Methanol

The companies names, cities and countries of used chemicals as followed: methanol (85%), benzene, distilled water and ethanol (70%) from CNW Technologies GmbH (Dusseldorf - Germany). Aluminum chloride from Sangon Biotech (Shanghai- China). Sodium nitrite, sodium bicarbonate and sodium hydroxide from Research Lab Fine Chem. Industries (Maharashtra - India). Dimethyl sulfoxide (DMSO) from Piochem (Giza - Egypt). Gentamycin, when agar from Himedia (Giza - Egypt). Folin-Ciocalteau from Sigma Aldrich (USA). The speacies was authenticated at fungi center, Al-Azhar University (Nasr city - Egypt).

All chemicals and solvents were analytical or HPLC grade. Water, methanol, pyridine, n-hexane, methoxylamine hydrochloride (97%), and bis (trimethylsilyl) trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane (TMCS) were purchased from CNW Technologies GmbH (Düsseldorf, Germany). Trichloromethane was obtained from Sinopharm Chemical Reagent Co.; Ltd. (Shanghai, China). L-2-chlorophenylalanine was obtained from Shanghai Hengchuang Biotechnology Co.; Ltd. (Shanghai, China).

Mesalazine was purchased from Shanghai Haoyuan Biomedical Technology Co., Ltd. (Shanghai, China); TNBS and formic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA); methanol and acetonitrile were purchased from CNW Technologies (Shanghai, China); 2-chloro-L-phenylalanine was purchased from Shanghai Hengbai Biotech Co., Ltd. (Shanghai, China); and enzyme-linked immunosorbent assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu, China).

LC-MS grade acetonitrile, methanol, ammonium acetate, and formic acid were purchased from CNW Technologies (Duesseldorf, Germany). 2-Chloro-L-phenylalanine was purchased from Shanghai HB Biotech Co., Ltd. (Shanghai, China). Deionized water was prepared on a Millipore Milli-Q water purification system (Billerica, MA, USA). Apigenin was obtained from China National Engineering Research Centre for Solid Preparation Manufacturing Technology (Beijing, China). Psoralen was purchased from China Food and Drug Control Institute (Beijing, China). Assay kits for aminotransferase (ALT) and aspartate aminotransferase (AST) were purchased from Nanjing Jiancheng Biological Engineering (Nanjing, China). RFH was purchased from Baining Pharmaceutical Co., Ltd. (Guangzhou, Guangdong).

Methanol and acetonitrile were purchased from CNW Technologies, acetic acid from Fisher Chemical, and ammonium acetate from Sigma-Aldrich (Shanghai, China). All reagents were of LC-MS grade. Ultra-high-performance liquid chromatography was performed using Q Exactive HFX High-Resolution Mass Spectrometry (Heraeus Fresco Model 17 Centrifuge; Thermo Fisher Scientific, Waltham, MA, USA). A scale (model BSA124S-CW) was purchased from Sartorius. A Ps-60al ultrasonic instrument was purchased from Shenzhen Redbond Electronics Co., Ltd. A rice cooker (Mb-wfs4029 Midea) was purchased from Midea Life Electric Appliance Manufacturing Co., Ltd. (Foshan, China).

4-6 weeks old healthy female KM mice (SPF grade) with body weights of 16-18 g were purchased from Ji’nan Pengyue Laboratory Animal Breeding Co., Ltd (Shandong, China). The mice were housed in a clean room at a temperature of 23 ± 2°C and humidity of 50 ± 5% with a 12 h alternating light and dark cycle. They were permitted free access to food and water. All animal experiments were performed according to the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals and were approved by the Animal Care and Use Committee of Shandong Province, China. The thyroxine tablets were purchased from Shandong Renhe Pharmaceutical Co., Ltd (China). PerCP-Cy™5.5 Hamster Anti-Mouse CD3e (Cat# 551163), FITC Rat Anti-Mouse CD4 (Cat# 553047), APC-H7 Rat anti-Mouse CD8a (Cat# 560182), PE Rat Anti-Mouse IFN-γ (Cat# 554412), Leukocyte Activation Cocktail, with BD GolgiPlug (Cat. No.550583) were purchased from BD Pharmingen™. All chemicals and solvents were analytical or HPLC grade water, methanol, acetonitrile, formic acid, pyridine, n-hexane, methoxylamine hydrochloride, N, O-Bis (trimethylsilyl) trifluoroacetamide (BSTFA) with 1% chlorosilane were purchased from CNW Technologies GmbH (Germany), L-2 chlorophenylalanine was from Shanghai Hengchuang Bio technology Co Ltd. (China).