Safety Assessment of Keratin and Keratin-Derived ... · Safety Assessment of Keratin and...

Safety Assessment of Keratin and Keratin-Derived Ingredients

as Used in Cosmetics Status: Draft Final Report for Panel Review Release Date: May 13, 2016 Panel Meeting Date: June 6-7, 2016

The 2016 Cosmetic Ingredient Review Expert Panel members are: Chairman, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Ronald A. Hill, Ph.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; James G. Marks, Jr., M.D.; Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is Lillian J. Gill, DPA. This safety assessment was prepared by Christina L. Burnett, Scientific Analyst/Writer and Bart Heldreth, Ph.D., Chemist CIR.

© Cosmetic Ingredient Review 1620 L St NW, Suite 1200◊ Washington, DC 20036-4702 ◊ ph 202.331.0651 ◊fax 202.331.0088

◊ [email protected]

__________________________________________________________________________________________ 1620 L St NW, Suite 1200, Washington, DC 20036

(Main) 202-331-0651 (Fax) 202-331-0088 (Email) [email protected] (Website) www.cir-safety.org

Commitment & Credibility since 1976

Memorandum

To: CIR Expert Panel Members and Liaisons From: Christina L. Burnett, Senior Scientific Writer/Analyst Date: May 13, 2016 Subject: Draft Final Safety Assessment on Keratin and Keratin-Derived Ingredients Enclosed is the Draft Final Report of the Safety Assessment of Keratin and Keratin-Derived Ingredients as Used in Cosmetics. (It is identified as kerati062016rep in the pdf document). At the March meeting, the Panel issued a tentative report with the conclusion that the 8 keratin-derived ingredients included in the report are safe in the present practices of use and concentration described in this safety assessment. Since March, no new unpublished data have been received. Additive unpublished data that was received before the March meeting has been incorporated into the report and highlighted in Tables 8 and 9. Information on the new final rule by FDA on the use of cattle materials in cosmetics is highlighted in |brackets| in the Use Section. Comments provided by Council on the draft report that was reviewed at the March meeting and on the tentative report have been considered. The comments and the unpublished data can be found in this report’s package (kerati062016pcpc1-2 and kerati062016data, respectively). The Panel should carefully review the abstract, discussion, and conclusion of this report and issue a Final Safety Assessment.

mailto:[email protected]

http://www.cir-safety.org/

SAFETY ASSESSMENT FLOW CHART

INGREDIENT/FAMILY __Keratin and Keratin-Derived Ingredients_______________________________

MEETING _____June 2016_______________________________________________________________________________________

Public Comment CIR Expert Panel Report Status

Priority List INGREDIENT

PRIORITY LIST

SLR

Dec 10, 2015

60 day public comment period

Draft Report

Table IDA TR

IDA Notice IDA

Draft TR

Table

Tentative Report April 13, 2106

60 day Public comment period

Draft FR

Table Different Conclusion

PUBLISH Final Report

DRAFT REPORT Mar 2016

DRAFT TENTATIVE REPORT

DRAFT FINAL REPORT June 2016

Issue TR

Issue FR

Table

Table

Table

Keratin History December 2015 – Scientific Literature Reviews announced for Keratin and Keratin-derived ingredients. March 2016 - The Panel issued a tentative report for public comment with the conclusion that the 8 keratin-derived ingredients are safe in cosmetics in the present practices of use and concentration as described in this safety assessment. Pesticide residues and heavy metals may be present in keratin source materials. The Panel stressed that the cosmetics industry should continue to use the necessary procedures to limit these impurities in the ingredients before blending into cosmetic formulations. The Panel was also concerned about the inherent risks of using animal- and human-derived ingredients in cosmetic products, namely the potential for transmission of infectious agents. The Panel stressed that these ingredients must be free of detectible infectious pathogens (e.g. Bovine Spongiform Encephalopathy (BSE), Human Immunodeficiency Virus (HIV), and Creutzfeld-Jacob disease (CJD)). Suppliers and users of these ingredients must assure that these ingredients are free from pathogenic viruses or infectious agents.

Distributed for comment only -- do not cite or quote

Keratin Data Profile –June 2016 – Writers: Christina Burnett and Bart Heldreth

In-U

se

Phys

ical

/Che

mic

al

Prop

ertie

s

Met

hod

of

Man

ufac

turi

ng

Com

posi

tion/

Impu

ritie

s

Tox

icok

inet

ics

Acu

te T

oxic

ity

Rep

eate

d D

ose

Tox

icity

Rep

ro. /

Dev

elop

. T

oxic

ity

Gen

otox

icity

Car

cino

geni

city

Irri

tatio

n/Se

nsiti

zatio

n –

Non

-Hum

an

Irri

tatio

n/Se

nsiti

zatio

n -

Hum

an

Ocu

lar/

Muc

osal

Phot

otox

icity

Cas

e St

udie

s

Keratin X X X X X X X X X Hydrolyzed Keratin X X X X X X X X X X X X

Hydrolyzed Hair Keratin X X

Hydrolyzed Oxidized Keratin

Hydrolyzed Sulfonated Keratin

Oxidized Keratin

Soluble Keratin X X X X X X Sulfonated Keratin

X indicates that data were available in the category for that ingredient.


Search Strategy for Keratin and Keratin-Derived Ingredients (Performed by Christina Burnett)

• SciFinder – February 19, 2014 o Search for “keratin toxicity” –6 hits, 0 ordered

• SciFinder – September 14, 2015 o Search keratin ingredients by name/CAS# (patents, excluded, limited to adverse effects, English

only) – 509 hits Further limited by category (Biology-substances in adverse effects-keratins) – 12 hits, 3

ordered • SciFinder – September 14, 2015

o Search for “keratin safety”(patents excluded, English only) – 48 hits, 3 ordered

Search Terms TOXLINE Hits (excluding PUBMED)

PUBMED Hits SCCS/SCCP Opinion

ECHA Hits NICNAS

(cosmetic keratin) + dermatitis

3, 1 relevant 15, 4 relevant NA NA NA

(cosmetic keratin) + irritation


(cosmetic keratin) + toxicity


keratin (generic) 1,113 (unmanageable)

31,173 (unmanageable)

1 (hydrolyzed hair keratin)

0 0

Total references ordered or downloaded: 16 Search updated April 15, 2016 = 0 relevant references found. .


Keratin March 31-April 1, 2016

Dr. Marks’ Team DR. MARKS: Okay. Next is keratin. This is our first review of these keratin and keratin derived ingredients, these proteins, and as always I asked my team members: Are these ingredients okay? They come from different sources. So, looking down the list of whatever it's not a large number, it's a small number for a change…8 ingredients. Are they all okay? DR. SHANK: Yes. DR. MARKS: And what needs do we have for them? DR. SHANK: None. DR. SLAGA: The same or no needs. DR. MARKS: That's what I had. The more (inaudible) that these are safe. DR. HILL: I had two needs. We don't have any direct data for the oxidized or sulfonated versions. So, for me, I thought if they stay in, they're insufficient without some toxicology data, and also analytical data concerned with molecular weight distributions because of the oxidation…and something about method of manufacture, because if they're doing sulfonation there are reactives that are probably used for that. And then I had a question mark here. Maybe that's covered by the Croda communication, and I need to re- research that. And then… DR. MARKS: So, let me get Tom and Ron to respond to that. Do you have any concerns about the oxidized keratin or the sulfonated keratin? DR. SHANK: I didn't. DR. SLAGA: No. DR. MARKS: No, okay. DR. HILL: If there's any…it goes to, again, the sensitization issue; nothing else in my mind, because if we get in that molecular weight sweet spot between… you know, so things long enough to crosslink IgE. When we have these artificially modified proteins, that's where I was concerned. DR. MARKS: So, IgE would be a type 1 reaction. DR. HILL: Yeah, that's what I was concerned about with making these modifications. DR. MARKS: Hmm. MS. BURNETT: We should note that oxidized and sulfonated keratin are not in use. DR. HILL: You're right. So, for me it would be a split conclusion because we don't have data. We're not going to likely get data. DR. MARKS: I wasn't concerned about type 1 reactions, and what we had on the sensitization data was both animal and human. They were all type 4. DR. HILL: And type 4 would be potential. I mean- DR. SLAGA: We have photosensitization.


DR. HILL: Pardon? DR. SLAGA: We even have photosensitization. DR. HILL: Yeah. DR. MARKS: Yeah, exactly. DR. HILL: Well, photo I wasn't even worried about. I don't know why we even had that, but type 4 I was also concerned about when you get down to low enough molecular weight, because then you can get potential absorption into the mucous membranes and like that. DR. MARKS: There are no alerts. DR. HILL: Because they're not in use. DR. MARKS: Well, I mean in keratin period. MR. BEST: I just had one…you said in this sensitization data it was found there's also a case study of severe allergic reaction, so I was sort of curious how to think about that. DR. MARKS: Yeah, one case may…if you look at allergic contact you get occasional case reports. Yeah, that's on page 12. DR. EISEMANN: And that product contained quaternaries collagen, not keratin, because they prick tested, there was some cross reactivity with hydrolyzed keratin. DR. EISEMANN: So. MS. BURNETT: The original reaction was not to the hydrolyzed keratin; it was to the prick test product. It was just the product data. I don't think they even…I pulled up DR. EISEMANN: So whether or not there was any clinically relevant response to hydrolyzed keratin in that person I'm not sure the paper stated. MS. BURNETT: No, it didn't. DR. HILL: That should be written then, too. (Inaudible) mention that. MS. BURNETT: I will rewrite that. DR. MARKS: Yeah, so even…so, the one case, and I had that highlighted actually, to me one case doesn't raise an alert that I would be worried about that ingredient in terms of the general public. DR. HILL: All right, thank you. The other information request I had…and I don't know if it's really in need or we can get it without much work. What we have on the bovine spongiform encephalopathy is a supplier has certified that their hydrolyzed keratin products are free of that, and then we don't have any information as to how they accomplished that in the processing. Wondering if we can find out … DR. SHANK: They don't use too much (inaudible). DR. EISEMANN: I don't know why they say…I mean, it's from wool, and wool is not considered… DR. HILL: I agree. DR. EISEMANN: So, they're just…


DR. HILL: So, we should write about that, too. DR. EISEMANN: They're just saying …you know, they probably have to…for (inaudible) purpose they may have to, for some countries, say anything that's animal derived, whether or not there's a BSE. DR. HILL: Some of this is derived from horn, and horn contains marrow. At least that's the information I got. There multiple sources: Bird feathers; animal horn was a common source at least internationally; and then the wool. So, there were at least three or four different sources, and I would think that the horn would be the place where you might have concern about BSE. I actually didn't. DR. SHANK: You could include that in the boilerplate. We've done that before. DR. MARKS: But with the horn in there, I didn't…with the horn? That's, like, a nail. DR. SHANK: I don't think it's a problem, but… DR. MARKS: Yeah. DR. SHANK: If you want to put it in the discussion, that… DR. HILL: If you took the whole horn and ground it up, the horn starts out living. There's marrow inside. DR. MARKS: Marrow. DR. HILL: It's a…oh, the horn. Horn (inaudible). DR. MARKS: It's a proximal nail fold essentially. DR. HILL: Okay. Cattle horn does not have a core. It's living, right? It grows out. DR. EISEMANN: I think horn or hoof - they're included as risk materials. DR. HILL: Yeah, okay, all right. So, we're good then. That's…it was brought up, and so I wondered why they responded in that way. Did we ask for information about that, because animal derived products just as a standard …is that what was going on there? MS. BURNETT: Just reported what was given to me in terms of impurities and composition. DR. MARKS: Yeah, I know. DR. EISEMANN: And some companies that …I mean, that means automatically we'll say that that type of information that's an animal derived ingredient. DR. HILL: Okay. Okay. And we do have something about that in the boilerplate, maybe not that specifically but… DR. MARKS: Well, the question is … DR. HILL: Do we need it? DR. MARKS: …do we need to put the boilerplate. My feeling is… I did with the sources of keratin that we have here. Can you find the boilerplate quickly, Christina? DR. MARKS: We'll see. We can MS. BURNETT: Going through the system is so slow.


