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The Development and Application of a HPTLC-Derived Database for the Identification of Phenolics in Honey
Authors:Ivan Lozada Lawag  Tomislav Sostaric  Lee Yong Lim  Katherine Hammer  Cornelia Locher
Affiliation:1.Cooperative Research Centre for Honey Bee Products Limited (CRC HBP), University of Western Australia, Agriculture North M085, Perth, WA 6009, Australia;2.Division of Pharmacy, School of Allied Health, University of Western Australia, Curnow Building M315, Perth, WA 6009, Australia;3.School of Biomedical Sciences, University of Western Australia, M Block QEII Medical Centre, Monash Ave., Perth, WA 6009, Australia
Abstract:
This study reports on the development and validation of a HPTLC-derived database to identify phenolic compounds in honey. Two database sets are developed to contain the profiles of 107 standard compounds. Rich data in the form of Rf values, colour hues (H°) at 254 nm and 366 nm, at 366 nm after derivatising with natural product PEG reagent, and at 366 nm and white light after derivatising with vanillin–sulfuric acid reagent, λ max and λ min values in their fluorescence and λ max values in their UV-Vis spectra as well as λ max values in their fluorescence and UV-Vis spectra after derivatisation are used as filtering parameters to identify potential matches in a honey sample. A spectral overlay system is also developed to confirm these matches. The adopted filtering approach is used to validate the database application using positive and negative controls and also by comparing matches with those identified via HPLC-DAD. Manuka honey is used as the test honey and leptosperine, mandelic acid, kojic acid, lepteridine, gallic acid, epigallocatechin gallate, 2,3,4-trihydroxybenzoic acid, o-anisic acid and methyl syringate are identified in the honey using the HPTLC-derived database.
Keywords:HPTLC   HPLC-DAD   Manuka honey   Leptospermum scoparium   database   biomarkers   phenolic compounds   phenolic determination
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