Sensitization and cross-reactivity patterns of contact allergy to diisocyanates and corresponding amines: investigation of diphenylmethane-4,4'-diisocyanate, diphenylmethane-4,4'-diamine, dicyclohexylmethane-4,4'-diisocyanate, and dicylohexylmethane-4,4'-diamine.

BACKGROUND: Isocyanates are used in polyurethane production. Dermal exposure to isocyanates can induce contact allergy. The most common isocyanate is diphenylmethane diisocyanate used for industrial purposes. The isomer diphenylmethane-4,4'-diisocyanate (4,4'-MDI) is used in patch testing. Diphenylmethane-4,4'-diamine (4,4'-MDA) is its corresponding amine. Concurrent reactions to 4,4'-MDI and 4,4'-MDA have been reported, as have concurrent reactions to 4,4'-MDI and dicyclohexylmethane-4,4'-diisocyanate (4,4'-DMDI). OBJECTIVES: To investigate the sensitization capacities and the cross-reactivity of 4,4'-MDI, 4,4'-MDA, 4,4'-DMDI, and dicyclohexylmethane-4,4'-diamine (4,4'-DMDA). METHODS: The guinea-pig maximization test (GPMT) was used. RESULTS: The GPMT showed sensitizing capacities for all investigated substances: 4,4'-MDI, 4,4'-MDA, 4,4'-DMDI, and 4,4'-DMDA (all p < 0.001). 4,4'-MDI-sensitized animals showed cross-reactivity to 4,4'-MDA (p < 0.001) and 4,4'-DMDI (all p < 0.05). 4,4'-MDA-sensitized animals showed cross-reactivity to 4,4'-DMDA (p = 0.008). CONCLUSION: All of the investigated substances were shown to be strong sensitizers. Animals sensitized to 4,4'-MDI showed cross-reactivity to 4,4'-MDA and 4,4'-DMDI, supporting previous findings in the literature. The aromatic amine 4,4'-MDA showed cross-reactivity to the aliphatic amine 4,4'-DMDA.

Is p-tert-butylphenol-formaldehyde resin (PTBP-FR) in TRUE Test® (Mekos test) sensitizing the tested patients?

BACKGROUND: In a population study using TRUE Test®, we noted late reactions to p-tert-butylphenol-formaldehyde resin (PTBP-FR) in 0.5% of subjects tested. OBJECTIVES: In order to explore possible test sensitization, differences in the contents of sensitizers within PTBP-FR in test preparations for TRUE Test® and Finn Chambers® were analysed. Subjects allergic to PTBP-FR and subjects with late reactions to PTBP-FR were retested in order to explore whether these groups reacted to different PTBP-FR sensitizers. PATIENTS/MATERIALS/METHODS: Four individuals with late reactions and 5 subjects with established allergy to PTBP-FR were retested with defined PTBP-FR sensitizers. PTBP-FR constituents in patches from TRUE Test® were analysed with high-performance liquid chromatography. Previously analysed samples of PTBP-FR constituents served as a reference. RESULTS: The pattern of reaction to PTBP-FR sensitizers was similar in both groups. Subjects with suspected sensitization had somewhat stronger reactions than controls. The concentrations of monomers, dimers and trimers were generally higher in the TRUE Test® resin than in reference substances. CONCLUSIONS: Retesting did not add information regarding causes of possible sensitization. Analysis showed that the resin used in TRUE Test® has a lower degree of polymerization or condensation, which may enhance its sensitizing properties. A follow-up of late reactions to PTBP-FR in TRUE Test® should be carried out.

Are gloves sufficiently protective when hairdressers are exposed to permanent hair dyes? An in vivo study.

BACKGROUND: The use of permanent hair dyes exposes hairdressers to contact allergens such as p-phenylenediamine (PPD), and the preventive measures are insufficient. OBJECTIVES: To perform an in vivo test to study the protective effect of gloves commonly used by hairdressers. PATIENTS/MATERIALS/METHODS: Six gloves from Sweden, Italy and Germany were studied: two vinyl, one natural rubber latex, two nitrile, and one polyethylene. The hair dye used for the provocation was a dark shade permanent dye containing PPD. The dye was mixed with hydrogen peroxide, and 8 PPD-sensitized volunteers were tested with the gloves as a membrane between the hair dye and the skin in a cylindrical open chamber system. Three exposure times (15, 30 and 60 min) were used. RESULTS: Eczematous reactions were found when natural rubber latex, polyethylene and vinyl gloves were tested with the dye. The nitrile gloves gave good protection, even after 60 min of exposure to the hair dye. CONCLUSIONS: Many protective gloves used by hairdressers are unsuitable for protection against the risk of elicitation of allergic contact dermatitis caused by PPD.

