A new humic acid stationary phase was prepared by immobilizing humic acid onto aminopropyl silica via an amide linkage formation and used, for the first time, for the separation and quantification of the tocopherol compounds in cold-pressed oil samples under normal-phase high-performance liquid chromatography conditions. Parameters affecting the chromatographic separation such as mobile phase composition and flow rate were optimized. By evaluating the calculations of capacity factor, asymmetry factor, resolution, selectivity factor, and theoretical plate number, the best separation was obtained with isocratic elution of n-hexane and isopropyl alcohol (99:1% v/v) at a flow rate of 1.0 mL/min. The effluent was monitored by a fluorescence detector set at excitation and emission wavelengths 295 and 330 nm, respectively. All compounds were separated in 20 min. The method was validated according to international guidelines and found to be linear in a wide concentration range, also the mean recovery of the compounds ranged from 97.9 to 99.2%, with a CV less than 2.7% in all cases. The results showed that the developed stationary phase is suitable for the separation and quantification of the tocopherol compounds in real oil samples.
The main goal of the present study was (i) to determine the formation of degradation products in cottonseed oil (CSO) blends during deep frying process by adsorption and high performance size exclusion chromatography techniques and (ii) to evaluate the impacts of food additives on total polar (TPC) and polymeric compound (PTAG) formation using a chemometric approach. In order to prepare the frying CSO blends; ascorbic palmitate, mixed tocopherols, dimethylpolysiloxane, lecithin and sesame oils were used as additives. To determine the real impacts of additives, a quarter-fraction factorial experimental design with two levels and five factors was used. The changes in TPC and PTAG data were carefully evaluated during 10 h of frying at 170 ± 5°C with normal distribution (ND) graphs and analyzed using a one-way analysis of variance (ANOVA), followed by Tukey's Post-hoc test (a = 0.05). The results indicated that the increasing values for TPC and PTAG during the frying processes for all blends, TPC and PTAG contents reached maximum levels of 16.37 and 6.01 % respectively, which are below the limit values stated by official authorities for the quality assessment of frying oils. The ANOVA test results were in good agreement with ND graphs and data indicated that the impact of mixed tocopherols was significant for TPC formation, meanwhile the impact of lecithin and ascorbic palmitate 9 dimethylpolysiloxane were significant for PTAG formation. Thus, the present study should be considered to be a very useful guide for developing new frying oil formulations based on CSO by using food additives.
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