Registration Dossier

Administrative data

Workers - Hazard via inhalation route

Systemic effects

Long term exposure
Hazard assessment conclusion:
no hazard identified
Most sensitive endpoint:
sensitisation (respiratory tract)
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
DMEL (Derived Minimum Effect Level)
Value:
60 ng/m³
Most sensitive endpoint:
sensitisation (respiratory tract)
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

Workers - Hazard via dermal route

Systemic effects

Long term exposure
Hazard assessment conclusion:
no hazard identified
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
low hazard (no threshold derived)
Most sensitive endpoint:
skin irritation/corrosion
Acute/short term exposure
Hazard assessment conclusion:
low hazard (no threshold derived)
Most sensitive endpoint:
skin irritation/corrosion

Workers - Hazard for the eyes

Local effects

Hazard assessment conclusion:
low hazard (no threshold derived)

Additional information - workers

Worker DMEL, acute short-term as well as long-term inhalation exposure:
Potential occupational exposure to levels of enzyme, which is toxicologically relevant, is unrealistic due to the stringent work practices and adherence to the voluntary Occupational Exposure Guidelines at or below the established ACGIH (www.acgih.org) exposure limit value of 60 ng/m3, based on pure enzyme protein for the benchmark enzyme subtilisin. This was established due to the endpoint of concern, which is risk of sensitisation by inhalation. Worker safety is assured through current proper work practices, engineering controls, and if needed of personal protective equipment. The industry has taken measures to minimize occupational exposure. Worker DMEL has been discussed and concluded by the involved industry in recent publications (Basketter et al. 2010, 2012a, 2012b) and a limit of 60 ng/m3, expressed in pure enzyme protein, was suggested (Basketter et al., 2010) in line with the established ACGIH threshold limit value.

References:
-Basketter DA, Broekhuizen C, Fieldsend M, Kirkwood S, Mascarenhas R, Maurer K, Pedersen C, Rodriguez C, Schiff HE (2010). Defining occupational and consumer exposure limits for enzyme protein respiratory allergens under REACH. Toxicology, 268(3):165-170.
-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.
-Basketter D, Berg N, Kruszewski FH, Sarlo K, Concoby B (2012b). The Toxicology and Immunology of Detergent Enzymes. J. Immunotoxicol., 9(3):320-326.

Worker DNEL, acute short-term as well as long-term dermal exposure:
The physico-chemical properties of a compound are decisive for the potential percutaneous penetration, in particular, factors like ionization, molecular size and lipophilicity. In general, non-ionized molecules easily penetrate the skin, with small molecules penetrating more easily than large molecules. Lipophilicity also facilitates penetration. Investigations of percutaneous absorption of peptides, proteins and other molecules of large size revealed that percutaneous absorption of proteins is extremely low and of no toxicological relevance (Basketter et al. 2008, Smith Pease et al. 2002, Basketter et al. 2012a, 2012b). This is further supported by the physico-chemical data of enzymes. They are proteins with molecular weight above 13,000 D (http://www.brenda-enzymes.info), they have a low logPow value (<0, i.e. low lipophilicity), indicating that they have no bioaccumulation potential and can be anticipated to be readily biodegraded. Thus, systemic exposure following enzyme exposure at occupational exposure levels is without toxicological significance.

References:

-Basketter DA, English JS, Wakelin SH, White IR(2008). Enzymes, detergents and skin: facts and fantasies. Br. J. Dermatol., 158(6):1177-1181.
-Smith Pease CK, White IR, Basketter DA(2002). Skin as a route of exposure to protein allergens. Clin. Exp. Dermatol., 27(4):296-300.
-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.
-Basketter D, Berg N, Kruszewski FH, Sarlo K, Concoby B (2012b). The Toxicology and Immunology of Detergent Enzymes. J. Immunotoxicol., 9(3):320-326.

