Chemical Constituents and Antibacterial Activity of Dipsacus as a Medicinal Plant of the Bai Ethnic Group - Raw Data

Compound 1 was confirmed as a new compound through SciFinder and named Cynoglossoside A.2. In vitro microdilution broth antimicrobial assayAfter removing the strain frozen at -80°C, gradually thaw it, revive and activate it, then inoculate it onto a solid culture medium plate. Place it in a constant temperature incubator at 37°C for 12–24 hours until colonies grow.

Use a loop to pick a single colony and inoculate it into a centrifuge tube containing broth medium. Incubate on a shaker at 37°C and 180 rpm for 8–12 hours for later use.Using the micro broth dilution method and the reference method, in vitro antibacterial activity screening was conducted on the extracted parts of DiDi Xiao ethyl acetate and the isolated and identified monomer compounds. Dilute the cultured bacterial solution to 5 × 105 CFU/mL, add 100 μ L of the diluted bacterial solution to each well of a 96 well plate, and add 96 μ L of the diluted bacterial solution and 4 μ L of the test sample solution (the test sample is dissolved in DMSO and prepared as a 10 mg/mL test sample solution) to the first row of wells of each sample.

Set the initial antibacterial concentration of the test sample to 200 μ g/mL, and use the half dilution method to obtain different antibacterial concentration gradients (3 replicates are set for each concentration). The positive control is vancomycin (Gram positive bacteria) or ciprofloxacin (Gram negative bacteria) with a final concentration of 2.0 μ g/mL, the negative control is DMSO (2 μ L) with the maximum antibacterial concentration gradient corresponding to the volume of the test solution added to each well, and the blank control is 100 μ L liquid culture medium.

After being placed in a constant temperature incubator at 37 ℃ for 24 hours, observe and compare with negative control, positive control, and blank control to determine the minimum inhibitory concentration (MIC) value.The five human pathogenic strains used in this experiment, namely methicillin-resistant Staphylococcus aureus ATCC 43300 (MRSA), Staphylococcus epidermidis ATCC 12228, Streptococcus agalactiae ATCC 13813, Klebsiella pneumoniae 13883, and Escherichia coli ATCC 35218, were all provided by the School of Life and Marine Sciences at Shenzhen University.

After identification by our research group, they were preserved in the Key Laboratory for Screening of Antipathogenic Plant Resources in Western Yunnan Province.IV. Experimental instruments, reagents and tools1. Separation and purification equipmentSilicone, GF254 thin layer board; D101 macroporous resin; Agilent 1260、 Shimadzu LC-20AT, Agilent analysis/preparation C18 chromatography column, rotary evaporator, constant temperature water bath, chromatography column device.2.

Structural identification instrumentBruker Avance II-400 nuclear magnetic resonance instrument (TMS internal standard); Agilent C6230 TOF-MS mass spectrometer; UV spectrophotometer, infrared spectrometer, polarimeter;3. Antibacterial experimental equipmentConstant temperature incubator, shaker, 96 well cell culture plate, pipette; Ultra pure water, vancomycin, ciprofloxacin reference standards; DMSO solvent, broth medium.4.

Data organization toolOffice Excel (Antibacterial Data Sheet); MestReNova (professional nuclear magnetic resonance analysis software); ChemDraw (structural drawing); Word document (storing experimental text and spectral data).V. Table Data Explanation (In vitro Antibacterial Activity Data Table. xlsx)1. Record the total number of recordsThere are a total of 14 sample records, including 11 monomeric compounds, ethyl acetate extraction sites, and 2 positive antibacterial drug controls;2.

Meaning of row labelsCompounds 1-11: isolated and obtained 11 monomeric natural products;Ethyl acetate extraction site: 70% acetone extract of DiDi Xiao, ethyl acetate extraction site;Vancomycin: Gram positive bacteria positive control drug;Ciprofloxacin: Gram negative bacteria positive control drug3. Column label meaningMRSA: Methicillin resistant Staphylococcus aureus ATCC 43300;Epidermal 12228: Staphylococcus epidermidis ATCC 12228;No milk 13813: Streptococcus agalactiae ATCC 13813;Pneumonia 13883: Klebsiella pneumoniae ATCC 13883;Large intestine 35218: Escherichia coli ATCC 35218;Cell value: Minimum inhibitory concentration (MIC);4.

Measurement units and identificationMIC unified unit: μ g/mL; '>200' represents that the sample has no antibacterial activity at the highest tested concentration of 200 μ g/mL; The symbol '-' indicates that the antibacterial drug is not tested on the corresponding strain.