2 2 azobis 2 amidinopropane dihydrochloride abap

Chemicals and solvents
for kinetic measurements
were purchased from Sigma-Aldrich, TCI, and Avanti Polar Lipids Inc.
and were used without any additional purification: 2,2′-azobis(2-amidinopropane)
dihydrochloride (ABAP; 97%, Sigma-Aldrich), Triton X-100 (polyethylene
glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether;
98%, Sigma-Aldrich), methyl linoleate (MeLin; 99%, Sigma-Aldrich),
1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC;
99%, Avanti Polar Lipids Inc.), 2,2,5,7,8-pentamethylchroman-6-ol
(PMHC; 99%, Sigma-Aldrich), 3,4′,5-trihydroxy-trans-stilbene (resveratrol; >99%, TCI), 3,5-dihydroxybenzyl alcohol
(99%,
Sigma-Aldrich), and 4-hydroxybenzyl alcohol (99%, Sigma-Aldrich).

Pepsin, trypsin, sialic acid (SA), 2′,7′-dichlorofluorescin diacetate (DCFH-DA), 2,2′-azobis (2-amidinopropane) dihydrochloride (ABAP), and sialic acid (SA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and penicillin–streptomycin were purchased from Shanghai Scientific & Technology Co., Ltd., Shanghai, China. Chromatography grade acetonitrile and tetrahydrofuran, analytical reagent of o-phenylenediamine hydrochloride (OPDH) and sulfuric acid were purchased from Sinopharm Chemical Reagent Co., Ltd, Shanghai, China. HepG2 and B16-F10 cells were kindly provided by Cell Bank, Chinese Academy of Sciences.
Dry EBN samples (Aerodramusfuciphagus) originated from Indonesia were kindly donated by Xiamen Yanzhiwu Sinong Food Co., Ltd. The contents of protein, carbohydrate, SA, and ash of EBN were 68.95%, 17.95%, 10.55%, and 2.15% (dry basis). EBN powder was prepared by a high-speed universal crusher (SS-1022, Shengshun, Jinhua, China) and then passed through a 60-mesh sieve. The EBN powder was stored in hermetically sealed glass bottles at room temperature before further analysis.

DHM, purity >98%, was purchased from Bomei Biotechnology Ltd. (Hefei, Anhui, China). Lipozyme TL IM (250 U/g) was obtained from Amano Enzyme Inc. (Nagoya, Aichi, Japan). Vinyl butyrate, vinyl hexanoate, vinyl octanoate, and vinyl laurate were obtained from Tokyo Chemical Industry Ltd. (Tokyo, Japan). ABTS and DPPH were purchased from Shanghai McLean Biochemical Technology Co., Ltd. (Shanghai, China). 2,2’-azobis (2-amidinopropane), dihydrochloride (ABAP), and 2’,7’-dichlorofluorescein diacetate (DCFH-DA) were obtained from Sigma–Aldrich (Shanghai) Trading Co., Ltd. (Shanghai, China). Thirty percent H₂O₂, L-ascorbic acid (Vc), tert-butylhydroquinone (TBHQ), trichloroacetic acid, 2-thiobarbituric acid, ammonium thiocyanate, n-butanol, isooctane, isopropanol, chloroform, glacial acetic acid, sodium thiosulfate, Tween 20, methyl tert-butyl ether (MTBE), and 1,1,3,3-tetraethoxypropane were provided by Shanghai Sinopharm Group Chemical Reagent Ltd. (Shanghai, China). An MTT assay kit was obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu, China). RPMI 1640 culture medium, dimethyl sulfoxide (DMSO), fetal bovine serum (FBS), phosphate buffer solution (PBS), 0.25% trypsin, penicillin, and streptomycin were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA).

Chemicals and reagents for synthesis: starting materials, reagents, and solvents were purchased from Sigma–Aldrich, Acros Organics and Combi-Blocks, and were used without any additional purification. The reaction progress was monitored by Thin Layer Chromatography performed on commercial Kieselgel 60, F254 silica gel plates with fluorescence-indicator UV254 (Merck, TLC silica gel 60 F254). For the detection of components, a UV light at λ = 254 nm or λ = 365 nm was used.
Chemicals and reagents for kinetic measurements: Initiator, α,α′-azobisisobutyronitrile, AIBN (>98% GC, Fluka Chemika), was recrystallized from methanol before use. 2,2’-Azobis(2-amidinopropane) dihydrochloride (ABAP; 97%), Triton X-100 (polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, 98%), methyl linoleate (LinMe; 99%), and 2,2,5,7,8-pentamethylchroman-6-ol (PMHC; ≥99%) were purchased from Sigma–Aldrich and were used as received.

Rice husk was provided by the Rice Research Institute of Guangdong Academy of Agricultural Science, China. Folin-Ciocalteu reagent, fluorescein disodium salt, 2,2′-azobis (2-amidinopropane) dihydrochloride (ABAP), 2′,7′-Dichlorofluorescin diacetate (DCFH-DA), (+)-catechin, gallic acid, and chromatographic grade of p-coumaric acid, caffeic acid, p–coumaric acid, ferulic acid were purchased from Sigma-Aldrich Ltd. (St. Louis, MO, USA). Chromatographic grade of acetonitrile used for HPLC analysis was obtained from Anpel Ltd. (Shanghai, China). Human liver cancer cell line HepG2 (ATCC HB-8065) were purchased from ATCC company (Manassas, VA, USA). WME medium, fetal bovine serum (FBS), Hank’s balanced salt solution (HBSS), Trypsin-EDTA solution and other cell culture reagents were purchased from Gibco Life Technologies Co. (Grand Island, NY, USA).

Ethanol, acetonitrile, acetone, mEthanol, phosphate-buffered saline (PBS), sodium carbonate, penicillin-streptomycin, sodium hydroxide, anhydrous sodium acetate, hydrochloric acid, trypsin EDTA, Williams’ Medium E, potassium persulfate, acetic acid, sulfuric acid, gentamicin, insulin, L-glutamine, Dulbecco’s modified eagle medium (DMEM) and advanced DMEM were obtained from Daejung chemicals and metals Co., Ltd, South Korea and Life Technologies (Grand Island, NY, USA). The Phenolic standards and other chemicals like ABTS (2,2′Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid), 2′,7′-Dichlorofluorescin diacetate or 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA, ≥ 97% purity), TPTZ (2,4,6-Tris(2-pyridyl)-s-triazine), caffeic acid, ferulic acid, 2,2 Diphenyl-1-picrylhydrazyl (DPPH), gallic acid, Folin–Ciocalteu reagent, quercetin dehydrate (≥ 98% purity), p-Coumaric acid, (+)-catechin hydrate, dimethyl sulfoxide (DMSO); 2,2′-azobis(2-amidinopropane) dihydrochloride (ABAP), and Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) were purchased from Sigma, South Korea. Analytical grade chemicals and reagents were used.