DR. MARKS: Okay. Well, why don't we…if there is the potential, just let us know, but I didn't think horn, feathers, wool would be… DR. HILL: Well, it seems so. If you take a whole feather…yeah. If you take a whole feather, isn't there a core of the shaft of the feather that …I mean, I don't know anything in birds. DR. EISEMANN: You know, although there may be some feather, I didn't get any information on feather direct here. It's primarily wool. DR. HILL: Wool. MS. BURNETT: For the ones that were missing sources, I've gone back and checked to make sure. Some were not provided, but most of the data was on sheep wool. There was…the prick test was potentially done on bovine, but that was not clear. And then one of these passed for chicken feathers. But the majority of the data are on sheep wool. DR. MARKS: And remind me, in the past when did we put a pesticide heavy metals? Do we need that for a wool source boilerplate? MS. BURNETT: I'm not sure. DR. HILL: I wouldn't think- DR. BERGFELD: I wouldn't think so. DR. HILL: I wouldn't think. I don't think (inaudible). DR. MARKS: Yeah, okay. So, tomorrow, I'll move that we find all these keratin ingredients safe and … MS. BURNETT: We do need to have something in the discussion about (inaudible). DR. MARKS: That they're safe? SPEAKER: No. MS. BURNETT: The discussion. I mean, that's the conclusion, but I do need to have a discussion. DR. MARKS: Well, I don't want to add the bovine… MS. BURNETT: What’s that? DR. MARKS: … unless there's a reason. DR. EISEMANN: I think you should discuss about human hair, whether or not you consider that to be safe, because, I mean, there is a hair keratin that's named that way so it couldn't be distinguished from keratin derived from other animals. So, I think you need to have a discussion point on that. DR. HILL: Right, because it says in our page 9 in the document, "...the species and tissue specific function types of the keratin source could significantly impact the composition of a resulting cosmetic ingredient. While the sequences of some keratin proteins have been elucidated with a definition like the protein derived from hair, wool, horn, nails or other similar tissues in animals, the composition of a keratin derived ingredient is virtually unknowable without submission from raw material suppliers." So, that's what flagged me to think about some of these issues that we have been discussing here. That's what's written on page 9, so I think if we just leave that hanging and don't put it to bed in the discussion, for me that would be problematic.


DR. MARKS: Does that help, Christina? MS. BURNETT: I was looking at the boilerplate. DR. HILL: I mean, in an ideal world we would get the full cross section as everybody using wool, or where is it coming from? I got the sense that probably horn was a common feed stock. MS. BURNETT: Most of the data I received was on the product second (inaudible) sheep or goat's wool. In terms of the boilerplate, we have a transition of infectious disease boilerplate that has …that discusses either animal-derived or human-derived cosmetic ingredients. DR. HILL: You said sheep or goat horn? MS. BURNETT: Wool. DR. HILL: Wool, okay. MS. BURNETT: Wool. Most of it was from wool, okay? Yeah. Would you like me to read the boilerplates? DR. MARKS: I guess it's where the sources we're concerned from..what animal source? I think mainly in terms of brain. MS. BURNETT: Main… I'm sorry? DR. MARKS: Brain. MS. BURNETT: (Inaudible). DR. MARKS: Does it specifically mention keratin tissue, such as horn, feather, wool? MS. BURNETT: The background discusses tallow amine, brain extract, brain lipid, cholesterol, pharmanectin, sphingolipids, and collagen. DR. MARKS: So, basically marrow and brain. Okay, so I think we can…I wouldn't put it in the discussion, because it's not relevant. What do you feel, Ron and Tom…Ron Hill, Ron Shank? Unless that is a consumer, unless that would come up, we could put it in the discussion and say we're not concerned, because the keratin tissue would not be expected to contain these infectious agents? Does that sound good? MR. BEST: Yeah. DR. MARKS: Yes, it came up probably for the consumer. It's not bad to put it in the discussion. MR. BEST: I'd say more information can't hurt. DR. MARKS: I'm sorry, Wilma, you were going to make another comment? DR. BERGFELD: I was going to add that comment just made. DR. MARKS: Yes. DR. BERGFELD: I thought that would be (inaudible). DR. SLAGA: Dr. Marks? DR. MARKS: Yes.


DR. SLAGA: In a previous discussion with the Panel when we were working tallow type ingredients, one point that came up I think maybe industry had made it… is that if you look in the Code of Federal Regulations it specifically says "No cosmetic shall be manufactured from, processed with, or otherwise contain prohibited cattle materials." So, if any manufacturer is making these types of ingredients from those types of prohibitive materials, it wouldn't be classified as a cosmetic. It wouldn't be something that's under our purview. DR. BERGFELD: Did you say cattle? DR. SLAGA: Yes, cattle. DR. BERGFELD: Only cattle. DR. SLAGA: Correct. So, only if we're talking about specifically bovine. DR. HILL: Do we know the source of the hoofs? MS. BURNETT: No. DR. HILL: Are they from sheep and goat, or could they be from cattle? Could they be bovine? MS. BURNETT: It says bovine, so I think DR. HILL: Oh, it did say ovine. Ovine? That would be sheep. MS. BURNETT: Bovine. DR. HILL: Oh, bovine. DR. EISEMANN: But hoofs aren't a risk material. DR. MARKS: Well, I know. That's why I'm saying in the discussion we could say it so there's no confusion with consumers if they think could it be contaminated. Well, Christina is asking for discussants, but Ron Shank, you're sort of smirking, would you like to…we've discussed this a lot, would you like to just not put it in the discussion because it's not relevant? DR. HILL: So, let me read from the highly authoritative source. DR. MARKS: Go ahead, Ron Shank, then Ron Hill can comment. DR. SHANK: I'm inclined to comment. If it has… if it sells it to the consumer, have that in, then I'd say fine. But you can carry this to extremes DR. MARKS: Exactly. DR. SHANK: …because you can say underlies protein. Do we have to read this if (inaudible)? I don't think we do. DR. SLAGA: I agree. DR. MARKS: So, I think the key is as important to the consumer, and we have a representative here. MR. BEST: I mean, generally speaking, more information is always better. I mean, it didn't go out to me and I didn't think about when I read it, actually. (Inaudible). The FDA has something on this. I mean, keratinocytes are (inaudible), but that's different, right?


DR. LANGE: They're different. MR. BEST: Totally different. DR. LANGE: That's your skin cells. MR. BEST: Okay, that's your skin cells. There you are. I mean, it didn't go out to me. I mean, it's not (inaudible) talk to somebody. I mean DR. MARKS: So, that would reinforce what you said, Ron Shank, not to really include it in the discussion. Ron Hill. DR. HILL: What I was going to say is BSE, to my understanding, is transmitted by a prion. I know it affects in the brain but is not my understanding it was confined to the brain as a prion, and I'm reading here, just to remind myself, "Horn is a covering of keratin and other proteins surrounding a core of live bone." So, if it's live bone, theoretically…you know, and then it depends on processing. DR. MARKS: Hmm. DR. HILL: And I know hooves have what amounts to a live bone core of the center from which everything grows. But, again, how did they obtain the… DR. BERGFELD: But it's a very small area of the hoof. DR. HILL: Yes. MR. BEST: I mean, going back on myself, it didn't come out to me, but of course I'm not an expert in this stuff …I mean, I could also see how it would be to have… maybe you could have some kind of boilerplate. When you talked about animal derived products that, like, you know, this is not a risk because the risk is confined to these yada yada kinds of things. I mean, that's …like I said, more information is always good. I mean, it doesn't necessarily open up any kind of can of worms, right? Just maybe somebody sees it and they think incorrectly, too, and go, wow, what about BSE, actually…right? And they don't know, and like me they go over to the keratinocytes and think it's a risk. So, actually got to mix…going back the other way, some kind of animal products or a disclaimer might be useful. DR. SHANK: We have that boilerplate, and in fact if anybody here has ever dipped a sheep, the wool contains pesticides. DR. MARKS: Yes, that's why I brought up earlier about the pesticide heavy metal. DR. SHANK: So, you could cover that with a boilerplate on pesticides, metals, prions. DR. MARKS: Okay. MS. BURNETT: I just looked up "lanolin" really quick. It was one of the first reports that were published, so we didn't really put anything about pesticides in there, so. But we can for this one. DR. MARKS: BSE and… DR. SHANK: It doesn't hurt to be inclusive. DR. MARKS: Well, I wasn't sure. We normally think about that, again, on plants and such. But that's why I brought up the issue of pesticides and heavy metals on wool. I'm not sure why I thought about that. Ron… DR. BERGFELD: In manufacturing, do they wash the wool before they use it?


DR. SHANK: I'm sure actually, they make… DR. BERGFELD: Would you say it's incorporated into the keratin? Are they protein, the pesticide? Or just layers it? DR. MARKS: Well, you know, obviously with plants, the pesticides are splurried on, and sometimes heavy metals, whereas I don't know they would be used on sheep. So after the initial thought, I okay, we don't need pesticides on it, because sheep aren't normally sprayed with pesticides, are they? MS. BURNETT: They treat them for fleas? DR. SHANK: Yes, they use pesticides. DR. MARKS: Okay. DR. SHANK: Sometimes they use an arsenical. DR. MARKS: On sheep. DR. SHANK: Yeah, they actually dip the animal. DR. MARKS: Okay, so then, good. DR. SHANK: Sheep dip. MS. BURNETT: Dip. DR. SHANK: Sheep. MS. BURNETT: I can think of something else. (Laughter) DR. SHANK: So, now you have… DR. MARKS: Pesticide boilerplate plates. MS. BURNETT: There you go, something like that. DR. MARKS: Okay, and it's interesting. I never thought of the hoof being connected to, actually, the bone, because I think of the structure of -a hoof is just a much more, what I say, glorified nail - fingernail or toenail - and I don't think about being connected to the bone per se. It's in proximity, but when you pull it out it's not like you're pulling marrow or tallow out. But… DR. HILL: Well, we'll put them in. DR. MARKS: And we'll see what the… I'll suggest that tomorrow to be put in the discussion, and we'll see how the Belsito team reacts. DR. HILL: I was more concerned about horn, because I didn't see hoof being mentioned as a source, but I did see horn being mentioned as a source in several places in the document. DR. MARKS: Yeah, and the horn I don't know about, you know. That may be your… DR. HILL: They say there's a live core, and that's what I was thinking, too, but I also knew that it varied between cows and sheep and goats and others.


DR. MARKS: BSE and the pesticide boilerplates in the discussion. Anything else we need in the discussion? We did a lot of discussion about these boilerplates, whether to include or not. It took longer than to decide they're safe. Do you need anything more, Christina, in the discussion? MS. BURNETT: No, I just want to go back to earlier. We are safe for all ingredients? DR. MARKS: Yes, that's correct. MS. BURNETT: Okay. DR. BERGFELD: Did you want to add a summary in the statement about where the keratins are derived from? I mean…and the majority being from sheep and goats? MS. BURNETT: I can …we have something in the beginning. I think we could reiterate that in the discussion. DR. BERGFELD: Yeah, you could repeat that. Then you could clarify all these boilerplates. MS. BURNETT: Okay. DR. HILL: And, again, the discretion of the group…ultimately, Dr. Hill does not agree with the sulfonated and oxidized ones being included with safe. DR. MARKS: Well, I expect you'll bring that up tomorrow, Ron. That will be good. And then we'll see what the discussion is with the Belsito team. Thanks, Wilma, for - I like the idea of the summary of the keratin sources. It would be interesting, Christina, if you learn in that more as to what the anatomy of a horn is. Does it actually go into the bone of the skull and tap into the marrow? That would be really interesting. MS. BURNETT: I will do a Google search real quick. DR. MARKS: Yeah, or tap into the brain. That would be even more…okay. No, I don't think so either, but… DR. HILL: But I think prions can move freely within an organism. I could be wrong, so. DR. MARKS: Okay. DR. BERGFELD: They'd have to be little. DR. HILL: They are very little. (Laughter) DR. MARKS: Okay, so tomorrow I'm going…this is the first review. Tomorrow I'm going to move that these ingredients are safe, and then when we get to the discussant points, I'll recommend we do the BSE and the pesticide boilerplates in the discussion, as well as a summary of the keratin sources. Okay? Any other comments about keratin?


