Rice bran FFA determination by diffuse reflectance IR spectroscopy

Rice bran with FFA levels above 0.1% cannot be used as a food ingredient due to oxidative off-flavor formation. However, extracting high FFA oil from bran by in situ methanolic esterification of rice bran oil to produce methyl ester biodiesel produces greater yields relative to low-FFA rice bran oil. Therefore, high-FFA bran could be exploited for biodiesel production. This study describes an FTIR spectroscopic method to measure rice bran FFA rapidly. Commercial rice bran was incubated at 37°C and 70% humidity for a 13-d incubation period. Diffuse reflectance IR Fourier transform spectra of the bran were obtained and the percentage of FFA was determined by extraction and acid/base titration throughout this period. Partial least squares (PLS) regression and a calibration/validation analysis were done using the IR spectral regions 4000-400 cm⁻¹ and 1731-1631 cm⁻¹. The diffuse reflectance IR Fourier transform spectra indicated an increasing FFA carbonyl response at the expense of the ester peak during incubation, and the regression coefficients obtained by PLS analysis also demonstrated that these functional groups and the carboxyl ion were important in predicting FFA levels. FFA rice bran changes also could be observed qualitatively by visual examination of the spectra. Calibration models obtained using the spectral regions 4000-400 cm⁻¹ and 1731-1631 cm⁻¹ produced correlation coefficients R and root mean square error (RMSE) of cross-validation of R=0.99, RMSE=1.78, and R=0.92, RMSE=4.67, respectively. Validation model statistics using the 4000-400 cm⁻¹ and 1731-1631 cm⁻¹ ranges were R=0.96, RMSE=3.64, and R=0.88, RMSE=5.80, respectively.     

Differentiation of mixtures of monovarietal olive oils by mid‐infrared spectroscopy and chemometrics

Fourier transform infrared (FT‐IR) spectroscopy in combination with chemometric techniques has become a useful tool for authenticity determination of extra‐virgin olive oils. Spectroscopic analysis of monovarietal extra‐virgin olive oils obtained from three different olive cultivars (Erkence, Ayvalik and Nizip) and mixtures (Erkence‐Nizip and Ayvalik‐Nizip) of monovarietal olive oils was performed with an FT‐IR spectrometer equipped with a ZnSe attenuated total reflection sample accessory and a deuterated tri‐glycine sulfate detector. Using spectral data, principal component analysis successfully classified each cultivar and differentiated the mixtures from pure monovarietal oils. Quantification of two different monovarietal oil mixtures (2–20%) is achieved using partial least square (PLS) regression models. Correlation coefficients (R²) of the proposed PLS regression models are 0.94 and 0.96 for the Erkence‐Nizip and Ayvalik‐Nizip mixtures, respectively. Cross‐validation was applied to check the goodness of fit for the PLS regression models, and R² of the cross‐validation was determined as 0.84 and 0.91, respectively, for the two mixtures.     

In‐process monitoring in industrial olive mill by means of FT‐NIR

A total of 287 olive lots and 161 olive oil samples were analyzed for fat content, moisture and free acidity, using a Fourier transform near‐infrared (FT‐NIR) instrument located in an industrial mill. Samples having a wide range of both reference values and olive lot sizes (from <0.5 to >4 t) were collected at three industrial mill plants, located in the same Italian region, which utilize different technological equipment for virgin olive oil production. Olive paste spectra were acquired in diffuse reflectance, while oil samples were measured in transmission. Calibration models for oil content and moisture of olives as well as free acidity of virgin olive oils were developed using partial least squares (PLS) regression, first derivative and straight line subtraction. Results of calibration and validation of the PLS models selected were good. The PLS results indicate good similarity between data obtained from FT‐NIR and reference laboratory methods, allowing a rapid and less expensive screening analysis. Unfortunately, the correlation between the oil yield values recorded for all olive lots at the industrial mills and the oil content predicted by FT‐NIR was not satisfactory (R² = 0.605).     

Characterization of Turkish Virgin Olive Oils Produced from Early Harvest Olives

Olives were collected from various districts of Turkey (North and South Aegean sub-region, Bursa-Akhisar, South East Anatolia region) harvested over seven (2001–2007) seasons. The aim of this study was to characterize the chemical profiles of the oils derived from single variety Turkish olives including Ayvalik, Memecik, Gemlik, Erkence, Nizip Yaglik and Uslu. The olive oils were extracted by super press and three phase centrifugation from early harvest olives. Chosen quality indices included free fatty acid content (FFA), peroxide value (PV) and spectrophotometric characteristics in the ultraviolet (UV) region. According to the FFA results, 46% (11 out of 24 samples) were classified as extra virgin olive oils; whereas using the results of PV and UV, over 83% (over 19 of the 24 samples) had the extra virgin olive oil classification. Other measured parameters included oil stability (oxidative stability, chlorophyll pigment, pheophytin-α), cis–trans fatty acid composition and color index. Oxidative stability among oils differed whereas the cis–trans fatty acid values were within the national and international averages. Through the application of two multivariate statistical methods, Principal component and hierarchical analyses, early harvest virgin olive oil samples were classified according to the geographical locations categorized in terms of fatty acid profiles. Such statistical clustering gave rise to defined groups. These data provide evidence of the variation in virgin olive oil quality, especially early harvest and cis–trans isomers of fatty acid profiles from the diverse agronomic conditions in the olive growing regions of Turkey.     

