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Description of key information

Enzymes are well documented not to be skin sensitisers in humans. Therefore, it can be concluded that enzymes should not be classified as skin sensitisers according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

From occupational data it is well known that active enzymes regardless of the catalytic activities are potential respiratory sensitisers. However, decades of expericence have shown that enzymes can be used safely by ensuring that exposure is limited, supported by DMEL for workers and consumers.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vitro
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
Skin sensitisation is referred to as allergic contact dermatitis (ACD) in a clinical setting. This is a cell-mediated type IV delayed hypersensitivity. The cellular mechanisms involved with ACD have been reviewed [1, 2]. To behave as a skin sensitiser, a substance must first penetrate the stratum corneum, partition into the epidermis and react there with proteins, probably on the surface of the Langerhans cells, to form a hapten-carrier conjugate. The skin sensitisation potential of enzymes has been reviewed in several publications indicating that enzymes should not be considered skin sensitisers [3-7; 13, 14]. In addition, there is an unequivocal statement from AMFEP (www.amfep.org) on this topic indicating that enzymes do not have skin sensitising potential. The lack of skin sensitising potential is substantiated by evidence from robust human experimental data and extensive in-use human studies performed with detergents containing enzymes [8-12; 14 -19]. Together, these studies confirm that the presence of enzymes in the detergents does not result in ACD, including those conducted with atopic individuals.
After review of the available evidence, it can be concluded that enzymes should not be classified as skin sensitisers according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.
This conclusion is based on the following considerations:
1. The results of predictive testing in humans demonstrate that enzymes do not have significant skin sensitisation potential.
2. In a clinical setting, enzymes have only very rarely been suggested as a possible cause of allergic contact dermatitis. Even in these few cases, a causal relationship has never been proven. More commonly, clinical studies have demonstrated that enzymes are not a cause of ACD.
3. ACD has never been reported where there has been extensive occupational enzyme exposure in the detergent enzyme industries that in the past has led to respiratory sensitisation and/or irritant dermatitis.
4. A few cases of contact dermatitis had occurred in occupational settings in response to irritating enzyme preparations (e.g. proteases), but this is a non-immunologic phenomenon (also known as irritant contact dermatitis) unrelated to allergic contact hypersensitivity, which is a cell-mediated delayed type hypersensitivity.
5. Contact urticaria has been reported in occupational settings but this is also a non-immunologic event or antibody mediated type I hypersensitivity; Contact urticaria (also known as protein contact dermatitis) is unrelated to allergic contact hypersensitivity, which is a cell-mediated delayed type hypersensitivity.
6. Over a 45-year period, billions of consumers have had skin exposure to enzymes but there is no evidence that this exposure has given rise to skin sensitization.

