title: Multiresponse Optimisation Applied to the Development of a TLC Autography for the Detection of Tyrosinase Inhibitors.
abstract: BACKGROUND
  Autographic methods are useful tools to detect bioactive compounds in complex matrixes. Experimental design and optimisation techniques were implemented for the development of an autographic assay suitable for the detection of tyrosinase inhibitors.
  
  
  OBJECTIVE
  To develop an autographic assay to detect tyrosinase inhibitors using gel entrapped enzyme, experimental design and response surface methodology (RSM) to optimise conditions with a minimum number of experiments.
  
  
  METHODS
  Gel entrapment was used for the assay and the effects of four factors on the sensitivity and the detection limit for known inhibitors of the enzyme were evaluated. The factors were: tyrosinase amount (TA), L-tyrosine amount (LTA), incubation time and incubation temperature.
  
  
  RESULTS
  The assay allowed the detection of kojic acid in an extract of Calamagrostis viridiflavescens (Poir.) Steud spiked with 0.1% w/w.
  
  
  CONCLUSIONS
  The developed assay is able to detect tyrosinase inhibitors present in complex matrixes in a reproducible way.
authors: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario. CONICET, Suipacha 531, S2002LRK, Rosario, Argentina.
authors: Paula
authors: P
authors: García
authors: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario. CONICET, Suipacha 531, S2002LRK, Rosario, Argentina.
authors: Ricardo L E
authors: RL
authors: Furlan
mesh_terms: D002855:Chromatography, Thin Layer / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D010936:Plant Extracts / Q000032:analysis / Q000494:pharmacology; D006109:Poaceae / Q000737:chemistry; D011753:Pyrones / Q000032:analysis / Q000494:pharmacology; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D010936:Plant Extracts; D011753:Pyrones; C011890:kojic acid; D014442:Monophenol Monooxygenase
keywords: Response surface methodology; TLC-bioautography; dermatological disorders; herbal drugs; tyrosinase inhibition