Dr. Belsito’s Team DR. BELSITO: Okay, now the keratins. So this is the first time we are looking at these ingredients from the keratin family of proteins. They are derived from many different sources including human hair, birds, sheep - hydrolyzed keratin most reported uses 667, maximum concentration 5 percent in hair tonics, dressings and other hair grooming aids. So this will be one where we need the infectious disease boilerplate, I don't know for hair from humans, so do we need the usual? I don't think so. DR. ANSELL: No, this would be illegal. DR. LIEBLER: Not from human hair and wing feathers. DR. BELSITO: I thought it was only in Europe that the hair was illegal. DR. ANSELL: No. DR. BELSITO: Right, isn't that the case? DR. ANSELL: It may well be. I'm looking into it. DR. BELSITO: Do you know if the concern for humans is ethical or scientific. So it is the EU that doesn't allow human hair and I don't know if it's an ethical thing because they also think they repeat human insult patch testing is unethical although since they can't use animal data anymore the cosmetic Europe continues to look at HRIPT and they both considered unethical. Which I find such a joke. DR. SNYDER: They feel better though. (Laughter) DR. ANSELL: I had no idea. I just heard that a couple of weeks ago, that they consider that unethical. DR. BELSITO: Oh yes, very strongly so. MS. BURNETT: So you don't test at all then, in Europe? Interesting. DR. BELSITO: They rely on HRIPT, they quote it and then they say we think this is unethical. MS. BURNETT: Interesting. I'm pulling up the SCCP opinion and see why this says that. Prohibits the use of cells, tissue or products of human origin. That's why. DR. BELSITO: Well, that's only in the EU so I thought we needed the animal and human source boilerplate and otherwise they were safe as used. DR. LIEBLER: So I agree with that and I have one chemistry method of manufacturing clarification - so two of the ingredients are oxidized keratin and hydrolyzed oxidized keratin and although we don't have any explicit method of manufacture for those, it is clear that those are produced as described in the last paragraph of your keratin description which is reducing agents (inaudible) reduced and the oxidizing agents include peracetic acid or performic acid so they will oxidize the cysteine to sulfonate residues. I'm pretty sure that's what oxidized is but since there are no suppliers to give you descriptions of their methods of manufacture then you don't have anything else to go on. But you might be able to point to that to complete the dossier on what oxidized keratin is. DR. BELSITO: And also Christina, on page 21 of the PDF the Table 8 journal, irritation studies, if you look down to the third study from the bottom on hydrolyzed keratin. MS. BURNETT: Um hum. DR. BELSITO: It is a thirty five day cumulative exposure. That sounds to me more like a sensitization study. And then, on page 12, photo toxicity, photosensitization, says very slight erythema was observed in


one of the six guinea pigs with a irradiation only at 24 hours, no erythema was observed in other animals with or without radiation and so that's simply a UV effect and just, I don't even know if it is worthy of mentioning in the discussion or I'm sure the authors concluded that there was no evidence of photo toxicity or photosensitization and if that is what they concluded I might get rid of that sentence that could confuse readers and just simply say no evidence of photo toxicity photosensitization. Do you see which line I'm talking about? MS. BURNETT: The last. DR. BELSITO: The last sentence. MS. BURNETT: A very slight… DR. BELSITO: A very slight erythema in one of six guinea pigs with a radiation only at 24 hours. No erythema or edema was observed in other animals with or without the radiation and that animal was only at 24 hours, which to me is more like minimal erythema dose and you just caused a sunburn effect from the light. MS. BURNETT: Okay, so you think I should just… DR. BELSITO: My take on this study was that there was no evidence of photo toxicity or photosensitization and I would remove that last line otherwise it seems confusing. MS. BURNETT: Um hum. DR. BELSITO: Anything else on this? So animal and human source boilerplate and inhalation boilerplate, safe as used. DR. LIEBLER: Right. MS. BURNETT: Pesticides, boilerplate? DR. BELSITO: Pardon? MS. BURNETT: Are pesticides boilerplates too possibly? There's some sheep. DR. BELSITO: No, I don't think so, that would be covered by the animal boilerplate. MS. BURNETT: Okay. That will be brought up just to let you know. DR. BELSITO: Pardon? MS. BURNETT: That will be brought up tomorrow. DR. BELSITO: Well, that's good. I didn't know sheep were plants but, I guess their wool can be contaminated with plant products. DR. BOYER: They also treat the animals. MS. BURNETT: Mostly for pesticides. They also treat them for fleas and fungus and stuff.


Full Panel Meeting DR. BERGFELD: All right, moving on then, Dr. Marks, keratin. DR. MARKS: So we have a draft report on eight keratin proteins. After our team reviewed these ingredients, we felt that we could move that these eight keratin proteins are safe as used. DR. BERGFELD: And that's a motion? DR. MARKS: Yes. DR. BERGFELD: Is there a second or a comment? DR. BELSITO: I believe, I am just checking to make sure that that was the same…yes. DR. BERGFELD: Yes, there is a second? DR. BELSITO: Safe as used. DR. BERGFELD: Any discussion or editorial changes? DR. MARKS: Yes. DR. BERGFELD: Go ahead, Jim. DR. MARKS: Actually we had a fair amount of discussion, whether we needed the BSE boilerplate in this discussion, could there be prions and keratins and since it comes from horn, we weren't quite sure, maybe you can clarify, Paul, we needed you yesterday in the discussion. Could there be any marrow in the horn that we need to put the BSE boilerplate…we also thought the pesticide boilerplate, as well as the summary of the keratin sources in the discussion but broadly interested is there any- DR. SNYDER: I don't think it's known but we should put in a boilerplate on infectious diseases. DR. MARKS: Okay. DR. BELSITO: So the animal and human pesticide boilerplates, inhalation boilerplates should also be added? And let me see, there was something else here. There was this very bizarre photo study where you saw one of six pigs with radiation only at 24 hours. I thought that was MED to the ultraviolet light, no evidence of photosensitization or phototoxicity and actually recommended that that sentence, which is the last sentence in the phototox/photosensitization PDF 12 be deleted as being misleading to the reader to think that there is some type of photo effect. DR. BERGFELD: That's fine. DR. BELSITO: And I didn't. I just raised the issue. It appears, as you know, that one of these components is keratin derived from human hair and the EU doesn't like that but that is more of an ethical concern of the EU on human derived materials and as an example, the EU does not accept human repeat insult patch testing. They believe that's unethical as well, although they do look at the results, they don't allow it to be used in European centers as a test for cosmetics. Just so you understand where the EU regulators are coming from with the human hair keratin as well. DR. BERGFELD: All right, so we have had a motion - yes, Ron? DR. HILL: I did not believe we had sufficient data to read across to the oxidized or sulfonated modified keratins and in the absence of information about the method of manufacture on those modifications, the molecular weight distributions of the resulting keratins, because there could be some small components; I


feel like those weren't covered. There are no reported uses of those so we aren't going to get the information and I am arguing for a split conclusion that would exclude those two from the safe. DR. BERGFELD: Dan? DR. LIEBLER: So yeah, I noticed the same thing, Ron, but I had a little bit of a different take on it. If you look at PDF page 9 under the method of manufacturing for keratin in the third paragraph, it describes possible reductive and oxidative work. This is all about the cysteine disulfides and the proteins and the oxidated workup describes peracetic acid or performic acid oxidations would get you to the sulfonates. That's I am sure where they come from. DR. HILL: Where would the sulfonates …oh you're saying the oxidation of that going all the way DR. LIEBLER: All the way DR. HILL: But that's not what this says. This says sulfonated and that is very different chemically DR. LIEBLER: Yeah I think that's an imprecise description. It should be clarified. DR. HILL: If it is truly sulfonated, then I can't see how that's covered in the safe DR. LIEBLER: That's what I am saying so I would keep those as insufficient and the rest fine. Anyway, I think that the information may be there to clarify this point. It depends on what the source of information is for these oxidated workups and is that associated with something that is called sulfonated or oxidized keratin because I think…you're not adding some other reagent to sulfonate something else. The most… DR. HILL: We don't know that. DR. LIEBLER: That's why I said I had a little different take on it. I agree we don't know for sure but I think what is going on here is simply imprecise communication and it may be that we don't get the information we need because there aren't any producers of this but I think that's where - since a lot of my research is all about mapping file redox modifications, this is… DR. HILL: Sure. DR. LIEBLER: I recognize it right away. It made perfect sense but we don't know for sure, I agree but there might be a relatively innocuous explanation and I expected the oxidized forms would be more innocuous than the nonoxidized forms. DR. HILL: I can't disagree with that and if the sulfonates are in fact generated from cystines and that's what we are talking about but because they appeared separately in the dictionary as ingredients, maybe that's just a gremlin, an artifact. I was uncomfortable including that. DR. BERGFELD: Well we have some point of contention and need for clarification. Can we have some guidance, Lillian? How would you like to proceed? Can we clarify that in house and still move forward? DR GILL: Certainly, this is the first time you're seeing this so we'll clarify it and ask Carol to find out whatever she can as well. We will look to see if the report, if we can make a clarification based on the information on the report. DR. BERGFELD: So we have an opportunity of moving forward with the suggestion that it will be clarified and be safe but we can also table it if we… DR. BELSITO: I don't think we need to table it. It's very early in the process so we can proceed. DR. BERGFELD: Proceed, so you wish to vote on it?


DR. MARKS: Yeah, I think we should issue it with a tentative report with the motion that I proposed and… DR. BELSITO: Some clarification of the sulfonation… DR. BERGFELD: All right, fine, it has been seconded so we can call for the vote with it, understanding that we need clarification of that particular step. All those in favor? Opposed? One opposed, thank you very much. DR. KLASSEN: May I add one point? DR. BERGFELD: Surely. DR. KLASSEN: Since sheep wool is a significant source for keratin, sheep are treated with pesticides DR. BELSITO: We added that to the boilerplate. DR. KLASSEN: You did add that? DR. BELSITO: Yes. DR. KLASSEN: Okay, thank you. DR. BERGFELD: Good, they put all the boiler plates in there, animal, human, pesticides, inhalation. DR. BELSITO: Yeah. DR. BERGFELD: All of them.


Safety Assessment of Keratin and Keratin-Derived Ingredients

as Used in Cosmetics Status: Draft Final Report for Panel Review Release Date: May 13, 2016 Panel Meeting Date: June 6-7, 2016

The 2016 Cosmetic Ingredient Review Expert Panel members are: Chairman, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Ronald A. Hill, Ph.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; James G. Marks, Jr., M.D.; Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is Lillian J. Gill, DPA. This safety assessment was prepared by Christina L. Burnett, Scientific Analyst/Writer and Bart Heldreth, Ph.D., Chemist CIR.

© Cosmetic Ingredient Review 1620 L St NW, Suite 1200◊ Washington, DC 20036-4702 ◊ ph 202.331.0651 ◊fax 202.331.0088

◊ [email protected]


ABSTRACT The Cosmetic Ingredient Review Expert Panel (the Panel) reviewed the safety of 8 keratin-derived ingredients, which function mainly as skin and hair conditioning agents in personal care products. The Panel reviewed relevant data provided in this safety assessment, and concluded that the 8 keratin-derived ingredients are safe in the present practices of use and concentration described in this safety assessment.

INTRODUCTION The keratin-derived ingredients detailed in this report are described by the International Cosmetic

Ingredient Dictionary and Handbook (Dictionary) to function mainly as skin and hair conditioning agents in personal care products.1 This report assesses the safety of the following 8 keratin-derived ingredients:

Hydrolyzed Keratin Hydrolyzed Hair Keratin Hydrolyzed Oxidized Keratin Hydrolyzed Sulfonated Keratin

Keratin Oxidized Keratin Soluble Keratin Sulfonated Keratin

This is the first Cosmetic Ingredient Review (CIR) report on ingredients derived from the keratin family of

proteins. However, the safety of several hydrolyzed proteins as used in cosmetics has previously been reviewed by the Panel. The Panel concluded that Hydrolyzed Collagen, Hydrolyzed Soy, Hydrolyzed Silk, Hydrolyzed Rice Protein, and Hydrolyzed Corn Protein are safe for use in cosmetics.2-7 Additionally, the Panel concluded that Hydrolyzed Wheat Gluten and Hydrolyzed Wheat Protein are safe for use in cosmetics when formulated to restrict peptides to a weight-average MW of 3500 Da or less.8

Keratin occurs naturally in epithelial cells and is essential for normal tissue structure and function. Much of the available published literature evaluated the ability of cosmetic or pharmaceutical ingredients to have a desired effect on naturally-occurring keratin in skin, hair, or other tissues. These studies were not relevant for assessing the safety of the keratin-derived ingredients included in this assessment.

The main sources of Keratin ingredients are sheep wool and bovine hoof (nail) or horn.9 Goat (cashmere) wool and bird (chicken) feathers may also be used. Human hair was once a major source of Keratin, but its use is now limited because of the European ban on human-sourced materials for cosmetics. These differing sources could potentially produce or result in keratin with unique properties, which may result in varying compositions and impurities within a single ingredient (e.g., Keratin from human hair may have some impurities that are different from Keratin obtained from bird feathers).

When the specific cosmetic ingredient names are discussed in this report, Keratin and the related ingredients will be capitalized, which is how they are presented in the Dictionary (see list above). When referring generally to the naturally-occurring protein from which these ingredients are derived, lower-case lettering will be used (i.e., keratin).

CHEMISTRY

Definition The definition, structures, and functions of the keratin-derived ingredients in this report are provided in

Table 1. The general term, keratin, refers to a broad category of proteins that result in intermediate filaments that

form the bulk of cytoplasmic epithelia and epidermal appendage structures (i.e., hair, wool, horn, nails and similar tissues in animals).10 Keratins can be classified into two distinct groups (“hard” and “soft”) based on their structure, function and regulation.