Determination of olive oil free fatty acid by fourier transform infrared spectroscopy

A new procedure for determining free fatty acids (FFA) in olive oil based on spectroscopic Fourier transform infrared-attenuated total reflectance spectroscopy measurements is proposed. The range of FFA contents of samples was extended by adding oleic acid to several virgin and pure olive oils, from 0.1 to 2.1%. Calibration models were constructed using partial least-squares regression (PLSR). Two wavenumber ranges (1775–1689 cm⁻¹ and 1480–1050 cm⁻¹) and several pretreatments [first and second derivative; standard normal variate (SNV)] were tested. To obtain good results, splitting of the calibration range into two concentration intervals (0.1 to 0.5% and 0.5 to 2.1%) was needed. The use of SNV as a pretreatment allows one to analyze samples of different origins. The best results were those obtained in the 1775–1689 cm⁻¹ range, using 3 PLSR components. In both concentration ranges, at a confidence interval of α = 0.05, no significant differences between the reference values and the calculated values were observed. Reliability of the calibration vs. stressed oil samples was tested, obtaining satisfactory results. The developed method was rapid, with a total analysis time of 5 min; it is environment-friendly, and it is applicable to samples of different categories (extra virgin, virgin, pure, and pomace oil).     

Comparison of fatty acid profiles and mid‐infrared spectral data for classification of olive oils

The composition of olive oils may vary depending on environmental and technological factors. Fatty acid profiles and Fourier‐transform infrared (FT‐IR) spectroscopy data in combination with chemometric methods were used to classify extra‐virgin olive oils according to geographical origin and harvest year. Oils were obtained from 30 different areas of northern and southern parts of the Aegean Region of Turkey for two consecutive harvest years. Fatty acid composition data analyzed with principal component analysis was more successful in distinguishing northern olive oil samples from southern samples compared to spectral data. Both methods have the ability to differentiate olive oil samples with respect to harvest year. Partial least squares (PLS) analysis was also applied to detect a correlation between fatty acid profile and spectral data. Correlation coefficients (R²) of a calibration set for stearic, oleic, linoleic, arachidic and linolenic acids were determined as 0.83, 0.97, 0.97, 0.83 and 0.69, respectively. Fatty acid profiles were very effective in classification of oils with respect to geographic origin and harvest year. On the other hand, FT‐IR spectra in combination with PLS could be a useful and rapid tool for the determination of some of the fatty acids of olive oils.     

Clarifications of the Carbonyl and Water Absorptions in Fourier Transform Near Infrared Spectra from Extra Virgin Olive Oil

The Fourier transform near infrared (FT-NIR) spectrum of extra virgin olive oils (EVOO) shows two minor carbonyl absorptions at 5280 and 5180 cm^–1 that has been used to assess their authenticity. To establish components absorbing at 5280 cm^–1, volatile aldehydes and ketones, triacylglycerol (TAG), diacylglycerols (DAG), free fatty acids (FFA), phenolics, and water are investigated and sometimes added to refined olive oil (ROO). Except TAG, the remaining carbonyls contribute to 5280 cm^–1 by broadening peak. Water absorption is demonstrated by its removal using Na₂SO₄ or deuterium oxide addition; FT-NIR spectral changes are reconstituted by water addition. Water absorption depends on being free or complexed with polar compounds in oil. The size of absorption is not related to abundance, but on unique absorption specificity of components; water shows the strongest absorption. Heat removes water and volatiles, leaving behind DAG, FFA, and phenolics, and makes it possible to differentiate absorption of water, volatile and non-volatile carbonyls. Cloudy olive oils are analyzed using FT-NIR methodology after warming for 3 min at 50 °C. FT-NIR index values are replaced by a new calibration model based on correlating gravimetric mass loss of water plus volatiles with spectral changes. The FT-NIR methodology is expanded to include EVOOs with 15.5% to 21% linoleic acid. Practical Applications: Testing for authenticity of EVOOs remains a challenge because adulterations continue to be a problem due to economic gains. Spectroscopy methods, specifically FT-NIR, are much preferred to targeted chemical methods because they measure all constituents in products and are non-destructive and fast. The current universal FT-NIR methodology assesses 13 different parameters: five major FAs, and the DAG and FFA contents. The FT-NIR index value measuring the content of moisture plus volatiles is now replaced by a gravimetric determination. The methodology identifies four major types of adulterants, high in oleic acid, linoleic acid, palm olein or ROO. The composition of olive oils makes it necessary to develop five oil-specific groups, but cloudy samples still need to be clarified by slight warming before measuring. The value of this universal FT-NIR methodology will increase after being adopted by commercial and in regulatory settings.     

