My research aim is to analyze the human risk of using popular professional non-ammonia based hair color products. My intent is to follow up with a second research paper aimed at comparing relative toxicity and carcinogenicity to popular professional ammonia based hair color products. I began by determining or verifying the constituencies of each product which was accomplished by gas chromatography and flame ionization detection. I then utilized Flame Atomic Absorption Spectroscopy to analyze the relative carcinogenicity of each product. Finally, I researched public information, product descriptions, and called manufacturer personnel to determine such qualitative factors as the social and environmental responsibility of the product manufacturers. My findings were significant especially in the case of the extremely high levels of deiethanolamine, a toxic and carcinogenic chemical which is especially dangerous when reacted with heat or acid, in one of the products tested.

1. Determination and Analysis the Toxicity and Carcinogenicity of the
Constituencies of Non-Ammonia based Hair Coloring Products using
Chromatography and Spectroscopy
March 2010
Executive Summary
The cultural movement towards environmental consciousness had led to a reemergence of non-
chemical, more natural, or even organic products. One product set that most warrants such a revolution
is that of hair color products which have long been dependent on ammonia as a key active ingredient for
years. This has led to several manufacturers developing non-ammonia based hair color products for the
professional salon industry that have varying degrees of toxic and carcinogenic properties. These new
products are marketed as safer and more natural although they still rely on oxidation to produce the
desired outcome of permanent hair color changes.
Hair color products have traditionally heretofore been reviewed and analyzed not just by the Food and
Drug Administration (FDA), but also private researchers. This magnifying glass has been the product of
two factors (1) hair colors are made of toxic chemical ingredients; and (2) hair colors are applied directly
to the scalp where pores and follicles allow absorption directly into the blood stream. Although the new
brands of non-ammonia based hair color products have been thoroughly reviewed by the FDA, they
have not undergone the same rigor of scrutiny by private independent researchers.
My research aim is to analyze the human risk of using popular professional non-ammonia based hair
color products. My intent is to follow up with a second research paper aimed at comparing relative
toxicity and carcinogenicity to popular professional ammonia based hair color products.
I began by determining or verifying the constituencies of each product which was accomplished by gas
chromatography and flame ionization detection. I then utilized Flame Atomic Absorption Spectroscopy
to analyze the relative carcinogenicity of each product. Finally, I researched public information, product
descriptions, and called manufacturer personnel to determine such qualitative factors as the social and
environmental responsibility of the product manufacturers.

2. My findings were significant especially in the case of the extremely high levels of deiethanolamine, a
toxic and carcinogenic chemical which is especially dangerous when reacted with heat or acid, in one of
the products tested.
Summary of Results
I determined Organic Color Systems to be the safest hair color product out of the four that I tested.
Organic Color Systems is manufactured in the United Kingdom by HerbUK. Being manufactured in the
United Kingdom, Organic Color Systems is subject to much more restrictive public safety regulations on
ingredients than that of manufacturers in the United States.
I determined that Mastey de Paris’ Teinture product to be the most highly toxic and carcinogenic. More
alarming, is the extremely high levels of deiethanolamine. In my opinion, this product should be
immediately removed from the market. I called Mastey de Paris which is located in California to share
my findings with them. They had refused to discuss Teinture or its ingredients list, and denied that their
product contained any deiethanolamine. I retested Teinture, using both tests, and verified that the test
results were internally reliable.
The table below exhibits a summary of my results.
Inoa Organic Color Mastey Ecocolors
(L’oreal) Systems Teinture
Passed Ammonia Test
Passed PPD Test
Passed MEA Test
Passed DEA Test
Lacks Carcinogenic Properties
Published Ingredients
No Animal Testing
Green Company Ethos
Ingredient/Product Guarantee
p-Phenylenediamine (PPD)
Para-phenylenediamine (PPD) (Figure 1) was first described in 18631 and by the
end of the 19th century the oxidative hair dye process had been invented.
