exposed to the test substance under semi-occlusive conditions for 3 minutes, 1 hour and 4 hours. After 3 minutes exposure with the 24% glutaraldehyde solution only erythema and edema effects up to grade 2 were observed. Following exposure to the test substance (24%) for 60 minutes severe erythema/edema and necrosis was reported. With 15% glutaraldehyde skin corrosion was found only after 4-hour exposure but severe skin irritation effects (erythema and edema up to grade 4) were found in the 1-hour exposure. In the lowest concentration tested (4% glutaraldehyde in water) no skin corrosion was observed but still severe skin irritation with a mean (3 animals, 24h-48h-72h) erythema/edema value of 2.8/3.0 was reported after 4 hours of exposure. 14 days after exposure the effects were still not revised. From these results it was concluded that the test substance at concentrations >= 15% is corrosive to the skin and at 4% it is still skin irritating.

The Bushy Run Research Center (1988) performed two studies with 50% glutaraldehyde. In accordance with OECD guideline 404 and in compliance with GLP, New Zealand White rabbits were exposed to the test substance under semi-occlusive conditions for 3 minutes (in only one of the two studies), 1 hour and 4 hours. After 3 minutes exposure only minor erythema effects were observed. Following exposure to the test substance for 60 minutes, minor to moderate erythema was reported and 4 hour exposure resulted in moderate to severe erythema and edema, and to necrosis. It was concluded that the test substance is corrosive to the skin.

BASF performed several studies investigating the irritating properties of glutaraldehyde. In one BASF study (1994), Vienna White rabbits were exposed to 50% glutaraldehyde for 3 minutes, 1 hour and 4 hours under occlusive conditions. After 3 minutes exposure only minor effects were seen, while after 1 hour exposure slight erythema and edema were observed. These effects remained until the end of the observation period and even extending beyond the application site. In addition scaling was observed. After four hours exposure, in addition to erythema and edema, necrosis was observed. Another BASF study (1979) was performed similar and similar effects were observed.

In four other studies (1983, 1975, 1970 and 1970), 25% glutaraldehyde was tested in Vienna White rabbits under occlusive conditions. In the first BASF study (1983), mild erythema and edema was observed when animals were exposed for 1 hour. At the end of the observation period after 8 days, two animals still displayed very slight erythema and all three animals showed scaling. In the second BASF study (1975) after a 20 hour exposure period, at reading time point 24 h, both animals displayed necrosis and slight to severe marginal edema. After 8 days, necrosis became severe in one case and had a leathery aspect; in the second case, necrosis was parchment-like. In the remaining BASF studies (1970), animals were exposed for 1, 5, and 15 minutes and for 20 hours under occlusive conditions. In one of these studies (1970) necrosis was observed when animals were exposed for 5 minutes or longer, while in the other study (1970) only the animals exposed for 20 hours showed necrosis. Finally, an in vitro skin corrosion and irritation test was performed (BASF, 2013) in accordance with OECD guideline 431 and 439 using the human reconstituted epidermis model EPIDERM. This study showed that glutaraldehyde has a corrosion potential in the Epiderm skin corrosion/irritation test.

The Carnegie-Mellon Institute of Research (1977) performed two tests exposing rabbits to either 25 or 50% glutaraldehyde. In the first test performed similar to the in vivo rabbit skin test method for corrosion, specified by the U.S. Dept. of Transportation (DOT), rabbits were exposed for 4 hours to the test substance under occlusive conditions. Two of the 6 rabbits exposed to 50% glutaraldehyde showed necrosis, while none of the test animals showed necrosis when exposed to 25% glutaraldehyde. In the second test, rabbits were exposed under open conditions for 24 hours to the test substance. In animals exposed to 50% glutaraldehyde two cases of marked capillary injection, one case of moderate erythema, one case of marked erythema and one case of moderate edema were reported (findings were graded 5 on a scale of 10). In animals exposed to 25% glutaraldehyde, three animals showed moderate erythema whereas the two remaining showed no signs of irritation (findings were graded 3 on a scale of 10).

The Mellon Institute (1964), exposed New Zealand White rabbits to 45% glutaraldehyde under open conditions for 24 hours. No irritation was observed.