“Hard” keratin filaments form ordered arrays and are the primary contributors to the tough structure of epidermal appendages. Hard keratins (derived from hair, wool, horn, nails or other similar tissues in animals) are utilized as cosmetic ingredients, rather than soft (derived e.g., from cytoplasmic epithelial keratins). These hard keratin proteins contain a much higher content of cysteine residues than the soft keratins in their non-helical domains, and, thus form tougher and more durable structures via intra/intermolecular disulfide bond formation. These hard keratins may be modified/extracted (i.e., hydrolyzed, oxidized, sulfonated, etc.) to produce other cosmetic ingredients (e.g., Hydrolyzed Oxidized Keratin). The structural subunits of these keratins comprise two chains, which differ by molecular weight and amino acid residue sequence (designated types I and II), that each contain non-helical end terminal domains, and a highly-conserved, central α-helical domain.


Chemical and Physical Properties Keratin is insoluble in water, and many keratin-derived materials have molecular weights between 9 and 60

kDa.11,12 Keratin proteins extracted from hair are classified into 3 broad groups: alpha, beta, and gamma.13 α-Keratin (found in hair fiber cortex) is low in sulfur content and has an average molecular weight of 60-80 kDa. β-Keratin is protective and forms the majority of the hair cuticle; this type of keratin is difficult to extract. γ-Keratin is globular, high in sulfur content, and has a molecular weight of ~15 kDa.

Molecular weights for Hydrolyzed Hair Keratin are reported to be around 400 Da.14 The molecular weights for Hydrolyzed Keratin have been reported to be as low as 150 Da, but may be around 1000 to 3000 Da.14-16

Method of Manufacturing Keratin

Keratins are difficult to solubilize compared to other proteins. After mechanical means of processing, such as grinding of materials like animal horn, one of the first known processes for extracting keratins involved harsh (caustic) conditions using lime. Eventually, a number of oxidative and reductive methods were developed for extracting keratin. Production of many of the ingredients in this assessment involves extraction of keratin via acid- or enzymatic- hydrolysis.17

Keratin may be produced by non-chemical and chemical methods. Using non-chemical methods, Keratin may be dissolved and converted from wool via steam explosion or from feathers via superheated water.11 Under conditions of steam explosion, wool is heated under pressure at a high temperature (with steam ~220ºC) for several minutes (~10 min), followed by explosive decompression. The wool is disrupted into solid and liquid phases consisting of oligopeptides, water soluble peptides, and free amino acids. Feather barbs are treated with superheated water (liquid ~220ºC) for 2 h and then cooled. The products are oligopeptides with molecular weights of 1.0 to 1.8 kDa.

For the chemical methods, wool Keratin is extracted by reduction or oxidation of disulfide bonds. Reducing agents include thioglycolic acid, dithiothreitol, or 2-mercaptoethanol, and oxidizing agents include peracetic acid or performic acid. These agents must work in combination with a protein denaturing agent, like urea, to break disulfide bonds and disrupt hydrogen bonding. These chemical extraction methods can directly result in some these modified ingredients, such as Oxidized Keratin or Sulfonated Keratin. Resultant keratins may have low or high sulfur content (low sulfur molecular weights between 45-60 kDa, high sulfur molecular weights between 11-28 kDa), or high glycine and tyrosine content (modular weight between 9-12 kDa). It has been reported that Keratin may be extracted from chicken feathers with reducing agents.18 Hydrolyzed Keratin Hydrolyzed Keratin may be prepared from sheep wool.19 The wool is first washed to remove soil and debris and then boiled to remove residual oils. Next, the wool is enzyme-hydrolyzed under mild conditions for 4-6 hours. When the target molecular weight is reached, the pH is adjusted to neutralize the enzyme. The resultant solution is a mixture of Hydrolyzed Keratin fractions with a molecular weight of ~ 1000 Da. The solution may be diluted to produce a 30% active material. A supplier reported that three Hydrolyzed Keratin products (MW = 310 Da, 320 Da, and 11,000 Da, respectively) are prepared by acidic, alkaline and/or enzymatic hydrolysis of sheep wool until the molecular weight reaches the target range.20 Further properties and specifications of these products are described in Table 2. Another supplier certified that their powdered and liquid Hydrolyzed Keratin products are derived from sheep or goat wool.21-23 Hydrolyzed Keratin and Hydrolyzed Hair Keratin

A supplier reported that Hydrolyzed Hair Keratin and Hydrolyzed Keratin are obtained through acid hydrolysis.15 Another supplier reported that Hydrolyzed Keratin is manufactured by enzymatic hydrolysis for a specific duration of time and at an elevated temperature (details not provided).24 The resultant hydrolyzed proteins have molecular weights in the 2000-4000 Da range and all contain di- and tri-peptides.

Soluble Keratin

A supplier reported that a Soluble Keratin powder product (MW = 30,000 Da) is prepared by extraction from sheep wool.25


Composition Keratins ubiquitously consist of central α-helical rod domains that are flanked by non-α-helical head and tail domains.26 However, the amino sequences are not highly conserved across various source species (e.g., equine versus human), or even among tissue-specific function-types within one species (e.g., hair versus nail versus skin). Indeed, the differences in amino acid sequences among keratins can be rather striking (e.g., many cysteine residues in hair keratin versus a very small number in epidermal keratin, to no cysteine residues in other types). Accordingly, the species and tissues from which Keratin is derived can significantly impact the composition of the cosmetic ingredient. The amino acid sequences of some keratin proteins have been elucidated. However, the definition of Keratin in the Dictionary is “the protein derived from hair, wool, horn, nails or other similar tissues in animals.” Thus, the composition of a keratin-derived ingredient cannot be known without composition data from raw material suppliers. Keratin Cysteine residues in Keratin protein molecules make up 7% to 20% of the total amino acid residues.11 Hydrolyzed Keratin Amino acid composition data on Hydrolyzed Keratin is described in Table 3.

Impurities Product specifications for Hydrolyzed Keratin and Soluble Keratin are presented in Table 4.

As with most non-synthetic raw materials, numerous environmental factors may profoundly affect the presence and concentration of impurities, dependent on the Keratin source. Accordingly, manufacturers should use best practices to ensure the lowest possible values of any impurities. Hydrolyzed Keratin A supplier has certified that their Hydrolyzed Keratin products are free of bovine spongiform encephalopathy (BSE) infectivity, or the prions that cause BSE.21-23

USE Cosmetic

The safety of the cosmetic ingredients included in this assessment is evaluated based on data received from the U.S. Food and Drug Administration (FDA) and the cosmetics industry on the expected use of these ingredients in cosmetics. Use frequencies of individual ingredients in cosmetics are collected from manufacturers and reported by cosmetic product category in FDA’s Voluntary Cosmetic Registration Program (VCRP) database. Use concentration data are submitted by Industry in response to surveys, conducted by the Personal Care Products Council (Council), of maximum reported use concentrations by product category.

According to 2016 VCRP data, Hydrolyzed Keratin has the most reported uses of the ingredients listed in this safety assessment in cosmetic products, with a total of 667; more than half of the uses are in rinse-off non-coloring hair products (Table 5).27 Keratin has the second greatest number of overall uses reported, with a total of 90; the majority of the uses are in non-coloring hair products. The results of the concentration of use survey conducted in 2015 by the Council indicate Hydrolyzed Keratin has the highest reported maximum concentration of use; it is used at up to 5% in hair tonics, dressings, and other hair grooming aids. Keratin is used at up to 0.075% in hair tonics, dressings, and other hair grooming aids.

Based on the VCRP data and the results of the Council’s concentration of use survey, Hydrolyzed Oxidized Keratin, Hydrolyzed Sulfonated Keratin, Oxidized Keratin, and Sulfonated Keratin are not in use.

Some of these ingredients may be used in products that can come into contact with the eye or mucous membranes. For example, Hydrolyzed Keratin is used in mascara at up to 0.2% and in bath soaps and detergents at up to 0.028%. Additionally, some of these ingredients were reported to be used in hair sprays and could possibly be inhaled. For example, Hydrolyzed Keratin was reported to be used in hair sprays at a maximum concentration of 0.059%. In practice, 95% to 99% of the droplets/particles released from cosmetic sprays have aerodynamic equivalent diameters >10 µm, with propellant sprays yielding a greater fraction of droplets/particles below 10 µm compared with pump sprays.28-31 Therefore, most droplets/particles incidentally inhaled from cosmetic sprays would be deposited in the nasopharyngeal and bronchial regions and would not be respirable (i.e., they would not enter the lungs) to any appreciable amount.28,30


The FDA has recently issued a new final rule on use of materials derived from cattle in cosmetics (21 CFR §700.27). Prohibited materials did not include hoofs or horns or the keratin that may be derived from these substances.

The Keratin ingredients described in this safety assessment, with the exception of Hydrolyzed Hair Keratin, are not restricted from use in any way under the rules governing cosmetic products in the European Union (EU).32 Hydrolyzed Hair Keratin is a substance prohibited in cosmetic products in the EU due to its human origin; however, the Scientific Committee on Consumer Products (SCCP) concluded that “the resulting risk, based on current scientific data, of the use [of amino acids obtained by hydrolysis of human hair] in cosmetic products for topical application…is negligible” when the human hair has undergone hydrolysis with concentrated HCl (> 20%) for 6 h at 100◦ C followed by activated carbon filtration, crystallization, and drying.33

Non-Cosmetic

Non-cosmetic uses of Keratin include use as a biopolymer in nanomaterials and in biomedical applications such as wound dressings, drug delivery, tissue engineering, and trauma and medical devices.10-13,34,35

TOXICOKINETICS Absorption, Distribution, Metabolism, Excretion

Keratin The tissue distribution of enzymatically and chemically fragmented Keratin (MW ~ 8000 Da and 33,000

Da, respectively) was studied in male ddY mice.35 The fragmented Keratin was radiolabeled with Na125I. The radiolabeled test materials were then injected via the tail vein at a dose of 2 mg/kg. The mice were killed at varying times following injection. Tissues were excised and weighed, and the radioactivity measured. The fragmented Keratin was found to be quickly eliminated from the plasma, taken up into the kidney, and gradually excreted in urine. Chemically fragmented Keratin was also observed to be taken up into the liver.

Dermal Penetration Hydrolyzed Keratin

A study of the efficacy of Hydrolyzed Keratin derived from wool stated that Hydrolyzed Keratin peptide can penetrate into the skin and increase moisturization.36 No further details regarding the penetration properties of the Hydrolyzed Keratin were provided.

TOXICOLOGICAL STUDIES

Acute Toxicity Acute toxicity studies are presented in Table 6.20,35,37-39 Hydrolyzed Keratin sourced from sheep wool and

hoof meal was non-toxic in oral studies in rats (i.e. the LD50 was greater than 40 ml/kg in a 20% solution). Keratin (MW = 8000 Da and 33,000 Da; buffalo horn/hoof source) and Hydrolyzed Keratin (MW = 310 Da; sheep wool source) were non-toxic in intravenous studies in mice.

Repeated Dose Toxicity No relevant published repeated dose toxicity studies on Keratin ingredients were identified in a literature search for these ingredients, and no unpublished data were submitted.

REPRODUCTIVE AND DEVELOPMENTAL TOXICITY No relevant published reproductive and developmental toxicity studies on Keratin ingredients were identified in a literature search for these ingredients and no unpublished data were submitted.

GENOTOXICITY In vitro genotoxicity studies are presented in Table 7.20,25,40-45 Hydrolyzed Keratin (MW ranging from 320

Da to 11,000 Da), Soluble Keratin (MW = 30,000 Da), and Keratin (MW not specified) that were mainly sourced from sheep wool were not mutagenic in Ames studies. In a comet assay, a Keratin peptide (MW = 1600 Da; synthesized from human hair) was not genotoxic in aqueous solutions, but genotoxicity was observed in organic solvent solutions. However, the negative controls also displayed genotoxicity in these solutions.


CARCINOGENICITY No relevant published carcinogenicity studies on Keratin ingredients were identified in a literature search

for these ingredients and no unpublished data were submitted.

IRRITATION AND SENSITIZATION Dermal Irritation

In vitro, animal, and human dermal irritation studies are presented in Table 8.18,20,24,25,36,37,45-56 No irritation was predicted in in vitro studies of sheep wool-sourced Hydrolyzed Keratin tested at 100% and Keratin tested at 5%. Keratin (concentration not reported; chicken feather source) and Hydrolyzed Keratin (tested neat; source not provided) were mainly not irritating in rabbit studies. Hydrolyzed Keratin was not irritating in human studies at concentrations up to 25% (sheep wool source), nor was Soluble Keratin at 10% (sheep wool source).

Ocular Irritation

Ocular irritation studies are presented in Table 9.20,24,25,37,45,47-51,54,57-60 No irritation was predicted in in vitro studies of Keratin (5%), Hydrolyzed Keratin (undiluted), or Soluble Keratin (5%), which were mainly sourced from sheep wool. Hydrolyzed Keratin was minimally to non-irritating in Draize ocular rabbit studies when tested neat (source not provided).