General Population - Hazard via inhalation route

Systemic effects

Long term exposure
Hazard assessment conclusion:
no hazard identified
Most sensitive endpoint:
sensitisation (respiratory tract)
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
DMEL (Derived Minimum Effect Level)
Value:
15 ng/m³
Most sensitive endpoint:
sensitisation (respiratory tract)
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

General Population - Hazard via dermal route

Systemic effects

Long term exposure
Hazard assessment conclusion:
no hazard identified
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
low hazard (no threshold derived)
Most sensitive endpoint:
skin irritation/corrosion
Acute/short term exposure
Hazard assessment conclusion:
low hazard (no threshold derived)
Most sensitive endpoint:
skin irritation/corrosion

General Population - Hazard via oral route

Systemic effects

Long term exposure
Hazard assessment conclusion:
no hazard identified
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

General Population - Hazard for the eyes

Local effects

Hazard assessment conclusion:
low hazard (no threshold derived)

Additional information - General Population

Consumer DMEL, acute short-term as well as long-term inhalation exposure:
Industry has documented that respiratory irritation or toxicity due to enzyme preparations is a very rare phenomenon, which will not occur at the low concentrations of enzymes found in consumer products (e.g. detergents). The risk to consumers is considered very low and regarded as toxicologically insignificant (Basketter et al. 2010, 2012a, 2012b). This is supported by the positive safety outcome of a clinical study of the highest reported consumer exposure level, 15 ng/m3, with spot cleaning by spray (Weeks et al. 2011, SDA 2005). Consumer DMEL has been discussed among the enzyme allergy specialists from enzyme and detergent manufacturers and it was concluded by the involved industry partners in a recent publication that the limit of 15 ng/m3 (Basketter et al. 2010).

References:

-Basketter DA, Broekhuizen C, Fieldsend M, Kirkwood S, Mascarenhas R, Maurer K, Pedersen C, Rodriguez C, Schiff HE (2010). Defining occupational and consumer exposure limits for enzyme protein respiratory allergens under REACH. Toxicology, 268(3):165-170.
-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.
-Basketter D, Berg N, Kruszewski FH, Sarlo K, Concoby B (2012b). The Toxicology and Immunology of Detergent Enzymes. J. Immunotoxicol., 9(3):320-326.

-Weeks JA, Harper RA, Simon RA, Burdick JD (2011). Assessment of sensitization risk of a laundry pre-spotter containing protease. Cutan. Ocul. Toxicol., 30(3):272-279.
-The Soap and Detergent Association (SDA) (2005). Risk assessment guidance for enzyme-containing products.

Consumer DNEL, acute short-term as well as long-term dermal exposure:
The physico-chemical properties of a compound are decisive for the potential percutaneous penetration, in particular, factors like ionization, molecular size and lipophilicity. In general, non-ionized molecules easily penetrate the skin, with small molecules penetrating more easily than large molecules. Lipophilicity also facilitates penetration. Investigations of percutaneous absorption of peptides, proteins and other molecules of large size revealed that percutaneous absorption of proteins is extremely low and of no toxicological relevance (Basketter et al. 2008, Smith Pease et al. 2002, Basketter et al. 2012a, 2012b). This is further supported by the physico-chemical data of enzymes. Trypsins are proteins with molecular weight above 11000 D (http://www.brenda-enzymes.info), they have a low logPow value (<0, i.e. low lipophilicity), indicating that they have no bioaccumulation potential and can be anticipated to be readily biodegraded. Thus, systemic exposure following enzyme exposure at occupational exposure levels is without toxicological significance.

References

-Basketter DA, English JS, Wakelin SH, White IR(2008). Enzymes, detergents and skin: facts and fantasies. Br. J. Dermatol., 158(6):1177-1181.
-Smith Pease CK, White IR, Basketter DA(2002). Skin as a route of exposure to protein allergens. Clin. Exp. Dermatol., 27(4):296-300.
-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.
-Basketter D, Berg N, Kruszewski FH, Sarlo K, Concoby B (2012b). The Toxicology and Immunology of Detergent Enzymes. J. Immunotoxicol., 9(3):320-326.

Consumer DNEL, acute short-term as well as long-term systemic oral exposure:

Proteins are digested into amino acids by gastric juices, digestive enzymes and pancreatic proteolytic enzymes in the lumen of the gastrointestinal tract [1]. As enzymes are simply a class of proteins, enzymes will undergo the same process as any food source based on proteins. Absorption of enzymes in toxicological significant amounts through the gastrointestinal tract is unlikely. Furthermore, enzymes have been used for decades in treatment of both adults and children with exocrine pancreatic insufficiency. Typical enzymatic drugs (e.g. Creon® from Solvay Pharmaceuticals or Pancrease Microtabs from Jansson/Cilaq) are a combination of the enzymes alpha-amylase, lipase and protease, which are also used in a wide range of industrial applications. These medical drugs are typically administered orally at therapeutic concentrations (i.e. at concentrations where a digestive effect can be expected). Clinical trials and crossover studies confirm the safe use of these compounds in patients, both in adults and children, confirming the low toxicity of the enzymes [3-13].

