Berberine

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).

QZWTT was provided by Liaoning Huarun Benxi Sanyao Co. Ltd(Benxi, China, lot no. 20200101). We used 26 chemical referencestandards in this study (Chengdu Pufeide BioTechnology Co. Ltd,Cheng du, China or Weikeqi Biological Technology Co. Ltd, Chengdu,China; Table S1). The acetonitrile, methanol (Merck, Germany) andformic acid (Fisher, USA) were of LC–MS grade, whereas the remain-ing chemicals were of analytically grade.

For preparing BBR-loaded PLs, the following materials were used: BBR in a base form C20H18NO4+ (Mw 336.36 g/mol, chemical purity > 98%) (Sichuan Weikeqi Biological Technology Co., Ltd., Chengdu, China), hydrogenated soy phosphatidyl choline, HSPC (>98%) (Lipoid GmbH, Ludwigshafen, Germany), distearoylphosphatidyl glycerol, DSPG (>98%)

All the medicinal materials were tested, and their quality met the relevant standards of the 2020 edition of the Chinese Pharmacopoeia. Standard reference compounds, including epiberberine (wkq20031807), coptisine (wkq20021306), palmatine (wkq20051201), berberine (wkq20032408), baicalin (wkq20010908), wogonoside (wkq20040108), baicalein (wkq20022608), wogonin (20030204), rhein (wkq20052209), aloe-emodin (wkq20032708), emodin (wkq20041308), chrysophanol (wkq20081705), and physcion (wkq20040101) were purchased from Weikeqi Biotechnology Co., Ltd (Chengdu, China). The purity of all standard products was higher than 98%. Chromatographic grade acetonitrile and formic acid were purchased from Thermo-Fisher (US).

Irinotecan (CPT-11, purity ≥99%) was purchased from Melonepharma Co., Ltd. (Dalian, China). Standard substances, puerarin, berberine, baicalin and liquiritin (purity ≥98%), were purchased from Weikeqi Biological Technology Co., Ltd (Chengdu, China). HPLC-grade methanol, acetonitrile and formic acid were purchased from Thermo Fisher Scientific (Thermo scientific, USA).

enzyme-linked immunosorbent assay (ELISA) kit was obtained from Multi Science Biotechnology Co., Ltd. (Hangzhou, China). Reference standards of astragaloside, heterophyllin B, atractylodin, harpagide, berberine, baicalin, puerarin, quercetin, protocatechuic acid and tanshinone II A were obtained from Sichuan Weikeqi Biological Technology Co., Ltd. (Sichuan, China).

Nuciferine, capsaicin, and evodiamine were purchased from Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Berberine and coptisine were purchased from Weikeqi Biotech Co., Ltd. (Chengdu, China). Camptothecin was purchased from Melone Pharma Co., Ltd. (Dalian, China).

Berberine (Mw 336.36 g/mol and chemical purity >98%; Sichuan Weikeqi Biological Technology Co., Ltd., Chengdu, China) was used as an active ingredient in base form (C20H18NO4+). Hydrogenated soy phosphatidyl choline (HSPC) (>98%) and distearoyl phosphatidyl glycerol (DSPG)

Irinotecan (CPT-11, purity ≥99%) was purchased from Melonepharma Co., Ltd. (Dalian, China). Gegen Qinlian pills were purchase from Guangxi Huahong Pharmaceutical Co., Ltd (Guangxi, China). Standard substances, puerarin, berberine, baicalin and liquiritin, alkaloid, Baicalein, palmatine (purity ≥98%), were purchased from Weikeqi Biological Technology Co., Ltd (Chengdu, China). loperamide (LPM, purity ≥98%) was purchased from Xian Janssen Pharmaceutical Co., Ltd.