Dermal Sensitization

Dermal sensitization studies are presented in Table 10.20,25,45,61-66 No sensitization was predicted in in vitro studies of Hydrolyzed Keratin (sheep and goat wool sources). No sensitization to Hydrolyzed Keratin was observed when tested up to 25% in a guinea pig maximization study (sheep wool source) or up to 5% in human repeat insult patch tests (HRIPTs; bovine source). No sensitization was observed in HRIPTs to Keratin (5%; sheep wool source) or Soluble Keratin (10%; sheep wool source).

Phototoxicity and Photosensitization The potential for phototoxicity of Hydrolyzed Keratin (MW = 320 Da; sheep wool source) was studied in 6 male Hartley guinea pigs.20 A 25% solution in water was applied to 2 clipped dorsal skin sites (2 cm2). Each site received 0.05 ml test material. One site was covered and the other site was irradiated for 2 h. The sites were evaluated at 24, 28, and 72 h post-irradiation. No further details were provided. No erythema or edema was observed with or without irradiation. The potential for photosensitization of Hydrolyzed Keratin (MW = 320 Da; sheep wool source) was studied in 6 male Hartley guinea pigs.20 The guinea pigs were induced on 2 clipped dorsal skin sites (2 cm2) with 0.05 ml of a 25% solution in water. One site was covered and the other irradiated for 2 h daily, 5 times per week for 2 weeks. Following a 2 week rest period, the test sites received the test material with the same site covered and the other site irradiated for 2 h. Sites were evaluated at 24, 48, and 72 h post-irradiation. No further details were provided. No evidence of phototoxicity or photosensitization was observed.

CLINICAL STUDIES Case Reports

Hydrolyzed Keratin A 22-year-old woman was reported to have a severe allergic reaction that included marked periorbital edema and swollen, sore, and itchy eyes and hands following use of a hair conditioner.67 Prick testing elicited a strong positive (10 mm) wheal-and-flare response to the hair conditioner, which contained steartrimonium hydrolyzed animal protein (also known as Steartrimonium Hydroxyethyl Hydrolyzed Collagen). Additional prick testing showed further reactions to the quaternary hydrolyzed protein as well as to shampoos and conditioners that contained gelatin keratin amino acids, Hydrolyzed Keratin, and/or hydrolyzed collagen. The allergic reaction to Hydrolyzed Keratin was only observed in the prick test. Patch tests using the European standard series and a series of 15 common bases of medicines and cosmetics were negative.

SUMMARY

The keratin-derived ingredients detailed in this report function mainly as skin and hair conditioning agents in personal care products. Keratin occurs naturally in epithelial cells and is essential for normal tissue structure and function.


According to 2016 VCRP data, Hydrolyzed Keratin has the most reported uses of the ingredients listed in this safety assessment in cosmetic formulations, with a total of 667; more than half of the uses are in rinse-off non-coloring hair formulations. Keratin has the second greatest number of overall uses reported, with a total of 90; the majority of the uses are in non-coloring hair formulations. The results of the concentration of use survey conducted in 2014 by the Council indicate Hydrolyzed Keratin has the highest reported maximum concentration of use; it is used at up to 5% in hair tonics, dressings, and other hair grooming aids. Keratin is used at up to 0.075% in hair tonics, dressings, and other hair grooming aids.

Non-cosmetic uses of Keratin include use as a biopolymer in nanomaterials and in biomedical applications such as wound dressings, drug delivery, tissue engineering, and trauma and medical devices.

In an intravenous tissue distribution study of enzymatically and chemically fragmented Keratin, fragmented Keratin was found to be quickly eliminated from the plasma, taken up into the kidney, and gradually excreted in urine. Chemically fragmented Keratin was also observed to be taken up into the liver.

Hydrolyzed Keratin peptide derived from wool may penetrate into the skin. Hydrolyzed Keratin was non-toxic in oral studies in rats (i.e., the LD50 was greater than 5 ml/kg in a

product with MW = 320 Da). Keratin (MW = 8000 Da and 33,000 Da) and Hydrolyzed Keratin (MW = 310 Da) were non-toxic in intravenous studies in mice. Hydrolyzed Keratin (MW ranging from 320 to 11,000 Da), Soluble Keratin (MW = 30,000 Da), and Keratin (MW not specified) were not mutagenic in Ames studies. In a comet assay, a Keratin peptide (MW = 1600 Da) was not genotoxic in aqueous solutions, but genotoxicity was observed in organic solvent formulations. However, the negative controls also displayed genotoxicity in these formulations. No irritation was predicted in in vitro studies of Hydrolyzed Keratin tested at 100% and Keratin tested at 5%. Keratin (concentration not reported) and Hydrolyzed Keratin (tested neat) were not irritating in rabbit studies. Hydrolyzed Keratin was not irritating in human studies at concentrations up to 25%, nor was Soluble Keratin at 10%.

No irritation was predicted in in vitro studies of Keratin (5%), Hydrolyzed Keratin (undiluted), or Soluble Keratin (5%). Hydrolyzed Keratin was minimally to non-irritating in Draize ocular rabbit studies when tested neat.

No sensitization was predicted in in vitro studies of Hydrolyzed Keratin. No sensitization to Hydrolyzed Keratin was observed when tested up to 25% in a guinea pig maximization study or up to 5% in HRIPTs. No sensitization was observed in HRIPTs to Keratin (5%) or Soluble Keratin (10%).

Hydrolyzed Keratin in a 25% solution was not phototoxic or photosensitizing in guinea pigs. No relevant published repeated dose toxicity, reproductive and developmental toxicity, or carcinogenicity

studies on Keratin ingredients were identified in a literature search for these ingredients and no unpublished data were submitted.

DISCUSSION

Keratin is a protein that occurs naturally in epithelial cells and is essential for normal tissue structure and function. The main sources for Keratin and Hydrolyzed Keratin are sheep wool and bovine hoof or horn. Goat wool, bird feathers, and human hair may also be used. Keratin ingredients derived from different sources are likely to have different compositions and impurities, which may result in varying compositions and impurities within a single ingredient (e.g., Keratin from human hair may have some impurities that are different from Keratin obtained from bird feathers).

Pesticide residues and heavy metals may be present in Keratin source materials. The Panel stressed that the cosmetics industry should continue to use the necessary procedures to limit these impurities in the ingredients before blending into cosmetic formulations.

The Panel was also concerned about the inherent risks of using animal- and human-derived ingredients in cosmetic products, namely the potential for transmission of infectious agents. The Panel stressed that these ingredients must be free of detectible infectious pathogens (e.g. Bovine Spongiform Encephalopathy (BSE), Human Immunodeficiency Virus (HIV), and Creutzfeld-Jacob disease (CJD)). Raw material suppliers and formulators of these ingredients must assure that these ingredients are free from pathogenic viruses and other infectious agents.

The Panel discussed the issue of incidental inhalation exposure in hair sprays. There were no inhalation toxicity data available. The Panel considered other pertinent data indicating that incidental inhalation exposures to keratin-derived ingredients in such cosmetic products would not cause adverse health effects, including data characterizing the potential for keratin-derived ingredients to cause ocular or dermal irritation or sensitization, and other effects. These ingredients are reportedly used at concentrations up to 0.059% in cosmetic products that may be aerosolized. The Panel noted that 95% – 99% of droplets/particles produced in cosmetic aerosols would not be respirable to any appreciable amount. The potential for inhalation toxicity is not limited to respirable


droplets/particles deposited in the lungs. In principle, inhaled droplets/particles deposited in the nasopharyngeal and thoracic regions of the respiratory tract may cause toxic effects depending on their chemical and other properties. However, coupled with the small actual exposure in the breathing zone and the concentrations at which the ingredients are used, the available information indicates that incidental inhalation would not be a significant route of exposure that might lead to local respiratory or systemic effects. A detailed discussion and summary of the Panel’s approach to evaluating incidental inhalation exposures to ingredients in cosmetic products is available at http://www.cir-safety.org/cir-findings.

CONCLUSION The CIR Expert Panel concluded the following keratin-derived ingredients are safe in cosmetics in the present practices of use and concentration as described in this safety assessment.

Hydrolyzed Keratin Hydrolyzed Hair Keratin Hydrolyzed Oxidized Keratin* Hydrolyzed Sulfonated Keratin*

Keratin Oxidized Keratin* Soluble Keratin Sulfonated Keratin*

*Not reported to be in current use. Were ingredients in this group not in current use to be used in the future, the expectation is that they would be used in product categories and at concentrations comparable to others in this group.


http://www.cir-safety.org/cir-findings

TABLES Table 1. Definitions, Structures, and functions of the ingredients in this safety assessment.1 Ingredient/CAS No. Definition & Structure Function Hydrolyzed Keratin 65997-21-9 73049-73-7

Hydrolyzed Keratin is the hydrolysate of keratin derived by acid, enzyme or other method of hydrolysis.

hair conditioning agents; nail conditioning agents; skin-conditioning agents-misc.

Hydrolyzed Hair Keratin 69430-36-0 73049-73-7 [peptones]

Hydrolyzed Hair Keratin is the hydrolysate of human hair keratin derived by acid, enzyme or other method of hydrolysis.

hair conditioning agents; skin-conditioning agents-misc.

Hydrolyzed Oxidized Keratin [1142948-22-8]

Hydrolyzed Oxidized Keratin is the hydrolysate of Oxidized Keratin obtained by acid, enzyme or other method of hydrolysis.


Hydrolyzed Sulfonated Keratin [1119233-83-8]

Hydrolyzed Sulfonated Keratin is the hydrolysate of Sulfonated Keratin derived by acid, enzyme or other method of hydrolysis.

hair conditioning agents; skin-conditioning agents-emollient; skin-conditioning agents-humectant

Keratin 169799-44-4 68238-35-7

Keratin is the protein derived from hair, wool, horn, nails or other similar tissues in animals.


Oxidized Keratin [143819-61-8]

Oxidized Keratin is the material derived chemically from Keratin by oxidation with hydrogen peroxide. This reaction converts some of the sulfur atoms in Cysteine and Cystine residues in keratin to the corresponding sulfonic acid grouping (cysteic acid).


Soluble Keratin Soluble Keratin [is] a water soluble nonhydrolyzed, native protein derived from Keratin.

hair conditioning agents

Sulfonated Keratin [1119232-93-7]

Sulfonated Keratin is the product obtained by the oxidative sulfitolysis of wool. film formers; hair conditioning agents; skin protectants


Table 2. Properties and specifications of Hydrolyzed Keratin products as reported by a supplier. 68-70 Specification Hydrolyzed Protein

(sheep wool source, powder) Hydrolyzed Protein (sheep wool source, solution)

Hydrolyzed Protein (goat wool source, solution)

Appearance Pale cream free flowing powder Clear to hazy amber liquid Clear to hazy amber liquid Color 8 Gardner max (10% solution) 11 Gardner max 11 Gardner max Odor Slight, characteristic Characteristic protein note Characteristic Moisture Content (1g-1h-105◦C) 8.0% max 30.0-35.0% 30.0-35.0% pH 5.0-7.0 (10% solution at 25◦C) 5.5-7.0 (direct) 5.5-7.0 (direct) Ash (800◦C) 4.0% max 4.0% max 4.0% max Nitrogen (Kjeldahl) 14.0% min 4.0% min 4.0% min Heavy Metals < 20 ppm < 20 ppm < 20 ppm Arsenic < 2 ppm < 2 ppm < 2 ppm Microbial content < 100 opg, no pathogens < 100 opg, no pathogens < 100 opg, no pathogens opg = organisms per gram


Table 3. Amino acid composition of commercialized keratin hydrolysate (g/100 g protein).71 Cysteic acid 0.3 Hydroxyproline 0 Aspartic acid 7.8 Threonine 6.1 Serine 8.1 Glutamic acid 17.0 Proline 7.6 Glycine 3.9 Alanine 7.6 Cystine 7.2 Valine 5.7 Methionine 1.0 Isoleucine 3.8 Leucine 8.1 Tyrosine 2.0 Phenylalanine 2.4 Lysine 3.2 Histidine 1.0 Arginine 9.0


Table 4. Specifications of keratin-derived products (sheep wool source).20,25 Average Molecular

Weight (Da) Concentration Heavy Metals Arsenic

Hydrolyzed Keratin Product 1

310 25% solution in water Not more than 10 ppm Not more than 1 ppm


320 25% solution in water Not more than 10 ppm Not more than 1 ppm


11,000 20% solution in water Not more than 10 ppm Not more than 1 ppm

Soluble Keratin Powder 30,000 NR Not more than 20 ppm lead

Not more than 2 ppm


Table 5. Frequency and concentration of use according to duration and type of exposure for Keratin ingredients.27,72