Reactions between oxidizable aromatic amines, such as PPD, toluene-2,5-diamine
(fig.2), aminophenol, resorcinol and hydrogen peroxide made it possible to make
a permanent coloring of hair. Since the 1960s the coloring of hair has been
performed not only by professionals at hairdressing salons, but also as a popular
home cosmetic procedure2. As early as 1939 Bonnevie suggested resorcinol, PPD and aminophenol as
part of a patch test standard series for diagnosing allergic contact dermatitis in patients sensitized by
1
Corbett JF. Hair coloring. Clin.Dermatol. 1988; 6: 93-101.
2
Hofmann AW. Organische Basen. Jahresberichte ueber die fortschritte der chemie 1863; 3: 422.

3. dyed furs, hair dyes, or through occupational exposure3. PPD is still used for coloring of human hair and
the sales of hair dye products containing aromatic amines are substantial.
Sensitization to PPD has, in the past, been considered so great a hazard that its use in hair dye was
banned in Germany in 1906, in Sweden in 1943, in France in 1951 and in Sweden again in 19644. The 6th
amendment of the European Union Cosmetic Directive made ingredient labeling of cosmetic products
mandatory. This was a major improvement in dermatotoxicologic safety as the content of possible
contact allergens is listed on the label to the benefit of primary and secondary prevention. Today, the
reported risk for the development of bladder cancer in past users of PPD requires many governments to
put limits on this dangerous chemical. PPD is permitted in the European Union at a concentration of 6%
and toluene-2,5-diamine is permitted at a concentration of 10%5.
PPD is particularly dangerous in any product whose prescribed application is on human skin6. PPD
belongs to the group of para-substitued benzenes. In essence cross reactions can only be studied in
animal experiments. In patients it is not possible to distinguish between simultaneous reactions and
cross-reactions. The clinical experience is, however, that PPD may cross react to para-substituted hair
dyes such as toluene-2,5-diamine, p-aminophenol, 2-nitro-PPD7 and to disperse orange 38. PPD is
generally not an effective screening agent for azo dyes9. However cross reactions or simultaneous
reactions have been described, especially to disperse orange 3, p-aminoazobenzene and p-
dimethylaminoazobenzene10. A patient reacted to a PPD-containing hair dye and had cross reaction to
3
Bonnevie P. Aetiologie und pathogenese der Ekzemkrankheiten. Klinische Studien über die Ursachen der Ekzeme
unter besonderer Berücksichtigung des Diagnostischen Wertes der Ekzemproben. 1939. Busch, Copenhagen /
Barth, Leipzig.
4
Fregert S. [Chemical demonstration of paraphenylene diamine in hair dyes]. Hautarzt 1972; 23: 393-394.
5
Council directive 76/768/EEC of 27 July 1976 on the approximation of the laws of the member states relating to
cosmetic products, amended. European Communities off.Journal L262 27.9. 1976.
6
Seidenari S, Mantovani L, Manzini BM, Pignatti M. Cross-sensitizations between azo dyes and para-amino
compound. A study of 236 azo-dye-sensitive subjects. Contact Dermatitis 1997
7
Fautz R, Fuchs A, van der Walle HB, Henny V, Smits L. Hair dye-sensitized hairdressers: the cross-reaction pattern
with new generation hair dyes. Contact Dermatitis 2002; 46: 319-324.
8
Goon AT, Gilmour NJ, Basketter DA, White IR, Rycroft RJ, McFadden JP. High frequency of simultaneous sensitivity
to Disperse Orange 3 in patients with positive patch tests to para-phenylenediamine. Contact Dermatitis 2003; 48:
248-250.
9
Hausen BM. Contact allergy to disperse blue 106 and blue 124 in black "velvet" clothes. Contact Dermatitis. 1993;
28: 169-173.
10
Seidenari S, Giusti F, Massone F, Mantovani L. Sensitization to disperse dyes in a patch test population over a
five-year period. Am.J Contact Dermat. 2002; 13: 101-107.

4. disperse red 1711. PPD is also described to cross-react12 with N-isopropyl-N-phenylenediamine (IPPD)13,
and local anesthetics14.
