Glycyrrhizic acid

Methanol and acetonitrile (chromatographic purity) was purchased from Fisher company (USA), formic acid (MS grade) was purchased from ACS company (USA), and distilled water was purchased from Guangzhou Watsonsfood and beverage company (Guangzhou, China). Reference standards of sophocarpine, matrine, febrifugine, berberine, palmatine, Tangeratin, nobiletin, liensinine, neferine, scopoletin, isoliquiritigenin, liquiritigenin, naringenin, naringin, hesperidin and glycyrrhizic acid, jatrorrhizine were purchased from Sichuan Weikeqi Biotechnology Co., Ltd. or Shanghai Yuanye Biotechnology Co., Ltd. XSNC were supplied by Shanxi Momentum Pharmaceutical Co., Ltd (Shanxi, China).

Liquiritin apioside, liquiritin, neoliquiritin, isoliquiritin apioside, isoliquiritin, ononin, naringin, liquiritigenin, poricoic acid A, poricoic acid B, poricoic acid C, glycyrrhetinic acid, dehydropachymic acid, pachymic acid, ginsenoside Re, ginsenoside Rg1, ginsenoside Rf, isoliquiritigenin, ginsenoside Rb1, ginsenoside Rg2, ginsenoside Rg3, ginsenoside Rg5, 20(S)-ginsenoside Rh1, notoginsenoside R1, ginsenoside Ro, ginsenoside Rc, formononetin, ginsenoside Rb2, glycyrrhizic acid, ginsenoside Rd, ginsenoside Rk1, and atractylenolide III were purchased from Chengdu DeSiTe Biological Technology Co., Ltd. (Chengdu, China) or Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, 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

14-O-benzoylmesaconine (PubChem CID 24832659), 14-O-benzoylaconine (PubChem CID 20055771) and 14-O-benzoylhypaconine (PubChem CID 78358526), liquiritin apioside (PubChem CID 10076238), liquiritin (PubChem CID 503737), liquiritigenin (PubChem CID 114829), isoliquiritin apioside (PubChem CID 6442433), isoliquiritin (PubChem CID 5318591), isoliquiritigenin (PubChem CID 638278), glycyrrhizic acid (PubChem CID 14982) and glycyrrhetic acid (PubChem CID 3230) were purchased from Chengdu Weikeqi Biological Technology Co., Ltd. (Chengdu, Sichuan, China). Hypaconitine (PubChem CID 441737), atenolol (PubChem CID 2249) and propranolol hydrochloride (PubChem CID 62882) were purchased from Dalian Meilun Biological Technology Co., Ltd.

LQ, LA, ILQ, ILA and GA were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, China), and their structures are shown in Fig. 1. Their purities were above 95% according to HPLC-UV analysis. All ionic liquids ([C2MIM]Br, [C4MIM]Ac, [C4MIM]Cl, [C4MIM]BF4, [C4MIM]NO3, [C4MIM]HSO4, [C4MIM]Br, [C6MIM]Br, [C8MIM]Br and [C10MIM]Br, where C2MIM = 1-ethyl-3-methylimidazolium, C4MIM = 1-butyl-3-methylimidazolium, C6MIM = 1-hexyl-3-methylimidazolium

Glycyrrhetinic  acid  and  glycyrrhizicacid (purity≥98%) were purchased from SichuanWeikeqi  Bio-technology  Co.  Ltd  (Sichuan,  China).4-methylumbelliferone (4-MU), 4-methylumbelliferone-b-D-glucuronide (4-MUG)

The kidney cells of channel catfish (CC-K) was gained from Prof. Lingbing Zeng

from the Yangtze River Fisheries Research Institute of the Chinese Academy of

Fishery Sciences (Wuhan, China). Curcumin, demethoxycurcumin,

bisdemethoxycurcumin, glycyrrhizic acid, glycyrrhetnic acid, taxol, rotenone, saponin,

sanguinarium chloride, quercetin, schisandrol B, schisandrin A, schisandrin B,

schisandrol A, schisantherin A, genistein, artemisinin, chlorogenic acid, toosendanin,

hyericin, baicalein, baicalin, ginkgolide A, ligustilide, dihydroartemisinin, oxymatrine

and matrine were obtained from Weikeqi Biotechnology Co., Ltd. (China), and the

purity of all natural products ≥ 98%. Rifampicin and erythromycin was purchased

from Beijing Solarbio Technology Co., Ltd. (Beijing, China).

The following programme was applied: 0–3.5 min, 79% B; 3.5–8.2 min, 79–98% B; 8.2–11 min, 98–79% B; and 11–20 min, 79% B. All of the chemical reference substances, including glycyrrhetinic acid (CAS: 471-53-4), glucopyranosiduronic acid (CAS: 34096-83-8), and glycyrrhizin (CAS: 1405-86-3), had > 98% purity and were commercially available (Weikeqi Biological Technology Co. Ltd, Sichuan, China).

Reference standard materials of daphnetin (DPH) (batch number 121025), daphnoretin (DR) (batch number 130305), 7-hydroxycoumarin (HDC) (batch number 120927), liquiritin (LQ) (batch number 111127), isoliquiritin (ILQ) (batch number 201010), liquiritigenin (LG) (batch number 201004), isoliquiritigenin (ILG) (batch number 101026), glycyrrhizin (GL) (batch number 111023), and glycyrrhetinic acid (GA) (batch number 110906) were all purchased from Sichuan Weikeqi Bio-tech Co., Ltd. (Chengdu, China). The chemical structures of these compounds are shown in Figure 3. Hesperidin (HP) (batch number 0721-200010) used as an internal standard (IS)

The extracts of LGWWJX formula and its single traditional Chinese herbs without adjuvants were provided by Sichuan Neo-Green Pharmaceutical Technology Development Co., Ltd. (Chengdu, Sichuan, China). The LGWWJX formula is listed in Table 1. Standards for LGWWJX, including liquiritin, schizandrin, and deoxyschizandrin were purchased from the National Institutes for Food and Drug Control (Beijing, China), and isoliquiritin and glycyrrhizinic acid were from Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, Sichuan, China). Rodent diets with 60 and 10 kcal % fat were purchased from Research Diets, Inc. (New Brunswick, NJ, USA) and Jiangsu Synergetic Pharmaceutical Bioengineering Co., Ltd.