Chemical Composition of Turkish Olive Oil––Ayvalik

Although large amounts of olive oil are produced in Turkey, not much information on its chemical composition is available in the literature to date. The aim of this study was to evaluate the chemical composition of commercial olive oils produced from the Ayvalik olive cultivar in Canakkale, Turkey. Five different samples corresponding to the olive oil categories of extra virgin (conventional, extra virgin olive oil (EVOO), and organic extra virgin olive oil (OGOO) production), virgin olive oil (OO-1), ordinary virgin olive oil (OO-2) and refined olive oil (RFOO) were evaluated. Olive oils were collected from two consecutive production years. According to the free fatty acids, the absorbance values (K₂₃₂ and K₂₇₀), and peroxide values of all the samples conformed to the European standards for olive oil. The level of oleic acid was in the range of 68–73%; while the linoleic acid content was significantly lower in the refined olive oils. The tocopherol and polyphenol content was in the lower range of some European olive oils. However, pinoresinol was a major phenolic compound (5–77 mg/kg depending on the oil category). Its content was markedly higher than in many other oils, which would be a useful finding for olive oil authentication purposes.     

Comparative study of the extraction and measurement of cottonseed free fatty acids

Crude oil was extracted from cottonseed by three different methods to study the influence of extraction technique on the free fatty acid (FFA) concentration. Extraction procedures that recovered more oil had higher levels of FFA. In addition, the highest concentration of FFA was found in oil recovered by Soxhlet reextraction of a meal initially defatted by a room-temperature extraction process. The FFA concentrations of oils recovered by Soxhlet extraction were highly correlated with the FFA concentration of oils recovered by the other extraction methods studied (R ²>0.96). Titration of oil and gas chromatography of silylated oil were compared as methods to determine FFA concentration. The methods compared well (R ²=0.998) with the titration method, giving ∼5% higher values for FFA than the chromatography method. Half of this difference appeared to be due to the oleic acid approximation used in the titration approach. The other half of the difference is likely due to the detection of other acidic components in crude oil.     

Detection of Extra Virgin Olive Oil Adulteration With Edible Oils Using Front‐Face Fluorescence and Visible Spectroscopies

Synchronous front‐face fluorescence and visible spectroscopies are utilized for the simple, rapid, and nondestructive quantification of extra virgin olive oil (EVOO) adulteration with corn, soybean, and sunflower oils. For each adulterant, 42 adulterated EVOO samples in the adulterant amount in the range 1.0–50 g/100 g were prepared. The partial‐least‐squares regression was executed for quantification. Both full (leave‐one‐out) cross‐validation and external validation were performed to evaluate the predictive ability. The plots of observed vs. predicted values exhibit high linearity. The coefficient of determination (R²) values are larger than 0.99. The root mean square errors of both cross‐validation and prediction are no more than 2%. The detection limits for the three seed oils using fluorescence and visible spectroscopies are in the range of 1.5–2.2% and 1.8–2.4%, respectively. The merit of this method is that both the front‐face fluorescence and visible spectroscopies are recorded toward neat oils, avoiding any sample pretreatment including dilution.     

Evaluation of Authenticity of Iranian Olive Oil by Fatty Acid and Triacylglycerol Profiles

For evaluation of the authenticity of Iranian olive oil, samples from many Iranian olive oil producers especially north of Iran in the production year 2007 were collected. The fatty acid and triacylglycerol compositions were measured. The most recent calculation methods including ∆ECN the difference between the actual and theoretical ECN42 (equivalent carbon number), triglyceride content and R of olive oils according to IOOC methods were applied. On the basis of our results, we were able to classify the olive oils into the extra virgin, virgin olive and olive oil categories. The important fatty acids are oleic, palmitic and linoleic acids and their main triacylglycerols are OOO, POO, OOL, PLO, SOS plus POP, and OLL, respectively. On the basis of the triacylglycerol results, experimental ECN48, ECN46, ECN50, ECN44 and ECN42 were obtained. By using the fatty acids results and a computer program, the theoretical ECN42 and ECN44 were calculated. Then R values, being the ratio of r ECN42/r ECN44 for authenticity of all olive oils and ∆ECN for determining categories of olive oils, were defined. The results of olive oil samples were in the accepted limits of Codex and IOOC. Finally we suggest that the R and ∆ECN can be used in identification of adulteration of olive oils and also they are useful from the point of view of authenticity and classification.     