# of Uses Max Conc of Use (%) # of Uses Max Conc of Use (%)

Keratin Hydrolyzed Hair Keratin Totals† 90 0.075 27 NR Duration of Use Leave-On 44 0.075 3 NR Rinse Off 46 NR 24 NR Diluted for (Bath) Use NR NR NR NR Exposure Type Eye Area 7 NR NR NR Incidental Ingestion NR NR NR NR Incidental Inhalation –Sprays 2; 18a 0.075a 1a NR Incidental Inhalation – Powders NR NR NR NR Dermal Contact 3 NR NR NR Deodorant (underarm) NR NR NR NR Hair - Non-Coloring 80 0.075 27 NR Hair-Coloring NR NR NR NR Nail 2 NR NR NR Mucous Membrane NR NR NR NR Baby Products NR NR NR NR

Hydrolyzed Keratin Soluble Keratind

Totals† 667 0.000034-5 1 NR Duration of Use Leave-On 249 0.000034-5 NR NR Rinse Off 417 0.0001-0.88 1 NR Diluted for (Bath) Use 1 <0.01 NR NR Exposure Type Eye Area 36 0.001-0.2 NR NR Incidental Ingestion NR NR NR NR

Incidental Inhalation –Sprays 26; 131a; 9b 0.000034-0.059; 0.0003-5a NR NR

Incidental Inhalation – Powders 9b 0.0025c NR NR Dermal Contact 40 0.001-0.21 NR NR Deodorant (underarm) 1a NR NR NR Hair - Non-Coloring 393 0.000034-5 1 NR Hair-Coloring 193 0.0001-0.5 NR NR Nail 10 0.002-0.04 NR NR Mucous Membrane 10 0.01-0.028 NR NR Baby Products NR NR NR NR

NR = Not reported. † Because each ingredient may be used in cosmetics with multiple exposure types, the sum of all exposure types may not equal the sum of total uses. a. It is possible these products may be sprays, but it is not specified whether the reported uses are sprays. b. Not specified whether a powder or a spray, so this information is captured for both categories of incidental inhalation. c. It is possible these products may be powders, but it is not specified whether the reported uses are powders. d. Listed in the VCRP as Soluble Animal Keratin.


Table 6. Acute toxicity studies Ingredient Concentration/Dose Animals Results Reference

Oral Hydrolyzed Keratin (MW = 320 Da; sheep wool source)

5 ml/kg of a 25% solution in water

5 male and 5 female Wistar rats LD50 > 5 ml/kg 20

Hydrolyzed Keratin (MW = 11,000 Da; sheep wool source)

2 g/kg of a 20% solution in water 3 male and 3 female Sprague-Dawley CD rats

LD50 > 2 g/kg 20

Hydrolyzed Keratin (hoof meal source)

10 g/kg of a 20% solution 5 male and 5 female Sprague-Dawley rats

LD50 > 10 g/kg 39


2.5, 5.0, 10.0, 20.0, or 40.0 ml/kg of a 20% solution

Five groups of 3 male and 2 female Wistar rats

LD50 > 40 ml/kg 38


5 g/kg of a 25% solution 5 male and 5 female Wistar rats LD50 > 5 g/kg 37

Intravenous Keratin (enzymatically and chemically fragmented, MW ~ 8000 Da and 33,000 Da, respectively; buffalo horn/hoof source)

300 mg/kg dissolved in 1/15 M sodium phosphate buffer

male ddY mice, number not reported

No significant changes observed in lungs, livers, kidneys, spleens; body weight changed similar to those of the control group

35

Hydrolyzed Keratin (MW = 310 Da; sheep wool source)

5 ml/kg of a 20% solution in water

15 male ddY mice LD50 > 5 ml/kg 20


Table 7. In vitro genotoxicity studies Ingredient Concentration/Dose Study Protocol Results Reference

Keratin (characterized as a peptide with a MW ~1600 Da, 13 amino acid sequence comprised of 2 cysteine residues; synthesized from human hair)

0.025 g/L to 0.5 g/L prepared in an aqueous solution with 100% phosphate buffer or in an organic solvent solution with 10% ethanol, 1.5% propylene glycol, 0.5% benzyl alcohol, and 88% phosphate buffer

Comet assay with human fibroblasts; cells incubated with test materials and controls for either 1 h or 72 h

In the aqueous solutions, Keratin peptide was not genotoxic; in the organic solvent solutions for the 1 h exposure, genotoxicity was observed in the negative controls as well as in the treated cells; for the 72 h exposure, genotoxicity was also observed, but the extent of DNA damage was not as great as observed in the 1 h exposure

40

Keratin (sheep wool source) 100, 333, 1000, 3333, and 5000 μg/plate

Bacterial reverse mutation assay (Ames test) Salmonella typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 and Escherichia coli strain WP2 uvr A, with and without metabolic activation

Non-mutagenic 45

Hydrolyzed Keratin (25% solution in water; MW = 320 Da; sheep wool source)

500 to 100,000 μg/plate Ames test using S. typhimurium strains TA 98, TA 100, TA 1535, TA 1537, and TA 1538 and in E. coli strain WP2 uvrA, with or without metabolic activation

Non-mutagenic 20

Hydrolyzed Keratin (MW = 3000 Da; 14% peptide content; source not provided)

up to 5000 μg/plate Ames test using S. typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 and in E. coli strain WP2 uvrA, with or without metabolic activation

Non-mutagenic 41

Hydrolyzed Keratin (20% solution in water; MW = 11,000 Da; sheep wool source)

50 to 5000 μg/plate

Ames test using S. typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 and in E. coli strain WP2 uvrA, with or without metabolic activation

Non-mutagenic 20

Hydrolyzed Keratin (powder product, sheep wool source; MW not provided)

1.5, 5.0, 15, 50, 150, 500, 1500, and 5000 μg/plate

Ames test using S. typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 and E. coli strain WP2 uvr A, with and without metabolic activation

Non-mutagenic 44

Hydrolyzed Keratin (solution, sheep wool source; MW not provided)

1.5, 5.0, 15, 50, 150, 500, 1500, and 5000 μ/plate


Non-mutagenic 43

Hydrolyzed Keratin (solution, goat wool source; MW not provided)

1.5, 5.0, 15, 50, 150, 500, 1500, and 5000 μ/plate


Non-mutagenic 42

Soluble Keratin (MW = 30,000 Da; sheep wool source)

50 to 5000 μg/plate. Ames test using S. typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 and in E. coli strain WP2 uvrA, with or without metabolic activation

Non-mutagenic 25


Table 8. Dermal irritation studies. Ingredient Concentration Method Results Reference

In Vitro Keratin (sheep wool source)

5% EpiDerm™ skin model Non-irritating 45

Hydrolyzed Keratin (source not provided)

100% (MW = 3000 Da) EpiDerm™ skin model Non-irritating 52


2 forms tested (MW = 2000-4000 Da), solution concentration not reported, powder undiluted

EpiDerm™ assay Non-irritating 24

Hydrolyzed Keratin (sheep wool source)

Undiluted powder product (MW not provided)

EpiDerm™ skin model Non-irritating 49


25, 50, 75, 100, and 125 μl (MW not provided)

Irritection® dermal assay Non-irritating 48

Hydrolyzed Keratin (goat wool source)


Irritection® dermal assay Non-irritating 47

Animal Keratin (chicken feathers source)

Not reported 6 male rabbits received test material in a cream daily for 8 weeks on a 4 cm2 area in an efficacy study ( no further studies)

No irritation was observed 18


20% solution Primary dermal irritation study in 6 female New Zealand white rabbits; 0.5 ml occluded for 24 h; intact and abraded skin; exposure area 2.5 cm2

Well defined erythema in both intact and abraded skin of 4 rabbits 24 h post-dosing while the remaining 2 rabbits had very slight erythema; very slight edema in 3 abraded and 3 intact sites, slight edema in 1 abraded and 2 intact sites, and moderate edema in 1 abraded site; irritation response declined during study with well-defined erythema and very slight edema remaining at abraded site and very slight erythema remaining at 5 abraded and 4 intact sites at 72 h post-dosing; PII = 2.0; mildly irritating

53


20% solution Draize primary dermal irritation study in 6 albino rabbits; 0.5ml occluded for 24 h; intact and abraded skin; exposure area 1 in2

Erythema in abraded skin of all 6 rabbits and edema in abraded skin of 4 rabbits; edema cleared in 2 of the rabbits by the 72 h observation period; mild irritant; PII = 0.75

56


25% solution Draize primary dermal irritation study in 6 New Zealand white rabbits; 0.5 ml occluded for 24 h; intact and abraded skin; exposure area 2.5 cm2

PII = 0.43; not a primary dermal irritant

37


25% solution in water (MW = 320 Da)

Draize primary dermal irritation study in 6 New Zealand white male rabbits; 0.5 ml occluded for 24 h; intact and abraded skin; exposure area 2.5 cm2

Very slight erythema in abraded skin of 2/6 rabbits after 24 h and 1/6 rabbits after 72 h; mild erythema in abraded skin of 1/6 rabbits after 72 h; PII = 0.2.; non-irritant

20


20% solution in water (MW = 11,000 Da)

Draize primary dermal irritation study in 3 male New Zealand white rabbits; 0.5 ml occluded for 4 h; intact skin; exposure area 2.5 cm2

No skin irritation noted; PII – 0.0; non-irritant

20


Neat (MW = 500 Da) Draize primary dermal irritation study in 6 New Zealand white rabbits; occluded for 24 h

PII = 2.15; not a primary irritant

51


Neat (MW not provided) Draize primary dermal irritation study in 6 male New Zealand white rabbits; occluded for 24 h

PII = 0.4; not a primary irritant 55


Neat (MW = 125,000 Da) Draize primary dermal irritation study in 6 female New Zealand white rabbits; occluded for 24 h



Neat (MW = 600 Da) Draize primary dermal irritation study in 6 New Zealand white rabbits; occluded for 24 h



Table 8. Dermal irritation studies. Ingredient Concentration Method Results Reference

Human – In Vivo Hydrolyzed Keratin (enzymatically hydrolyzed; non-specific wool source)

Hand cream containing 3% Keratin peptide (as supplied, 0.3% active; MW = < 1000 Da or 6-8 amino acids)

Efficacy study in 16 female volunteers on undisturbed hand skin and on sodium lauryl sulfate disturbed hand skin; exposure was once a day for 2.5 weeks for a total of 12 applications; exposure area = 9 cm2; amount not reported

No adverse effects 36


25% solution in water (as supplied, MW = 310 Da)

24 h occlusive human patch test in 40 healthy subjects and 10 allergic subjects; 0.5 ml test material applied by Finn chambers

Non-irritant 20


25% solution in water (as supplied, MW = 320 Da)

24 h occlusive human patch test in 24 health subjects; 0.2 ml test material applied by adhesive tape

Non-irritant 20


20% solution in water (as supplied, MW = 11,000 Da)

24 h occlusive human patch test in 23 subjects; 0.03 g test material applied by Finn chambers

1 subject had a mild erythema reaction at 30-60 min post-patch removal; no erythema in any subjects at 24 h post-patch removal; non-irritant

20

Soluble Keratin (sheep wool source)

10% active in distilled water (MW = 30,000 Da)

24 h occlusive human patch test in 20 subjects; test material applied by Finn chambers

Non-irritant 25


Table 9. Ocular irritation studies. Ingredient Concentration Method Results Reference

In Vitro Keratin (sheep wool source)

5% tested at 1%, 5%, and 10% in distilled water

HET-CAM method Practically no irritation potential 45



Irritection® ocular assay Minimal irritant 48

Hydrolyzed Keratin (goat wool source)


Irritection® ocular assay Minimal irritant 47


Undiluted powder product (MW not provided)

EpiOcular assay Non-irritating 49


1%, 5% and 10% (MW = 3000 Da)

HET-CAM method Practically no irritation potential at 1% and 5%. Slight irritation potential at 10%.

