Milli q50 sp reagent water system

Sodium selenite (Na₂SeO₃, 97.0%) was purchased from Shanghai Tyrael Chemical, Co., Ltd. Ammonium phosphate dibasic ((NH₄)₂HPO₄, 99.0%) was obtained from Shanghai Macklin Biochemical Co., Ltd. Calcium nitrate tetrahydrate (Ca(NO₃)₂·4H₂O, 99.0%) was purchased from Sinopharm Chemical Reagent Co., Ltd. Silver nitrate (AgNO3, 99.8%) was obtained from Tianjin Tiangan Chemical Technology Development Co., Ltd. Ammonium solution ( NH₃·H₂O, 25–28%) was purchased from Ron Reagent Co., Ltd. Ultrapure water was prepared by a Milli-Q50 SP Reagent Water System (Millipore Corporation, MA, USA). The chemicals used in this study were all analytical grade procured from commercial sources, without further purification.

DNts were friendly provided by Shanghai Hutchison Pharmaceuticals (Shanghai, China) at July 2010. Standard compounds (emodin, aleo-emodin, rhein, chrysophanol, physcion) purchased from National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). ANIT were purchased from Sigma-Aldrich (St. Louis, USA). The purity of each compound was determined to be above 98% by HPLC analysis. Ammonium acetate, formic acid, acetonitrile and methanol (HPLC-grade) were purchased from Fisher Scientific (Nepean, Ont., Canada). Ultrapure water was prepared by a Milli-Q50 SP Reagent Water System (Millipore Corporation, MA, USA) for the preparation of samples and buffer solutions. Serum levels of ALT, AST, ALP, T-Bil, D-Bil and TBA were determined using a commercially available test kits (SHINO-TEST Corporation, Japan) with HITACHI 7080 Automatic Analyzer system (Japan). All other chemicals and solvents used were of analytical grade.

Chitosan with the viscosity of 50–800 mPa·s and the average degree of deacetylation of 90% was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). 2-(Dimethylamino) ethyl 2-methacrylate (DMAEMA) glutaraldehyde, and KPS were purchased from J&K Scientific Ltd. (Beijing, China). DMAEMA was purified by distillation under reduced pressure before use. Pyraclostrobin was obtained from Jiangsu Anpon Electrochemical Co., Ltd. (Huaian, China). Solvent oil S-200 was kindly provided by Jiangsu Hualun Chemical Industry Co., Ltd. (Yangzhou, China). Hydrochloric acid (HCl), acetone and other chemicals were all of analytical reagent and used as received without further purification. Ultrapure water used in all experiments was prepared using a MilliQ-50 SP reagent water system (Millipore Corporation, Bedford, MA, USA).

Radix Paeoniae Rubra (Chi Shao in Chinese), Salvia miltiorrhiza Bge (Dan Shen in Chinese), Trichosanthis Fructus (Gua Lou in Chinese) and Puerariae Lobatae Radix (Ge Gen in Chinese) were purchased from Lvye Co., Ltd. (Beijing, China) and identified by Professor Xiaohe Xiao, a taxonomist at China Military Institute of Chinese Materia Medica. HPLC-grade acetonitrile and formic acid were purchased from Merck Company Inc. (Merck, Darmstadt, Germany). Ultrapure water was prepared by a Milli-Q50 SP Reagent Water System (Millipore Corporation, MA, USA) for the preparation of samples and buffer solutions. Standard compounds which were purchased from National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China) were identified by Ultra Performance Liquid Chromatography (UPLC). ANIT was purchased from Sigma Chemical Co. (St. Louis, MO) and dissolved in olive oil to a final concentration of 2% before use. Kits for assaying the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), direct bilirubin (DBIL), alkaline phosphatase (ALP) and γ-glutamyl transpeptidase (γ-GT) were purchased from Mindray bio-medical electronics Co., Ltd. (Shenzhen, China). All other reagents or drugs used were of analytical grade.

Folin-Ciocalteu reagents, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) were purchased from Sigma Chemical (St. Louis, MO, USA). HPLC grade acetonitrile and formic acid were purchased from Fisher Scientific (Pittsburgh, PA, USA). Ultra-pure water was prepared using a Milli-Q50 SP Reagent Water System (Millipore Corporation, Billerica, MA, USA). Other reagents (analytical grade) were purchased from Sinopharm Chemical Reagent Co. Ltd. (Beijing, China).
Twenty Acer truncatum Bunge trees, whose tree-age (around 15 years old) and growth environment were approximatively identical, were randomly selected in Bajiajiaoye Park, Haidian District (N 40°00′57.21′′; E 116°19′43.36′′ with altitude 39–41 m) and authenticated by associate professor Zhonghua Liu, Beijing Forestry University, Beijing, China. A. truncatum leaves (ATL) were uniformly collected from the selected trees in 20 April 2015 and taken back to the laboratory immediately. The leaves were cleaned with distilled water and air-dried, the dried leaves were ground and passed through a 250 × 250-μm sieve, and the powder was stored at −20 °C in a refrigerator for extraction.

HPLC grade acetonitrile was purchased from Fisher Scientific (Pittsburgh, PA, USA). Ultra-pure water was prepared using a Milli-Q50 SP Reagent Water System (Millipore Corporation, Billerica, MA, USA). Other reagents (analytical grade) were purchased from Sinopharm Chemical Reagent Co. Ltd. (Beijing, China). Authentic standard ginsenosides Rh₂, 20(S)-Rg₂, F₁, Rd, Rb₁, Rb₂, Rf, 20(S)-Rg₁, Rb₃, Rc, 20(S)-Rh₁, 20(R)-Rh₁, 20(R)-Rg₂, 20(S)-Rg₃, 20(R)-Rg₃, CK, 20(S)-PPT and Re were purchased from Lyle Biological (Luoyang, China), and F₄ and Rg₅ were bought from Mansite Biological (Chengdu, China). Samples of 3-year-old ginseng flowers were collected on 25 June 2016 and purchased in Huanren County (Benxi, Jilin, China). All samples were morphologically authenticated as the flowers of P. ginseng Meyer by Dr. Zhong-hua Liu (Beijing Forest University, Beijing, China). Voucher specimens of P. ginseng Meyer were deposited in the Pharmacognosy Laboratory, Shenyang Pharmaceutical University (Shenyang, China). Ginseng flowers were air-dried in the shade, ground and stored at −20 °C for further use.