Comparing Different Chemometric Approaches to Detect Adulteration of Cold-Pressed Flaxseed Oil with Refined Rapeseed Oil Using Differential Scanning Calorimetry.

Flaxseed oil is one of the best sources of n-3 fatty acids, thus its adulteration with refined oils can lead to a reduction in its nutritional value and overall quality. The purpose of this study was to compare different chemometric models to detect adulteration of flaxseed oil with refined rapeseed oil (RP) using differential scanning calorimetry (DSC). Based on the melting phase transition curve, parameters such as peak temperature (T), peak height (h), and percentage of area (P) were determined for pure and adulterated flaxseed oils with an RP concentration of 5, 10, 20, 30, and 50% (w/w). Significant linear correlations (p ≤ 0.05) between the RP concentration and all DSC parameters were observed, except for parameter h1 for the first peak. In order to assess the usefulness of the DSC technique for detecting adulterations, three chemometric approaches were compared: (1) classification models (linear discriminant analysis-LDA, adaptive regression splines-MARS, support vector machine-SVM, and artificial neural networks-ANNs); (2) regression models (multiple linear regression-MLR, MARS, SVM, ANNs, and PLS); and (3) a combined model of orthogonal partial least squares discriminant analysis (OPLS-DA). With the LDA model, the highest accuracy of 99.5% in classifying the samples, followed by ANN > SVM > MARS, was achieved. Among the regression models, the ANN model showed the highest correlation between observed and predicted values (R = 0.996), while other models showed goodness of fit as following MARS > SVM > MLR. Comparing OPLS-DA and PLS methods, higher values of R2X(cum) = 0.986 and Q2 = 0.973 were observed with the PLS model than OPLS-DA. This study demonstrates the usefulness of the DSC technique and importance of an appropriate chemometric model for predicting the adulteration of cold-pressed flaxseed oil with refined rapeseed oil.

The Effects of Cellular Membrane Damage on the Long-Term Storage and Adhesion of Probiotic Bacteria in Caco-2 Cell Line.

Adhesion is one of the main factors responsible for the probiotic properties of bacteria in the human gut. Membrane proteins affected by cellular damage are one of the key aspects determining adhesion. Fluid-bed-dried preparations containing probiotic bacteria were analyzed in terms of their stability (temperature of glass transition) and shelf life in different conditions (modified atmosphere, refrigeration). Imaging flow cytometry was utilized to determine four subpopulations of cells based on their physiological and morphological properties. Lastly, adhesion was measured in bacteria cultured in optimal conditions and treated with heat shock. The results show that the subpopulations with no or low levels of cell membrane damage exhibit the ability to adhere to Caco-2 cells. The temperature of protein denaturation in bacteria was recorded as being between 65 °C and 70 °C. The highest glass transition temperature (Tg) value for hydroxypropyl methylcellulose (used as a coating substance) was measured at 152.6 °C. Drying and coating can be utilized as a sufficient treatment, allowing a long shelf-life (up to 12 months). It is, however, worth noting that technological processing, especially with high temperatures, may decrease the probiotic value of the preparation by damaging the bacterial cells.

DSC Phase Transition Profiles Analyzed by Control Charts to Determine Markers for the Authenticity and Deterioration of Flaxseed Oil during Storage.

An approach of implementing X-bar and R control charts as a statistical control tool to monitor the changes in the melting profile of fresh and stored flaxseed oils by differential scanning calorimetry (DSC) was used. Phase transition melting profiles were collected after 0, 2, 4, and 6 months of storing flaxseed oils, originating from five different cultivars. Four peaks at around -36, -30, -25, and -12 °C were identified using the deconvolution analysis procedure, which enabled the data to be collected at peak temperature (T), peak height (h), the peak area (A), and the percentages of the area (P A), as well as the ratio calculated from these parameters. Control charts obtained for the second peak of the melting profile showed a significant decrease of peak height (h2) from 0.50 to 0.39 W/g and the percentage of the area (P A2) from 50 to 38%, within the storage time (p ≤ 0.05); thus, they were considered to be indicators of oil deterioration. Strong negative correlations of the unstable parameters of DSC with chemical indicators of the oils' oxidative stability (PV, p-AV, TOTOX) were found. For DSC parameters, related to the first peak (h1, A1) and the third peak (h3, A3), changes were statistically not significant within storage (p > 0.05); thus, they can be used as markers of flaxseed oil authenticity. The study demonstrated that X-bar and R control charts could effectively monitor changes in the specific peaks and calculated ratios from the DSC melting profile of fresh and stored flaxseed oils, serving as reliable indicators of oil deterioration.