Analysis of Hexavalent Chromium in Cement Samples From Countries Within and Outside the EU: A Study From the International Contact Dermatitis Research Group.

Background: Allergic contact dermatitis (ACD) caused by hexavalent chromium (Cr(VI)) is often severe and difficult to treat. The content of Cr(VI) in cement can be reduced by, for example, addition of iron(II) sulfate. Since 2005 the content of Cr(VI) in cement is regulated in the EU Directive 2003/53/EC and must not exceed 2 ppm. Since this regulation came into force, ACD caused by cement has markedly been reduced. Objective: To investigate Cr(VI) and total chromium content in samples of cement from countries within and outside the EU. Methods: The members of the International Contact Dermatitis Research Group (ICDRG) were invited to participate in the study with the aim to collect cement samples from geographically different areas. The content of Cr(VI) in the samples was estimated by the diphenyl carbazide spot test, atomic absorption spectroscopy was used to assess the total chromium content. Results: Forty-five cement samples were analyzed, containing amounts of Cr(VI) from <0.1 to >70 ppm. Twenty-one samples contained >2 ppm Cr(VI), 24 contained less. Four of 17 samples from within the EU contained >2 ppm Cr(VI), that is, higher amounts than stipulated in the EU directive, as compared with 17 samples from countries outside the EU. Conclusion: In countries outside the EU, significantly more cement samples contained >2 ppm Cr(VI).

High frequency of false-positive reactions in attempted patch testing with acrylate/methacrylate mixes.

BACKGROUND: Although acrylate/methacrylate allergy has been frequently reported, until now patch testing with this group of allergens has been unwieldy, requiring the application of large supplementary series in most centres. OBJECTIVES: To formulate and evaluate two mixes of acrylate/methacrylate allergens in three centres (Malmö, Singapore, and Leuven). PATIENTS/MATERIALS/METHODS: All patients tested with the baseline series during the study period were also patch tested with the mixes. Mix 1 consisted of: triethyleneglycol diacrylate (TREGDA) 0.1% wt/wt, 2-hydroxyethyl methacrylate (2-HEMA) 1.0% wt/wt and ethyleneglycol dimethacrylate 1.0% wt/wt in petrolatum. Mix 2 consisted of: TREGDA 0.1% wt/wt and 2-HEMA 2.0% wt/wt in pet. The separate components of the two mixes were also tested simultaneously. RESULTS: There were 25 (5 males; 20 females) positive reactions to mix 1 with 16 in Malmö, 8 in Singapore, and 1 in Leuven. Positive reactions to mix 2 were seen only in Malmö, in 8 female patients. Thus, the positive reaction rate for mix 1 was 8.3% overall (Malmö 7.7%, Singapore 18.6%, and Leuven 2.1%), and that for mix 2 was 2.7% overall (Malmö 3.8%, Singapore 0%, and Leuven 0%). Of the 16 positive reactions to mix 1 in Malmö, only 4 were considered to be true allergic reactions, as the component allergen testing gave totally negative results in 12/16. For mix 2, only 3/8 positive reactions were considered to be true allergic reactions, as the component testing was negative in 5/8. Many doubtful (10-20%) and positive but non-allergic reactions were recorded, leading to early termination of the study. CONCLUSIONS: Although this was an unsuccessful attempt to formulate an acrylate/methacrylate mix, our experience will be useful for those embarking on future attempts to do this.

Sensitization to dimethyl fumarate with multiple concurrent patch test reactions.

