Reaction products of sodium glucoheptonate with manganese sulfate and sodium hydroxide

Endpoint summary

Description of key information

Skin irritation: according to OECD 404, rabbit, undiluted, 4 h, semiocclusive, not irritating
Eye irritation: according to OECD 405, rabbit, 1 % solution in distilled water, not irritating

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

Study period:

January 15, 2016 - March 10, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well documented GLP Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

Exception to guideline 404, for not testing weight of evidence analysis by the test facility.

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Sainath Agencies, Hyderabad, India
- Age at study initiation: 4.5 to 5.5 months
- Weight at study initiation: Minimum: 2.487, Maximum: 2.595
- Housing: individually in stainless steel wire meshed cages were used.
- Diet (e.g. ad libitum): Teklad certified Global High Fiber Rabbit pellet Feed manufactured by Harlan, USA ad libitum
- Water (e.g. ad libitum): UV sterilised drinking water filtered through Kent Reverse Osmosis water filtration system ad libitum
- Acclimation period: 6 to 8 days

Animal Identification
Each rabbit was uniquely numbered on the ear using a tattoo machine. Appropriate labels were attached to the cages indicating the study number, test item code, sex, dose, type of study, cage number and animal number.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 22 °C
- Humidity (%): 63 to 65%
- Air changes (per hr): minimum 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 h artificial light and 12 h darkness, light hours being 06:00 - 18:00 h

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 mg
- Concentration (if solution): 1 %

Duration of treatment / exposure:

4 hours

Observation period:

The skin reactions of each rabbit were observed at 1, 24, 48 and 72 h post patch removal.

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 6 cm²
- % coverage:
- Type of wrap if used: gauze patch that was secured at the margins by non-irritating tape for a period of 4 h

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the 4 h exposure period, the residual test item was removed with cotton soaked in distilled water.
- Time after start of exposure: 4 hours

SCORING SYSTEM:
Irritation was scored according to OECD 404.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

There were no signs of systemic adverse effect in any treated rabbits.
Erythema was evident at 1 h in all rabbits, which resolved by 24 h post patch removal in all rabbits. The mean dermal irritation scores at 24, 48 and 72 h post-patch removal for individual animals were 0.00, 0.00, 0.00 for erythema and 0.00, 0.00, 0.00 for oedema, respectively.

Mean Dermal Irritation Score

The mean dermal irritation scores at 24, 48 and 72 h post-patch removal for animal Nº 1, 2 and 3 were 0.00, 0.00, 0.00 for erythema and 0.00, 0.00, 0.00 for oedema, respectively (Table 2).

Narrative Description of Skin Reactions

Following the 4 h exposure period (day 0), the skin of each rabbit was observed at 1, 24, 48 and 72 h post patch removal.

At 1 h post patch removal, the treated skin site revealed very slight erythema (barely perceptible) in all three rabbits (Table 1).

At 24 h post patch TIA, the treated skin site recovered completely and appeared normal in all three rabbits throughout the experimental period (Table1).

The control skin sites of all three rabbits were normal with no erythema and no oedema was observed throughout the experimental period (Table 1).

Clinical Observations other than Dermal Irritation

No clinical signs were observed in any rabbit throughout the experimental period (Table 2).

Interpretation of Results

The mean scores of erythema (0.00) and oedema (0.00) observed at the 24, 48 and 72 h post patch removal observation time-points indicated that the DABQUEL COMPLEX MnP is a non-irritant under the described experimental conditions.

Table 1: Dermal Irritation Scores 

Control Site: Untreated

Sex: Male                          

Rabbit N°	Site of Application	Observations after Patch Removal
		Erythema				Oedema
		Hour				Hour
		1	24	48	72	1	24	48	72
1	Right	0	0	0	0	0	0	0	0
2	Right	0	0	0	0	0	0	0	0
3	Right	0	0	0	0	0	0	0	0

 

Treated Site: 500 mg DABQUEL COMPLEX MnP (pulverised and moistened with 0.5 mL distilled water)

Rabbit N°	Site of Application	Observations after Patch Removal
		Erythema				Oedema
		Hour	Hour
		1	24	48	72	1	24	48	72
1	Left	1	0	0	0	0	0	0	0
2	Left	1	0	0	0	0	0	0	0
3	Left	1	0	0	0	0	0	0	0

