title: Safety evaluation of styrax liquidus from the viewpoint of genotoxicity and mutagenicity.
abstract: BACKGROUND
  Styrax liquidus is a resinous exudate (balsam) obtained from the wounded trunk of the Liquidambar orientalis Mill. (Hamamelidaceae). Styrax has been used for treatment of various ailments in Turkish folk medicine such as skin problems, peptic ulcers, nocturnal enuresis, parasitic infections, antiseptic or as expectorant.
  
  
  OBJECTIVE
  In spite of frequent use of styrax in Turkish folk medicine as well as once as a stabilizer in perfumery industry, negative reports have been noticed by the international authority for restriction its use based on some limited evidences from an in vitro study. The aim of the present study was to evaluate the genotoxic and cytotoxic potential of styrax and its ethanolic extract using in vivo and in vitro assays, as well as an antimutagenic assay and also to determine its phenolic constituents with chromatographic analysis.
  
  
  METHODS
  In vitro mutagenicity and antimutagenicity of styrax and its ethanolic extract were evaluated by Ames test performed on Salmonella TA98 and TA100 strains with and without metabolic activation (10- 30,000µg/plate). The genotoxicity was also studied in vivo by chromosomal aberrations assay on bone marrow of Balb C mice with different its concentrations (500-2000mg/kg body weight). Cytotoxicity has been evaluated by the MTT assay using L929 cell line. Its phenolic constituents were determined by HPLC analysis.
  
  
  RESULTS
  Genotoxicological investigations of styrax or its ethanolic extract showed that none of the tested concentrations induced a significant increase in the revertant number of TA98 and TA100 strains with or without metabolic activation, indicating no mutagenicity to the tested strains. Also results indicated that up to 2000mg/kg body weight, styrax is not genotoxic in mammalian bone marrow chromosome aberration test in vivo. In cytotoxicity study, the IC50values of styrax and its ethanolic extract were found to be 50.22±1.80 and 59.69±11.77µg/mL, respectively. Among the studied reference standards the major phenolic acids in styrax balsam was found to be p-coumaric acid (2.95mg/g), while in its ethanolic extract not only p-coumaric acid (11.46mg/g), but also gallic acid (1.60mg/g) were found to the main components.
  
  
  CONCLUSIONS
  The findings of the present study provide scientific basis to the safety of styrax from the viewpoint of genotoxicity risk, and in fact, it was found to be beneficial against genotoxicity.
authors.affiliation: Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: mohammad.saz@yeditepe.edu.tr.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Charehsaz
authors.affiliation: Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: renginreis@gmail.com.
authors.forename: Rengin
authors.initials: R
authors.lastname: Reis
authors.affiliation: Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: sinem_helvacioglu@hotmail.com.
authors.forename: Sinem
authors.initials: S
authors.lastname: Helvacioglu
authors.affiliation: Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: hande.sipahi@yeditepe.edu.tr.
authors.forename: Hande
authors.initials: H
authors.lastname: Sipahi
authors.affiliation: Department of Pharmacognosy, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: etil.ariburnu@yeditepe.edu.tr.
authors.forename: Etil
authors.initials: E
authors.lastname: Guzelmeric
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: ebru.acar@yeditepe.edu.tr.
authors.forename: Ebru Turkoz
authors.initials: ET
authors.lastname: Acar
authors.affiliation: Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: gamzecck@yahoo.co.uk.
authors.forename: Gamze
authors.initials: G
authors.lastname: Cicek
authors.affiliation: Department of Pharmacognosy, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: yesilada@yeditepe.edu.tr.
authors.forename: Erdem
authors.initials: E
authors.lastname: Yesilada
authors.affiliation: Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Yeditepe University, Kayışdağı, Atasehir, 34755 Istanbul, Turkey. Electronic address: Ahmet.aydin@yeditepe.edu.tr.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Aydin
mesh_terms: D000818:Animals; D029201:Hamamelidaceae / Q000737:chemistry*; D020128:Inhibitory Concentration 50; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009152:Mutagenicity Tests; D010936:Plant Extracts / Q000627:therapeutic use / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D010936:Plant Extracts
keywords: Ames test; Genotoxicity; In vivo chromosomal aberration test; Styrax