Nutrition International Inc. (1982) exposed New Zealand White rabbits to 50 and 25% glutaraldehyde under occlusive conditions for 24 hours. The EPA 40 CFR 183 guideline was followed and the observation period was 14 days. All treated animals (50% glutaraldehyde) showed erythema, edema and eschar formation almost lasting over the whole observation period. Animals exposed to 25% glutaraldehyde showed eschar formation in all animals within 72 hours after application. This condition persisted throughout the 14 day of observation indicating no reversibility.

In a publication by Ballantyne et al. (2001), New Zealand Whites were exposed under occlusive conditions to 1, 2, 5, 10, 45, 50% glutaraldehyde for 3 minutes, 1 hour or 4 hours and observed for 3 weeks. At 50% glutaraldehyde there was severe local inflammation (erythema and edema) with tissue injury (alopecia, scab formation, necrosis) with gradual decreased severity at concentrations down to 1%, which was essentially a marginal (threshold) effect concentration for irritancy. At the 10% and 25 % concentrations moderate erythema, minor edema and punctate necrosis was observed. It was concluded that glutaraldehyde is corrosive to the skin. Ballantyne et al. (1997) also investigated possible differential toxicity in acidic unbuffered glutaraldehyde and alkaline buffered glutaraldehyde. For the unbuffered glutaraldehyde erythema was very slight and disappeared within 7 days. The effects were fully reversible. Local skin irritation was relatively minor and slightly more marked for the buffered glutaraldehyde.

Uetmitsu et al. (1976) exposed rabbits (strain not specified) under occlusive conditions for 24 hours to 25, 7, 2, 0.7, and 0.2% glutaraldehyde. Test concentrations from 0.2 -7% did not result in irritating effects. At 25% glutaraldehyde only mild effects were observed.

In Hess et al. (1989), ICR mice were exposed to 0.25 or 2% glutaraldehyde under open application for 2 consecutive days. Treatment with the test substance resulted in multiple brownish lesions with local thickening and hardening, epithelial erosion and collagen degeneration.

Conclusion

Although some studies did not observed corrosive effects, the majority of the available studies indicate that glutaraldehyde is corrosive to the skin. Furthermore, based on all available data it can also be concluded that 1 and 4 hour exposure is corrosive to the skin, while 3 minute exposure only causes irritating effects. Concentrations of glutaraldehyde of 15% and more induce skin corrosion. 4% glutaraldhyde in water is not corrosive but still skin irritating.

Eye irritation:

BASF performed three studies exposing Vienna White rabbits to 50 or 25% glutaraldehyde. The test substance was not washed out and the observation period lasted 8 days. In one BASF study (1994) performed in accordance to Fed. Reg. 38, No. 187, § 1500.42, p. 27019 of Sept. 27, 1973., severe lesions were observed which were not reversible within 8 days after application of 0.1 mL 50% glutaraldehyde. In the two other studies (1970, both) exposing the rabbits to 0.05 mL of 25% glutaraldehyde also severe irreversible lesions to the eye were observed. In another study (BASF, 2013) the objective was to assess the potential for acute eye irritation of the test material after single topical application using the reconstructed human cornea model EpiOcular. In this study the test substance showed an eye irritation potential in the EpiOcular eye irritation test. However, this test method did not allow for the evaluation of serious eye damage.

The Dow Chemical company (1987) exposed three male and female New Zealand white rabbits to 0.1 mL of a 45% glutaraldehyde solution. Severe corneal injury, iritis and severe conjunctival irritation were observed in each of six rabbits. Ocular effects included a purulent discharge, conjunctival necrosis, adhesion of the nictitating membrane and cornea and pannus. Corneal opacity and conjunctival swelling were so severe that complete scoring of the cornea and iritis was not possible for most of the observation period. Adhesion of the nictitating membrane to the cornea as well as the purulent discharge further interfered with the ocular examinations. One rabbit died 17 days after dosing but did not appear related to treatment. In the surviving animals, all eyes were severely affected through 21 days of observation.

