Treatment effects of ischemic stroke by berberine, baicalin and jasminoidin from Huang-Lian-Jie-Du-Decoction (HLJDD) explored by integrated metabolomics approach Qian Zhang1, Xiaowei Fu1, Junsong Wang2 *, Minghua Yang1, and Lingyi Kong1 * 1State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing, 210009, P.R. China 2Center for Molecular Metabolism, Nanjing University of Science and Technology, 200 Xiao Ling Wei Street, Nanjing, 210094, P.R. China * Correspondence and requests for materials should be addressed to Lingyi Kong (email:
[email protected]) or Junsong Wang (email:
[email protected]) 1 Supplementary Methods Herbal Materials and Samples Preparations HLJDD, composed of Rhizoma Coptidis (Coptis chinensis Franch, Ranunculacea), Radix Scutellariae (Scutellaria baicalensis Georgi, Labiatae), Cortex Phellodendri (Phellodendron chinensis Schneid, Rutaceae) and Fructus Gardeniae (Gardenia jasminoides Ellis, Rubiaceae), in a ratio of 3:2:2:3 (w/w/w/w), reaching a total weight of 1.0 kg, was extraction with 70 % ethanol (1:10, w/v) under reflux three times for 1h each. The extract solution was combined and freeze-dried in a vacuum to collect HLJDD extract (264.8 g, yield: 26.48 %). All herbs were purchased from Jiangsu Medicine Company (Nanjing, China, Drug GMP certificate: SUJ0623; Drug Manufacturing Certificate: SUY20110051), and authenticated by Professor Mian Zhang, Department of Medicinal Plants, China Pharmaceutical University, Nanjing, China. Berberine (purity > 98.0%) and jasminoidin (purity > 98.0%) were purchased from Qingdao Jieshikang Biotech Co., Ltd. (Qingdao, China), and baicalin (purity > 98.0%) was purchased from Dalian Meilun Biotech Co., Ltd. (Dalian, China). Sodium 3-trimethylsilyl-1-(2, 2, 3, 3-2H4) propionate (TSP) was provided by Sigma Chemical Co.