Table 2: Clinical Observations and Body Weight (kg) of Individual Rabbit

Sex: Male

Rabbit N°	Clinical Observations made on Day				Body Weights (kg)
	0	1	2	3	Before Treatment	At Termination
1	1	1	1	1	2.595	2.670
2	1	1	1	1	2.571	2.600
3	1	1	1	1	2.487	2.660

Key: 0 = Day of dermal application

Clinical Sign: 1 = Normal

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of this study, and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015), DABQUEL COMPLEX MnP is not classified as a skin irritant

Executive summary:

In a GLP-compliant acute dermal irritation study according to OECD TG 404, three adult male New Zealand White rabbits were dermally exposed to 500 mg DABQUEL COMPLEX MnP (pulverised and moistened with 0.5 mL distilled water), for 4 h (day 0), applied to approximately 6 cm² area of skin. Initially one rabbit was tested with a single patch applied evenly to the intact skin for a period of 4 h. Based on the observations at 24 h post patch removal, two additional rabbits were tested simultaneously to confirm the irritation response. The control skin site of each rabbit was untreated. The treated and the control sites were covered with a gauze patch that was secured at the margins by non-irritating tape for a period of 4 h. At the end of the 4 h exposure period, the residual test item was removed with cotton soaked in distilled water. The skin reactions of each rabbit were observed at 1, 24, 48 and 72 h post patch removal. Irritation was scored according to OECD 404. There were no signs of systemic adverse effect in any treated rabbits. Erythema was evident at 1 h in all rabbits which resolved by 24 h post patch removal in all rabbits. The mean dermal irritation scores at 24, 48 and 72 h post-patch removal for individual animals were 0.00, 0.00, 0.00 for erythema and 0.00, 0.00, 0.00 for oedema, respectively. Based on the results of this study, an indication of the classification for DABQUEL COMPLEX MnP is as follows: Globally Harmonized System of Classification and Labeling of Chemicals (GHS 2015): Not classified as a skin irritant

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

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:

January 15, 2016 - March 10, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well documented GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Exception to OECD guideline 405, for not testing weight of evidence analysis by the test facility.

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Sainath Agencies, Hyderabad, India
- Age at study initiation: 4 to 5 months
- Weight at study initiation: Minimum: 2.386, Maximum: 2.505
- Housing: individually in stainless steel wire meshed cages
- Diet (e.g. ad libitum): Teklad certified Global High Fiber Rabbit pellet Feed manufactured by Harlan, USA ad libitum
- Water (e.g. ad libitum): UV sterilised drinking water filtered through Kent Reverse Osmosis water filtration system ad libitum
- Acclimation period: 8 - 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 22 °C
- Humidity (%): 64 to 65%
- Air changes (per hr): Minimum 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 h artificial light and 12 h darkness, light hours being 06:00 - 18:00 h except light was kept ON at the time of the subcutaneous injection during night hours. Photoperiod was maintained through automatic timer.

Animal Identification
Each rabbit was serially numbered on the ear using a tattoo machine on day 1 of acclimatisation. Appropriate labels were attached to the cages indicating the study number, test item code, sex, dose, type of study, cage number and animal number.

Vehicle:

other: distilled water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): 1 % solution in distilled water

VEHICLE
- Amount(s) applied (volume or weight with unit): distilled water, test item solved in it
- Concentration (if solution): 1 %

Duration of treatment / exposure:

8 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with 0.9 % normal saline
- Time after start of exposure: 24 after treatment

SCORING SYSTEM: according to OECD 405

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

Conjunctival redness was evident at 1, 24 and 48 h in rabbit N° 1, which was resolved by 72 h and at 1 and 24 h in rabbit N° 2 and 3, which was resolved by 48 h post TIA. Conjunctival chemosis was evident at 1 and 24 h in rabbit N° 1, which was resolved by 48 h post TIA and at 1 h post TIA in rabbit N° 2 and 3, which was resolved by 24 h post TIA. The animal mean eye irritation scores of the 24, 48 and 72 h post-TIA observation were 0.00, 0.00, 0.00 for corneal opacity, 0.00, 0.00, 0.00 for iris effects, 0.67, 0.33, 0.33 for conjunctival redness and 0.33, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively (Table 1). Examination with fluorescein dye and cobalt blue filter at 24 h post TIA revealed no corneal epithelium damage in all rabbits.

