Agilent column 19091s 433 hp 5ms

According to Kabran’s approach [42 ], the phytochemical components were identified using the silylation method. Separated, dried on anhydrous MgSO₄, and then concentrated under vacuum, the organic fractions were processed. Then, after being heated at 37 °C for 30 min, 3 mg of the resulting fraction was combined with 200 L of N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA). The samples were then analyzed using a gas chromatograph connected to a mass spectrophotometer (GC-MS; Agilent Technologies Model 5973 with an Agilent column 19091S-433 HP-5MS equipped with a single quadrupole mass spectrometer, operated using electron ionization (EI)) to determine the composition of 0.1 L of the samples (Agilent Technolo-gies, Santa Clara, CA, USA). Helium served as the carrier gas and had an average pressure range (psi) of 0.9 mL/s. The oven was set to a temperature range of 60 to 300 °C. The retention time of the silylated compounds was compared with those of the standards acquired from the database to identify them.

The determination of phytochemical compounds was carried out according to the silylation method described by Kabran and al. [31 ]. In short, 50 g of each sample (treated with petroleum ether and 250 mL of 2 N (HCl) was heated under reflux for two hours. After cooling, the hydrolysate was treated with 3 × 250 mL of ethyl acetate. The organic fractions were grouped, dried on anhydrous MgSO4, and then concentrated under vacuum. Afterward, 200 μL of N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) was added to 3 mg of the resulting fraction and then heated at 37 °C for 30 min. Next, 0.1 μL of the sample was injected for analysis using a gas chromatograph coupled to a mass spectrophotometer (Brand Agilent Technologies Model 5973 with an Agilent column 19091S-433 HP-5MS, 30 m long, 0.25 mm inside diameter, and 0.25 μm film thickness of the stationary phase, (Helsinki, Finland) in positive mode. Helium was used as a carrier gas, with a typical pressure range (psi) of 0.9 mL/sec. The oven temperature program was set to 60–300 °C for 10 min and then maintained at 300 °C for 20 min. The detector temperature was set to 250 °C and the injector temperature to 260 °C. Identification of the silylated compounds was conducted by comparing the retention times with those of the standards obtained from the database.