Monoethanolamine (MEA)
Monoethanolamine, also called 2-aminoethanol or Ethanolamine (often
abbreviated as ETA or MEA), is an organic based, non-toxic compound that
is both a primary amine (due to an amino group in its molecule) and a
primary alcohol (due to a hydroxyl group)15. Monoethanolamine is organic
and found throughout nature but is most abundantly found in coconuts.
Like other amines, monoethanolamine acts as a weak base. If not reacted
with an acid like Hydrogen Peroxide to lower it pH level16, some forms of Ethanolamine are toxic,
flammable, corrosive, and colorless with an odor similar to that of ammonia. However, in hair colors;
Ethanolamine is reacted with the acid Peroxide and can be combined with other ingredients to eliminate
the odor and substantially neutralizes the toxicity, corrosiveness, or odor17.
Ethanolamine is not a dangerous chemical in low exposure limits and if not reacted with ethylene oxide
(C2H4O). In fact, the only effects that have been determined are mild skin irritation only if it is applied
directly to the skin in a pure form and allowed to sit for periods of time more than 1.5 hours18. Because
ethanolamine has a high alkalinity, it will cause a burning sensation if directly ingested but only in its
pure form19.
Diethanolamine (DEA)
Deiethanolamine is an anime that hydrophilic character of the
alcohol groups. Diethanolamine is products by reacting
monoethanolamine with ethylene oxide. Studies show that DEA
11
Ho SG, White IR, Rycroft RJ, McFadden JP. Allergic contact dermatitis from para-phenylenediamine in Bigen
powder hair dye. Contact Dermatitis 2004; 51: 93-94.
12
Shmunes E. Purpuric allergic contact dermatitis to paraphenylenediamine. Contact Dermatitis 1978; 4: 225-229.
13
Herve-Bazin B, Gradiski D, Duprat P, Marignac B, Foussereau J, Cavelier C, Bieber P. Occupational eczema from
N-isopropyl -N'-phenylparaphenylenediamine (IPPD) and N-dimethy-1,3 butyl-N'-phenylparaphenylenediamine
(DMPPD) in tyres. Contact Dermatitis 1977; 3: 1-15.
14
Sosted H, Johansen JD, Andersen KE, Menne T. Severe allergic hair dye reactions in 8 children. Contact
Dermatitis 2006; 54: 87-91.
15
Le Coz CJ, Schneider GA. Contact dermatitis from tertiary-butylhydroquinone in a hair dye, with cross-sensitivity
to BHA and BHT. Contact Dermatitis 1998
16
Kligman AM. The identification of contact allergens by human assay. II. Factors influencing the induction and
measurement of allergic contact dermatitis. J.Invest Dermatol. 1966
17
Bowling JC, Scarisbrick J, Warin AP, Downs AM. Allergic contact dermatitis from trideceth-2-carboxamide
monoethanolamine (MEA) in a hair dye. Contact Dermatitis 2002
18
Hathaway GJ, Proctor NH, Hughes JP, and Fischman ML [1991]. Proctor and Hughes’ chemical hazards of the
workplace. 3rd ed. New York,NY: Van Nostrand Reinhold.
19
Gosselin RE, Smith RP, Hodge HC [1984]. Clinical toxicology of commercial products. 5th ed. Baltimore, MD:
Williams & Wilkins.

5. inhibits the absorption of choline, which is required for brain development and maintenance.20 DEA is
more potent in regard to systemic toxicity and respiratory track irritation than any ammonia derivatives.
Exposure to DEA resulted in body and organ weight changes, clinical-and histo-pathological changes
indicative for mild blood, liver, kidney and testicular systemic accumulative toxicity.21 Also, A 2009 study
found that Deiethanolamine has potential acute, sub-chronic and chronic toxicity properties. 22
Diethanolamine, which can only be derived by reacting ethanolamine with ethylene oxide, is an
extremely dangerous substance having a high likelihood of converting into its corresponding
nitrosamines which are highly carcinogenic23. While diethanoline is used in many shampoos and soaps,
they are safe in very low concentrations and provide a form consistency to products when they are
diluted. However, even in moderately low volumes, diethanolamine will cause accumulative toxicity in
the liver, several types of cancer; irritation of the eyes, skin, nose, throat; eye burns, corneal necrosis,
skin burns, lacrimation (discharge of tears), and subtle but periodic coughing, sneezing.