Lipidomic characteristics of three edible cold-pressed oils by LC/Q-TOF for simple quality and authenticity assurance.

Distinguishing oil samples from each other is challenging but it is crucial for ensuring food quality, and for detecting and preventing the possible adulteration of these products. Lipidomic profiling is believed to provide sufficient information to get fit-to-purpose confidence of oil identification as well as to deliver oil-specific lipid features which could be used as targets for routine authenticity testing of camelina, flax, and hemp oil in food control laboratories. Conducted di- and triacylglycerol profiling by LC/Q-TOFMS yielded successful differentiation of the oils. A marker panel consisting of 27 lipids (both DAGs and TAGs) useful for quality verification and authenticity assurance of the oils was established. Moreover, sunflower, rapeseed, and soybean oils were analysed as potential adulterants. We identified 6 lipid markers (DAGs 34:6, 35:2, 40:1, 40:2, 42:2, and TAG 63:1) which can be used for revealing the adulteration of camelina, hemp, and flax seed oils with these oils.

Influence of Seeds' Age and Clarification of Cold-Pressed Raspberry (Rubus idaeus L.) Oil on the DSC Oxidative Stability and Phase Transition Profiles.

After cold-pressing, small particles of seed residue remain in raspberry seed oil (RSO), even after passing it through cold filtration. The removal of the remaining seed residue is rather an alternative option to improve the visual properties of RSO. This study investigated the influence that the seeds' age (0, 10, 20 months) and clarification process after pressing has on the oxidative stability and phase transition of RSO by means of differential scanning calorimetry (DSC). The results proved that the oil centrifugation process reduces the DPPH radical scavenging activity and oxidative stability measured by p-anisidine value (p-AnV) and DSC oxidation induction time (OIT) at 120 °C of all RSO samples, regardless of the age of the seeds (p ≤ 0.05). No significant differences were observed on the DSC melting and crystallization properties at 1 °C/min after the oil clarification by centrifugation (p > 0.05). The storage time of raspberry seeds, i.e., 10 and 20 months after expiry date, influenced the quality deterioration of RSO, as measured by higher p-AnV, lower DPPH, and OIT values (p ≤ 0.05). The results presented provide new information about oil production processing, suggesting that producers should reconsider giving up the clarification process of oil, since it lowers all quality parameters.

Authenticity Assessment from Sesame Seeds to Oil and Sesame Products of Various Origin by Differential Scanning Calorimetry.

The aim of this study was to conduct thermal characterization of sesame seeds and oils from various geographical origins (Ethiopia, India, Nigeria, Sudan, Turkey), different method of extraction (hexane and cold-pressing), and different types of derived products (halva and tahini). Thermal characterization was investigated using differential scanning calorimetry (DSC), which showed that origin of the seeds has no influence on the melting profile of sesame oil (peak temperature and enthalpy). Method of extraction (hexane and cold-pressing) influenced the peak temperatures of the resulting oils (p ≤ 0.05). The addition of 20% of palm olein to pure sesame oil influenced the significant changes in thermodynamic parameters such as peak temperature (Tm2), which was lowered from −5.89 °C to −4.99 °C, peak half width (T1/2), elevated from 3.01 °C to 4.52 °C, and the percentage of first peak area (% peak 1) lowered from 87.9 to 73.2% (p ≤ 0.05). The PCA method enabled to distinguish authentic and adulterated sesame oils of various origins. There were no significant differences in thermal properties among the products (halva, tahini) and the authentic sesame oil (p > 0.05). The obtained results showed DSC feasibility to characterize sesame oil and sesame products in terms of authenticity.

Shotgun Lipidomic Analysis for Differentiation of Niche Cold Pressed Oils.

