STUDY Identification and Quantification of para -Phenylenediamine in a Temporary Black Henna Tattoo Ronald R. Brancaccio, Lance H. Brown, Young Tae Chang, Joshua P. Fogelman, Erick A. Mafong, and David E. Cohen Background: Temporary black henna tattoos are very popular as body adornment. Although contact allergy to natural henna is unusual, the inclusion of hair dye, p-phenylenediamine (PPD), increases the risk of contact sensitization. Objective: This study was performed to identify the presence and concentration of PPD in a black henna tattoo mixture to which our patient developed contact allergy. Methods: The presence of PPD in a black henna tattoo mixture, various samples of commercially available henna powders, and several hair dye products was qualitatively and quantitatively detected using high performance liquid chromatography (HPLC). Results: This study demonstrated that PPD was present in the black henna tattoo mixture at a concentration of 15.7%, which is significantly higher than commercial hair dye preparations. Conclusion: The presence of PPD in black henna tattoo mixtures in high concentration poses a health hazard and a risk of allergic contact sensitization with potential long-term consequences. Copyright 2002, Elsevier Science (USA). All rights reserved. T HE ART OF BODY ADORNMENT by body piercing and tattooing has become increasingly popular in our modern culture. Temporary henna tattoos are readily available worldwide, last several weeks on the skin, and offer a self-limited, convenient alternative to a permanent tattoo. Since ancient times, henna has been used to paint the skin and dye hair. Although delayed 1-3 and immedi- ate 4,5 hypersensitivity to henna has been reported, it is considered a rare allergen. The addition of p-phenylenediamine (PPD), which is widely recognized as a sensitizer, increases the risk of allergic contact dermatitis from these henna tattoo mix- tures and a number of cases have been reported. 6-11 We identified the presence of PPD both qualitatively and quan- titatively in a black henna tattoo mixture by high-perfor- mance liquid chromatography (HPLC). Case Report A 37-year-old woman painted 2 black henna tattoos on her left upper arm and lower back. Within 24 to 48 hours, a pruritic dermatitis developed within the designs of both tattoos. Oral prednisone and topical clobetasol proprionate cream were given, and resolution occurred in 2 weeks (Fig 1). Ten years before presentation, the patient developed an allergic reaction in her scalp following the use of a hair dye, which also required oral prednisone for treatment. Previ- ously, she had worked as a hairdresser, but did not recall any problems with exposure to hair dyes. She denies any recent occupational exposure to PPD and currently works as a management consultant. Patch testing was performed on the inner aspects of her arms using Finn Chambers on Scanpor tape (Allerderm, Laboratories, Petaluma, CA). The materials tested in- cluded the black henna tattoo mixture, reconstituted with tap water to a paste consistency as directed by the inserted package instructions; PPD 1% pet. (Chemotechnique, Che- motechnique Diagnostics, AB, Malmo ¨, Sweden); lawsone 1.0% in petrolatum; and 3 different preparations of com- mercially available henna powders, each at a 1.0% and 10.0% aq. concentration. Within 7 hours, the patch test sites of PPD and the black henna tattoo mixture became so severely pruritic that the patient removed them from her left arm. At 24 hours after application, strongly positive 3+ (erythema, edema, and vesicles) reactions were noted at both sites (Fig 2). These reactions persisted and remained strongly positive by the second reading at 1 week. The patch test results to lawsone and the henna mixtures on the right arm were negative at 48 hours and at 1 week. No reactions were noted in 10 control patients patch tested with lawsone and the 3 henna samples. Materials and Methods of HPLC A stock solution of PPD (Aldrich Chemical Co, Milwaukee, WI) was made in dimethylsulfoxide (DMSO), 10 mg/mL (Fisher Scientific, Pittsburgh, PA). Dilutions were made in acetonitrile (Fisher Scientific, Pittsburgh, PA) to give 10, 5, From the Ronald O. Perelman Department of Dermatology, New York University Medical Center and the Department of Chemistry, New York University, New York, NY. Address reprint requests to Ronald R. Brancaccio, MD, 67 Perry St, New York, NY 10014. Copyright 2002, Elsevier Science (USA). All rights reserved. 1046-199X/02/1301-0004$35.00/0 doi:10.1053/ajcd.2002.30466 American Journal of Contact Dermatitis, Vol 13, No 1 (March), 2002: pp 15-18 15