Hp 5ms30

To identify the phytochemicals, present in Fenugreek seeds extract, GCMS-EI analysis of the hydroethanolic extract was conducted through Agilent Technologies 7000 A. Throughout the analysis the triple Quadrupole acquisition method was practiced. GCMS was available with a non-polar column i. e embedded with film that contain 95% Dimethylpolysiloxane and 5% phenyl (Agilent HP-5MS30 m length × 250 μm distance across × 0.25 μm film thickness). An electron ionization energy with 70eV was utilized for the detection of various compounds. As a carrier gas, ultra speckless helium gas (99.99%) was consumed (flow rate 1.5 ml/min) with 3 ml/min of septum purge flow. The infusion volume was 2 μL with 10:1 of split ratio. The pressure was 1.9952 psi and, the total run time was 71.429 min. Sample preparation was conducted by dissolving 1 g of extract in 20 ml of ethanol and then the solution was strained through Whatman filter paper No. 1 to remove stocky particles. Throughout the procedure analytical grade chemicals were used. The chromatograms were analyzed via Nist library.

IPEC-J2 cells (5 × 10⁵ cells per dish) were seeded in 10 cm dishes for GC–MS analysis as described by our previous study (Xiao et al., 2016 ). Briefly, cells were washed with phosphate buffer saline (PBS) and treated by 0.25% trypsin. And then, cells were collected and pelleted at 1,000 × g for 5 min. After being quenched using 500 L of prechilled 50% (vol/vol) methanol, cells were centrifuged at 1,000 × g for 5 min and then removed and added 500 L of prechilled 100% (vol/vol) methanol. Cells were measured by an Agilent 7890B-5977A GC–MS equipped with HP-5 ms (30 m × 250 m × 0.25 m) capillary column (Agilent J&W, Santa Clara, CA, USA). All metabolites were previously validated using authentic standards (Sigma, St. Louis, MO, USA). The data are expressed relative to the control cells.