The fast-growing food industry is bringing significant number of new products to the market. To protect consumers' health and rights, it is crucial that food control laboratories are able to ensure reliable quality testing, including product authentication and detection of adulterations. In our study, we applied a fast and eco-friendly method based on shotgun-lipidomic mass spectrometry for the authentication of niche edible oils. Comprehensive lipid profiles of camelina (CA), flax (FL) and hemp (HP) seed oils were obtained. With the aid of principal component analysis (PCA), it was possible to detect and distinguish each of them based on their lipid profiles. Lipidomic markers characteristic ofthe oils were also identified, which can be used as targets and expedite development of new multiplexed testing methods.

Assessment of Hemp Seed Oil Quality Pressed from Fresh and Stored Seeds of Henola Cultivar Using Differential Scanning Calorimetry.

Cold-pressed hemp (Cannabis Sativa L.) seed oil has become very popular amongst consumers and researchers, due to its manifold application in food and medicine industry. In this study, oils pressed from stored and fresh hemp seeds of the Henola cultivar were analyzed. Determination of the acid value (AV) and color of oil (a* parameter) revealed significant differences between the two groups of oils (fresh and stored seeds) in contrast to the peroxide value (PV), p-anisidine value (p-AV), and fatty acid composition. On the other hand, isothermal and non-isothermal assessments of the thermo-oxidative stability by differential scanning calorimetry (DSC) showed no significant differences in oxidation induction time (OIT) as well as in onset temperature (Ton) between two groups of oils (p > 0.05). The DSC isothermal test (OIT 160) showed significant correlations with mono- and polyunsaturated fatty acids as well as with values of AV and a* (p ≤ 0.05), in contrast to the non-isothermal test, for which correlations were not significant (p > 0.05). However, the best distinction of both groups of oils was obtained analyzing all results together (DSC, fatty acid and tocochromanols composition, color, and oxidative stability results) by principal component analysis (PCA).

The Effect of Freezing Sheep's Milk on the Meltability, Texture, Melting and Fat Crystallization Profiles of Fresh Pasta Filata Cheese.

Sheep's milk is produced in smallholdings, which hinders the continuity of production. Therefore, freezing during periods of high production can be a solution. Herein, we examined the effect of freezing on sheep's milk and a mixture of sheep and cow's milk (70:30, v/v) on the quality of fresh pasta filata cheeses produced from the milk. Frozen/thawed sheep's milk contributes little to the development of innovative and reformulated cheeses. This was due to 24% higher hardness and greater extensibility and cutting force, as well as lower stretching and elasticity. Although their flowability increased (Oiling-off from 3 to 12%), the meltability (tube test, and Schreiber test) decreased. Additionally, the use of frozen milk caused consumer dissatisfaction. The consumer penalty analysis of the just-about-right showed that freezing of the milk caused the loss of the refreshing, elasticity and shininess of pasta filata cheeses.

Phase Transitions and Structural Changes in DPPC Liposomes Induced by a 1-Carba-Alpha-Tocopherol Analogue.

Steady-state emission spectroscopy of 1-anilino-8- naphthalene sulfonate (ANS) and 1,6-diphenyl-1,3,5-hexatriene (DPH), fluorescence anisotropy, and DSC methods were used to characterize the interactions of the newly synthesized 1-carba-alpha-tocopherol (CT) with a 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) membrane. The DSC results showed significant perturbations in the DPPC structure for CT concentrations as low as 2 mol%. The main phase transition peak was broadened and shifted to lower temperatures in a concentration-dependent manner, and pretransition was abolished. Increasing CT concentrations induced the formation of new phases in the DPPC structure, leading to melting at lower temperatures and, finally, disruption of the ordered DPPC structure. Hydration and structural changes of the DPPC liposomes using ANS and DPH fluorescent probes, which are selectively located at different places in the bilayer, were studied. With the increased concentration of CT molecules in the DPPC liposomes, structural changes with the simultaneous formation of different phases of such mixture were observed. Temperature studies of such mixtures revealed a decrease in the temperature of the main phase transition and fluidization at decreasing temperatures related to increasing hydration in the bilayer. Contour plots obtained from concentration-temperature data with fluorescent probes allowed for identification of different phases, such as gel, ordered liquid, disordered liquid, and liquid crystalline phases. The CT molecule with a modified chromanol ring embedded in the bilayer led to H-bonding interactions, expelling water molecules from the interphase, thus introducing disorder and structural changes to the highly ordered gel phase.

