A method for identifying causative chemicals of allergic contact dermatitis using a combination of chemical analysis and patch testing in patients and animal groups: application to a case of rubber boot dermatitis

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Abstract
A 63year‐old woman developed allergic contact dermatitis from rubber boots, Initial investigation, by patch testing in the patient and chemical analysis of the causative rubber boots, revealed that mercaptobenzothiazole (MBT) and dibenzothiazyl disulfids (MBTS) were the causative chemicals Subsequent investigations were performed by patch testing in animal groups. An extract of the causative rubber boots. MBT and MBTS were used for sensitization of guinea pigs by the guinea pia maximization test(GPMT). 3 animal groups. A (with the boot extract). B(with MBT)and (with MBTS) were Successfully prepared, The boot extract was fractionated by column chromatography and thin‐layer chromatography (TLC). Each Fraction was subjected to patch testing in the animal groups. Positive reactions in all groups would show that the active Tractions contained MBT‐type compounds. whereas a positive reaction in group A hut negative ones in group B and C would show that the active fractions did not contain my MBT‐type compounds. Each fraction was then analyzed by gas chromatography (GC).GC mass spectrometry (GC‐MS), direct inlet‐MS (DI‐MS) and high‐performance liquid chromatopgraphy (HPLC). By this investigation, we found not only Known allergens (MBT. MBTS). but also unknown allergens. S‐substituted MBT‐type compounds and Styrenated phenol (SP). Thus, SP was Shown to be a candidate as a human sensitizer even though the patient did not react to it.