References:
1) Kimber I. (1994). Cytokines and regulation of allergic sensitization to chemicals, Toxicology, 93(1):1-11.
2) Scheper RJ, Mary B, von Blomberg E. (1992). Cellular mechanisms in allergic contact dermatitis. Textbook of Contact Dermatitis 11-27.
3) Association Internationale de la Savonnerie et de la Detergence (AISE)/AMFEP. (1995). Enzymes: Lack of skin sensitisation potential.
4) Basketter DA, English JS, Wakelin SH, White IR (2008). Enzymes, detergents and skin: facts and fantasies. Br. J. Dermatol., 158 (6):1177-1181.
5) Basketter D, Berg N, Broekhuizen C, Fieldsend M, Kirkwood S, Kluin C, Mathieu S, Rodriguez C (2012a). Enzymes in Cleaning Products: An Overview of Toxicological Properties and Risk Assessment/Management. Regul. Toxicol. Pharmacol., 64(1):117-123.
6) HERA. Human and environmental risk assessment on ingredients of household cleaning products -alpha-amylases, cellulases and lipases. Edition 1.0. November 2005.
7) HERA. Human and environmental risk assessment on ingredients of household cleaning products - Subtilisins (Proteases). Edition 2.0. February 2007.
8) Bannan EA, Griffith JF, Nusair TL, Sauers JL, Jackson EM. (1992). Skin testing of laundered fabrics in the dermal safety assessment of enzyme containing detergents. J. of Toxicol. – Cutan. Ocul. Toxicol. 11(4):327-339.
9) Griffith JF, Weaver JE, Whitehouse HS, Poole RL, Newmann EA, Nixon GA. (1969). Safety Evaluation of Enzyme Detergents Oral and Cutaneous Toxicity, Irritancy and Skin Sensitization Studies. Food Cosmet. Toxicol. 7(6):581-593.
10) Rodriguez C, Calvin G, Lally C, LaChapelle JM. (1994). Skin effects associated with wearing fabrics washed with commercial laundry detergents. J. of Toxicol. – Cutan. Ocul. Toxicol. 13(1):39-45.
11) Cormier EM, Sarlo K, Scott LA, MacKenzie DP, Payne NS, Carr GJ, Smith LA, Cua-Lim F, Bunag FC, Vasunia K. (2004) Lack of type 1 sensitization to laundry detergent enzymes among consumers in the Philippines: results of a 2-year study in atopic subjects. Ann. Allergy Asthma Immunol. 92(5):549-557.
12) White IR, Lewis J, el Alami A. (1985). Possible adverse reactions to an enzyme-containing washing powder. Contact Dermatitis, 13(3) 175-179.
13) Basketter D, Berg N, Kruszewski FH, Sarlo K, Concoby B (2012b). The Toxicology and Immunology of Detergent Enzymes. J. Immunotoxicol., 9(3):320-326.
14) Andersen PH, Bindslev-Jensen C, Mosbech H, Zachariae H, Andersen KE. (1998). Skin symptoms in patients with atopic dermatitis using enzyme-containing detergents. A placebo-controlled study. Acta Derm. Venereol., 78(1):60-62.
15) Belsito DV, Fransway AF, Fowler JF Jr, Sherertz EF, Maibach HI, Mark JG Jr, Mathias CG, Rietschel RL, Storrs FJ, Nethercott JR. (2002). Allergic contact dermatitis to detergents: a multicenter study to assess prevalence. J Am Acad Dermatol. 46(2):200-206.
16) Lee MY, Park KS, Hayashi C, Lim HH, Lee KH, Kwak I, Laurie RD. (2002). Effects of repeated short-term skin contact with proteolytic enzymes. Contact Dermatitis, 46(2):75-80.
17) Pepys J, Wells ID, D'Souza MF, Greenberg M. (1973). Clinical and Immunological Responses to Enzymes of Bacillus Subtilis in Factory Workers and Consumers. Clin. Allergy, 3(2):143-160.
18) Peters G, Johnson GQ, Golembiewski A. (2001). Safe use of detergent enzymes in the workplace. Appl. Occup. Environ. Hyg., 16(3):389-395.
19) Zachariae H, Thomsen K, Rasmussen OG. (1973). Occupational enzyme dermatitis. Results of patch testing with Alcalase. Acta Derm. Venereol. 53(2):145-148
Endpoint:
skin sensitisation: in vivo (LLNA)
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
Skin sensitisation is referred to as allergic contact dermatitis (ACD) in a clinical setting. This is a cell-mediated type IV delayed hypersensitivity. The cellular mechanisms involved with ACD have been reviewed [1, 2]. To behave as a skin sensitiser, a substance must first penetrate the stratum corneum, partition into the epidermis and react there with proteins, probably on the surface of the Langerhans cells, to form a hapten-carrier conjugate. The skin sensitisation potential of enzymes has been reviewed in several publications indicating that enzymes should not be considered skin sensitisers [3-7; 13, 14]. In addition, there is an unequivocal statement from AMFEP (www.amfep.org) on this topic indicating that enzymes do not have skin sensitising potential. The lack of skin sensitising potential is substantiated by evidence from robust human experimental data and extensive in-use human studies performed with detergents containing enzymes [8-12; 14 -19]. Together, these studies confirm that the presence of enzymes in the detergents does not result in ACD, including those conducted with atopic individuals.
After review of the available evidence, it can be concluded that enzymes should not be classified as skin sensitisers according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.
This conclusion is based on the following considerations:
1. The results of predictive testing in humans demonstrate that enzymes do not have significant skin sensitisation potential.
2. In a clinical setting, enzymes have only very rarely been suggested as a possible cause of allergic contact dermatitis. Even in these few cases, a causal relationship has never been proven. More commonly, clinical studies have demonstrated that enzymes are not a cause of ACD.
3. ACD has never been reported where there has been extensive occupational enzyme exposure in the detergent enzyme industries that in the past has led to respiratory sensitisation and/or irritant dermatitis.
4. A few cases of contact dermatitis had occurred in occupational settings in response to irritating enzyme preparations (e.g. proteases), but this is a non-immunologic phenomenon (also known as irritant contact dermatitis) unrelated to allergic contact hypersensitivity, which is a cell-mediated delayed type hypersensitivity.
5. Contact urticaria has been reported in occupational settings but this is also a non-immunologic event or antibody mediated type I hypersensitivity; Contact urticaria (also known as protein contact dermatitis) is unrelated to allergic contact hypersensitivity, which is a cell-mediated delayed type hypersensitivity.
6. Over a 45-year period, billions of consumers have had skin exposure to enzymes but there is no evidence that this exposure has given rise to skin sensitization.