Nutrition International Inc. (1982) exposed New Zealand White rabbits to 0.25, 0.5, 1, and 1.5% glutaraldehyde in several studies. All concentrations were once tested with an observation period of 72 hours and no wash out. In addition, a study was performed with 1.5% glutaraldehyde comparing effects between rinsing after 30 seconds after application or not rinsing. In this study effects were observed for 21 days. At all concentrations from 0.25 to 1% the test substance was not considered to be irritating to the eye. In all three studies with 1.5% glutaraldehyde irritating effects were observed.

The Bushy Run Research Center (1981) exposed six rabbits to 0.1 mL or 0.01 mL of 0.5, 0.2, or 0.1% glutaraldehyde. Treatment with 0.1 mL of a 0.5% glutaraldehyde resulted in slight redness of the eyelids, slight injection of the conjunctivae and in white discharge from 6 hours to one or two days. Similar effects were produced from 0.01 mL of GA 0.5%, but were only observed at 6 hour in half of the treated eyes. Treatment with 0.1 mL of a GA 0.2% resulted in slight redness of the eyelids, injection of the conjunctivae and in white discharge in 3 cases at time point 6 hours. These effects persisted through 24 hours. No effects resulted from treatment with 0.01 mL of the test material. No effects were seen in the eyes following treatment with 0.1 mL and 0.01 mL 0.1% glutaraldehyde.

In a publication by Ballantyne et al. (2001), New Zealand Whites were exposed to various concentrations of glutaraldehyde ranging from 0.1 to 45 % in volumes ranging from 0.001 to 0.1 mL. Rabbits were observed for up to three weeks. There was a clear concentration-effect relationship for ocular and periocular inflammation and injury at any given installation volume, from severe at 45 % to no effect at 0.1%. Where different volumes of the same concentration were instilled (5, 2,1, 0.5, or 0.2%), the effects were, in general, less market for the smaller volumes. With respect to corneal injury (at 0.1 mL instilled volume) responses were severe at 5 and 45%, mild at 2%, threshold at 1% and no corneal effect occurred at 0.5%. For conjunctival inflammation the threshold concentration was 0.2%, and the no-effect concentration 0.1%. At 1 to 2%, conjunctival hyperemia and chemosis were marked and at 5% and higher they were severe and persistent. Overall, the no effect concentration for eye irritation was 0.1% Ballantyne et al. (1997) also investigated possible differential toxicity in acidic unbuffered glutaraldehyde (2.2%) and alkaline buffered glutaraldehyde (2.2%). The rabbit eye irritation studies conducted with 0.1 mL test material instilled in the conjunctival sac of the eye showed slightly more marked conjunctival reactions with buffered glutaraldehyde. Corneal injury was marked and persistent with buffered glutaraldehyde and only slight and transient with unbuffered glutaraldehyde. With 0.01 mL applied on the cornea surface, no corneal injury occurred but conjunctival reaction was more marked with unbuffered glutaraldehyde when compared to buffered glutaraldehyde.

The Carnegie-Mellon Institute of Research (1977) performed two tests exposing rabbits to either 50, 5, and 1% (test 1) and 25, 5, and 1% (test 2) glutaraldehyde. In both tests, severe corneal injury, iritis, swelling and necrosis of the eyelids were reported for 0.005 mL undiluted test material.

Severe corneal injury, iritis, swelling and necrosis of the eyelids were also reported for 0.5 mL of the 5% dilution of test material. Minor corneal injury were seen at the 1% dilution.

The Mellon Institute (1964), exposed New Zealand White rabbits to 45% and 1% glutaraldehyde. Instillation of 0.005 mL of undiluted test material and instillation of 0.5 mL of a 1% aqueous solution of the test material resulted in severe necrosis.

Uemitsu et al. (1976) exposed rabbits to 0.1 mL of 0.2, 2, and 25% glutaraldehyde. At the test concentrations 0.2 and 2% only irritating effects were observed, while 25% glutaraldehyde cause irreversible effects on the iris and cornea.

Stonehill et al. (1963) applied 0.1 mL of 2% glutaraldehyde to the eye of 5 rabbits and severe reaction consisting of inflammation, lacrimation and edema was observed, which was reversible within 7 to 8 days.

TFB-Toxikologia (1986) performed an in vitro HET-CAM with 1, 10, and 25% glutaraldehyde and concluded that glutaraldehyde is a severe irritant at all tested concentrations.