References

1) Niesink RJM, de Vries J, Hollinger MA. (1996). Toxicology, Principles and Applications CRC Press, Inc. and Open University of The Netherlands.

2) Barra E, Stolarczyk A, Socha J, Oralewska B, Kowalska M, Skoczen M, Wawer Z. (1998). Efficacy of enzyme supplementation in children with cystic fibrosis. Pediatria Polska, 73:177-182.

3) Borowitz D, Goss CH, Stevens C, Hayes D, Newman L, O'Rourke A, Konstan MW, Wagener J, Moss R, Hendeles L, Orenstein D, Ahrens R, Oermann CM, Aitken ML, Mahl TC, Young KR Jr, Dunitz J, Murray FT. (2006). Safety and preliminary clinical activity of a novel pancreatic enzyme preparation in pancreatic insufficient cystic fibrosis patients. Pancreas, 32(3):258-263

4) Borowitz D, Goss CH, Limauro S, Konstan MW, Blake K, Casey S, Quittner AL, Murray FT. (2006). Study of a novel pancreatic enzyme replacement therapy in pancreatic insufficient subjects with cystic fibrosis. J. Pediatr., 149(5):658-662.

5) Domínguez-Muñoz JE, Iglesias-García J, Iglesias-Rey M, Figueiras A, Vilariño-Insua M. (2005). Effect of the administration schedule on the therapeutic efficacy of oral pancreatic enzyme supplements in patients with exocrine pancreatic insufficiency: A randomized, three- way crossover study. Aliment Pharmacol Ther., 21(6):993-1000.

6) Halm U, Löser C, Löhr M, Katschinski M, Mössner J. (1999). A double-blind, randomized, multicentre, crossover study to prove equivalence of pancreatin minimicrospheres versus microspheres in exocrine pancreatic insufficiency. Aliment Pharmacol Ther., 13(7), 951-957.

7) Heubi JE, Boas SR, Blake K, Nasr ARH, Woo MS, Graff GR, Hardy KA, Maro-Galvez R, Latino M, Lee C. (2008). Zentase, a novel pancreatic enzyme product (Pep), is effective in mild, moderate, and severe exocrine pancreatic insufficiency (Epi). Gastroenter., 134:A583-A584.

8) Keller J, Layer P. (2006). Are monolithic enteric-coated enzyme preparations effective in pancreatic exocrine insufficiency? A multicentre, double blind, placebo controlled cross-over trial. Gastroenter., 130:A517.

9) Konstan MW, Stern RC, Trout JR, Sherman JM, Eigen H, Wagener JS, Duggan C, Wohl ME, Colin P.(2004). Ultrase MT12 and ultrase MT20 in the treatment of exocrine pancreatic insufficiency in cystic fibrosis: Safety and efficacy. Aliment Pharmacol Ther., 20(11-12):1365-1371.

10) Konstan MW, Liou TG, Strausbaugh S, Ahrens RC, Kanga JF, Graff GR, Moffett KS, Millard S, Nasr SZ, Vezina M, Spenard J, Grondin J. (2008). Efficacy and safety of Ultrase (R) MT20 in treating pancreatic insufficiency in cystic fibrosis. Gastroenter., 134:A228-A229.

11) Laake K. (1980). ENZYMIC DRUGS. Side Effects of Drugs Annual., 222-225.

12) Patchell CJ, Desai M, Weller PH, Macdonald A, Smyth RL, Bush A, Gilbody JS, Duff SA.(2002). Creon 10,000 Minimicrospheres vs. Creon 8,000 microspheres - an open randomised crossover preference study. J. Cyst. Fibros., 1(4):287-291.

13) Saeed Z, Wojewodka G, Marion D, Guilbault C, Radzioch D. (2007). Novel pharmaceutical approaches for treating patients with cystic fibrosis. Curr. Pharm. Des., 13 (31):3252-3263.