5975 series msd

The policosanol and sterol composition of the tested oils was determined by GC-MS as described by Harrabi et al. 15) . GC-MS analyses were performed using a capillary HP5MS column (30 m×0.25 mm I.D., 0.25 μm film thickness; Agilent Technologies) with gas chromatography (Agilent Technologies 7820A) coupled directly to the mass detector (Agilent Technologies 5975 series MSD) . Tocopherols were analysed by HPLC according to the method described by Deiana et al. 16) . The separation of tocopherol isomers was accomplished on an Atlantic column (4.6×150 mm, 5 μm) .
The fatty acid composition of the seed oil was determined by gas chromatography (GC) . Fatty acid methyl esters were analysed by GC using a HP 6890 chromatograph equipped with a flame ionisation detector (FID) on a capillary column, CP-Sil 88 (50 m length, 0.25 mm id, 0.2 μm film thickness; Varian) .

The concentrations of the remaining fatty acids and accumulated keto-fatty acid in the reaction medium (e.g., ricinoleic acid, 12ketooleic acid) were determined as previously described [7] . The reaction medium was mixed with ethyl acetate containing 0.1 or 0.5 g/l palmitic acid as an internal standard. The organic phase was harvested after vigorous vortexing and then subjected to derivatization with N-methyl-N-(trimethylsilyl) trifluoroacetamide (TMS)/pyridine (1:3 (v/v)). The TMS derivatives were analyzed using a 5975 series MSD and Agilent 7890A gas chromatograph. The derivatives were separated on a nonpolar capillary column (30 m length, 0.25 µm film thickness; HP-5MS; Agilent Technologies, Palo Alto, CA, USA). A linear temperature gradient was programmed as 90°C, 5°C/min to 280°C, with an injection port temperature of 230°C. Samples were injected at a split ratio of 1:5. Mass spectra were obtained by electron impact ionization at 70 eV. Scan spectra were obtained within the 100-600 m/z range. Selected ion monitoring was used for the detection and fragmentation analysis of the reaction products [6] .

The selection of the most representative compound in each plant (i.e. to follow the evolution of the maceration during the experimental design) was carried out after analysis by GC-MS of the corresponding essential oil previously obtained by ASE. This extraction was performed using a Dionex extractor (ASE 350) obtained from Vertex Technics (Barcelona, Spain) in accordance with the procedure described by Rodríguez-Solana et al. 17 The volatile profile of the methanolic extracts obtained was determined using an Agilent 7820A gas chromatograph (Agilent, Santa Clara, CA, USA) equipped with an Agilent 5975 series MSD and a nonpolar HP-5MS column (5% diphenyl, 95% dimethylpolysiloxane, length 30 m, inner diameter 0.25 mm, film thickness 0.25 mm) with a ramp temperature and operating in the electron impact mode (70 eV) and with transfer line and ion source temperatures maintained at 230 ∘ C. The injector temperature was maintained at 250 ∘ C, whereas that of the quadrupole was 150 ∘ C. Carrier gas used was H 2 (from a Hydrogen generator AD-180 Series; CINEL, Padova, Italy) at a flow of 1.5 mL min -1 . The amount of sample injected was 0.5 𝜇L (in splitless mode). The oven temperature was programmed as: 50-220 ∘ C (2.5 ∘ C min -1 ), 220-300 ∘ C (10 ∘ C min -1 ). The identification procedure was as described by Rodríguez-Solana et al. 17