The ethanol extract of grown in Hainan province (China) was analysed

The ethanol extract of grown in Hainan province (China) was analysed by GC and GC/MS. we analyzed the bioactivity of the extract, including antioxidant and anti-tumor activities. 2.?Materials and Methods 2.1. Materials was obtained from the Spice and Beverage Research Institute at the Xing TNFSF8 Long Tropical Botanical Garden of Chinese Academy of Tropical Agricultural Sciences (Hainan province, China). Leaves of was dried (a relative humidity between 2.8% and 3.4%) and ground into fine powder (70 mesh) using a mechanical grinder. Mouse leukemic L1210 cells was obtained from the Shanghai Cellular Institute of China Scientific Academy, Human Colon Carcinoma LoVo Cell Collection kindly offered from your Pharmacological Laboratory in Shanghai Jiao Tong University or college. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma Chemical Organization GSK-J4 manufacture (St. Louis, MO, USA). Methotrexate (MTX) was purchased from Shanghai Hualian Pharmacy Co Ltd. All other solvents and chemicals were of analytical grade. 2.2. Extraction of bioactive compounds The bioactive compounds of ground leaf were extracted using ethanol as solvent. Extraction was carried out using a shaking incubator at room heat for 2 h followed by filtration through Whatman No.1 filter paper. The residue was re-extracted twice in the same manner and the three filtrates were combined. The ethanolic extract was concentrated using a rotary evaporator at 55 C to near dryness. The extraction rate was 4.2%. 2.3. GC-MS analyses The ethanolic extracts compositions were analysed by GC-MS performed using a Trace GC/MS gas chromatograph coupled to an ion trap detector [7]. The fused-silica column was a SLB-5MS silica column (Supelco, GSK-J4 manufacture Bellafonte, PA, USA) GSK-J4 manufacture (30 m 0.25 mm film thickness 0.25 cm). GC-MS data were obtained using the following conditions: carrier gas helium (He 99.999%); circulation rate 1.0 mLmin?1; the split ratio 1/70 (v/v). An aliquot of 100 mg of distilled oils were diluted with 1 mL acetone, as were the extracts, and 1.0 L was injected into the GC-MS system. The oven heat plan was: 70 C for 13 min, from 70 to 280 C at 6 C min?1, and keeping 280 C for 16 min. The injector, transfer ion and series snare temperature ranges had been 250, 280 and 200 C, respectively. The electron influence (70 eV) spectra had been documented at 1 scan/s using a filament emission current of 10 A. The id of volatile substances was structured both on evaluation from the linear retention indexes (RI) computed using the Truck der Dool and Kratzs formula with those reported in the books [8] and by the complementing of mass spectra from the compounds using the guide mass spectra of two libraries (Wiley5 and Nist05) in conjunction with the program of GC-MS and Adams collection [9]. For the main chromatographic peaks, id was confirmed using authentic criteria. 2.4. Nuclear magnetic resonance The 400 MHz 1H-NMR spectra of most sample batches had been analyzed with an AMX-II 600 MHz spectrometer (Bruker Musical instruments, Inc.), 32 scans had been gathered into 64 K data factors more than a spectral width of 4789 Hz (12 ppm) using the transmitter offset at 5.00 ppm, yielding an electronic resolution of 0.15 Hz per stage. 1H-NMR high-temperature spectra at 353 K had been assessed at 300.13 MHz utilizing a QMP probe. 32 scans had been gathered into 64 K data factors more than a spectral width of 6172.84 Hz (20.56 ppm). A turn position of 30 was utilized. The acquisition period was 5.31 s, accompanied by a relaxation hold off of just one 1 s. An exponential series broadening home window function of 0.3 Hz was found in the data handling. 13C-NMR spectra had been documented at 25 C utilizing a Bruker Avance 600 NH3 spectrometer working at 14 T and built with a 5 mm broad-band probe. For the quantitative 13C-NMR research the next experimental.