Acetonitrile (ACN) from RCI Labosan Ltd. (Bangkok, Thailand), ethanol (EtOH) and methanol (MeOH) from Merk (Darmstadt, Germany) were both in HPLC grade without further purification. Berberine, used as internal standard (IS) for quantification of piperine, was bought from Sichuan Weikeqi Biological. Milli-Q water from Millipore water purification system (Millipore, Milford, MA, USA) was used in the preparation of nanoparticles and all other solutions.

The TWMM was provided by Beijing HanDian Pharmaceutical Co., Ltd. (Beijing, China). Chlorogenic acid, linarin, luteolin, jaspolyside, tolbutamide, stachydrine, magnoflorine, calycosin-7-O-β-d-glucoside, stepharanine, jatrorrhizine, epiberberine, columbamine, ononin, berberine, palmatine, berberrubine, calycosin, formononetin, 2α-hydroxyoleanic acid, hydroxytyrosol, salidroside, luteolin-7-O-glucoside, verbascoside, specnuezhenide, oleuropein, physcion, oleonuezhenide, astragaloside IV, astragaloside II, isoastragaloside II, astragaloside I, luteolin, formononetin, and tolbutamide (purity ≥ 98%) were purchased from Shanghai Yuanye Pharmaceutical Technology Co., Ltd and Sichuan Weikeqi Pharmaceutical Technology Co., Ltd. Acetonitrile and formic acid both in HPLC grade were obtained from Fisher Scientific (Fair Lawn, NJ, USA). Ultrapure water was prepared using a Milli-Q water purification system (Millipore, Bredford, MA, USA). Other reagents were all analytical grade.

Berberine and curcumin of purity >98% were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Chengdu, Sichuan, China) and Zhengzhou Stars Shine Technology Co., Ltd. (Zhengzhou, Henan, China) respectively. Detection kits for low-density lipoprotein cholesterol (LDL-c), low-density lipoprotein cholesterol (HDL-c), alanine aminotransferase (ALT)

rat liver microsome, pooled human liver microsome and MgCl2 were from Sigma-Aldrich Co. Berberine was used as internal standard (IS) and was bought from Sichuan Weikeqi Biological. HPLC grade acetonitrile (ACN) from RCI Labosan Ltd. (Bangkok, Thailand) and methanol (MeOH) from Merck (Darmstadt, Germany)

BBR was provided by Weikeqi Biological Technology Co., Ltd. (Sichuan, China). Metformin (MET), carboxymethyl cellulose sodium (CMC-Na), and STZ were purchased from Sigma-Aldrich (California, USA). BestarTM qPCR RT kit and Bestar SYBY Green qPCR Master mix were purchased from DBI (Ludwigshafen, Germany). All test assay kits and the CORT, ACTH, CRH, Orenxin A, and OX2R enzyme-linked immunosorbent assay (ELISA) kits were supplied by NanJing JianCheng Bioengineering Institute

Magnetic microbeads (MBs) (mean diameter 1 μm) modified by streptavidin (SA–MBs) were purchased from BioMag Scientific Inc. (China). Berberine, epiberberine, coptisine, columbamine, palatine, jatrorrhizine, and sinomenine were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (China). The cyanine dye MTC was synthesized according to the methods of Hamer32 and Ficken.

Piperine (purity>97%) was purchased from Aldrich Chem. Co. (Milwaukee, WI, USA).

Berberine was used as internal standard (IS) and was bought from Sichuan Weikeqi Biological

Technology Co. (Sichuan, China). Formic acid was obtained from BDH Laboratory Supplied Ltd.

(Kampala, Ukraine).

Acetonitrile was of HPLC grade and supplied by Merck Company, Inc. (Merck, Darmstadt, Germany). Ultrapure water was prepared by a Milli-Q50 SP Reagent Water System (Millipore Corporation, MA, USA). All other reagents were of analytical grade. Authentic standards of Phe, Mag, Jat, Pal, Ber, Ob1 and Ob2 were provided by Sichuan WeiKeqi Bio-Tech Co., Ltd (Sichuan, China).