benzofurans and hydroquinone

Two new reagents, NDTE (2,5-dihydroxyphenylacetic acid, 2,5-bis-tetrahydropyranyl ether p-nitrophenyl ester) and HLTE (homogentisic gamma-lactone tetrahydropyranyl ether), are described for the chemical derivatization of primary and/or secondary amines to form an electrochemically active product. These reagents undergo reaction with the aforementioned analytes to form a product possessing the hydroquinone moiety, thus allowing for reversible electrochemical detection at mild oxidation potentials. The reactivity of each reagent was demonstrated by using N-ethylbenzylamine (EBzA) and the dipeptide isoleucine leucine methyl ester as model analytes. The investigation included the isolation and identification of the intermediates and final products from derivatization of EBzA. These isolated standards were subsequently characterized with respect to electrochemical properties by means of cyclic voltammetry. In LC-EC experiments, the concentration limit of detection (CLOD) of the purified EBzA product was determined to be 5 nM (100 fmol) at a detection potential of +200 mV vs Ag/AgCl ([Cl-] = 3 M). The CLOD values obtained by LC-EC after derivatization of aqueous solutions of EBzA and Ile-Leu-OMe with NDTE were 25 nM (250 fmol) and 250 nM (2.5 pmol), respectively.

Topics: Amines; Benzofurans; Chromatography, Liquid; Electrochemistry; Hydroquinones; Indicators and Reagents; Kinetics; Pyrones

Normal human melanocytes were amplified and cultured in a new defined culture medium without phorbol esters or cholera toxin. The medium decreased considerably the doubling time and increased the possible passage number. Melanocytes were co-seeded with normal human keratinocytes into 24 well culture dishes to screen potentially active modulators of melanogenesis. For the assay, the co-cultures were exposed to the compounds under investigation in the presence of 14C-thiouracil and 3H-leucine. Control cultures contain L-tyrosine or kojic acid, modulators which served as internal calibration standards. Changes in the rate of melanin synthesis were measured on the basis of 14C-thiouracil incorporation into newly synthesized melanin. A reduction or increase in 3H-leucine incorporation was taken as an indication of cytotoxicity or induction of proliferation, respectively. The NHK-NHM co-culture screening assay provides a useful tool to compare the activity of known modulators of melanogenesis and to perform structure-activity studies with newly identified modulators to improve their activity. The efficacy of particularly interesting new compounds was further evaluated on reconstructed pigmented epidermis after repeated topical application. The same model was used to assess the anti-pigmenting effect of sunscreens on UV-induced pigmentation. Integration of melanocytes from different ethnic origin resulted in pigmented epidermis reflecting different skin phenotypes, Caucasian, Asian and African.

Topics: 1-Methyl-3-isobutylxanthine; 3T3 Cells; Animals; Arbutin; Asian People; Benzofurans; Black People; Cells, Cultured; Coculture Techniques; Culture Media; Epidermal Cells; Ethnicity; Humans; Hydroquinones; Hydroxybenzoates; Indoles; Keratinocytes; Melanins; Melanocytes; Mice; Phenotype; Pyrones; Skin Pigmentation; Skin, Artificial; Theophylline; Thiophenes; Ultraviolet Rays; White People