Mean Eye Irritation Scores

The animal mean eye irritation scores observed for corneal opacity (0.00), iritis (0.00), conjunctival redness (0.33 to 0.67) and conjunctival chemosis (0.00 to 0.33) following grading at 24, 48 and 72 h post TIA.

Narrative Description of Eye Irritation

Treated Eye

At 1 h post-TIA, the treated eye of all three rabbits revealed conjunctival redness [some blood vessels definitely hyperaemic (injected); score of 1] and conjunctival chemosis [some swelling above normal (includes nictitating membranes); score of 1].

At 24 h post-TIA, the treated eye revealed conjunctival redness [some blood vessels definitely hyperaemic (injected) in all three rabbits; score of 1] and conjunctival chemosis [some swelling above normal (includes nictitating membraes) in rabbit N° 1, score of 1].

At 48 h post-TIA, the treated eye revealed conjunctival redness [some blood vessels definitely hyperaemic (injected) in rabbit N°1; score of 1) while rabbit N°2 and 3 recovered completely and appeared normal throughout the experimental period.

At 72 h post-TIA, the treated eye of rabbit N° 1 recovered completely and appeared normal.

Corneal opacity and iritis were not observed in any of the rabbits throughout the experimental period.

Examination with fluorescein dye and cobalt blue filter [corneal epithelium damage showing as green fluorescein staining] revealed no (area) corneal epithelium damage in all three rabbits at 24 h post TIA.

 

Control Eye

No abnormalities were detected in the control eye of each rabbit during the course of this study (Table 3). No disruption of corneal epithelium was observed during the examination with fluorescein dye and cobalt blue filter.

Clinical Observations other than Eye Irritation

Other than eye irritation, no signs of systemic toxicity including clinical observation and body weight were observed in the rabbits throughout the experimental period.

Interpretation of Results

Conjunctival redness was evident at 1, 24 and 48 h in rabbit N° 1 which was resolved by 72 h post-TIA and at 1 and 24 h in rabbit N° 2 and 3 which resolved by 48 h post TIA. Conjunctival chemosis was evident at 1 and 24 h in rabbit N° 1 which resolved by 48 h post-TIA. Conjunctival chemosis was evident at 1 and 24 h in rabbit N° 1 which resolved by 48 h post-TIA and at 1 h post-TIA in rabbit N° 2 and 3 which resolved by 24 h post-TIA. The individual animal mean eye irritation scores of the 24, 48 and 72 h post-TIA observation were 0.00, 0.00, 0.00 for corneal opacity, 0.00, 0.00, 0.00 for iris effects, 0.67, 0.33, 0.33 for conjunctival redness and 0.33, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively (TABLE 1). Examination with fluorescein dye and cobalt blue filter at 24 h post TIA revealed no corneal epithelium damage in all rabbits.

 

Conclusion   

Based on the results of this study, an indication of the classification for DABQUEL COMPLEX MnP is as follows:

Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015): Not classified as an eye irritant

Table 1: Mean Eye Irritation Scores

Sex: Female

Rabbit N°	Mean Score at 24, 48 and 72 Hours
	Opacity: Degree of Density	Iris Lesion	Conjunctivae
			Redness	Chemosis
1	0.00	0.00	0.67	0.33
2	0.00	0.00	0.33	0.00
3	0.00	0.00	0.33	0.00

Table 2: Clinical Observations (Non Ocular) and Body Weight (kg) of Individual Rabbit 

A. Clinical Observation (Non Ocular)                                                                         

Sex: Female

Rabbit N°	Observations made on Day
	0	1	2	3
1	1	1	1	1
2	1	1	1	1
3	1	1	1	1

Key:  0 = Day of treatment

          1 = Normal

B. Body Weight Record

Rabbit N°	Body Weight (kg)
	Initial (Day 0)	Termination (72 h)
1	2.386	2.412
2	2.505	2.613
3	2.456	2.591

Table 3 :Individual Scores of Eye Reactions Post Application 

Control Eye                                                                                                                   

Sex: Female

Rabbit N°	1				2				3
Site of Application	Left				Left				Left
Reaction	Hour				Hour				Hour
	1	24	48	72	1	24	48	72	1	24	48	72
Opacity: Degree of Density	0	0	0	0	0	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctivae (Redness)	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctivae (Chemosis)	0	0	0	0	0	0	0	0	0	0	0	0

