Qp 2010 ultra gcms spectrometer

Melting points were recorded on a Koffler hot-stage apparatus (Electrothermal 9100, Dubuque, IA, USA) and were uncorrected. GC-MS analysis were carried out on a QP-2010 Ultra GCMS spectrometer (Shimadzu, Kyoto, Japan) equipped with a flame ionization detector (FID). 1D and 2D NMR spectra were recorded on a Unity INOVA 500 MHz spectrometer (Varian, Palo Alto, CA, USA) using standard pulse programs. Chloroform-d (CDCl₃) and methanol-d₄ (CD₃OD) were used as NMR solvents, and TMS was utilized for internal referencing. Solvents used for extraction and isolation were of analytical grade and obtained from R & M chemicals (Edmonton, AB, Canada). Kieselgel 60 (0.040–0.063 mm) and Lichroprep RP-18 (40–63 µm) were purchased from Merck (Darmstadt, Germany) while Sephadex LH-20 was purchased from Sigma (St. Louis, MO, USA). Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), trypsin–EDTA, penicillin/streptomycin were purchased from GIBCO (Lifetechnologies, Grand Island, NY, USA). Trypan blue and MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide) were purchased from Sigma. A SpectraMax Plus (Molecular Devices, Sunnyvale, CA, USA) microplate reader was used in the bioassays.

GC-MS analysis of the ethanolic extract of A. sessilis was performed using a QP-2010 Ultra GC-MS spectrometer (Shimadzu, Kyoto, Japan) fused with a BPx5 column (30 × 0.25 μm ID × 0.25 μm df). The oven temperature was programmed from 50°C at 0 min and increased to 300°C and remained constant for 10 min. The stem extract was diluted in methanol. Helium gas (99.999%) was used as a carrier gas with the following conditions: total flow: 11.8 mL/min, column flow: 0.8 mL/min, linear velocity: 32.4 cm/s, purge flow: 3.0 mL/min, and split ratio: 10. Mass spectra were taken with ion source temperature and interface temperature of 200°C and 250°C, respectively. The mass scan parameters had a start time of 2.5 min, with end time 93.0 min. The acquisition (ACQ) parameters involved the following conditions: scan event time: 0.10 s, scan speed: 10000, and mass range: 40 m/z to 700 m/z. The relative percentage of each compound was calculated by comparing its average peak area to the total amount of areas.