title: Human percutaneous absorption of a direct hair dye comparing in vitro and in vivo results: implications for safety assessment and animal testing.
abstract: Although in vitro skin absorption studies often detect small residues of applied test material in the epidermis/dermis, it is uncertain whether the residue is within the living skin. We studied the dermal absorption of a hair dye hydroxyanthraquinone-aminopropyl methyl morpholinium methosulphate (HAM) in human skin in vivo and in vitro. In vivo, skin (back and scalp) received 0.5% HAM in a commercial formulation at 20microg/cm2 After 0.5 or 48h, skin was tape stripped, followed by cyanoacrylate biopsies (CAB). Sebum from scalp sites was collected for 48h. In vitro, skin was treated with 20mg/cm2 dye for 0.5h, penetration determined after 24h. In vivo, at 0.5h, total recovery (back) was 0.67microg/cm2 (tape strips+CAB). Fluorescence microscopy showed HAM in the hair follicle openings (HFO). At 0.5h, scalp tape strips contained 1.80microg/cm2, HFO 0.82microg/cm2. At 48h, HFO contained 0.21microg/cm2, sebum 0.80microg/cm2. In vivo, skin residues were in the non-living skin and eliminated via desquamation and sebum secretion. In vitro, the SC contained 1.50microg/cm2, epidermis/dermis 0.86microg/cm2, receptor fluid<0.04microg/cm2, a total of 0.90microg/cm2 was considered to be bioavailable. In vitro epidermis/dermis residues were nearly identical to those located in non-living skin in vivo. In conclusion, in vitro percutaneous penetration studies may produce seemingly bioavailable material , which raises the need for a Threshold of Skin Absorption (TSA) addressing a negligible dermal absorption in order to avoid unnecessary in vivo toxicity studies on substances that produce no significant human systemic exposure.
authors.affiliation: Department of Dermatology and Allergy, Clinical Research Centre for Hair and Skin Physiology, Charité-Universitätsmedizin, 10117-Berlin, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Lademann
authors.forename: H
authors.initials: H
authors.lastname: Richter
authors.forename: U
authors.initials: U
authors.lastname: Jacobi
authors.forename: A
authors.initials: A
authors.lastname: Patzelt
authors.forename: F
authors.initials: F
authors.lastname: Hueber-Becker
authors.forename: C
authors.initials: C
authors.lastname: Ribaud
authors.forename: F
authors.initials: F
authors.lastname: Benech-Kieffer
authors.forename: E K
authors.initials: EK
authors.lastname: Dufour
authors.forename: W
authors.initials: W
authors.lastname: Sterry
authors.forename: H
authors.initials: H
authors.lastname: Schaefer
authors.forename: J
authors.initials: J
authors.lastname: Leclaire
authors.forename: H
authors.initials: H
authors.lastname: Toutain
authors.forename: G J
authors.initials: GJ
authors.lastname: Nohynek
mesh_terms: D000826:Animal Testing Alternatives; D000818:Animals; D000880:Anthraquinones / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D006202:Hair Dyes / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity*; D018859:Hair Follicle / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008856:Microscopy, Fluorescence; D009025:Morpholines / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity*; D012629:Sebum / Q000378:metabolism; D012869:Skin Absorption / Q000502:physiology*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000880:Anthraquinones; D006202:Hair Dyes; D009025:Morpholines; C529916:hydroxyanthraquinone-aminopropyl methyl morpholinium methosulfate