Magnesium 2,3,4,5,6,7-Hexahydroxyheptanoate

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: due to read-across

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)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: SA Dabeer, 4140Z0925
- Expiration date of the lot/batch: September 25, 2018
- Purity test date: 2015-11-12

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature; keep container tightly closed in a dry cool and well ventilated place.
- Stability under test conditions: Test item characterization (identity, purity, strength and composition), stability, method of synthesis and location of documents for the synthesis is the responsibility of the Sponsor.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation, India
- Age at study initiation: 16 to 18 weeks
- Weight at study initiation: Minimum: 2.613, Maximum: 2.824
- 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: 7 to 9 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 23 °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): 0.5 mL

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
- 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:

mean

Time point:

24 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

mean

Time point:

48 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

mean

Time point:

72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

48 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

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. No sign of skin irritation was observed throughout the experimental period.

Other effects:

There were no signs of systemic adverse effect in any treated rabbits.

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 removal, the treated skin site recovered completely and appeared normal in all three rabbits throughout the experimental period (TABLE 1).

The control skin sites of all three rabbits were normal with no erythema and no oedema 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 (APPENDIX 1).

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 CaL is a non-irritant under the described experimental conditions (TABLE 2).

Acute Dermal Irritation Study of DABQUEL COMPLEX CaL in Rabbits

TABLE1: 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: 0.5 mL DABQUEL COMPLEX CaL (undiluted)

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

Note: Refer section 2.12 for Irritation Scores

Acute Dermal Irritation Study of DABQUEL COMPLEX CaL in Rabbits

APPENDIX1: 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	2.824	2.887
2	1	1	1	1	2.613	2.692
3	1	1	1	1	2.739	2.801

Key:               0 = Day of dermal application

Clinical Sign:  1 = Normal

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

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

Executive summary:

In an acute dermal irritation study, three adult male New Zealand White rabbits were dermally exposed to 0.5 mL DABQUEL COMPLEX CaL (undiluted), 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. No sign of skin irritation was observed throughout the experimental period.

Based on the results of this study, an indication of the classification for DABQUEL COMPLEX CaL 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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: due to read-across

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)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: SA Dabeer, 4140Z0925
- Expiration date of the lot/batch: September 25, 2018
- Purity test date: 2015-11-12

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature; keep container tightly closed in a dry cool and well ventilated place.
- Stability under test conditions: Test item characterization (identity, purity, strength and composition), stability, method of synthesis and location of documents for the synthesis is the responsibility of the Sponsor.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation
- Age at study initiation: 16 to 17 weeks
- Weight at study initiation: Minimum: 2.561, Maximum: 2.741
- 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 23 °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:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Conjunctival redness was evident at 1 h in all three rabbits which resolved by 24 h post TIA. Conjunctival chemosis was evident at 1 h in rabbit N° 1 which was 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, 0.00, 0.00, 0.00 for conjunctival redness and 0.00, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively.

Mean Eye Irritation Scores

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

Narrative Description of Eye Irritation

Treated Eye

At 1 h post TIA, the treated eyes 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 membranes) in rabbit N° 1; score of 1].

At 24 h post TIA, the treated eyes of all three rabbits recovered completely and appeared normal 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.

Corneal opacity and Iritis were not observed in all three rabbits throughout the experimental period.

Control Eye

No abnormalities were detected in the control eye of each rabbit during the course of this study (Table 3). No corneal epithelium damage 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 h in all three rabbits which resolved by 24 h post TIA. Conjunctival chemosis was evident at 1 h in rabbit N° 1 which was 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, 0.00, 0.00, 0.00 for conjunctival redness and 0.00, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively.

Conclusion   

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

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

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits 

TABLE1: 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	0.00	0.00
2	0.00	0.00	0.00	0.00
3	0.00	0.00	0.00	0.00

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits 

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	2.561	2.598
2	2.720	2.939
3	2.741	2.813

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits 

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	0	0	0	1	0	0	0	1	0	0	0
Conjunctivae (Chemosis)	1	0	0	0	0	0	0	0	0	0	0	0

Acute Eye Irritation Study ofDABQUEL COMPLEX CaLin Rabbits

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.

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits

 Table 5:Details of Injections and Applications 

Rabbit N°	On Day	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	11:30 am and 8:50 pm	8:50 pm	12:25 pm	12:30 pm
	1	8:42 am	-	-	-
2 and 3	0	11:47 am to 11:48 am and 8:49 pm to 8:50 pm	8:49 pm to 8:50 pm	12:42 pm to 12:43 pm	12:47 pm to 12:48 pm
	1	8:53 am to 8:54 am	-	-	-

Key : - = Not applicable

Note: Day 0 = Day of treatment

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

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

Executive summary:

In an acute eye irritation study, 3 adult male New Zealand White rabbits were given a single ocular application of 0.1 mL DABQUEL COMPLEX CaL in the right eye of the rabbit 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 in all three rabbits which 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, 0.00, 0.00, 0.00 for conjunctival redness and 0.00, 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 CaL 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)

Additional information

There are no skin and eye irritation studies available for Magnesium glucoheptonate. Therefore data on irritation potential of Calcium glucoheptonate have been taken into account to assess irritating properties of Magnesium glucoheptonate. Additionally, data on structurally similar gluconic acid, its derivatives and organic and inorganic magnesium compounds have been used to draw a conclusion on irritating properties of Magnesium glucoheptonate.