BACKGROUND: Chairs and sofas imported from China to Europe were shown to contain dimethyl fumarate (DMF), a sensitizing, volatile chemical. Many of the sensitized patients also had positive patch test reactions to acrylates. OBJECTIVES: To analyse the occurrence and strength of DMF sensitization and the appearance of concomitant reactions. METHODS: Patch testing with DMF in concentrations of 0.1-0.00001% was carried out in 37 patients. Diethyl fumarate (DEF), diethyl maleate (DEM), dimethyl maleate (DMM), ethyl acrylate (EA), methyl acrylate (MA), and methyl methacrylate (MMA) were also tested with a dilution series at equimolar concentrations. RESULTS: The lowest concentration of DMF eliciting a reaction varied between 0.0001% and 0.1% and all but four patients reacted concurrently to DEF. DEM elicited positive patch test reactions in 21/37 patients and DMM reactions were seen in all 9 patients tested. EA elicited positive reactions in 13/37 patients and a positive MA reaction was seen in 7/37 patients, 2 of whom also reacted to MMA. CONCLUSIONS: The strength of the sensitization to DMF showed variation and concurrent reactions were common. Concurrent reactions to (meth)acrylates were seen in patients, who reacted to lower (0.001% or less) DMF concentration probably elicited by cross-reactivity.

Can simultaneous contact allergies to phenyl glycidyl ether and epoxy resins of the bisphenol A/F-types be explained by contamination of the epoxy resins?

BACKGROUND: Simultaneous contact allergies to epoxy resins based on diglycidyl ether of bisphenol A (DGEBA-R) or epoxy resins of the bisphenol F-type and the reactive diluent phenyl glycidyl ether (PGE) have been reported. The reason might be cross-reactivity, exposure to an epoxy resin system with PGE as a component, or contamination by PGE in the epoxy resin. AIMS: To study contamination by PGE, 20 commercial epoxy resins were analysed for the presence of PGE. To study contact allergy to PGE and its relation to epoxy resins by inserting PGE in the standard series. RESULTS: Among 2227 patients, 7 reacted to PGE. Of 23 (30%) patients, 7 with contact allergy to DGEBA-R and 7/19 (37%) with contact allergy to an epoxy resin of the bisphenol F-type reacted to PGE. All 7 patients with contact allergy to PGE reacted both to the DGEBA-R and to the epoxy resin of the bisphenol F-type. PGE was found in 90% of the investigated resins. The amounts of PGE ranged between 0.004% w/w and 0.18% w/w. CONCLUSION: Most probably, the presence of PGE as a contaminant in epoxy resins is of minor importance for the sensitization, but possibly the contamination of PGE might elicit contact dermatitis in individuals with a high reactivity to PGE.

Occupational airborne allergic contact dermatitis from methacrylates in a dental nurse.

BACKGROUND: There are very few reports of airborne allergic contact dermatitis from methacrylates. OBJECTIVES: To report a dental nurse with facial eczema supposedly caused by airborne methacrylates present in the work environment. METHODS: Patch testing with serial dilutions of several methacrylates and work provocations in environments containing methacrylates was performed. RESULTS: Patch testing with serial dilutions of several methacrylates disclosed a high patch test reactivity. Repeated provocations when working with methacrylates resulted in facial eczema that resolved out of work. Attempts to collect the sensitizers using air pumps for the collection of vapors and filters for the collection of air-born aerosols failed. CONCLUSIONS: The clinical presentation was that of a facial dermatitis due to airborne exposure to methacrylates. It seems likely that 1 or several of these allergens caused the dermatitis.

How to optimize patch testing with diphenylmethane diisocyanate.

We have previously shown that patch test preparations of polymeric diphenylmethane diisocyanate (PMDI) are more stable than preparations of diphenylmethane-4,4'-diisocyanate (4,4'-MDI). This study was conducted to (i) investigate whether PMDIs yield as many positive reactions as 4,4'-MDI, (ii) study concurrent reactions to 4,4'-MDI and 4,4'-diaminodiphenylmethane (4,4'-MDA), and (iii) follow the course of positive reactions during 4 weeks. It was shown that PMDIs detect as many positive reactions as 4,4'-MDI. Thus, they are better patch test agents being more stable than preparations of 4,4'-MDI. We recommend that PMDIs with a monomer content of at least 35% is used in 2.0% petrolatum (pet.) (i.e. monomer patch test concentration approximately 0.7%). It was shown that reactions to 4,4'-MDI and PMDIs appear late and we recommend readings on both day (D) 3/4 and D7. 4,4'-MDA was shown to be a good marker for 4,4'-MDI and patch testing with 4,4'-MDA in 0.25% pet. can be used instead of PMDI. Concomitant reactions to 4,4'-MDI and 4,4'-MDA are probably not caused by conversion of 4,4'-MDI into 4,4'-MDA by reaction with water. Another explanation is a path of reactions leading to ureas and MDI conjugates with skin constituents, which are hydrolysed into 4,4'-MDA. This complex process depends upon several factors and might explain why positive MDI reactions appear after D7.