57


2 forms tested (MW = 2000-4000 Da), solution concentration not reported, powder undiluted

EpiOcular assay Non-irritating 24


Product (MW = 30,000 Da) diluted in distilled water to 5%

HET-CAM method Average score 0.75; non-irritant 25

Animal Hydrolyzed Keratin (hoof meal source)

20% solution 0.1 ml instilled into the right eye of 6 female New Zealand white rabbits; eyes assessed 24, 48, and 72 h post-dosing

Slight conjunctival redness noted in 1 rabbits at 24 h post-dosing; non-irritating

59


20% solution Unreported amount instilled into eyes of 6 albino rabbits; eyes assessed 24, 48, and 72 h and 7 days post-dosing

Conjunctival effects observed in 3 rabbits that cleared by the 48 h observation period; mild, transient irritant

54


25% solution 0.1 ml instilled into one eye of 6 New Zealand white rabbits; eyes assessed 24, 48, and 72 h and 4 and 7 days post-dosing

Non-irritating 37


25% solution in water diluted by 1%, 5%, 10%, 15%, and 25% v/v in saline (MW = 310 Da)

Dilutions instilled into one eye each of groups of 3 male domestic rabbits while other eye received saline; eyes observed immediately after instillation and at 1 and 24 h after instillation

Redness of the eye was not observed in 1 % and 5% dilutions. Redness of the eye was observed immediately after instillation, but not observed at 1 and 24 h after instillation in 10 %, 15%, and 25% dilutions. Considered a non-irritant at 1.25% active and a mild irritant at 2.5% active

20



Ocular irritation study in 6 male New Zealand white rabbits; material instilled in one eye and other eye served as control, eyes observed at 24, 48, and 72 h after instillation; no further details provided

Ocular irritation score = 0; non-irritating

20



Cumulative ocular irritation study in 6 male New Zealand white rabbits; test material instilled into one eye 3 times a day for 4 days; other eye served as control

Hyperemia of the iris observed in 3/6 rabbits for 4 days and in 1/6 rabbits for 3 days; hyperemia of the conjunctivae observed in 4/6 rabbits for 4 days and in 1/6 rabbits for 1 day; minimal irritant

20


Neat (MW not provided) Draize ocular irritation study in 6 New Zealand white rabbits; unrinsed eyes

Slight ocular irritant 60


Neat (MW = 500 Da) Draize ocular irritation study in 6 New Zealand white rabbits; unrinsed eyes

Minimal ocular irritant 51


Neat (MW = 125,000 Da) Draize ocular irritation study in 6 female New Zealand White rabbits; unrinsed eyes

Non-irritating 58


Neat (MW = 600 Da) Draize ocular irritation study in 6 New Zealand white rabbits; unrinsed eyes

Non-irritating 50


Table 10. Dermal sensitization studies. Ingredient Concentration Method Results Reference

In vitro Hydrolyzed Keratin

(powder product, sheep wool source)

0.39-1600 μg/ml in DMSO ARE-Nrf2 Luciferase Test Method Predicted to be non-sensitizing

62

Hydrolyzed Keratin (solution, sheep wool

source)

0.39-1600 μg/ml in DMSO ARE-Nrf2 Luciferase test method Predicted to be non-sensitizing

63

Hydrolyzed Keratin (solution, goat wool source)

0.39-1600 μg/ml in DMSO ARE-Nrf2 Luciferase test method Predicted to be non-sensitizing

61

Animal Hydrolyzed Keratin (sheep

wool source) 25% solution in water (MW = 320 Da)

Guinea pig maximization test in 3 male and 3 female Harley guinea pigs

No reactions observed in animals at 24 and 48 h after challenge patch removal; non-sensitizer

20



35 day cumulative exposure study in 8 Hartley male guinea pigs; exposure area 2.5 cm2 on clipped dorsal skin

Very slight erythema in 2/8 guinea pigs after day 13 and 1/8 guinea pigs after day 16; mild erythema in 2/8 guinea pigs after day 13, 1/8 guinea pigs after day 14 and 1/8 after day 15; minimally irritant

20

Human Keratin (sheep wool source) 5% HRIPT with 51 subjects; exposure site

was 1 inch2 No dermal irritation or sensitization

45


Not reported (MW = 3000 Da) HRIPT with 51 subjects; semi-occlusive

No dermal irritation or sensitization

64

Hydrolyzed Keratin (non-specific bovine source)

0.1% (prick test) and 5% (patch test

HRIPT with 500 patients; prick tests in 25 subjects with scalp dermatitis to the test materials (no further details provided)

No positive patch test reactions and no positive prick test reactions

65

Multiple Hydrolyzed Proteins including Hydrolyzed Keratin (source not provided)

Not reported Sensitization study of protein hydrolysates in hair care products in 3 groups of patients. Group 1 was comprised of 11 hairdressers with hand dermatitis, group2 was comprised of 2160 consecutive adults with suspected allergic respiratory disease, and group 3 was comprised of 28 adults with atopic dermatitis. Subjects submitted to scratch and/or prick tests.

No adverse reactions to Hydrolyzed Keratin were observed.

66


10% active in distilled water HRIPT with 50 subjects; occlusive; 0.2 ml applied

Non-sensitizer 25


REFERENCES 1. Nikitakis J and Lange B. International Cosmetic Ingredient Dictionary and Handbook. 16th ed.

Washington, DC: Personal Care Products Council, 2016.

2. Elder RL (ed.). Final Report on the Safety Assessment of Hydrolyzed Collagen. JACT. 1985;4(5):199-221.

3. Andersen FA, Bergfeld WF, Belsito DV, Klaassen CD, Marks JG, Shank RC, Slaga TJ, and Snyder PW. Final Report of the Safety Assessment of Cosmetic Ingredients Derived from Zea Mays (Corn). Int J Toxicol. 2011;30(Suppl. 1):17S-39S.

4. Andersen FA (ed.). Annual Review of Cosmetic Ingredient Safety Assessments - 2004/2005. Int J Toxicol. 2006;25(Suppl. 2):1-89.

5. Andersen FA (ed.). Amended Final Report on the Safety Assessment of Oryza Sativa (Rice) Bran Oil, Oryza Sativa (Rice) Germ Oil, Rice Bran Acid, Oryza Sativa (Rice) Bran Wax, Hydrogenated Rice Bran Wax, Oryza Sativa (Rice) Bran Extract, Oryza Sativa (Rice) Extract, Oryza Sativa (Rice) Germ Powder, Oryza Sativa (Rice) Starch, Oryza Sativa (Rice) Bran, Hydrolyzed Rice Bran Extract, Hydrolyzed Rice Bran Protein, Hydrolyzed Rice Extract, and Hydrolyzed Rice Protein. Int J Toxicol. 2006;25(Suppl 2):91-120.

6. Johnson WJ, Bergfeld WF, Belsito DV, Hill RA, Klaassen CD, Liebler DC, Marks JG, Shank RC, Slaga TJ, Snyder PW, and Gill LJ. Safety Assessment of Silk Protein Ingredients as Used in Cosmetics. 1620 L St NW, Suite 1200, Washington, DC 20036, Cosmetic Ingredient Review. 2015.

7. Burnett CL, Heldreth B, Belsito DV, Hill RA, Klaassen CD, Liebler DC, Marks JG, Shank RC, Slaga TJ, Snyder PW, and Gill LJ. Safety Assessment of Soy Proteins and Peptides as Used in Cosmetics. 1620 L St, NW, Suite 1200, Washington, DC 20036, Cosmetic Ingredient Review. 2015.

8. Burnett CL, Heldreth B, Boyer I, Bergfeld WF, Belsito DV, Hill RA, Klaassen CD, Liebler DC, Marks JG, Shank RC, Slaga TJ, Snyder PW, and Gill LJ. Safety Assessment of Hydrolyzed Wheat Protein and Hydrolyzed Wheat Gluten as Used in Cosmetics. 1620 L St, NW, Suite 1200, Washington, DC 20036, Cosmetic Ingredient Review. 2014.

9. Chahal S. 2016. Letter from Dr. Surinder Chahal concerning the CJR review of Keratin and Hydrolyzed Keratin. Unpublished data submitted by Personal Care Products Council.

10. Rouse JG and Van Dyke ME. A review of keratin-based biomaterials for biomedical applications. Materials. 2010;3:999-1014.

11. Tonin C, Aluigi A, Varesano A, and Vineis C. Keratin-based nanofibres. Chapter: 7. Kumar A.InTech; 2010:139-158.

12. O'Neil MJ (ed). The Merck Index. 15th ed. The Royal Society of Chemistry, 2013.

13. Hill P, Brantley H, and Van Dyke M. Some properties of keratin biomaterials: Kerateines. Biomaterials. 2010;31:585-593.

14. Personal Care Products Council. 7-3-2012. Information on Hydrolyzed Keratin, Hydrolyzed Silk Protein, Hydrolyzed Elastin, Hydrolyzed Soy Protein, Hydrolyzed Milk Protein and Hydrolyzed Vegetable Protein. Unpublished data submitted by Personal Care Products Council. 25 pages.

15. Personal Care Products Council. 2012. Molecular Weights and Hydrolysis Method: Hydrolyzed Brazil Nut Protein, Hydrolyzed Hair Keratin, Hydrolyzed Keratin and Hydrolyzed Sweet Almond Protein. Unpublished data submitted by the Personal Care Products Council.


16. Personal Care Products Council. 5-14-2012. Method of manufacture and Molecular Weights: Hydrolyzed Protein Ingredients. Unpublished data submitted by Personal Care Products Council. 8 pages.

17. Wortmann FJ, Sendelbach G, and Popescu C. Fundamental DSC investigations of α-keratinous materials as basis for the interpretation of specific effects of chemical, cosmetic treatments on human hair. J Cosmet Sci. 2007;58:311-317.

18. Mohd Amir SNK, Gupta A, Tuan Chik SMS, and Ahmad K. Development of anti-ageing cream from chicken feathers. Res J Pharm Biol Chem Sci. 2014;5(6):389-393.

19. Arch Personal Care Products LP. 2012. Hydrokeratin AL-30 (Hydrolyzed Keratin) Manufacturing procedure. Unpublished data submitted by Personal Care Products Council. 8 pages.

20. Personal Care Products Council. 1-28-2016. Hydrolyzed Keratin. Unpublished data submitted by Personal Care Products Council.

21. Active Concepts. 2014. Certificate of origin (AC Keratin Hydrolysate SD - powder Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

22. Active Concepts. 2015. Certificate of origin (AC Keratin Hydrolysate 30- solution Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

23. Active Concepts. 2015. Certificate of origin (AC Cashmere Hydrolysate - solution Hydrolyzed Keratin from goat wool). Unpublished data submitted by Personal Care Products Council.

24. Anonymous. 2012. Summaries of Dermal and Ocular Irritation Tests of Hydrolyzed Protein Ingredients (including proteins hydrolyzed to amino acids). Unpublished data submitted by Personal Care Products Council. 4 pages.

25. Personal Care Products Council. 2016. Soluble Keratin. Unpublished data submitted by Personal Care Products Council.

26. Strnad P, Usachov V, Debes C, Gräter F, Parry DAD, and Omary MB. Unique amino acid signatures that are evolutionarily conserved distinguish simple-type, epidermal and hair keratins. J Cell Sci. 2011;124(24):4221-4232.

27. Food and Drug Administration (FDA). Frequency of use of cosmetic ingredients. FDA Database. 2016. Washington, DC: FDA.

28. Bremmer HJ, Prud'homme de Lodder LCH, and Engelen JGM. Cosmetics Fact Sheet: To assess the risks for the consumer; Updated version for ConsExpo 4. 2006. Report No. RIVM 320104001/2006. pp. 1-77.

29. Rothe H, Fautz R, Gerber E, Neumann L, Rettinger K, Schuh W, and Gronewold C. Special aspects of cosmetic spray safety evaluations: Principles on inhalation risk assessment. Toxicol Lett. 2011;205(2):97-104.

30. Rothe H. Special Aspects of Cosmetic Spray Evalulation. 9-26-2011. Unpublished data presented at the 26 September CIR Expert Panel meeting. Washington, D.C.

31. Johnsen MA. The Influence of Particle Size. Spray Technology and Marketing. 2004;14(11):24-27.

32. European Union. Regulation (EC) No. 1223/2009 of the European Parliament and of the Council of 30 November 2009 on Cosmetic Products. 2009. Internet site accessed September 13, 2013. http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2009:342:0059:0209:en:PDF


http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2009:342:0059:0209:en:PDF

33. Scientific Committee on Consumer Products (SCCP). Opinion on amino acids obtained by hydrolysis of human hair. 2005. http://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp_o_005.pdf. Date Accessed 2-4-2016. Report No. SCCP/0894/05.

34. Singaravelu S, Ramanathan G Raja MD, Barge S, and Sivagnanam UT. Preparation and characterization of keratin-based biosheet from bovine horn waste as wound dressing material. Materials Letters. 2015;152:90-93.

35. Noda J, Imai T, Kida K, and Otagiri M. The physicochemical and biopharmaceutical properties of fragmented keratin as a new drug carrier. Biol Pham Bull. 1996;19(3):466-473.

36. Barba C, Mendez S, Roddick-Lanzilotta A, Kelly R, Parra JL, and COderech I. Wool peptide derivatives for hand care. J Cosmet Sci. 2007;58:99-107.

37. Consumer Product Testing Co. 1983. Primary dermal irritation in rabbits; primary ocular irritation in rabbits; acute oral toxicity in rats (25% Hydrolyzed Keratin from hoof meal). Unpublished data submitted by the Personal Care Products Council.

