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  • Separation of the antioxidant compound quercitrin from Lindera obtusiloba Blume and its antimelanogenic effect on B16F10 melanoma cells.

Separation of the antioxidant compound quercitrin from Lindera obtusiloba Blume and its antimelanogenic effect on B16F10 melanoma cells.

Bioscience, biotechnology, and biochemistry (2013-01-08)
Chung-Oui Hong, Hyun Ah Lee, Chae Hong Rhee, Se-Young Choung, Kwang-Won Lee
ABSTRACT

Considering the growing evidence of the presence of antioxidant compounds in plant extracts, the objectives of this study were to identify antioxidant compounds in Lindera obtusiloba Blume (Lauraceae) and to evaluate their antimelanogenic activities on B16F10 melanoma cells. Organic solvent fractions were separated from L. obtusiloba extracts (LOE). The ethyl acetate fraction (LOE-E) was significantly active against oxidative damage induced by tert-butyl hydroperoxide in primary rat hepatocytes. Two single purified compounds, quercitrin (quercetin-3-O-α-L-rhamnopyranoside) and afzelin (kaempferol-3-O-α-L-rhamnoside), were identified by HPLC and NMR. These compounds were evaluated for antioxidant activities by 1,1-diphenyl 2-picrylhydrazyl (DPPH) radical scavenging assay and ferric reducing antioxidant power (FRAP) assay, and for their antimelanogenic activities by tyrosinase inhibitory assay melanin formation inhibition assay and Western bolt analysis for the signaling pathway. The significant effects of quercitrin on antioxidant and antimelanogenic activities, and signal modulation of ERK and MITF in B16F10 melanoma cells were observed. This is the first report to identify quercitrin in L. obtusiloba and its whitening effect.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tyrosinase from mushroom, lyophilized powder, ≥1000 unit/mg solid
Sigma-Aldrich
tert-Butyl hydroperoxide solution, 70 wt. % in H2O
Sigma-Aldrich
tert-Butyl hydroperoxide solution, 5.0-6.0 M in decane
Sigma-Aldrich
tert-Butyl hydroperoxide solution, 5.0-6.0 M in nonane
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Ethyl acetate, natural, ≥99%, FCC, FG
Sigma-Aldrich
Ethyl acetate, ≥99%, FCC, FG
Quercitrin, primary reference standard
Sigma-Aldrich
Ethyl acetate, ReagentPlus®, ≥99.8%
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Ethyl acetate, anhydrous, 99.8%
Supelco
Ethyl acetate, analytical standard
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Ethyl acetate, ACS reagent, ≥99.5%
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Ethyl acetate, puriss., meets analytical specification of Ph. Eur., BP, NF, ≥99.5% (GC)
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Ethyl acetate, ACS reagent, ≥99.5%
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Ethyl acetate, biotech. grade, ≥99.8%
Sigma-Aldrich
Ethyl acetate, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)
Sigma-Aldrich
Ethyl acetate, suitable for HPLC, ≥99.8%
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Ethyl acetate, suitable for HPLC, ≥99.7%
Sigma-Aldrich
Ethyl acetate, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%