Contact allergy to acrylates/methacrylates in the acrylate and nail acrylics series in southern Sweden: simultaneous positive patch test reaction patterns and possible screening allergens.

In a recent study we showed that all our dental personnel/patients were detected with 2-hydroxyethyl methacrylate (2-HEMA) and 2,2-bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (bis-GMA). We studied 90 patients tested to the acrylate and nail acrylics series at our department over a 10 year period to see whether screening allergens could be found. Patch testing with an acrylate and nail acrylics series was performed. Among the 10 acrylate/methacrylate-allergic occupational dermatitis patients tested to the acrylate series, the most common allergens were triethyleneglycol diacrylate (TREGDA, 8), diethyleneglycol diacrylate (5), and 1,4-butanediol diacrylate (BUDA, 5). All 10 of these patients would have been picked up by a short screening series combining TREGDA, 2-hydroxypropyl methacrylate (2-HPMA), and BUDA or 1,6-hexanediol diacrylate (HDDA). Among the 14 acrylate/methacrylate-allergic nail patients, the most common allergens were ethylene glycol dimethacrylate (EGDMA, 11), 2-HEMA, (9), and triethyleneglycol dimethacrylate (9). Screening for 3 allergens i.e. 2-HEMA plus EGDMA plus TREGDA, would have detected all 14 nail patients. A short screening series combining 2-HEMA, EGDMA, TREGDA, 2-HPMA, bis-GMA, and BUDA or HDDA would have picked up all our past study patients (dental, industrial, and nail) with suspected allergy to acrylate/methacrylate allergens.

Is it possible to improve the patch-test diagnostics for isocyanates? A stability study of petrolatum preparations of diphenylmethane-4,4'-diisocyanate and polymeric diphenylmethane diisocyanate.

We have previously shown that the concentration of diphenylmethane-4,4'-diisocyanate (4,4'-MDI) in commercial test preparations was so low that patch testing with the same was not reliable. The stability of 4,4'-MDI in petrolatum (pet.) was compared with pet. preparations of polymeric diphenylmethane diisocyanate (PMDI), which consists of a complex mixture of monomeric isomers and oligomers of MDI. Preparations of 4,4'-MDI and PMDI were stored under 3 different conditions, i.e. at room temperature, refrigerated and frozen. They were analysed continuously during 1 year with regard to the content of 4,4'-MDI, 3-ring oligomers and 4-ring oligomers using liquid chromatography-mass spectrometry. PMDI preparations kept frozen were stable for a year. All other preparations failed to fulfil the requirements of stability, i.e. +/-20% of the initial concentration. Storage in a freezer prolonged the lifetime for 4,4'-MDI. The decrease in concentration for preparations kept at room temperature and refrigerated was less rapid in PMDI preparations than in 4,4'-MDI preparations. PMDI preparations are better suited for patch testing patients exposed to MDI because they are more stable and homogeneous than 4,4'-MDI preparations. They better reflect possible allergens that workers are exposed to because products used in industry contain both monomers and oligomers.

Contact allergy to (meth)acrylates in the dental series in southern Sweden: simultaneous positive patch test reaction patterns and possible screening allergens.