Skin irritation

Skin irritation of Calcium glucoheptonate

The skin irritation potential of Calcium glucoheptonate (CAS 1821694-25-0) was tested in vivo in rabbits (Patel, 2016a, OECD 404). For this acute dermal irritation study, three adult male New Zealand White rabbits were dermally exposed to 0.5 mL DABQUEL COMPLEX CaL (undiluted, Calcium complexed content 6.10 %), 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. No sign of skin irritation was observed throughout the experimental period.

Gluconic acid and its derivatives

In addition to this, primary dermal irritation tests with 0.5 ml of the 50% solution of gluconic acid (pH: 1.8) in 12 albino rabbits 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, cited in OECD SIDS, 2004).

Moreover, 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.

Inorganic magnesium compounds

Magnesium sulfate

A skin irritation study (mice) on magnesium sulfate (in propylene glycol) was performed as a preliminary study for a skin sensitisation study (CIR, 2014a). Two young adult mice were used, and the ears were treated with anhydrous magnesium sulfate (50 %) on 3 consecutive days. The test sites were evaluated for irritation at approximately 3 h to 4 h after the last application. The test system, procedures, and techniques used in the skin irritation study were identical to those used during days 1 to 3 in the skin sensitisation study. Skin irritation of the ears was not observed in any of the animals tested with 50% anhydrous magnesium sulfate in the skin irritation study and in the following skin sensitisation study. Additionally, there was no evidence of macroscopic abnormalities of the surrounding area. Body weights and body weight gain reported for test animals remained in the same range when compared to controls. The slight body weight loss reported for some animals was not considered toxicologically significant. There were no symptoms of systemic toxicity, and none of the animals died.

Additionally, the 2014 Cosmetic Ingredient Review Expert Panel (CIR, 2014a) aknowledged that Magnesium sulfate "is being used in cosmetics up to 25 % (in paste masks - rinse-off products) and 11% (in hair sprays - leave-on products)". "Cosmetic products containing magnesium sulfate may be applied to the skin and hair, or, incidentally, may come in contact with the eyes and mucous membranes. Products containing these ingredients may be applied as frequently as several times per day and may come in contact with the skin or hair for variable periods following application. Daily or occasional use may extend over many years". In dilutions, i.e for baths use, the upper concentration level can be 49 %.

Magnesium chloride

The efficacy of bathing atopic subjects in a salt rich in magnesium chloride from deep layers of the Dead Sea (Mavena® Dermaline Mg Dead Sea salt, Mavena AG, Belp, Switzerland) was examined (Proksch et al., 2005). The Dead Sea salt has a high magnesium content of approximately 120 g/kg. Volunteers with atopic dry skin submerged one forearm for 15 min in a bath solution containing 5% Dead Sea salt. The second arm was submerged in tap water as control. Before the study and at weeks 1–6, transepidermal water loss (TEWL), skin hydration, skin roughness, and skin redness were determined.

There was one subgroup with a normal and one subgroup with an elevated TEWL before the study. Bathing in the Dead Sea salt solution significantly improved skin barrier function compared with the tap water-treated control forearm in the subgroup with elevated basal TEWL. Skin hydration was enhanced on the forearm treated with the Dead Sea salt in each group, which means the treatment moisturized the skin. Skin roughness and redness of the skin as a marker for inflammation were significantly reduced after bathing in the salt solution. This demonstrates that bathing in the salt solution was well tolerated, improved skin barrier function, enhanced stratum corneum hydration, and reduced skin roughness and inflammation. According to the authors, the favorable effects of bathing in the Dead Sea salt solution are most likely related to the high magnesium content. Magnesium salts are known to bind water, influence epidermal proliferation and differentiation, and enhance permeability barrier repair.

Magnesium containing minerals

Hectorite, Magnesium Aluminum Silicate and Sodium Magnesium Silicate were not irritating in 24-hour dermal irritating studies under occlusive conditions in rabbits and guinea pigs except one study in which Magnesium Aluminium Silicate induced primary irritation index of 0.1 (CTFA, 1970a, cited in CIR, 2003). The substance was classified as a weak primary skin irritant (CTFA 1970a, CIR, 2003).

Organic Magnesium compounds

Magnesium Ascorbyl Phosphate (7.5, 15 and 30 mg/kg bw) was not irritating in a cream applied to skin of guinea pigs for 10 days (CIR, 2005). Additionally, Magnesium Ascorbyl Phosphate functions as an antioxidant in cosmetics and was reported being used in 37 formulations over a wide concentration range (0.001% to 3%). Magnesium stearate (100 %) was not irritating to rabbit skin under oclussive conditions during 4 and 24 hours (CIR, 1982). Magnesium stearate is used in a considerable number of cosmetic formulations up to 25 % (CIR, 1982).

