阿魏酸/天然阿魏酸

Chinese medicine reference materials, ferulic acid (batch No. wkq20010902) and chlorogenic acid (batch No. wkq20042702) were purchased from Weikeqi Biotechnology Co., Ltd. (Sichuan, Chengdu); nodakenin(batch No.111821–201102), psoralen(batch No.110739–200814), and isoperatorin(batch No.110827–201109) were purchased from China Institute for Food and Drug Control.

The solvent flow rate was 0.8 ml/min. The column temperature was 25 °C. The detection wavelength was 275 nm. Ferulic acid (FA, Sichuan Weikeqi Biotech Ltd, Chengdu, China) was recommended as a representative compound in AS by the Chinese Pharmacopeia. The content of FA was calculated based on the absorbance at 275 nm.

All 10 standard compounds used in this study (DL-Arginine, Adenine, Uracil, liquiritin, Ferulic acid, Liquiritigenin, Z-Ligustilide, Isoliquiritigenin, Senkyunolide A, and Isoimperatorin) with a purity >98% were all purchased from Weikeqi Biotechnology Co., Ltd (Chengdu, China). Methotrexate (MTX) was purchased from Shanghai Xinyi Pharmaceutical Co., Ltd (Shanghai, China) and was used as the positive control.

D-(-)-Fructose, gallic acid, 5-hydroxymethylfurfural, loganic acid, chlorogenic acid, gentiopicroside, ferulic acid, isoorientin, liquiritigenin, liquiritin, nodakenin, rutin, 20-hydroxyecdysone, senkyunolide I, hydroxysafflor yellow A, quercetin, salicylic acid, kaempferol, isoliquiritigenin, amentoflavone, E-ligustilide, kaempferide, isoimperatorin, levistolide A, Z-ligustilide (purity > 98%) were purchased from Weikeqi Biotechnology Co., Ltd (Chengdu, China). CFA was purchased from Chondrex, Inc (Woodinville, USA).

Quercetin (QE), gallic acid (GA), Folin-Ciocalteu reagent, 2,2′-azobis-amidinopropane (ABAP), and dichlorofluorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich Inc. (St. Louis, USA). Protocatechuic acid (PA), ferulic acid (FA), epicathin (Epi), catechin (CE), QE glycosides, isorhamnetin (IS) glycosides and kaempferol (KA) glycosides were purchased from Weikeqi Biological (Chengdu, China). Human liver cancer cells HepG2 were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Rat interleukin (IL)-1β, IL-6, tumor necrosis factor alpha (TNF-α) enzyme-linked immunosorbent assay (ELISA) kit was obtained from Multi Science Biotechnology Co., Ltd. (Hangzhou, China). Reference standards of ferulic acid, atractylodin, tanshinone, astragaloside, hyperoside, loganin, gallic acid, morroniside, allantoin, oleanic acid, ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, paeoniflorin, vaccarin, pyrrolidinium, leonurine, and catalpol were obtained from Sichuan Weikeqi Biological Technology Co., Ltd. (Sichuan, China).

The reference standards of isorhmnetin (3), diosmetin (6), isochlorogenic acid A (7), kaempferol (10), quercetin (14), isochlorogenic acid C (15), apigenin (19), forsythin (20), chlorogenic acid (23), liquiritin (25), luteolin-7-O-glucoside (26), astragalin (28), luteolin (29), buddleoside (31), quercetin-3-O-glucoside (33), amygdalin (37), caffeic acid (38), forsythoside A (39), glycyrrhizic acid (40), ferulic acid (42), rutin (44), hyperoside (50), and platycodin D (51) were purchased from Sichuan Weikeqi Biological Technology, Co., Ltd. (Chengdu, China). Mulberry leaf, chrysanthemum, Fructus Forsythiae, licorice, Semen Armeniacae Amarum, Platycodi Radix and Phragmitis Rhizoma

Twelve reference standards (purity>98%) were pur-chased from Weikeqi (Sichuan, China). These standardswere  isoscopoletin,  scopoletin,  7-methoxycoumarin,7-hydroxycoumarin,  isomeranzin,  bergapten,  tan-geretin, nobiletin, hesperidin, limonin, ferulic acidand  bergamottin.  Chromatographic-grade  methanoland dichloromethane were purchased from WeikeqiBio-Technology (Sichuan, China). Formic acid and ACN(purity>99.9%) of HPLC-grade were provided by MerckKGaA (Darmstadt, Germany) and Thermo Fisher Scien-tific Inc. (NYSE:TMO), respectively. CO2(purity>99.9%)was provided by Messer Gas (Guangzhou, China).