Conclusion

Overall it can be concluded that glutaraldehyde causes severe lesion to the eye.

Respiratory irritation

An acceptable respiratory sensory test conducted with mice exposed to glutaraldehyde (25% aqueous solution) vapor was published (Werley et al., 1995). The peripheral sensory irritation (PSI) study was conducted with male ND4 Swiss Webster mice for determination of the respiratory irritating potential of glutaraldehyde. Mice were exposed to concentrations of 1.6, 4.0, 4.7, 5.6, 7.5, 17.7, and 36.7 ppm glutaraldehyde for 30 minutes. The respiratory rate (RR) decreased in all glutaraldehyde treated groups almost immediately following exposure starting; a plateau was reached within 5 to 10 minutes. The decrease in RR was related to prolongation of the expiratory phase of the respiratory cycle; this resulted in a notching indicative of peripheral sensory irritation of the respiratory tract. A clear effect-concentration relationship was evident. A RD50 (concentration resulting in 50% decrease in RR) of 13.86 ppm (95% confidence limits: 9.86 – 23.58 ppm) was calculated. The authors concluded that glutaraldehyde was a moderately potent peripheral sensory irritant in mice.

Human data on irritation

Skin

Pianiello et al (1997) found that although 135 nurses exposed to glutaraldehyde were more likely to report skin symptoms, the prevalence of skin symptoms was not correlated with personal glutaraldehyde exposure. A similar conclusion was drawn by Water et al (2003), where skin symptoms were 3.6 times more likely to be reported by exposed workers. Norbeaeck et al (1988) studied the prevalence of certain symptoms (eye, skin and airway symptoms, headache, nausea, and fatigue) among hospital workers with and without exposure to glutaraldehyde during cold sterilization work. Exposure measurements revealed that the present exposure to glutaraldehyde was intermittent and well below the Swedish occupational exposure limit. In spite of this low exposure, the exposed group exhibited a significantly increased frequency of skin and airway symptoms, as well as headache, in comparison with the unexposed group. A dose-response relationship between the frequency of exposure and the number of symptoms could also be demonstrated. Reifenrath et al (1985) performed an 8-week irritancy test by applying a 10% aqueous solution of glutaraldehyde to the ankle and heel area of 12 volunteers. Irritation and one case of sensitization resulted from glutaraldehyde application to areas of thin stratum corneum (anterior ankle) but not from applications to thick stratum corneum (posterior ankle).

Eye

In a study performed by the Union Carbide Corporation in 1991, the ocular and respiratory irritation experienced by 12 subjects exposed to glutaraldehyde was investigated. The test panel was placed in a closed room and exposed to a vapour of the test substance each day for 8 hours. The panel was exposed to Cidex on the first two exposure days and to Sporicidin on the last exposure day (fifth). Both substances contain glutaraldehyde. Burning, itching of the eyes and watery eyes were reported by 1 or 2 subjects. For both test materials, lachrymation, palpebral conjuctiva redness, and bulbar conjunctiva redness were not observed. Nayebzadeh et al (2007) observed a higher prevalence of headache and itchy eyes among health care personnel who were responsible for handling glutaraldehyde and also those with potential exposure to glutaraldehyde who worked in the same location where unsafe work practices were observed. Pianiello et al (1997) found that although 135 nurses exposed to glutaraldehyde were more likely to report eye symptoms, the prevalence of eye symptoms was not correlated with personal glutaraldehyde exposure.

