Schizandrin

Ginsenosides Rb₁, Rb₂, Rc, Rd, Re, Rg₁, Rh₁ and compound K were purchased from ChromaDex (Irvine, CA, USA). Ophiopogonin D was purchased from TAUTO (Shanghai Tauto Biotech Co. Ltd., Shanghai, China). Schizandrin, 5-methoxyflavone and digoxin were obtained from Sigma-Aldrich Chemicals (St. Louis, MO, USA). LC/MS grade acetonitrile as obtained from J. T. Baker. Inc. (Phillipsburg, NJ, USA). Triple-deionized water (Millipore, Bedford, MA, USA) was used in the study. The root parts of Panax ginseng and Radix ophiopogonis, and the fruit parts of Fructus schisandrae were purchased from Zheng-Yuan herbal and traditional Chinese medicine store, Taipei, Taiwan. After comparing with the specimens in the National Research Institute of Chinese Medicine of Taiwan, the purchased herbs were the same as the specimens. Five different commercial pharmaceutical products of Sheng-Mai-San were included: SunTen Pharmaceutical Co., Ltd. (Taipei, Taiwan), Kaiser Pharmaceutical Co., Ltd. (Tainan, Taiwan), Chuang-Song-Zong Pharmaceutical Co., Ltd. (Kaohsiung, Taiwan), ShengFoong Co., Ltd. (Taipei, Taiwan), and ShengChang Pharmaceutical Co., Ltd. (Taipei, Taiwan). When presenting the analysis results, numeral codes were used to represent the names of these manufacturers. This study didn't be funded by any of the pharmaceutical herbal products of these manufacturers.

2,000 cells of MDA-MB-231 (purchased from ATCC) which are cultured in RPMI media containing 10% fetal bovine serum (FBS) and 1% antibiotics, were seeded in 96 well plates and incubated for 12 hr. Vanillic acid, L-Malic acid, Schizandrin and Syringin (Eleutheroside B) were purchased from Sigma. After 12 hr, they were dissolved in DMSO and treated in 5% FBS-containing media dose dependently. After 24, 48 and 72 hr, MTT reagent (5mg/ml, Sigma) was added to each well, and the plates were incubated in 37^°C for 2 hr to check the cell viability. Purple-colored formazan dissolved in DMSO was analyzed spectrophotometrically at 570nm using ELISA plate reader. All the experiments were repeated three times.