Comprehensive Thermal Characteristics of Different Cultivars of Flaxseed Oil (Linum usittatissimum L.).

The aim of this study was to describe the thermal properties of selected cultivars of flaxseed oil by the use of the differential scanning calorimetry (DSC) technique. The crystallization and melting profiles were analyzed depending on different scanning rates (1, 2, 5 °C/min) as well as oxidative induction time (OIT) isothermally at 120 °C and 140 °C, and oxidation onset temperatures (Ton) at 2 and 5 °C/min were measured. The crystallization was manifested as a single peak, differing for a cooling rate of 1 and 2 °C/min. The melting curves were more complex with differences among the cultivars for a heating rate of 1 and 2 °C/min, while for 5 °C/min, the profiles did not differ, which could be utilized in analytics for profiling in order to assess the authenticity of the flaxseed oil. Moreover, it was observed that flaxseed oil was highly susceptible to thermal oxidation, and its stability decreased with increasing temperature and decreasing heating rate. Significant negative linear correlations were found between unsaturated fatty acid content (C18:2, C18:3 n-3) and DSC parameters (OIT, Ton). Principal component analysis (PCA) also established a strong correlation between total oxidation value (TOTOX), peroxide value (PV) and all DSC parameters of thermo-oxidative stability.

A Chemometric Approach to Oxidative Stability and Physicochemical Quality of Raw Ground Chicken Meat Affected by Black Seed and Other Spice Extracts.

The effects of black seed (Nigella sativa), allspice, bay leaf, caraway, cardamom, clove, and nutmeg extracts on the quality of raw ground chicken legs stored at 4 °C were investigated. During 12 days of storage, conjugated diene (CD) content, thiobarbituric acid reactive substances (TBARS), oxidation induction time (IP) by DSC (differential scanning calorimetry), hexanal content by GC-SPME-MS, thiol group (SH) content were determined. Moreover, microbial growth, pH and color of the samples were investigated. Sensory analysis was also realized. All extracts increased oxidative stability and safety of meat, significantly changed the color of the samples, stabilized the pH and increased their sensory scores (except color of samples with bay leaf and black seed) when comparing to control. Black seed, allspice and clove extracts showed high antioxidant capacity in lipid (CD = 0.23%, 0.28%, and 0.37%, respectively; TBARS = 0.55, 0.50, and 0.48 mg/kg, respectively) and protein fraction (SH content = 47.9, 52.1 and 52.7 nmol/g, respectively), although the ABTS•+ radical scavenging activity of black seed (33.1 µM/g) was significantly lower than the cloves (2496 µM/g) and allspice (815 µM/g). In the sensory analysis the highest scores were ascribed to the sample with cardamom followed by cloves. Principal component analysis (PCA) indicated complex and inseparable interrelationship among lipid and protein oxidation processes and the relationship of the protein oxidation on the lightness of meat. The results enabled to discriminate the meat samples, showing a great impact of the extracts on the final quality of raw chicken meat with black seed being potent antioxidant active additive.

Tocopheryl Succinate-Induced Structural Changes in DPPC Liposomes: DSC and ANS Fluorescence Studies.

Recent studies show that alpha-tocopheryl succinate (TS) exhibits selective toxicity against cancer cells. In this study, we investigated the effect of TS's presence on the physico-chemical and structural properties of DPPC liposomes using fluorescence parameters (intensity, lifetime, and position of emission maximum) of 1-anilino-8-naphtalene sulphonate (ANS), differential scanning calorimetry (DSC) and zeta potential methods. Increasing the TS presence in the DPPC gel phase produced ANS fluorescence enhancement with a hypsochromic shift of the maximum. The zeta potential measurements show an increase in the negative surface charge and confirmed that this process is connected with the hydrophobic properties of dye, which becomes located deeper into the interphase region with a progressing membrane disorder. Temperature dependence studies showed that an increase in temperature increases the ANS fluorescence and shifts the ANS maximum emission from 464 to 475 nm indicating a shift from hydrophobic to a more aqueous environment. In the liquid crystalline phase, the quenching of ANS fluorescence occurs due to the increased accessibility of water to the ANS located in the glycerol region. The DSC results revealed that increasing the presence of TS led to the formation of multicomponent DSC traces, indicating the formation of intermediate structures during melting. The present results confirmed that TS embedded into the DPPC membrane led to its disruption due to destabilisation of its structure, which confirmed the measured biophysical parameters of the membrane.

