Model qp 2010

The reagents were commercially purchased from Sigma-Aldrich (St. Louis, MO, USA) or TCI (Tokyo, Japan). If necessary, solvents were purified and/or dried prior to use. All anhydrous reactions were carried out under a dry atmosphere of nitrogen. Melting points (m.p.) were measured on Thomas-Hoover melting point apparatus (Thomas Scientific, Swedesboro, NJ, USA) and not corrected. ¹H, ¹³C-NMR and HMBC spectra were measured on a Varian 400 MHz spectrometer (Agilent Technologies, Santa Clara, CA, USA) in DMSO-d₆, CDCl₃, or (CD₃)₂CO. Chemical shifts (δ) are in ppm relative to tetramethylsilane, and coupling constants (J) are in Hz. DIP-MS (EI) was measured on an Agilent 7890A-5975C GC/MSD (Agilent Technologies). GC/MS (EI) was determined on a SHIMADZU QP 2010 model (Shimadzu, Kyoto, Japan) and FAB-MS was determined on a JEOL JMS-700 Mstation (JEOL, Tokyo, Japan). Fraction collection was performed on an EYELA fraction collector DC-1500 (Tokyo Rikakikai, Tokyo, Japan). An analytical TLC was performed on pre-coated silica gel 60 F₂₅₄ plates (Merck, Kenilworth, NJ, USA). Solvent systems for TLC and column chromatography were ethyl acetate/n-hexane mixtures and 10% methanol in dichloromethane. Column chromatography was carried out on Merck silica gel 9385 (Merck).

GC–MS system QP2010 model (Shimadzu®) equipped with Mass selective Detector and split–a split-less system of injection. The instrument was fitted with capillary column RTx-5MS (cross bond 5% diphenyl—95% dimethylpolysiloxane) with 30 m × 0.25 mm with 0.25 μm film thickness. At the rate of 1.2 ml/min, helium was being used as carrier gas. The temperature program of the column was started at 150 °C (1 min) then programmed at 4 °C/min to 150 °C (10 min). The temperature of the injector was 275 °C while the detector was at 250 °C. 0.2 µl volume was injected in split mode. A split ratio was 1:50, and the mass spectra were operated electron ionization at 70 eV in Selected Ion Monitoring (SIM) mode were maintained. The run time of the machine was 40 min. The relative percentage of the plant extract compounds was expressed in percentage with normalization of peak area.

Chromatographic analysis was carried out using GC-MS (Model QP-2010, Shimadzu Co., Kyoto, Japan) in electron impact ionization (EI) mode. The ionization voltage was 70 eV, and the temperatures of the ion source and injector were 230°C and 250°C, respectively. The mass spectrometer was scanned from 50 to 400 m/z. The separation was done by a capillary column, DB-WAX (60 m length × 0.25 mm diameter, 0.25 μm film thickness, Agilent J&W, USA). The program of oven temperature was initially started at 40°C (isothermal for 3 min), which was ramped to 180°C (isothermal for 5 min) at 2°C/min. Subsequently, it increases to 200°C (isothermal for 10 min) at 4°C/min and to 220°C (isothermal for 5 min) at 5°C/min. Finally, it reaches to 250°C (isothermal for 10 min) at 5°C/min. Helium was used as the carrier gas at a flow rate of 1 ml/min, and the sample injector volume was 1 μL using 1 : 100 split ratio.