# Thermal and Physical Properties of Crude Palm Oil with Higher Oleic Content
## 2 Material and Methods  
### Physicochemical Characterization  
Commercial crude oils obtained from two different oil palms--_Elaeis guinensis_ and the interspecific hybridization of _Elaeis guinensis_\(\times\)_Elaeis oleifera_, here so-called African (APO) and hybrid palm oil (HOPO), respectively--were kindly supplied by Denpasa (Santa Barbara do Para, Brazil). The oil samples were characterized regarding standard quality parameters, following the AOCS official methods [22]: iodine value and unsaponifiable matter (Cd 1c-85 and Ca 6a-40). Total caroten content, expressed as \(\beta\)-carotene, was determined by measuring the absorbance at 446 nm of samples homogenized and diluted in hexane (Spectrophotometer Cary 5000, Varian, Palo Alto, CA, USA) [23]. All analyses were performed in triplicate. Acylglycerols were quantified by the Agilent 7890B gas chromatography system, following the AOCS official method (Santa Clara, CA, USA) (Cd 11b-91). \(\alpha\)- and \(\beta\)-carotene quantification: The chromatographic analysis was carried out in HPLC (Shimadzu, Kyoto, Japan) equipped with a quaternary pump (LC-20 AD), a degasser, a UV-Vis DAD (SPD-M20A) detector, and an autoinjector (SIL-20A). The oven was set at 40 \({}^{\circ}\)C, and chromatograms were obtained at 450 nm. The compound separation was performed using a C30 column (Develosil, Hinode, Japan, 250 mm \(\times\) 4.6 mm; 5 \(\upmu\)m particle size). Elution gradient was a mixture of acetonitrile and methanol (0.05% triethylamine-TEA) and Tetrahydrofuran (THF) with a flow rate of 1.2 mL min\({}^{-1}\), in which the gradient only changed the concentrations of methanol and acetonitrile. The channels of the mobile phase were acetonitrile (A), methanol (0.05% TEA) (B), and tetrahydrofuran (C). Gradient proceeded as follows: (C) remained isocratic at 5% during all the analysis, and all the compositions varied linearly; from 0-10 min, (A) concentration varied from 95 to 65%, and (B) channel went from 0 to 30%; 10-20 min compositions varied to 55% (A) and 40% (B) and remained isocratic up until 60 min of the run; 60-65 min, the composition was set back to 65% (A) and 30% (B) and 65. After 65 min, (A) raised to 95% and (B) went down to 0%. Samples consisted of approximately 50 mg of crude oils diluted in 1 mL of acetone. The injection volume was 10 \(\upmu\)L. The quantification was performed using Sigma-Aldrich \(\alpha\)- and \(\beta\)-carotene standards (St. Louis, MO, USA). Tocols quantification: Tocopherols and toctorienols quantification was performed by UHPLC without sample pre-treatment, as previously reported [24]. The chromatographic analysis was performed using a Waters Acquity SQD/UPLC system (Milford, MA, USA), equipped with a PDA detector, a single-quadrupole mass spectrometer system with Electrospray Ionization interface, automatic injector, a quaternary pump, and a column oven. An Acquity UPLC BEH C18 column, 1.7 \(\upmu\)m, 2.1 mm \(\times\) 100 mm, was used (Milford, MA, USA). Instrument control and data acquisition were performed using the MassLynx software. Chromatographic separation was achieved by gradient elution using different proportions of Solvent A (methanol: water: ammonium hydroxide 99:1:0.1 \(v/v/v\)) and Solvent B (isopropanol). The flow rate was 0.2 mL min\({}^{-1}\) up to 6 min, followed by 0.15 mL min\({}^{-1}\) from 6 to 10 min, and 0.2 mL min\({}^{-1}\) until 15 min. The elution gradient was as follows: 0-6 min 100% A; 60-8 min 100% B; 8.01-15 min 100% A. The injection volume was 5 \(\upmu\)L, and the column temperature was 25 \({}^{\circ}\)C. Calibration curves were performed using commercial standards of \(\alpha\), \(\beta\), \(\gamma\), and \(\delta\) toctopherols (Merck, Kenilworth, NJ, USA), and \(\alpha\), \(\gamma\), and \(\delta\) toctorienols (Chromadex, Los Angeles, CA, USA). Main fatty acids analysis: The oleic, linoleic, and linolenic determination were performed by \({}^{1}\)H nuclear magnetic resonance (NMR).