Method for Testing
The determination of the constituencies of each hair color product was performed by both gas
chromatography and flame ionization detection to ensure that the results were consistent where
applicable. The determination of carcinogens in each hair color product was performed by Flame
Atomic Absorption Spectroscopy. The determination of carcinogens does not take into account
survivable dietary intake levels which may, in extreme cases, offset the carcinogenic effect.
The determination of the “soft factors”, such as “Green Company Ethos” and “Published Ingredients”
were performed by reviewing the respective company’s website, marketing materials, product
information packages, and in some cases; calling the company directly.
Flame Atomic Absorbtion Spectroscopy Results
ASTM ASTM ASTM ASTM ASTM
D3539 D3557 D516 D4658 D4658
Fe2+ Pb2+ Cd2+ S2- SO4
2-
Iona 0.608 0.003 0.005 0.044 0.059
Organic Color Systems 0.098 BDL BDL 0.019 0.018
Mastey Teinture 0.709 0.004 0.008 0.079 0.084
Ecocolors 0.702 0.003 0.006 0.084 0.077
20
University of North Carolina at Chapel Hill. 2009. Study Shows Ingredient Commonly Found In Shampoos May
Inhibit Brain Development.
21
Gamer AO, Rossbacher R, Kaufmann W, van Ravenzwaay B (2008). "The inhalation toxicity of di- and
triethanolamine upon repeated exposure". Food Chem Toxicol 46 (6): 2173–83.
22
Libralato G, Volpi Ghirardini A, Avezzù F (2009). "Seawater ecotoxicity of monoethanolamine, diethanolamine
and triethanolamine". J Hazard Mater. PMID 20022426
23
Diethanolamine(DEA): A Carcinogenic Ingredient in Cosmetics & Personal
Products:http://www.preventcancer.com/consumers/cosmetics/diethanolamine.htm

6. The results are based on the American Standard Test Method (ASTM). Five ions namely: Fe2+, Pb2+, Cd2+,
S2-, and SO42- were detected with concentrations varying concentrations. BDL was used to notate
“Below Detectable Level”.
The results for Iona and Organic Color Systems indicated much lower carcinogenic levels than Mastey
Teinture and Ecocolors particularly with the important Cd2+ ion. No permissible limits have been quoted
specifically in hair by any regulatory authority for any of the ions. However, permitted Dietary
Reference Intake level for Fe2+ in the human body is 45mg/kg. Permissible levels for Pb are 1.5μg/m324
and 50 μg/m325. In air the permissible levels for Cd are 5ppb26. Though the metals observed in the
sample are in traces, there is a high tendency for accumulation due to regular use of these dyes which
may lead to the hazardous effects associated with each of the identified elements. I have therefore
concluded Iona to be carcinogenic and Organic Color Systems to be non-carcinogenic.
No permissible limits have been quoted specifically in hair by regulatory authorities for any of the ions
or elements hence comparison with such values was not possible. The presence of radioactive isotopes
could serve as a possible explanation for the observation that the negative health impact of certain hair
color products is felt many years after initial use27.
Conclusions
The importance of private research on hair color products cannot be overstated.
The regulations enforcing more restrictive limitations on professional hair color product limitations that
are employed by countries such as the United Kingdom may have a positive effect on consumer health.
Products used exclusively by beauticians in beauty salons and labeled "For Professional Use Only,"28 are
not required to include the ingredients. However, consumers and professionals would be well served to
perform due diligence on product ingredients and exercise extreme caution and skepticism of products
that do not disclose their ingredients.
The motivations of businesses refuse to disclose their product’s ingredient list should be brought into
serious question as, at least in this study; serious dangers could be concealed in the process.
24
United States Environmental Protection Agency
25
United States Occupational Safety and Health Administration
26
United States Environmental Protection Agency
27
Allan, P. 2005. United States World Health Report on Harmful/Toxic Toiletries and Cancer Causing
Chemicals. Washington DC: Jonsson Cancer Centre, p. 1-4.
28
Federal Food, Drug & Cosmetic Act (1938 as amended).