The residue was dissolved in methanol, and then filtered through a 0.22?μm filter membrane (Millipore, USA). Standards were as follows: rotocatechualdehyde, hydroxysafflor yellow A, ferulic acid, naringin, salvianolic acid B, salvianolic acid A and cryptotanshinone (Weikeqi, China).

The reference standards ginkgolide M (3), apigenin-7-O-glucoside (7), bilobalide (8), ginkgolide C (9), quercetin (14), luteolin (15), apigenin (16), kaempferol (17), isorhamnetin (18), ferulic acid (19), ginkgolide A (20), myricetin (22), genistein (23), chlorogenic acid (24), caffeic acid (26), luteolin-7-O-glucoside (33), kaempferol-3-O-glucoside (34), kaempferol-7-O-glucoside (35), quercetin-7-O-rhamnoside (40), isoquercitrin (45) and rutin (65) were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, China), and their structures are shown in Fig. 1. Purities of all the reference standards were over 98% according to HPLC-DAD analysis.

Purified water was purchased from Wahaha(Zhejiang, Hangzhou, China). The reference standards astragalin, gallic acid, quinic acid, chlorogenic acid, ferulic acid were purchased from the Sichuan Weikeqi Biological Technology Co., Ltd (Beijing,China); adenosine, cordycepin, deoxyuridine, hypoxanthine, adenine, uridine, guanosine, ginsenoside Rg1, ginsenoside Rb1, ginsenoside Rf, ginsenoside Rb2 were purchased from Shanghai Shifeng Biotechnology Co., Ltd(Shanghai, China)

Phellodendri Chinensis Cortex was purchased from Shanxi HongSen Traditional Chinese Medicine Co., Ltd. (Xian, China); Asini Corii Colla was purchased from Shandong Jishui Ejiao Co., Ltd. (Heze, China). Zingiberone, ferulic acid, palmatine, jatrorrhizine, berberine hydrochloride, gallic acid, ellagic acid, zingerone and ligustilide had a purity of 98%, as determined by HPLC or GC, and were provided by Chengdu Weikeqi Biotechnology Co. Ltd. (Chengdu, China). Methanol and acetonitrile were purchased from Merck & Co. Inc. (Darmstadt, Germany).

The standards of oleanolic acid, ursolic acid, stigmasterol, β-Sitosterol, and the internal standard danshensu (IS1) were purchased from National Institute for Food and Drug Control (Beijing, China). The standards of coniferin, ferulic acid, and the internal standard icariin (IS2) were supplied from Sichuan Weikeqi Biological Technology Co., Ltd. (Si chuan, China). All standards, which the purity was more than 98.0%, were suitable for UHPLC–MS/MS analysis.

Standards of chicoric acid, and ferulic acid (purity?≥98%) were purchased from Weikeqi Biological Technology Co., Ltd (Sichuan, China). Chromatographic grade methanol, ethyl acetate, and acetonitrile were purchased from Tedia Company Inc. (Fairfield, USA).

Folin-Ciocalteureagent,gallic acid, catechinand otherpolyphenolicstandardswere purchasedfrom SichuanWeikeqiBiologicalTechnologyCo., Ltd. (China).1,1-Diphenyl-2-picrylhydrazyl(DPPH),2,2’-azinobis(3-ethyl-benzothiazoline-6-sulfonicacid)diammoniumsalt(ABTS),2,4,6-tris(2-pyridyl)-s-triazine(TPTZ),trolox,α-amylase,α-glucosidaseand 4-nitrophenyl-α-D-gluco-pyranoside(α-PNPG)wereobtainedfromSigma–Aldrich(USA).

Thestandards of syringic acid, ferulic acid,p-coumaric acid, isoquercitrin,hyperoside, avicularin, quercitrin, and the IS of danshensu (IS2) werefrom Sichuan Weikeqi Biological Technology Co., Ltd. (Sichuan,China). The structure of analytes and ISs is presented in Figure 1.Acetonitrile, methanol, acetic acid, and formic acid of HPLC gradewere obtained from Thermo Fisher Scientific (Fair Lawn, NJ, USA)

Tetraethtylamine (TEA, no. 134473), 4-aminopyridine (4-AP, no. 275875), BaCl2 (no. 342920), glyburide (Gli, no. Y0001511), ethylene glycol-bis (2-aminoethylether)-N, N, N′, N′-tetraacetic acid (EGTA, no. E3889), and 9, 11-dideoxy-9α, 11α-methanoepoxy prostaglandin F2α (U46619, no. D8174) were purchased from Sigma. Astragaloside IV (no. wkq16042601) and ferulic acid (no. wkq17022303) were provided by Weikeqi Biological Technology Co., Ltd. Formononetin (no. DST201025-011) and ligustalide (no. DST200610-007) were procured from Lemeitian medicine.