title: Optimization of extraction conditions for osthol, a melanogenesis inhibitor from Cnidium monnieri fruits.
abstract: BACKGROUND
  Coumarin derivatives have been reported to inhibit melanin biosynthesis.
  
  
  OBJECTIVE
  The melanogenesis inhibitory activity of osthol, a major coumarin of the fruits of Cnidium monnieri Cusson (Umbelliferae), and optimized extraction conditions for the maximum yield from the isolation of osthol from C. monnieri fruits were investigated.
  
  
  METHODS
  B16F10 melanomas were treated with osthol at concentration of 1, 3, and 10 μM for 72 h. The expression of melanogenesis genes, such as tyrosinase, TRP-1, and TRP-2 was also assessed. For optimization, extraction factors such as extraction solvent, extraction time, and sample/solvent ratio were tested and optimized for maximum yield of osthol using response surface methodology with the Box-Behnken design (BBD).
  
  
  RESULTS
  Osthol inhibits melanin content in B16F10 melanoma cells with an IC50 value of 4.9 μM. The melanogenesis inhibitory activity of osthol was achieved not by direct inhibition of tyrosinase activity but by inhibiting melanogenic enzyme expressions, such as tyrosinase, TRP-1, and TRP-2. The optimal condition was obtained as a sample/solvent ratio, 1500 mg/10 ml; an extraction time 30.3 min; and a methanol concentration of 97.7%. The osthol yield under optimal conditions was found to be 15.0 mg/g dried samples, which were well matched with the predicted value of 14.9 mg/g dried samples.
  
  
  CONCLUSIONS
  These results will provide useful information about optimized extraction conditions for the development of osthol as cosmetic therapeutics to reduce skin hyperpigmentation.
authors.affiliation: a College of Pharmacy, Chungbuk National University , Cheongju , Korea ;
authors.forename: Seon
authors.initials: S
authors.lastname: Beom Kim
authors.affiliation: b RNS Inc. , Daejeon , Korea ;|c College of Pharmacy, Chungnam National University , Daejeon , Korea ;
authors.forename: CheongTaek
authors.initials: C
authors.lastname: Kim
authors.affiliation: a College of Pharmacy, Chungbuk National University , Cheongju , Korea ;
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.affiliation: a College of Pharmacy, Chungbuk National University , Cheongju , Korea ;
authors.forename: Yang
authors.initials: Y
authors.lastname: Hee Jo
authors.affiliation: d Traditional and Biomedical Research Center, Daejeon University , Daejeon , Korea.
authors.forename: Hak
authors.initials: H
authors.lastname: Joo Choi
authors.affiliation: a College of Pharmacy, Chungbuk National University , Cheongju , Korea ;
authors.forename: Bang Yeon
authors.initials: BY
authors.lastname: Hwang
authors.affiliation: c College of Pharmacy, Chungnam National University , Daejeon , Korea ;
authors.forename: Sang
authors.initials: S
authors.lastname: Kyum Kim
authors.affiliation: a College of Pharmacy, Chungbuk National University , Cheongju , Korea ;
authors.forename: Mi
authors.initials: M
authors.lastname: Kyeong Lee
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid; D031048:Cnidium / Q000737:chemistry*; D003374:Coumarins / Q000302:isolation & purification* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005638:Fruit / Q000737:chemistry*; D019746:Intramolecular Oxidoreductases / Q000378:metabolism; D008543:Melanins / Q000096:biosynthesis; D008546:Melanoma, Experimental / Q000201:enzymology; D051379:Mice; D010088:Oxidoreductases / Q000378:metabolism; D008517:Phytotherapy; D010936:Plant Extracts / Q000302:isolation & purification* / Q000494:pharmacology; D010946:Plants, Medicinal; D062327:Skin Lightening Preparations / Q000302:isolation & purification* / Q000494:pharmacology; D012880:Skin Pigmentation / Q000187:drug effects; D013997:Time Factors
chemical_list: D003374:Coumarins; D008543:Melanins; D010936:Plant Extracts; D062327:Skin Lightening Preparations; D010088:Oxidoreductases; C064056:tyrosinase-related protein-1; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase; C046627:osthol
keywords: Cosmetics; TRP-1; tyrosinase