JOURNAL OF COSMETIC SCIENCE 378 oxidative stress. Bacteria such as P. acnes and M. furfur act as immune stimulators through the production of proinfl ammatory cytokines, which are involved in development of the infl ammatory process. P. acnes is an aerotolerant, anaerobic, gram-positive rod-shaped bacterium related to vari- ous skin conditions. It is commonly found on the skin (in pores and hair follicles) and plays an important role in acne development. M. furfu r is a lipophilic fungus that affects the hair and causes dandruff. Dandruff is gen- erally characterized by the presence of fl akes on the scalp and hair and is often accompa- nied by itching. The scalp represents a unique environment, with thick terminal hair, large numbers of sweat and sebaceous glands, and high relative humidity, creating favor- able conditions for microbial colonization (3,4). Acnes and dandruff are predicated by three major factors: colonization, sebum production, and individual predisposition. Plants’ secondary metabolites present in herbal drugs, cosmetic ingredients, and food are useful in the prevention and treatment of many diseases (5). Cnidium offi cinale belongs to the family Umbelliferae, and its roots have medicinal properties. In Asia, it has been tradition- ally used as an herbal treatment and is used to treat headaches, abdominal pain, and blood circulation. Senkyunolide A is a known major antioxidant constituent in C. offi cinale. In the present study, we investigated the antibacterial and anti-infl ammatory properties of C. offi cinale hexane (COH) extract and senkyunolide A (SA) against P. acnes and M. furfur. MATERIAL S AND METHODS PLANT MA TERIALS AND EXTRACTION C. offi c inale root was collected from local market, and C. offi cinale roots were dried and ground to powder. The powder was extracted for 5 d using 70% ethanol, and then the extract was fi ltered. The crude extract was sequentially partitioned with n-hexane (Hex). The extract and Hex fractions were stored in tightly sealed collection bottles at -20°C until further analysis. SA SA (CFN 995 94, ChemFaces, Wuhan, China) was analyzed by high-performance liquid chromatography (HPLC) to determine its purity. The mobile phase for HPLC consisted of acetonitrile (A) and water (B), using an optimized gradient elution of 50% B for 0–10 min and 50–5% B for 16–30 min. The fl ow rate was 1 ml min-1. The detection wavelength was maintained at 278 nm. MICROORGANISMS AND CULTURE P. acnes (ATCC 6919, American Type Culture Collection, Manassas, VA) was incubated in a reinforced clostridial medium (RCM) for 48 h at 37°C, under anaerobic conditions in a jar with a gas pack. M. furfur (ATCC 14521) was incubated in modifi ed Leeming– Notman broth (mLNB) for 96 h at 27°C.

ANTIBACTERIAL ACTIVITY OF SENKYUNOLIDE A 379 DETERMINATION OF ANTIBACTERIAL ACTIVITY PAPER DISK DIFFUS ION METHOD The antibacterial activities of the samples were evaluated using a disc diffusion assay. In brief, Advantec paper discs of 8 mm diameter were impregnated with 50 μL of the solu- tion impregnating ethanol extract and fractions at a concentration of 50 mg ml-1 and evaporated at room temperature for 24 h (6). Then 1 ml of bacteri a (107 cfu ml-1) incubated for 24 h was dispensed into petri dishes, and the medium was poured and solidifi ed. And then discs of COH and SA were placed on the RCM (P. acnes) and mLNB agar plates (M. furfur). The antibacterial activity was evaluated by measuring the zones of inhibition on the agar plate. All experiments were carried out in triplicate. MINIMUM INHIBITORY CON CENTRATION (MIC) The MICs were determin ed using the broth dilution method recommended by the Tenover (7). The cultures were prepared from 48-h and 96-h broth cultures of P. acnes and M. furfur, respectively. A 15-ml conical tube was prepared by dispensing 3 ml of broth media, sample (concentration ranging from 1.00 to 20.00 mg ml-1), and an appropriate amount of cell suspension (107 cfu 3 ml) into each tube. The tubes were incubated at 37°C for 48 h and 27°C for 96 h for P. acnes and M. furfur, respectively. At the end of the incubation period, the tubes were evaluated for the presence or absence of growth. All experiments were performed in triplicate. DETERMINATION OF ANTI-INFL AMMATORY ACTIVITY PANCREATIC LIPASE INHIBITI ON ACTIVITY Porcine pancreatic lipase (PPL, type II) activity was measured using p-nitrophenyl lau- rate (p-NPL) as a substrate. The method used for measuring the PPL activity was modi- fi ed from that previously described by Zheng et al. (8) and Bustanji et al. (9). The reaction mixture consisted of 50 mM Tris-HCl buffer (930 μl, 150 mM NaCl, 1 mM EDTA, and 10 mM MOPS, pH 7.4), PPL 5 mg ml-1 (30 μl), and 5 mM p-NPL (20 μl). The reaction was started by adding p-NPL as a substrate, all in a fi nal volume of 1,000 μL. After incu- bation at 35°C for 15 min, the amount of p-nitrophenol released during the reaction was measured at 405 nm using a UV-visible spectrophotometer. All experiments were carried out in triplicate. LIPOXYGENASE INHIBITION ACTIVITY L ipoxygenase (soybean) enzyme acti vity was measured according to the method of Lycka- nder and Malterud (10) with a minor modifi cation. Tris-HCl buffer (2,800 μl, pH 9.0) was added to 100 μl of sample at different concentrations, nordihydroguaiaretic acid