Hplc grade solvent

¹H and ¹³C-NMR were detected on a Varian 400 MHz spectrometer in CD₃OD with Tetramethylsilane (TMS) as the internal standard. HR-ESI-MS data were obtained on a 6538 Ultra High Definition (UHD) Accurate-Mass Q-TOF LC/MS system (Agilent, Santa Clara, CA, USA). High performance liquid chromatography (HPLC) was performed on a Hitachi Elite Chromaster system including a 5110 pump, 5210 autosampler, 5310 column oven, a 5430 diode array detector, and operated by EZChrom Elite software. Luna C18 (2) column (5 µm, 4.6 × 250 mm) for analysis and Luna C18 (2) column (10 µm, 10 × 250 mm) for HPLC preparation were purchased from Phenomenex Inc (Torrance, CA, USA). The HPLC grade solvents were purchased from Sigma (Sigma, St. Louis, MO, USA). All analytical solvents were purchased from Tansoole (Shanghai, China). Silica gel (250 mesh; Qingdao Haiyang Chemical Co., LTD, Qingdao, China) was used as normal phase, whereas YMC Pack ODS-A (50 µm; YMC) was used as reversed phase column material. MCI gel CHP20P (75–150 µm; Mitsubishi Chemical Corp, Tsukuba, Japan) and Sephadex LH-20 (GE Healthcare, Uppsala, Sweden) were also used for column chromatography.

Ricobendazole, oxfendazole and HPLC-grade solvents were purchased from Sigma-Aldrich (Milan, Italy) and used without further purification. HPLC enantioseparations were performed by using stainless-steel Chiralpak IG (250 mm × 10 mm I.D.) columns (Chiral Technologies Europe, Illkirch, France). The HPLC apparatus for semipreparative separations consisted of a Perkin-Elmer (Norwalk, CT, USA) 200 LC pump equipped with a Rheodyne (Cotati, CA, USA) injector, a 5000 μL sample loop, a Perkin-Elmer LC 101 oven and a Waters 484 detector (Waters Corporation, Milford, MA, USA). Data were processed using Clarity software (DataApex, Prague, Czech Republic).

All HPLC grade solvents were obtained from Sigma-Aldrich (Seelze, Germany) except for ethanol (99.5%) which was obtained from Kemetyl AB (Haninge, Sweden). Liquid carbon dioxide was purchased from AGA Gas AB (Stenungsund, Sweden). Diethyl amine p.a. (DEA) from Fluka (Buchs, Switzerland) and formic acid p.a. (FA) from Riedel-de Haën (Seelze, Germany) was used.
The compounds used were synthesized at AstraZeneca R&D (Gothenburg, Sweden) or at our partner contract research laboratories.

Perilla leaves were purchased from a local market in Korea. The flavonoids used were obtained from Indofine Chemical Company (Hillsborough, NJ, USA) and Extrasynthese (Genay Cedex, France). p-Coumaric acid, eugenol and isoeugenol were purchased from Sigma-Aldrich (St. Louis, MO, USA). Caffeic acid was obtained from Indofine Chemical Company. SAM was purchased from Sigma-Aldrich. HPLC grade solvents and other reagents were obtained from Sigma-Aldrich, Duchefa Biochemie (Haarlem, The Netherlands), and Samchun Chemicals (Seoul, Korea).

Escherichia coli LPS (O128:B12, Sigma-Aldrich, Castle Hill, Australia), sulfanilic acid, N-(1-Naphthyl) ethylenediamine (NED), 85% orthophosphoric acid sodium nitrite (NaNO₂), and HPLC grade solvents were obtained from Sigma Aldrich (St. Louis, MO, USA). The mouse TNFα ELISA kit was purchased from BD biosciences (Sparks, MD, USA). Penicillin–streptomycin solution, Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), sodium pyruvate, and l-glutamine were from Life Technology Australia (Mulgrave, VIC, Australia). The two cell lines, RAW264.7 mouse macrophages and 3T3 Swiss albino (ATCC^® CCL92™) cell lines, were purchased from the American Type Culture Collection (ATCC^®, Manassas, VA, USA).