Gas chromatograph model 7820a

FA contents of control CB (100:0 system), CB:OG mixed systems (50:50, 40:60, 30:70 and 20:80 systems), pure OG (0:100 system), as well as of HORSO were determined (in triplicate) by saponification and bimethylation using C13:0 as internal standard. Fatty acid methyl ester (FAME) was analyzed by gas chromatography on an Agilent gas chromatograph (Model 7820A, CA-USA) fitted with a GC-28 Agilent DB-23 capillary column (60 m × 250 µm × 0.25 μm), and a flame ionization detector was used. Injector and detector temperatures were 250 and 260 °C, respectively. The temperature profile of the oven was as follows: initially 50 °C (held for 1 min), increasing 25 °C/min up to 175 °C and 4 °C/min up to 230 °C (held for 16 min). FAs were identified by comparing retention times with a FA standard (47015-U Supelco PUFA No.2 Animal Source, Sigma-Aldrich Co., St. Louis, MO, USA). FAs were expressed as mg of fatty acid/g sample. Three replicates were performed with samples prepared on two different days.
Based on the FAME results, the atherogenic index (AI) and thrombogenic index (TI) were computed as follows [30 (link)]:

The FA contents of OPO, PS, CB, CFP, M1, and M3, as well as of the baked PP-CB, PP-CFP, PP-M1, and PP-M3 counterparts, were determined by saponification and methylation, as previously described [9 (link),28 (link)]. Both the laminating fat and PP samples were lyophilized beforehand. Fatty acid methyl ester (FAME) was analyzed on an Agilent gas chromatograph (Model 7820A, CA, USA), fitted with a GC-28 Agilent DB-23 capillary column, and a flame ionization detector was used. Results are expressed as mg FA/g sample.