Acetonitrile and methanol

Chinese bayberry leaves of ‘Biqi’ cultivar were hand-harvested randomly in June, 2020 in Cixi (Zhejiang, China). Sorghums of ‘Hongzhenzhu’ cultivar were provided by Shandong Academy of Agricultural Science (Jinan, China). GSPs (95%) were purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Acetonitrile and methanol of HPLC grade were purchased from Merck KgaA (Darmstadt, Germany). Acetic acid of HPLC grade was purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Other chemical reagents of analytical grade were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).

The chemicals and sources for purchase were Standard AmB. Eighty percent Pure HPLC grade (Sigma-Aldrich, St. Louis, MO, USA), AmB, USP grade was (Biobasic Inc, Markham, ON, Canada), mannitol, sodium dihydrogen phosphate dihydrate, sodium acetate (Ajax Finechem Pty Ltd., New South Wales, Australia), simulated intestinal fluid (SIF) powder (Biorelevant, London, UK), dimethyl sulfoxide (DMSO) (BD Biosciences, New South Wales, Australia), HPLC grade organic solvents, including acetonitrile and methanol (Merck, Darmstadt, Germany), phosphate buffer solution (PBS) tablets, and D-α-tocopheryl polyethylene glycol succinate (VitE-TPGS) (Sigma-Aldrich, NSW, Australia), Alexa Fluor 488 conjugate and 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) (Invitrogen, Carlsbad, CA, USA), glacial acetic acid, hydrochloric acid and sodium chloride (Chem-supply (Gillman, SA, Australia), CaCo-2 (c2bbe clone, ATCC^® CRL-2102™) cells (American Type Culture Collection (ATCC), Manassas, VA, USA), Hanks balanced salt solution (HBSS), phosphate buffered solution (PBS), Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), and Penicillin–Streptomycin (PS) (Thermo Fisher, Victoria, Australia). Soluplus^® was kindly supplied by BASF (Ludwigshafen, Germany). High purity Milli Q water (Merck Millipore, NSW, Australia) were used throughout the study.

The chemical reference standards isoquercitrin, luteolin, naringenin, and kaempferol were provided by Chengdu Herbpurify biotechnology CO., LTD (Chengdu, China); Phlorizin, Phloretin, and Trilobatin were purchased from Sichuan Wei Keqi Biotechnology Co., Ltd (Sichuan, China); eriodictyol and hyperoside were provided by Chengdu Push biotechnology CO., LTD (Chengdu, China); quercetin and apigenin were provided by Chengdu Alfa biotechnology CO., LTD (Chengdu, China); baicalein and wogonin were obtained from Chengdu Desite biotechnology CO., LTD (Chengdu, China). The purities of all chemical reference standards were above 98% by HPLC-DAD.
Acetonitrile and methanol of chromatography grade were provided by MERCK (Darmstadt, Germany); The ultrapure water was produced by a milli-Q water purification system (Millipore, Milford, MA, United States); formic acid of LC-MS grade and all other reagents of analytical grade were purchased from Aladdin Industrial Corporation.
P. freyniana was collected from Tong-Dao country of Huaihua (109.86 longitude, 26.03 latitude), Hunan province and were identified by Professor Wei Cai (Hunan Provincial Key Laboratory of Dong Medicine, Hunan University of Medicine). The voucher specimen was deposited at School of Pharmaceutical Sciences, Hunan University of Medicine.

QFGBG (Produced by Pharmaceutical Factory of Heilongjiang Institute of Traditional Chinese Medicine, batch number: 07210501, which meets the standards on pages 979–980 of Chinese Pharmacopoeia I, 2020), Cyclophosphamide injection (jiangsu hengrui Pharmaceutical Co., Ltd.; batch number: 20092225), Levamisole hydrochloride tablets (Renhetang Pharmaceutical Co., Ltd.; batch number: 201001), Watsons water, Formic acid (Fisher Company), Acetonitrile and methanol (Merck, Germany), UltraSYBR One Step RT-qPCR Kit (CW0659, CoWin Biosciences, China), themouse serum IL-4 (EK0405), IFN-γ (EK0375) ELISA kit (Boster, China), Mouse ACTB Endogenous Reference Genes Primers, 10 μM (B661302-0001, Sangon Biotech, China).
Nitrogen generator (Hangzhou Dacker, DFNW-5LB), Cryogenic centrifuge (Thermofisher, 17R), Upright white photo microscope (Nikon Japan, Eclipse Ci-L), Scanning software (3 dhistech Hungary, CaseViewer2.4), Panoramic slice scanner (3DHISTECH Hungary, PANNORAMIC, DESK/MIDI/250/1000), Ultra-high performance liquid chromatograph (AB Sciex ExionLC AD, United States), Mass spectrometer (AB SCIEX Triple-TOFTM 5600+, United States), Infifinite M200 PRO (Tecan, Switzerland), Bioer Line gene 9600 flfluorescence Quantitative PCR instrument (Hangzhou Bioeri, China).