Seed-Roasting Process Affects Oxidative Stability of Cold-Pressed Oils.

The oxidative stability of vegetable oils mainly depends on their fatty acid composition, their degree of unsaturation, and the presence of compounds with antioxidant activity. This paper reports on the effects of the process of roasting oil seeds, prior to pressing them, on the basic characteristics of the oils produced and their oxidative stability. The differential scanning calorimetry (DSC) technique was used to study the process of oxidation of the oil samples in an oxygen-flow cell. Chromatographic analysis revealed that roasting the seeds increased the levels of chlorophyll and ß-carotene in all the cold-pressed oils. Similar results were observed for the oil's antioxidant activity, measured by the scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical method. Our results also indicated that roasting seeds prior to pressing them for oil had a positive effect on the oil's stability, as determined by the DSC method. This manifested in both the extension of oxidation induction time and the final oxidation time.

Disruptive effect of tocopherol oxalate on DPPC liposome structure: DSC, SAXS, and fluorescence anisotropy studies.

α-Tocopherol oxalate (TO), a tocopherol ester derivative, was investigated for its effect on the structural changes of fully hydrated 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) liposomes, as a function of concentration and temperature, by applying differential scanning calorimetry (DSC), small angle X-ray scattering (SAXS), and DPH fluorescence anisotropy methods. The DSC and DPH anisotropy data indicated that TO embedded into DPPC membrane lowered the enthalpy (ΔHm) and temperature (Tm) of the main phase transition as well its cooperativity. Fluidization of the membrane at a lowered temperature was accompanied by formation of mixed structures of tocopherol-enriched domains. SAXS studies showed the formation of various ordered structures in DPPC gel-phase during incorporation of TO into the bilayer, as evidenced by the existence of lamellar phases with repeat distances (d) of 6.13 and 6.87 nm, assigned to TO-enriched domains and a lamellar, liquid-ordered DPPC phase with d = 8.45 nm at increasing TO concentrations with lowering and broadening of the Bragg peaks, and diffuse scattering, characteristic of a fluid Lα phase, were observed. In DPPC fluid-phase, the increasing presence of TO at low concentrations resulted in the appearance of a liquid-ordered phase with repeat d = 6.9 nm coexistent with a lamellar structure with d = 9.2 nm, assigned to liquid-disordered structures. An increasing repeat distance observed with raising the TO amount in the DPPC bilayer evolved from an increasing interlamellar water layer of increasing thickness. Presence of TO facilitated penetration of water molecules into the acyl chain region which decreased van der Waals interactions in the bilayer. The DSC, SAXS, and fluorescence anisotropy data established that TO exhibited pronounced disruptive activity in DPPC membranes compared to α-tocopherol. The driving force of the observed action was attributed to electrostatic and dipole interactions of the acidic moiety with the polar head group of phospholipids in the interface region of the bilayer.

Quantitative determination of titin and nebulin in poultry meat by SDS-PAGE with an internal standard.

The method of quantitative determination of titin and nebulin in chicken meat by SDS-PAGE electrophoresis technique was developed by application of ß-galactosidase as the internal standard. The method was tested first on marker protein samples of known concentrations (myosin, transferrin, glutamic dehydrogenase) and then the method was used in the determination of titin and nebulin content in chicken meat. The method demonstrated high accuracy, confirmed by correlation coefficient 0.91÷0.99. Two gel analysis techniques, i.e. scanning and densitometry were also compared. By the use of marker proteins as well as titin and nebulin, higher accuracy and precision were achieved in scanning than in the densitometric technique. Recoveries of three marker proteins were between 93 and 102% for the technique of the scanning of the gel and between 99 and 116% for the densitometry.