Pigeon pea seeds were provided

by Honghe Taifu Agriculture and Forestry Technology Development

Co., Ltd. (Honghe, China). Cajaninstilbene acid was separated and

purified in our laboratory with a purity higher than 98%. Other

standards were bought from Weikeqi Biological Technology Co., Ltd.

(Chengdu, China).

Orientin, vitexin, isovitexin, genistin, luteolin, genistein, apigenin, isorhamnetin, biochanin A, and pinostrobin were purchased from Weikeqi Biological Technology Co., Ltd. (Chengdu, China). Cajaninstilbene acid was obtained by the separation and purification from pigeon pea leaves by column chromatography in our laboratory. All standards were identified with the purity higher than 98%.

Five standard compounds, including rutin, quercetin, kaempferol, caffeic acid, and luteolin 7-glucoside, were purchased from the China National Institutes for Food and Drug Control. Six standard compounds, including isoquercitrin, isorhamnetin, catechin, epicatechin, nuciferine, and chlorogenic acid, were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Sichuan, China). The purity of all the regents was HPLC ≥ 98%.

Orientin (CAS Registry Number 28608-75-5), vitexin (CAS Registry Number 3681-93-4), isovitexin (CAS Registry Number 38953-85-4), genistin (CAS Registry Number 529-59-9), luteolin (CAS Registry Number 491-70-3), genistein (CAS Registry Number 446-72-0), apigenin (CAS Registry Number 520-36-5), isorhamnetin (CAS Registry Number 480-19-3), and biochanin A (CAS Registry Number 491-80-5) were purchased from Weikeqi Biological Technology Co., Ltd. (Chengdu, China). Cajaninstilbene acid (CAS Registry Number 87402-84-4) was obtained by the separation and purification from pigeon pea leaves by column chromatography in our laboratory.

characterization of these compounds were described previously [54]. Norwogonin and 8?methoxykaempferol were purchased from ChemFaces (Wuhan, China). Isorhamnetin, eupatilin, icaritin, fisetin, and morin were purchased from Sichuan Weikeqi Biotechnology Company (Chengdu, Sichuan, China).

Eight I. tinctoria hairy root lines (I–VIII) were successfully induced via the genetic transformation of A. rhizogenes LBA9402 strain in our laboratory [6]. Eight ITHRCs (I–VIII) originating from different hairy root lines were harvested by filtration after 4 weeks of cultivation, and their moisture contents were predetermined for the further quantitative analysis. The standard compounds epigoitrin (EPI), isatin (ISA), indole-3-carboxaldehyde (INC), tryptanthrin (TRY), indigo (ING), indirubin (INR), rutin (RUT), neohesperidin (NEO), buddleoside (BUD), liquiritigenin (LIQ), quercetin (QUE), isorhamnetin (ISR), kaempferol (KAE), and isoliquiritigenin (ISL) were purchased from Weikeqi Biological Technology (Sichuan province, 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.

Gallic acid, Folin-Ciocalteu reagent, 2, 2’-azobis-amidinopropane

(ABAP), and dichlorofluorescin diacetate (DCFH-DA) were

purchased from Sigma-Aldrich Inc. (St. Louis, USA). Isorhamnetin,

kaempferol, quercetin and their glycosides, syringic acid, p-cumaric

acid, epicatechin and catechin were purchased from Weikeqi

Biological (Chengdu, China). Alpha-amylase, Pepsin, Pancreatin, Bile

Extracts, Lipase, and Viscozyme L were purchased from SigmaAldrich, Ltd. (St. Louis, MO, USA). Human cancer cells HepG2, MCF7, MDA-MB-231 and Caco-2 were purchased from Sun Yat-Sen

University.

Mature seeds of I. tinctoria were generously provided by Rongquan Medicine Plant Co. Ltd., Daqing, Heilongjiang Province, China. FL standards including RUT, NEO, BUD, LIQ, QUE, ISR, KAE and ISL were purchased from Weikeqi Biological Technology Co. Ltd. (Sichuan province, China). Other reagents of either analytical or optical grades were obtained from Beijing Chemical Reagents Co. (Beijing, China). To produce I. tinctoria aseptic plantlets, seeds were surface sterilized with 70% (v/v) ethanol for 45 s and 4% (v/v) sodium hypochlorite solution for 8 min, and then rinsed 5 times with sterilized water.

Daidzein, genistein, myricetin, kaempferide, isorhamnetin, kaempferol, isoquiritigenin and hesperetin (all purities?>?98%) were purchased from Weikeqi Biological Technology Co., Ltd. (Sichuan, China). α-CD, β-CD, γ-CD, Me-β-CD, HP-β-CD and SBE-β-CD were purchased from Binzhou Zhiyuan Biotechnology Co., Ltd. (Shandong, China).

GBE was purchased from Pizhou Futong Biochemical Technology Co., Ltd. (Xuzhou, China). The reference standards quercetin, kaempferol, isorhamnetin and genistein were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, China). Purities of all the reference standards were over 98% according to HPLC-DAD analysis.

Institute for the Control of Pharmaceutical and Biological Products (Beijing, P.R. China), while ginkgolide B and isorhamnetin were purchased from Weikeqi Biotechnology Co., Ltd. (Sichuan, China). The purity of these reference standards was determined by LC–MS and not less than 99.5%.

Substrates including tectorigenin (1), formononetin (2), genistein (3), daidzein (4), apigenin (5), luteolin (6), naringenin (7), eriodictyol (8), hesperetin (9), kaempferol (10), isorhamnetin (11), 3,4-dichloroaniline (13), 3,4-dichlorobenzenethiol (14), emodin (15), tectoridin (16), puerarin (17), vitexin (18), orientin (19), and mangiferin (20) were purchased from Sichuan Weikeqi Bio-Technology CO., LTD. (Chengdu, China) and Chengdu Must Bio-Technology Co., Ltd (Chengdu, China) while the compound phloretin (12) as a gift was obtained from Dr. Peng Wu, Guangzhou University of Chinese Medicine.

Isorhamnetin (98% purity), purchased from Sichuan Weikeqi Biological Technology Co., Ltd (Sichuan, China), was

dissolved in dimethyl sulfoxide (DMSO) as a stock solution,

stored at , and diluted with medium before each experiment.

The reference standards of apigenin, luteolin, quercetin, isorhamnetin,ILG, ILQ, and liquiritigenin (purity >98.5%, HPLC) were purchasedfrom Sichuan Weikeqi Biological Technology, Co., Ltd. (Chengdu,China). Acetonitrile and methanol of HPLC grade were purchasedfrom Fisher Scientific (Waltham, MA, USA).

bilobalide, genkwanin and apigenin were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China), while ginkgolide B and isorhamnetin were from Weikeqi Biotechnology Co. Ltd (Sichuan, China). The purity of these reference standards was assumed to be 99.5% as stated by the suppliers.

Ginkgolic acid standard (C17:1) CAS # 111047-30-4 (HPLC ≥ 98%), quercetin CAS # 117-39-5 (HPLC ≥ 98%), kaempferide CAS # 520-18-3 (HPLC ≥ 98%), and isorhamnetin CAS # 480-19-3 (HPLC ≥ 98%) were purchased from Sichuan Weikeqi Biotechnology Co. (Sichuan, China).

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).

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