N methylmorpholine

EGCG was purchased from Bio Verde Inc. (Kyoto, Japan). 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMT-MM) and N-methylmorpholine (NMM) were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan) and Nacalai Tesque Inc. (Kyoto, Japan), respectively. Gelatin extracted from porcine skin in acidic conditions (Type A gelatin) was purchased from Sigma-Aldrich (St. Louis, MO, USA). EGCG-GS was prepared by an aqueous synthesis method reported previously [23 (link)]. In brief, gelatin (100 mg) was dissolved in warm Milli-Q water (5 mL) at 50 °C. A solution with NMM (27.5 µL), EGCG (0.07, 0.7 or 6.7 mg), and DMT-MM (69.2 mg) was stirred for 24 h at room temperature in the dark. The products were purified by dialysis (Spectra/Por7 MWCO 1000; Spectrum Labs, Rancho Dominguez, CA, USA) in Milli-Q water in the dark. The same conditions but without EGCG, DMT-MM, and NMM were used to prepare the gelation solution. After dialysis, the resulting solution was diluted to 10 mL with Milli-Q water and was poured in φ5-mm silicon tubes, followed by lyophilization with DC800 (Yamato Co., Ltd., Tokyo, Japan) to produce the EGCG-GSs or GS. To fabricate vhEGCG-GSs and vhGS, EGCG-GSs and GS were treated by vacuum heating using ETTAS AVO-250NS (AS ONE, Osaka, Japan) at 150 °C for 24 h with a gauge pressure of −0.1 MPa. All sponges were stored at 4 °C in the dark until use.

Formic acid, free cholesterol enzymatic test kit, trimethylamine, pyridine, sodium sulphate, palmitic acid, and palladium/carbon were purchased from FUJIFILM WAKO Chemicals (Osaka, Japan). Acetone, chloroform, ethanol, filipin complex, 1-hydroxytriazole, Ham′s F-12 medium, iodine, pivaloyl chloride, serine, and toluene were purchased from Nacalai tesque (Kyoto, Japan). Acetonitrile, methanol-d₄, and chloroform-d were purchased from Kanto Kagaku (Tokyo, Japan). Acetyl chloride, dichloromethane, 1, 3-dicyclohexylcarbodiimide, imidazole, N-methylmorpholine, L-serine benzyl ester hydrochloride, and phosphorus trichloride were purchased from Tokyo chemical industry Co. (Tokyo, Japan). NBD-F was purchased from DONJINDO laboratories (Kumamoto, Japan). Choline tosylate was purchased from Toronto Research Chemicals (North York, ON, Canada). Foetal bovine serum was purchased from Gibco (Grand island, NY, USA). SPC and Lyso-SM (d17:1) were purchased from Avanti polar lipids, Co. Ltd. (Alabaster, AL, USA). Ultrapure water was prepared with PURELAB ultra (Organo Co. Ltd., Tokyo, Japan) and used in all MS analyses.