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Multiresponse Optimisation Applied to the Development of a TLC Autography for the Detection of Tyrosinase Inhibitors
Author(s) -
García Paula,
Furlan Ricardo L. E.
Publication year - 2015
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.2562
Subject(s) - tyrosinase , chemistry , kojic acid , chromatography , detection limit , response surface methodology , incubation , enzyme , biochemistry
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. Objectives 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. Conclusion The developed assay is able to detect tyrosinase inhibitors present in complex matrixes in a reproducible way. Copyright © 2015 John Wiley & Sons, Ltd.