38. Food and Drug Research Laboratories, Inc. 1976. Acute oral toxicity in rats (20% Hydrolyzed Keratin from hoof meal). Unpublished data from the Personal Care Products Council.

39. Toxicol Laboratories Limited. 1986. Single dose oral toxicity study in the rat (20% Hydrolyzed Keratin from hoof meal). Unpublished data submitted by the Personal Care Products Council.

40. Fernandes MM, Lima CF, Loureiro A, Gomes AC, and Cavaco-Paulo A. Keratin-based peptide: Biological evaluation and strengthening properties on relaxed hair. Int J Cosmet Sci. 2012;34:338-346.

41. BioReliance. 2004. Bacterial reverse mutation assay Hydrolyzed Keratin (MW - 3,000 Da). BioReliance Study Number AA88DT.502.BTL. Unpublished data submitted by Personal Care Products Council. 15 pages.

42. Active Concepts. 2016. Bacterial reverse mutation test (AC Cashmere Hydrolysate- solution Hydrolyzed Keratin from goat wool). Unpublished data submitted by Personal Care Products Council.

43. Active Concepts. 2016. Bacterial reverse mutation test (AC Keratin Hydrolysate 30 - solution Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

44. Active Concepts. 2016. Bacterial reverse mutation test (AC Keratin Hydrolysate SD- powder Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

45. Anonymous. 2016. Summary of keratin safety studies. Unpublished data submitted by Personal Care Products Council.

46. Toxicol Laboratories Limited. 1986. Primary skin irritation study: Hydrolyzed Keratin (MW - 125,000 Da) Study Ref. No. 63/8601. Unpublished data submitted by Personal Care Products Council.

47. Active Concepts. 2007. AC Cashmere Hydrolysate irritation analysis (AC Cashmere Hydrolysate - solution Hydrolyzed Keratin from goat wool). Unpublished data submitted by Personal Care Products Council.

48. Active Concepts. 2015. Dermal and ocular irritation tests (AC Keratin Hydrolysate 30 - solution Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.


http://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp_o_005.pdf

49. Active Concepts. 2012. Dermal and ocular irritation tests (AC Keratin Hydrolysate SD - powder Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

50. Consumer Product Testing Co. 1981. Primary dermal irritation in rabbits; primary ocular irritation in rabbits, acute oral toxicity in rats: Hydrolyzed Keratin (MW ~ 600 Da). Experiment Reference No: 81103-S. Unpublished data submitted by Personal Care Products Council.

51. Consumer Product Testing Co. 1984. Primary dermal irritation in rabbits; primary ocular irritation in rabbits; acute oral toxicity in rabbits: Hydrolyzed Keratin (MW ~ 500 Da) Experiment Reference No. 84229-1. Unpublished data submitted by Personal Care Products Council.

52. Consumer Product Testing Co. 2004. The MatTek Corporation EpiDerm™ skin model in vitro toxicity testing system: Hydrolyzed Keratin (MW ~ 3,000 Da). Experiment Reference Number: V04-0032-1. Unpublished data submitted by Personal Care Products Council.

53. Toxicol Laboratories Limited. 1986. Primary skin irritation study (20% Hydrolyzed Keratin from hoof meal). Unpublished data submitted by the Personal Care Products Council.

54. Food and Drug Research Laboratories, Inc. 1976. Rabbit eye irritation study (20% Hydrolyzed Keratin from hoof meal). Unpublished data submitted by the Personal Care Products Council.

55. Anonymous. 2001. Summary: Study of primary cutaneous irritation of a pharmaceutical or cosmetic compound in the male rabbit. Unpublished data submitted by Personal Care Products Council.

56. Food and Drug Research Laboratories, Inc. 1976. Primary skin irritation study with rabbits (20% Hydrolyzed Keratin from hoof meal). Unpublished data submitted by the Personal Care Products Council.

57. Consumer Product Testing Co. 2004. The Hen's egg test -- utilizing the chorioallantoic membrane (HET-CAM): Hydrolyzed Keratin (MW - 3,000 Da) Experiment Reference Number: V04-0033-1. Unpublished data submitted by Personal Care Products Council.

58. Toxicol Laboratories Limited. 1986. Eye irritation study: Hydrolyzed Keratin (MW - 125,000 Da) Study Ref. No. 64/8601. Unpublished data submitted by Personal Care Products Council.

59. Toxicol Laboratories Limited. 1986. Eye irritation study (20% Hydrolyzed Keratin from hoof meal). Unpublished data submitted by the Personal Care Products Council.

60. Anonymous. 2001. Study of ocular irritation of a pharmaceutical or cosmetic compound in the male rabbit. Unpublished data submitted by Personal Care Products Council.

61. Active Concepts. 2015. OECD TO 4420: In vitro skin sensitization (AC Cashmere Hydrolysate - solution Hydrolyzed Keratin from goat wool). Unpublished data submitted by Personal Care Products Council.

62. Active Concepts. 2015. OECD TO 442D: In vitro skin sensitization (AC Keratin Hydrolysate SD - powder Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

63. Active Concepts. 2015. OECO TG 4420: In vitro skin sensitization (AC Keratin Hydrolysate 30 - solution Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.


64. Consumer Product Testing Co. 2004. Repeated insult patch test: Hydrolyzed Keratin (MW ~ 3,000 Da). Experiment Reference Number: C04-0181.01. Unpublished data submitted by Personal Care Products Council.

65. McFadden JP, Rycroft RJG, White IR, Wakelin SH, and Basketter DA. Hydrolyzed protein shampoo additives are not a common contact allergen. Contact Dermatitis. 2000;43:243-243.

66. Niinimaki A, Niinimaki M, Makinen-Kiljunen S, and Hannuksela M. Contact urticaria from protein hydrolysates in hair conditioners. Allergy. 1998;53:1078-1082.

67. Freeman S and Lee MS. Contact urticaria to hair conditioner. Contact Dermatitis. 1996;35:195-196.

68. Active Concepts. 2014. Product specification (AC Keratin Hydrolysate SD- powder Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

69. Active Concepts. 2015. Product specification (AC Keratin Hydrolysate 30- solution Hydrolyzed Keratin from sheep wool). Unpublished data submitted by Personal Care Products Council.

70. Active Concepts. 2015. Product specification (AC Cashmere Hydrolysate- solution Hydrolyzed Keratin from goat wool). Unpublished data submitted by Personal Care Products Council.

71. Arnaud JC and Bore P. Ion-exchange chromatography for the analysis of protein derivatives and specific amino acids. Cosmet Sci Tech. 1985;4:223-244.

72. Personal Care Products Council. 4-9-2015. Concentration of Use by FDA Product Category: Hydrolyzed Keratin and other Keratin Ingredients. Unpublished data submitted by Personal Care Products Council.


2016 RAW FDA VCRP DATA 05A - Hair Conditioner HYDROLYZED HAIR KERATIN 16 05F - Shampoos (non-coloring) HYDROLYZED HAIR KERATIN 8 05G - Tonics, Dressings, and Other Hair Grooming Aids HYDROLYZED HAIR KERATIN 1 05I - Other Hair Preparations HYDROLYZED HAIR KERATIN 2 02B - Bubble Baths HYDROLYZED KERATIN 1 03A - Eyebrow Pencil HYDROLYZED KERATIN 1 03C - Eye Shadow HYDROLYZED KERATIN 1 03D - Eye Lotion HYDROLYZED KERATIN 1 03F - Mascara HYDROLYZED KERATIN 31 03G - Other Eye Makeup Preparations HYDROLYZED KERATIN 2 04E - Other Fragrance Preparation HYDROLYZED KERATIN 1 05A - Hair Conditioner HYDROLYZED KERATIN 110 05B - Hair Spray (aerosol fixatives) HYDROLYZED KERATIN 20 05C - Hair Straighteners HYDROLYZED KERATIN 6 05D - Permanent Waves HYDROLYZED KERATIN 3 05E - Rinses (non-coloring) HYDROLYZED KERATIN 5 05F - Shampoos (non-coloring) HYDROLYZED KERATIN 93 05G - Tonics, Dressings, and Other Hair Grooming Aids HYDROLYZED KERATIN 123 05H - Wave Sets HYDROLYZED KERATIN 1 05I - Other Hair Preparations HYDROLYZED KERATIN 32 06A - Hair Dyes and Colors (all types requiring caution statements and patch tests) HYDROLYZED KERATIN 124 06B - Hair Tints HYDROLYZED KERATIN 38 06C - Hair Rinses (coloring) HYDROLYZED KERATIN 21 06D - Hair Shampoos (coloring) HYDROLYZED KERATIN 2 06E - Hair Color Sprays (aerosol) HYDROLYZED KERATIN 5 06H - Other Hair Coloring Preparation HYDROLYZED KERATIN 3 08B - Cuticle Softeners HYDROLYZED KERATIN 2 08C - Nail Creams and Lotions HYDROLYZED KERATIN 1 08E - Nail Polish and Enamel HYDROLYZED KERATIN 4 08G - Other Manicuring Preparations HYDROLYZED KERATIN 3 10A - Bath Soaps and Detergents HYDROLYZED KERATIN 9 10B - Deodorants (underarm) HYDROLYZED KERATIN 1 12A - Cleansing HYDROLYZED KERATIN 2 12D - Body and Hand (exc shave) HYDROLYZED KERATIN 9 12F - Moisturizing HYDROLYZED KERATIN 8 12J - Other Skin Care Preps HYDROLYZED KERATIN 4


03A - Eyebrow Pencil KERATIN 1 03D - Eye Lotion KERATIN 1 03F - Mascara KERATIN 5 05A - Hair Conditioner KERATIN 28 05B - Hair Spray (aerosol fixatives) KERATIN 2 05C - Hair Straighteners KERATIN 1 05E - Rinses (non-coloring) KERATIN 1 05F - Shampoos (non-coloring) KERATIN 15 05G - Tonics, Dressings, and Other Hair Grooming Aids KERATIN 17 05H - Wave Sets KERATIN 1 05I - Other Hair Preparations KERATIN 15 08B - Cuticle Softeners KERATIN 1 08G - Other Manicuring Preparations KERATIN 1 12F - Moisturizing KERATIN 1 05F - Shampoos (non-coloring) SOLUBLE ANIMAL KERATIN 1


Personal Care iProducts CouncilCommitted to Safety,Qualify & Innovation

Memorandum

TO: Lillian Gill, D.P.A.Director - COSMETIC INGREDIENT REVIEW (CJR)

FROM: Beth A. Lange, Ph.D. •7n / <7Industry Liaison to the CIR Expert Panel trcL

DATE: March 24, 2016

SUBJECT: Comments on the Draft Report: Safety Assessment of Keratin and KeratinDerived Ingredients as Used in Cosmetics (prepared for the March 31-April 1,2016 CIR Expert Panel meeting)

Key IssuesThe Introduction implies that these ingredients may differ by source and states: “These variations

of the source material with one ingredient name are currently not well-defined in thissafety assessment.” Information provided by industry indicates the source of theingredients (consistent with Dr. Chahal’s letter, primarily sheep wool but also from hoofsand goat wool). This information is not always presented in the tables (reference 18 and23 both from sheep’s wool) and there is no mention of source of the ingredients tested inthe text of many sections, such as Genotoxicity, Irritation and Sensitization and theSummary.

The statement in the Summary that “An allergic reaction has been reported to a hair conditionercontaining Hydrolyzed Keratin” is misleading. The case report section indicates that thehair conditioner to which the subject reacted contained steartrimonium hydrolyzed animalprotein. Hydrolyzed animal protein is a former INCI name for Hydrolyzed Collagen. Thecase report section indicates that prick testing of this patient with quaternary hydrolyzedproteins and Hydrolyzed Keratin resulted in reactions. It is not stated if normal use ofproducts containing Hydrolyzed Keratin resulted in reactions in this patient.

Additional ConsiderationsAlthough it states that capitaLized names are used for cosmetic ingredients and lower-case names

were used for the proteins, it is not clear if this was consistently applied throughout thereport. For example, was a cosmetic ingredient used in the iv kinetic study of Keratin inmice (reference 33)?

Tables 3,4, 6, 7, 8, 9 and 10 - Please include the source for all entries in these tables.

1620 L Street, N.W., Suite 1200 Washington, D.C. 20036 202.331.1770 I 202.331.1969 (fax) www.personalcarecouncil.org


Table 6 - In the second row, the dose should be reported as 2 g/kg (rather than the incorrect 2000g/kg) to be consistent with the LD50 which is reported as >2 g/kg.

Table 7- Did reference 38 really test “formulations”? If this is not a finished product that couldbe used by a consumer, for the purposes of the CIR report, it would clearer to call thesemixtures or solutions.


Safety Assessment of Keratin and Keratin-Derived ... · Safety Assessment of Keratin and...

Documents

Transcript of Safety Assessment of Keratin and Keratin-Derived ... · Safety Assessment of Keratin and...