Contact allergy to dental allergens is a well-studied subject, more so among dental professionals than dental patients. 1632 subjects had been patch tested to either the dental patient series or dental personnel series at the department of Occupational and Environmental Dermatology, Malmö, Sweden. Positive patch tests to (meth)acrylate allergens were seen in 2.3% (30/1322) of the dental patients and 5.8% (18/310) of the dental personnel. The most common allergen for both groups was 2-hydroxyethyl methacrylate (2-HEMA), followed by ethyleneglycol dimethacrylate (EGDMA), triethyleneglycol dimethacrylate, and methyl methacrylate. 47 (29 dental patients and 18 dental personnel) out of these 48 had positive patch tests to 2-HEMA. All 30 subjects who had a positive reaction to EGDMA had a simultaneous positive reaction to 2-HEMA. One dental patient reacted only to 2,2-bis[4-(2-hydroxy-3-methacryloxypropoxy) phenyl]propane (bis-GMA). From our data, screening for (meth)acrylate contact allergy with 2-HEMA alone would have picked up 96.7% (29/30) of our (meth)acrylate-allergic dental patients and 100% (18/18) of our (meth)acrylate-allergic dental personnel. The addition of bis-GMA in dental patients would increase the pick-up rate to 100%.

Sensitizing capacity of some trimers in p-tert-butylphenol-formaldehyde resin.

Contact allergy to p-tert-butylphenol formaldehyde resin (PTBP-F-R) is not rare. This resin consists of a large number of substances, most of which are still unknown. For diagnostic and preventive reasons the chemical identity of the sensitizers should be known, as well as their sensitizing capacities, cross-reaction patterns and presence in the environment. The aims of this study were to investigate the sensitizing capacities and potential cross-reacting patterns for 4-tert-butyl- 2,6-bis-(5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzyloxymethyl)-phenol (XIII), 4-tert-butyl-2- (5 - tert - butyl - 2 - hydroxy-benzyloxymethyl) - 6 - (5 - tert - butyl - 2 - hydroxy - 3 - hydroxymethyl-benzyloxy methyl)-phenol (XIVa) and 7,15,23-tri-tert-butyl-25,26,27-trihydroxy-2,3,10,11,18,19-hexahomo-3,11,19-trioxacalix(3)arene (XVIII) by the guinea pig maximization test. 4-tert-Butyl-2,6-bis-hydroxymethyl-phenol, 4-tert-butylbenzene-1,2-diol, 4-tert-butyl-2-hydroxymethyl-phenol, 4-tert-butyl-phenol, 4-tert-butyl-2-(5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzyloxymethyl)-6-hydroxymethyl-phenol, 4-tert-butyl-2-[5-tert-butyl-3-(5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzyloxymethyl) - 2 -hydroxy-benzyloxymethyl] - 6 - (5 - tert-butyl- 2 -hydroxy- 3 -hydroxymethyl-benzyloxymethyl)- phenol and were used as potential cross-reacting substances. In this study it is strongly indicated that the linear trimer XIII has a sensitizing capacity in the guinea pig which was significant when compared to the controls (p = 0.024). No cross-reactions were detected in animals induced with the linear trimer XIII. The linear trimer XIVa and the cyclic trimer XVIII failed to induce sensitization.

Sensitizing capacity of two monomeric aldehyde components in p-tert-butylphenol-formaldehyde resin.

Contact allergy to p-tert-butylphenol-formaldehyde resin is not rare. This resin consists of a large number of substances, most of which are unknown. For diagnostic and preventive reasons, the chemical identity of the sensitizers should be known as well as their sensitizing capacities, cross-reaction patterns and presence in the environment. The aim of this study was to investigate the sensitizing capacities and cross-reaction patterns for 5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde and 5-tert-butyl-2-hydroxy-benzaldehyde in the guinea pig maximization test. 2,6-Dimethylol p-tert-butylphenol, p-tert-butylcate chol, 2-methylol p-tert-butylphenol, p-tert-butylphenol, 4-tert-butyl-2-(5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzyloxymethyl)-6-hydroxymethyl-phenol and 4-tert-butyl-2-(5-tert-butyl-2-hydroxy-benzyloxymethyl)-phenol were used as potential cross-reacting substances. 5-tert-Butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde was shown to be a sensitizer (p = 0.041). In animals induced with this compound no cross-reactions to the putative cross-reacting substances were seen. In contrast, 5-tert-butyl-2-hydroxy-benzaldehyde failed to induce sensitization and no cross-reactions were detected.

Contact allergy to o-cresol--a sensitizer in phenol-formaldehyde resin.

