Phome 10009134

Column chromatography was performed using silica gel (Kieselgel 60, 70–230 and 230–400 mesh, Merck, Darmstadt, Germany), Sephadex LH-20 (GE Healthcare, Uppsala, Sweden), and C-18 (ODS-A 12 nm S-150, S-75 μm; YMC Co., Kyoto, Kansai, Japan) resins. Thin-layer chromatography (TLC) was performed using pre-coated silica gel 60 F254 and RP-18 F254S plates (both 0.25 mm, Merck). Spots in the TLC were visualized by spraying with 10% aqueous H₂SO₄ solution followed by heating to 300℃ in dried air. Nuclear magnetic resonance (NMR) spectra were recorded using the JEOL ECA 500 spectrometer (¹H, 500 MHz; ¹³C, 125 MHz) (JEOL, Tokyo, Japan) (Supplementary Materials). AUDA (10007927), soluble epoxide hydrolase (10011669) and PHOME (10009134) were purchased from Cayman (Cayman, Ann Arbor, MI, USA).

ESI–mass spectra were obtained on a Shimadzu LC MS-2020-EV. Nuclear magnetic resonance (NMR) spectra were obtained using a Bruker Fourier 300 (¹H-300 MHz, ¹³C-75 MHz) and Jeol JNM-AL 400 (¹H-400 MHz, ¹³C-100 MHz) spectrometer using TMS as an internal standard; chemical shifts were expressed as δ values. CC was carried out using 230–400 mesh silica gel (Kieselgel 60, Merck, Darmstadt, Germany) and RP-18 (ODS-A, 12nm, S-150 mM, YMC). The thin layer chromatography (TLC) was carried out glass plates pre-coated with silica gel 60 F₂₅₄ and RP-18 F₂₅₄ (20 × 20 cm, Merck). Compounds were visualized by dipping plates into 10% (v/v) H₂SO₄ reagent and then heated at 110 °C for 5–10 min. The solvents used for material separation, MeOH, n-hexane, CHCl₃, EtOAc, n-BuOH, were purchased from SK chemical (Suwon, Gyeonggi-do, Korea). AUDA, sEH (10011669) and PHOME (10009134) were purchased from Cayman (Cayman, MI, USA.).

NMR spectra was obtained on Bruker Advance 600 and 300 MHz (Bruker; Rheinstetten, Germany) spectrometer. Column chromatography was performed using silica gel (Kieselgel 60; 70–230; and 230–400 mesh; Merck; Darmstadt, Germany) and YMC RP-18 resins. Thin layer chromatography (TLC) was performed using pre-coated silica-gel 60 F₂₅₄ and RP-18 F_254S plates (both 0.25 mm; Merck; Darmstadt, Germany). Spots were visualized by spraying with 10% aqueous H₂SO₄ solution followed by heating. Methanol; acetonitrile (Sigma-Aldrich; St. Louis, MO, USA) and trifluoroacetic acid (Alfa Aesar; Ward Hill, MA, USA) used in this study were of HPLC grade and the distilled water was prepared by the ultra-pure water manufacturing device (Optimos; Sinhan Science Tech, Daejeon, Korea). The chromatographic system was Shimazdu prominence system equipped with a SPD-20A dual UV-Vis detector and installed with a Shimadzu LC solution software (Ver. 1.25; Shimadzu; Kyoto, Japan). The quantitative analysis for three selaginellins was carried out on Optimapak C18 (250 mm × 4.6 mm I.D.; 5 μm) column. Soluble epoxide hydrolases (human recombinant, 10011669) and PHOME (10009134) were purchased from Cayman (Ann Arbor, MI, USA).