References:
1) Kimber I. (1994). Cytokines and regulation of allergic sensitization to chemicals, Toxicology, 93(1):1-11.
2) Scheper RJ, Mary B, von Blomberg E. (1992). Cellular mechanisms in allergic contact dermatitis. Textbook of Contact Dermatitis 11-27.
3) Association Internationale de la Savonnerie et de la Detergence (AISE)/AMFEP. (1995). Enzymes: Lack of skin sensitisation potential.
4) Basketter DA, English JS, Wakelin SH, White IR (2008). Enzymes, detergents and skin: facts and fantasies. Br. J. Dermatol., 158 (6):1177-1181.
5) Basketter D, Berg N, Broekhuizen C, Fieldsend M, Kirkwood S, Kluin C, Mathieu S, Rodriguez C (2012a). Enzymes in Cleaning Products: An Overview of Toxicological Properties and Risk Assessment/Management. Regul. Toxicol. Pharmacol., 64(1):117-123.
6) HERA. Human and environmental risk assessment on ingredients of household cleaning products -alpha-amylases, cellulases and lipases. Edition 1.0. November 2005.
7) HERA. Human and environmental risk assessment on ingredients of household cleaning products - Subtilisins (Proteases). Edition 2.0. February 2007.
8) Bannan EA, Griffith JF, Nusair TL, Sauers JL, Jackson EM. (1992). Skin testing of laundered fabrics in the dermal safety assessment of enzyme containing detergents. J. of Toxicol. – Cutan. Ocul. Toxicol. 11(4):327-339.
9) Griffith JF, Weaver JE, Whitehouse HS, Poole RL, Newmann EA, Nixon GA. (1969). Safety Evaluation of Enzyme Detergents Oral and Cutaneous Toxicity, Irritancy and Skin Sensitization Studies. Food Cosmet. Toxicol. 7(6):581-593.
10) Rodriguez C, Calvin G, Lally C, LaChapelle JM. (1994). Skin effects associated with wearing fabrics washed with commercial laundry detergents. J. of Toxicol. – Cutan. Ocul. Toxicol. 13(1):39-45.
11) Cormier EM, Sarlo K, Scott LA, MacKenzie DP, Payne NS, Carr GJ, Smith LA, Cua-Lim F, Bunag FC, Vasunia K. (2004) Lack of type 1 sensitization to laundry detergent enzymes among consumers in the Philippines: results of a 2-year study in atopic subjects. Ann. Allergy Asthma Immunol. 92(5):549-557.
12) White IR, Lewis J, el Alami A. (1985). Possible adverse reactions to an enzyme-containing washing powder. Contact Dermatitis, 13(3) 175-179.
13) Basketter D, Berg N, Kruszewski FH, Sarlo K, Concoby B (2012b). The Toxicology and Immunology of Detergent Enzymes. J. Immunotoxicol., 9(3):320-326.
14) Andersen PH, Bindslev-Jensen C, Mosbech H, Zachariae H, Andersen KE. (1998). Skin symptoms in patients with atopic dermatitis using enzyme-containing detergents. A placebo-controlled study. Acta Derm. Venereol., 78(1):60-62.
15) Belsito DV, Fransway AF, Fowler JF Jr, Sherertz EF, Maibach HI, Mark JG Jr, Mathias CG, Rietschel RL, Storrs FJ, Nethercott JR. (2002). Allergic contact dermatitis to detergents: a multicenter study to assess prevalence. J Am Acad Dermatol. 46(2):200-206.
16) Lee MY, Park KS, Hayashi C, Lim HH, Lee KH, Kwak I, Laurie RD. (2002). Effects of repeated short-term skin contact with proteolytic enzymes. Contact Dermatitis, 46(2):75-80.
17) Pepys J, Wells ID, D'Souza MF, Greenberg M. (1973). Clinical and Immunological Responses to Enzymes of Bacillus Subtilis in Factory Workers and Consumers. Clin. Allergy, 3(2):143-160.
18) Peters G, Johnson GQ, Golembiewski A. (2001). Safe use of detergent enzymes in the workplace. Appl. Occup. Environ. Hyg., 16(3):389-395.
19) Zachariae H, Thomsen K, Rasmussen OG. (1973). Occupational enzyme dermatitis. Results of patch testing with Alcalase. Acta Derm. Venereol. 53(2):145-148
Endpoint conclusion
Endpoint conclusion:
no study available

Respiratory sensitisation

Link to relevant study records
Reference
Endpoint:
respiratory sensitisation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

From occupational data it is well known that active enzymes regardless of the catalytic activities are potential respiratory sensitisers. All enzymes must therefore be classified as respiratory sensitisers, “H334: Hazard Category 1: May cause allergy or asthma symptoms or breathing difficulties if inhaled” in accordance with the CLP Regulation.

For enzyme protein respiratory allergens, a DMEL for workers and consumers has been summarized and discussed in a recent publication (Ref. 1). The conclusion is drawn from a thorough review of existing occupational and consumer data on exposure by inhalation from the involved industrial partners in combination with medical data. As no valid animal models exist to test and rank respiratory sensitisers, the human surveillance data is the core of such evaluation. Any sub-categorization based on relative potency is not feasible (Ref. 2).

REFERENCES

1.Basketter DA, Broekhuizen C, Fieldsend M, Kirkwood S, Mascarenhas R, Maurer K, Pedersen C, Rodriguez C, Schiff HE: Defining occupational and consumer exposure limits for enzyme protein respiratory allergens under REACH. Toxicology. 268:165-170, 2010.

2. Basketter D.A., Kimber I. (2011) Assessing the potency of respiratory allergens: Uncertainties and challenges. Regul. Toxicol. Pharmacol., 61, 365-372.


Migrated from Short description of key information:
From occupational data it is well known that active enzymes regardless of the catalytic activities are potential respiratory sensitisers.

Justification for classification or non-classification

Alpha-amylase should not be classified as a skin sensitiser.

Alpha-amylase is classified as a respiratory sensitiser.