BACKGROUND: In patients hypersensitive to phenol formaldehyde resin (PFR) it is, for therapeutic and preventive reasons, important to know the identity of the primary sensitizing substances, their sensitizing capacity, as well as their cross-reaction patterns. When elucidating the issue of cross reactivity in patients with contact allergy to simple methylol phenols (MP), o-cresol was shown to be a contact sensitizer. Besides cross reactivity, contamination of one or more MP(s) in o-cresol as well as o-cresol being a sensitizer of its own in PFR were possible explanations of the simultaneous positive patch test reactions to MP and o-cresol. OBJECTIVE: The aim of this study was to investigate if the simultaneous allergic reactions to PFR and o-cresol could be explained by the presence of this substance in PFR. METHODS: Patch testing, high-pressure liquid chromatography (HPLC), nuclear magnetic resonance spectrometry (NMR), gas chromatography (GC), and mass spectrometry (MS) were used. RESULTS: o-Cresol was isolated from the specific PFR used in our standard patch test series and identified. The concentration in the resin was 0.066% wt/wt. CONCLUSION: The current study establishes o-cresol as a contact sensitizer in a PFR. The observed reactions to o-cresol could be on the basis of cross reactivity or primary sensitization.

Low-molecular-weight contact allergens in p-tert-butylphenol-formaldehyde resin.

BACKGROUND: p-tert-Butylphenol-formaldehyde resin (PTBPFR) is a contact allergen that is included in most standard patch test series. This resin consists of a large number of substances most of which are unknown. OBJECTIVE: The aim of this study was to investigate if allergens in PTBPFR are molecules mainly of low (MW < 250), medium, or high molecular weight (MW > 1,000); to isolate and identify some of the low molecular weight components of the resin; and for new substances to determine if these were allergens. METHODS: Gel permeation chromatography, patch testing, high-pressure liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectrometry were used. RESULTS: Patch test reactions indicated allergens in low-, medium-, and high-molecular-weights fractions. The strongest patch test reactivity was seen to a medium molecular weight fraction constituting only 6% wt/wt of the resin for 4 of the patients. Two patients reacted positively to low molecular weight fractions, and one of these patients reacted only to these fractions. The following substances were isolated: 4-tert-butyl-[1,2]benzoquinone, 4-tert-butyl-2,6-bis-hydroxymethyl-phenol, 4-tert-butylbenzene-1,2-diol, 4-tert-butyl-2-hydroxymethyl-phenol, 5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde, 4-tert-butyl-2-hydroxymethyl-6-methoxymethyl-phenol, and p-tert-butylphenol. Patients reacted positively to 5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde and 4-tert-butyl-2-hydroxymethyl-6-methoxymethyl-phenol but not to 4-tert-butyl-[1,2]benzoquinone. CONCLUSION: Two new allergens in PTBPFR were found: 5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde and 4-tert-butyl-2-hydroxymethyl-6-methoxymethyl-phenol.

Contact allergy to the monomers in p-tert-butylphenol-formaldehyde resin.

In many adhesive formulations p-tert-butylphenol-formaldehyde resin (PTBP-F-R) is used as a binder. Contact allergy to this resin is not rare. In patients hypersensitive to PTBP-F-R, and butylphenol derivatives therein, it is for diagnostic and preventive reasons necessary to know the nature of the primary sensitizing substances, as well as the cross-reaction patterns for these. The aim of this study was to investigate contact allergy to monomers in PTBP-F-R and potential cross-reacting substances. 12 patients hypersensitive to PTBP-F-R were patch tested with 2 monomers, the raw materials formaldehyde and p-tert-butylphenol, and 3 closely related substances. High pressure liquid chromatography (HPLC) was used to investigate the purity of the test substances. It was shown that the monomers 2-methylol p-tert-butylphenol and 2,6-dimethylol p-tert-butylphenol could elicit allergic reactions in humans hypersensitive to PTBP-F-R. No simultaneous reactions or cross-reactions were shown to formaldehyde, p-tert-butylphenol, p-tert-butylcatechol, 2(3)-tert-butyl-4-hydroxyanisole (BHA) or 3,5-di-tert-butyl-4-hydroxytoluene (BHT). It was also shown that low amounts of contaminants in the test substances, if not taken into account, could influence the conclusions drawn from the test results obtained.