Treated Eye

Rabbit N°	1				2				3
Site of Application	Right				Right				Right
Reaction	Hour				Hour				Hour
	1	24	48	72	1	24	48	72	1	24	48	72
Opacity: Degree of Density	0	0	0	0	0	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctivae (Redness)	1	1	1	0	1	1	0	0	1	1	0	0
Conjunctivae (Chemosis)	1	1	0	0	1	0	0	0	1	0	0	0

Table 4: Individual Corneal Observations 24 h Post Application using Fluorescein Dye Staining 

Control Eye                                                                                                                   

Sex: Female

Rabbit N°	Control Eye	Fluorescein Staining Response	Details of Corneal Damage Observed in Control Eye
1	Left	Negative	No corneal epithelium damage was observed.
2	Left	Negative	No corneal epithelium damage was observed.
3	Left	Negative	No corneal epithelium damage was observed.

Treated Eye

Rabbit N°	Treated Eye	Fluorescein Staining Response	Details of Corneal Damage Observed in Treated Eye
1	Right	Negative	No corneal epithelium damage was observed.
2	Right	Negative	No corneal epithelium damage was observed.
3	Right	Negative	No corneal epithelium damage was observed.

 Table 5:Details of Injections and Applications 

Rabbit N°	On Days	Administration of Systemic Analgesics (Subcutaneous Injection)		Administration of Topical Anaesthetic (1 to 2 drops) 0.5% Proparacaine Hydrochloride	Time of Test Item Application (0.1 mL)
		Buprenorphine Hydrochloride (0.01 mg/kg body weight)	Meloxicam (0.5 mg/kg body weight)
1	0	11:50 am & 8:51 pm	8:51 pm	12:45 pm	12:50 pm
	1	8:37 am and 8:52 pm	8:52 pm	-	-
	2	8:38 am and 8:48 pm	8:48 pm	-	-
	3	8:55 am	-	-	-
2 and 3	0	11:48 am to 11:49 am & 8:49 pm to 8:50 pm	8:49 pm to 8:50 pm	12:43 pm to 12:44 pm	12:48 pm to 12:49 pm
	1	8:56 am to 8:57 am and 8:43 pm to 8:44 pm	8:43 pm to  8:44 pm	-	-
	2	8:45 am to 8:46 pm	-	-	-

Key : - = Not applicable

Note: Day 0 = Day of treatment

Interpretation of results:

GHS criteria not met

Conclusions:

Examination with fluorescein dye and cobalt blue filter at 24 h post TIA revealed no corneal epithelium damage in all rabbits. Based on the results of this study, an indication of the classification for DABQUEL COMPLEX MnP is as follows:
Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015): Not classified as an eye irritant.

Executive summary:

In a GLP-compliant acute eye irritation study according to OECD TG 405, 3 adult male New Zealand White rabbits were given a single ocular application of 0.1 mL DABQUEL COMPLEX MnP in the right eye, while the contralateral eye remained untreated and served as the control. Initially, one rabbit was tested. Based on the results obtained at 24 h post Test Item Application (TIA), the irritation response was confirmed by testing two additional rabbits simultaneously. Observations were made at 1, 24, 48 and 72 h post TIA. General health status was also checked. Conjunctival effects were evident at 1 h, 24 h and 48 h which resolved by 72 h post TIA. The animal mean eye irritation scores of the 24, 48 and 72 h post TIA observation were 0.00, 0.00, 0.00 for corneal opacity, 0.00, 0.00, 0.00 for iris effects, 0.67, 0.33, 0.33 for conjunctival redness and 0.33, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively. Examination with fluorescein dye and cobalt blue filter at 24 h post TIA revealed no corneal epithelium damage in all rabbits. The control eye did not show any abnormal reaction during the study. Moreover, there were no signs of systemic toxicity in any animal observed. Based on the results of this study, an indication of the classification for DABQUEL COMPLEX MnP is as follows: Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015): Not classified as an eye irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