Conclusion

Taken together, the structurally similar analogue Calcium glucoheptonbate was not irritating to skin in the in vivo study according to OECD 404 (Patel, 2016a). Most of organic and inorganic magnesium compounds are used in a variety of cosmetic formulation in considerable amounts. Magnesium compounds as used in cosmetics are mostly not irritating in in vivo skin irritation animal studies and in humans. Furthermore, Magnesium salts improve skin barrier fuction, enhance skin hydration and reduced inflammation in patients with atopic dry skin (Proksch et al., 2005). Based on these data, Magnesium glucoheptonate is considered as not irritating to skin.

Eye irritation

Eye irritation of Calcium glucoheptonate

The eye irritation potential of Calcium glucoheptonate (CAS 1821694-25-0) was tested in vivo in rabbits (Patel, 2016b, OECD 405). For this acute eye irritation study, 3 adult male New Zealand White rabbits were given a single ocular application of 0.1 mL DABQUEL COMPLEX CaL (undiluted, Calcium complexed content : 6.10%) in the right eye of the rabbit 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 in all three rabbits which 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, 0.00, 0.00, 0.00 for conjunctival redness and 0.00, 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.

Gluconic acid and its derivatives

Eye irritation potential of structurally similar analogue gluconic acid and its derivatives was evaluated by the 2014 Cosmetic Ingredient Review Expert Panel (CIR, 2014). The results of the following studies originates also from the OECD SIDS report (2004) on gluconate category.

In an in vitro study, the ocular irritation potential of a 50% aq. solution of gluconic acid was evaluated in vitro in enucleated rabbit eyes (TNO, 1984; cited in OECD SIDS, 2004). 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.

In an in vivo study, a 50% aq. solution of gluconic acid was 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.

The Expert Panel concluded that monosaccharides, disaccharides, and their related Ingredients, including calcium gluconate and gluconic acid, are safe for humans at concentrations as used in cosmetics (CIR, 2014). The substances in this category are not irritating to eyes. The same conclusion is done in the OECD SIDS report for gluconates and the derivatives: "These compounds are neither irritant to the eye or the skin nor show sensitizing properties".

Inorganic magnesium compounds

Magnesium sulfate

The 2014 Cosmetic Ingredient Review Expert Panel (CIR, 2014a) aknowledged that Magnesium sulfate "is being used in cosmetics up to 25 % (in paste masks - rinse-off products) and 11% (in hair sprays - leave-on products)". "Cosmetic products containing magnesium sulfate may be applied to the skin and hair, or, incidentally, may come in contact with the eyes and mucous membranes. Products containing these ingredients may be applied as frequently as several times per day and may come in contact with the skin or hair for variable periods following application. Daily or occasional use may extend over many years". In dilutions, i.e for baths use, the upper concentration level can be 49 %. The upper concentration of Magnesium sulfate in cosmetic products contacting eye area is set to 2.1 %. Magnesium sulfate is considered as not irritating to eyes as used in cosmetics.

Magnesium containing minerals

Magnesium Aluminum Silicate and Sodium Magnesium Silicate caused minimal eye irritation in a Draize eye irritation test. In a primary eye irritation study in rabbits, Hectorite was moderately irritating without washing and practically nonirritating to the eye with a washout (CIR, 2003).

Organic Magnesium compounds

Magnesium Ascorbyl Phosphate is used in cosmetics (eye lotion) at concentrations of 0.05 - 3 % (CIR, 2005). Magnesium aspartate is well tolerated in humans at concentrations used in eye creams (0.005 -0.05 %) (CIR, 2013). Undiluted Magnesium stearate was not irritating to rabbits eyes. Magnesium stearate is used in eye liner and eye shadow formulations up to 25 % (CIR, 1982).

Conclusion

In conclusion, the structurally similar analogue Calcium glucoheptonbate was not irritating to eyes in the in vivo study according to OECD 405 (Patel, 2016b). Most of organic and inorganic magnesium compounds are used in a variety of cosmetic formulation contacting eyes (eye lotion, eye shadow, eye pencil) at considerable amounts (up to 25 %). Based on these data, Magnesium glucoheptonate is considered as not irritating to eyes.

Justification for selection of skin irritation / corrosion endpoint:
GLP guideline study conducted with a structurally similar analogue Calcium glucoheptonate.

Justification for selection of eye irritation endpoint:
GLP guideline study conducted with a structurally similar analogue Calcium glucoheptonate.

Justification for classification or non-classification

Based on the results of the in vivo irritation studies with Calcium glucoheptonate (CAS 1821694 -25 -0) and weight-of-evidence on non-irritancy of magnesium containing compounds, Magnesium glucoheptonate (CAS 1821694 -25 -0) is in accordance with European Regulation (EC) No. 1272/2008, does not meet criteria for classification and labelling as a skin or eye irritant.