Respiratory

Pianiello et al (1997) found that although 135 nurses exposed to glutaraldehyde were more likely to report throat symptoms, the prevalence of throat symptoms was not correlated with personal glutaraldehyde exposure. Waters et al (2003) investigated work practices and glutaraldehyde exposure in relation to lung function. For this purpose, a questionnaire was given to 76 nurses. Exposed nurses (n = 38) also completed lung function tests and visual analogue scales before and after a work session in which glutaraldehyde exposure occurred. None of the other symptoms were significantly associated with glutaraldehyde exposure. There were significant cross-shift reductions in FVC and FEV(1) in the exposed group. However, no evidence of a dose-response relationship for respiratory symptoms or lung function was found. In a study performed by the Union Carbide Corporation in 1991, the ocular and respiratory irritation experienced by 12 subjects exposed to glutaraldehyde was investigated. The test panel was placed in a closed room and exposed to a vapour of the test substance each day for 8 hours. The panel was exposed to Cidex on the first two exposure days and to Sporicidin on the last exposure day (fifth). Both substances contain glutaraldehyde. Dryness of nose, and throat was reported by 1 to 2 subjects. In addition, significantly increased frequency of airway symptoms was also described in the study by Norbaeck et al (1988). Katagiri et al (2006) measured the environmental glutaraldehyde levels during the disinfection of endoscopes and also investigated the subjective symptoms of the workers engaged in that work. In 6 hospitals in the Tokyo and Kanagawa area, 8 rooms for endoscope washing and disinfecting the endoscopy equipment were surveyed. The geometric mean environmental GA levels in the 8 rooms were 1.3 to 19.6 ppb. The personal exposure levels at the time of replacing the antiseptic solution containing GA in two of the disinfecting rooms were 94.2 and 84.9 ppb. Subjective symptoms such as ophthalmic, nasal, respiratory, pharyngeal symptoms and nausea were more prevalent among workers than controls as evidenced from the questionnaire survey.

In addition to the above mentioned studies, the Dutch Expert Committee (2005) reported work-related irritation symptoms of the eye, skin, nose, throat and/or lower respiratory tract be related to glutaraldehyde exposure.

Direct observations

The following case studies were identified of clinical cases regarding glutaraldehyde exposure and irritation. Wiggins et al (1998) described a case of a woman suffering from upper respiratory tract irritation, skin rash and recurrent epistaxis (noose bleeding). A visit to the workplace revealed inadequate personal protective measures and inadequate local ventilation. The patient's symptoms resolved with the implementation of personal protective measures and engineering controls. A similar case was described by Benson et al (1984) in which a 38-year-old woman suffered from nasal irritation, sore eyes and skin rashes, related to the use of glutaraldehyde for the disinfection of fibrooptic endoscopes. Prigent et al (1996) described the case of a 7-year-old child consulted for major, dry an painful necrosis of the pulp of the greater right toe. The 20% glutaraldehyde solution which had been daily applied on plantar warts for 8 weeks was the only agent which could be the cause. Murray et al (1985) described the case of a patient subjected to anesthesia who developed eye injury during induction of anesthesia. By the six postoperative hour bilateral bulbar and palpebral conjunctival inflammation was prominent and accompanied by tearing, swelling of the eyelids, burning pain, and photophobia. The inflammation was completely resolved on the third postoperative day and no visual disturbance ensued. Ünal et al (2006) described the case of 6 patients subjected to cataract surgery. A 2% glutaraldehyde solution was used inadvertently by the operating room staff who cleaned and sterilized reusable ocular instruments before autoclaving. Six eyes of 6 patients developed toxic anterior segment syndrome (TASS) after uneventful phacoemulsification cataract surgery with implantation of a 3-piece acrylic intra ocular lenses performed by 2 ophthalmologists on the same day. Clinical findings included corneal edema, Descemet´s membrane folds, anterior chamber reaction, fibrin formation, and irregular, dilated, and unreactive pupils. None of the affected corneas improved. Additional surgical procedures were required and included penetrating keratoplasty, trabeculectomy, and glaucoma tube implantation.

Justification for selection of skin irritation / corrosion endpoint:
Test performed equivalent to OECD 404 with different concentrations

Justification for selection of eye irritation endpoint:
Test performed according to guideline and with the highest concentration of glutaraldehyde (50%).

Effects on skin irritation/corrosion: corrosive

Effects on eye irritation: corrosive

Effects on respiratory irritation: irritating

Justification for classification or non-classification

Glutaraldehyde is corrosive to the skin and results in severe damage in the eye; further, glutaraldehyde is a moderately potent peripheral sensory irritant. Therefore, according to Annex VI of EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008 and based on the available data, glutaraldehyde has to be classified as Skin Corr. 1B: H314; Causes severe skin burns and eye damage, Eye Damage 1:H318; Causes Serious Eye Damage and STOT SE 3: H335; May cause respiratory irritation.