In a GLP-compliant acute dermal irritation study according to OECD TG 404, three adult male New Zealand White rabbits were dermally exposed to 500 mg DABQUEL COMPLEX MnP (pulverised and moistened with 0.5 mL distilled water), for 4 h (day 0), applied to approximately 6 cm² area of skin. Initially one rabbit was tested with a single patch applied evenly to the intact skin for a period of 4 h. Based on the observations at 24 h post patch removal, two additional rabbits were tested simultaneously to confirm the irritation response. The control skin site of each rabbit was untreated. The treated and the control sites were covered with a gauze patch that was secured at the margins by non-irritating tape for a period of 4 h. At the end of the 4 h exposure period, the residual test item was removed with cotton soaked in distilled water. The skin reactions of each rabbit were observed at 1, 24, 48 and 72 h post patch removal. Irritation was scored according to OECD 404. There were no signs of systemic adverse effect in any treated rabbits. Erythema was evident at 1 h in all rabbits which resolved by 24 h post patch removal in all rabbits. The mean dermal irritation scores at 24, 48 and 72 h post-patch removal for individual animals were 0.00, 0.00, 0.00 for erythema and 0.00, 0.00, 0.00 for oedema, respectively. Based on the results of this study, an indication of the classification for DABQUEL COMPLEX MnP is as follows: Globally Harmonized System of Classification and Labeling of Chemicals (GHS 2015): Not classified as a skin irritant

This test result is supported by the primary dermal irritation tests with 0.5 mL of 50% solution of gluconic acid (pH: 1.8) in 12 albino rabbits. It was demonstrated, that – as all the effects have cleared up after a 72 hours observation period – the test substance is not a dermal irritant (TNO, 1984).

The 2014 Cosmetic Ingredient Review Expert Panel acknowledged that the group of monosaccharides, disaccharides, and their related Ingredients, including calcium gluconate and gluconic acid, are safe for humans at concentrations as used in cosmetics. Based on the clinical experience of the Panel, there is little concern that these ingredients are irritants or sensitizers.

These results are also valid for manganese glucoheptonate, as this metal complex is composed of metal manganese and the glucoheptonate-residual. This residual is expected to act in the same manner as gluconic acid and its derivatives.

Eye irritation

In a GLP-compliant acute eye irritation study according to OECD TG 405, 3 adult male New Zealand White rabbits were given a single ocular application of 0.1 mL DABQUEL COMPLEX MnP in the right eye, while the contralateral eye remained untreated and served as the control. Initially one rabbit was tested. Based on the results obtained at 24 h post Test Item Application (TIA), the irritation response was confirmed by testing two additional rabbits simultaneously. Observations were made at 1, 24, 48 and 72 h post TIA. General health status was also checked. Conjunctival effects were evident at 1 h, 24 h and 48 h, which resolved by 72 h post TIA. The animal mean eye irritation scores of the 24, 48 and 72 h post TIA observation were 0.00, 0.00, 0.00 for corneal opacity, 0.00, 0.00, 0.00 for iris effects, 0.67, 0.33, 0.33 for conjunctival redness and 0.33, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively. Examination with fluorescein dye and cobalt blue filter at 24 h post TIA revealed no corneal epithelium damage in all rabbits. The control eye did not show any abnormal reaction during the study. Moreover, there were no signs of systemic toxicity in any animal observed. Based on the results of this study, an indication of the classification for DABQUEL COMPLEX MnP is as follows: Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015): Not classified as an eye irritant.

This test result is supported by the test results available for gluconic acid and its derivatives (SIDS, 2004 and CIR, 2014). The following observations were made: The ocular irritation potential of a 50% aq. solution of gluconic acid was evaluated in vitro in enucleated rabbit eyes. The test material was applied to four eyes and observed over a period of 4 h following application. Slight corneal swelling and slight permeability of the superficial epithelial cells were not considered to be of any toxicological significance.

Moreover, in vivo test results showed a 50% aq. solution of gluconic acid to be not irritating to rabbit eyes. A 50% solution of gluconic acid (pH 1.8; 0.1 ml) was instilled into the conjunctival sac of one eye in nine New Zealand white rabbits; the contralateral eye served as an untreated control. The eyes of three animals were rinsed after 2 sec, and of another three animals after 4 sec; the eyes of the remaining three animals were not rinsed. The eyes were examined for irritation 1, 24, 48, and 72 h and 7 days after instillation. Slight redness and conjunctival swelling were observed initially; however, no signs of irritation were observed after 72 h.

These results are also valid for manganese glucoheptonate, as this metal complex is composed of metal manganese and the glucoheptonate-residual. This residual is expected to act in the same manner as gluconic acid and its derivatives.

Justification for classification or non-classification

Based on the results of the available studies manganese glucoheptonate does not meet criteria for classification and labelling as a skin or eye irritant in accordance with European Regulation (EC) No 1272/2008.