Pure lycopene from tomato preserves extra virgin olive oil from natural oxidative events during storage

In this study, changes in an extra virgin olive oil treated with lycopene during storage were analyzed. Pure lycopene (0.5 and 1.0 mg) obtained from tomato was added to two separate bottles, each containing 100 mL of extra virgin olive oil, while another bottle containing the same oil was stored without any treatment. Samples enriched with pure lycopene showed PV remarkably lower than the sample without lycopene, and the total phenol contents were higher in treated oil samples than in the reference sample. In addition, a good correlation (r ²=0.969) between total phenol content and antioxidant power calculated as Trolox Equivalent Antioxidant Capacity (TEAC) was observed. The concentration of added lycopene decreased very slowly; after about 8 mon its residual value was over 60% with respect to the initial concentration.     

Application of FTIR Spectroscopy for the Determination of Virgin Coconut Oil in Binary Mixtures with Olive Oil and Palm Oil

Rapid Fourier transform infrared (FTIR) spectroscopy combined with attenuated total reflectance (ATR) was applied for quantitative analysis of virgin coconut oil (VCO) in binary mixtures with olive oil (OO) and palm oil (PO). The spectral bands correlated with VCO, OO, PO; blends of VCO and OO; VCO and PO were scanned, interpreted, and identified. Two multivariate calibration methods, partial least square (PLS) and principal component regression (PCR), were used to construct the calibration models that correlate between actual and FTIR-predicted values of VCO contents in the mixtures at the FTIR spectral frequencies of 1,120–1,105 and 965–960 cm⁻¹. The calibration models obtained were cross validated using the “leave one out” method. PLS at these frequencies showed the best calibration model, in terms of the highest coefficient of determination (R ²) and the lowest of root mean standard error of calibration (RMSEC) with R ² = 0.9992 and RMSEC = 0.756, respectively, for VCO in mixture with OO. Meanwhile, the R ² and RMSEC values obtained for VCO in mixture with PO were 0.9996 and 0.494, respectively. In general, FTIR spectroscopy serves as a suitable technique for determination of VCO in mixture with the other oils.     

Olive oil phenolics protect LDL and spare vitamin E in the hamster

An animal feeding trial was conducted to investigate whether olive oil phenolics can act as functional antioxidants in vivo. To this end, hamsters were exposed for a period of 5 wk to a dietary regime with either a phenol-rich extra virgin olive oil or extra virgin olive oil from which phenols were removed by ethanol/water-washing. The original oil used in the high olive phenol diet was also used for the preparation of the low phenol diet in order to keep the FA compositions exactly the same. In addition, the vitamin E content was kept identical in both diets. This careful preparation of the diets was undertaken in order to prevent these factors from influencing the antioxidative status in plasma and LDL. Removal of olive oil phenols was shown to reduce both the vitamin E level in plasma and the resistance of LDL to ex vivo oxidation. The results of this study support the idea that extra virgin olive oil phenols improve the antioxidant defense system in plasma by sparing the consumption of vitamin E under normal physiological circumstances.     

Accelerated oxidation: Comparative study of a new reactor with oxidation stability instrument

Accelerated oxidation tests, such as the determination of the induction period, increase the lipid oxidation rate by exposing a food to elevated temperatures, in the presence of excess quantities of air or oxygen. In addition to the well‐founded oxidative stability index (OSI) method, an innovative and promising technique is the oxidation test (OXITEST) reactor. A new analytical method was developed with OXITEST to oxidize vegetable oils. At a preliminary stage of the investigation, the induction periods of sunflower and extra‐virgin olive oil obtained by the OXITEST reactor were plotted against temperature, on the basis of the Arrhenius law; the activation energy and the frequency factor of lipid oxidation were calculated and resulted in 98.61 kJ/mol and 2.33×10¹⁰ s^–1, respectively, for sunflower oil and 106.48 kJ/mol and 6.27×10¹⁰ s^–1, respectively for extra‐virgin olive oil. The new oxidative technique was employed to determine the induction periods of vegetable oils; the results obtained were well correlated with those achieved with OSI technology, with a Pearson correlation coefficient r = 0.9785 (p <0.05) for oilseeds and palm oil and r = 0.9501 (p <0.05) for extra‐virgin olive oils.     

Identification of adulterants in olive oils

The application of discriminant analysis for identifying and quantifying adulterants in extra virgin olive oils is presented. Three adulterants were used (sunflower oil, rapessed oil, and soybean oil) and were present in the range 5–95%. Near-infrared spectroscopy and principal components analysis were used to develop a discriminant analysis equation that could identify correctly the type of seed oil present in extra virgin olive oil in 90% of cases. Partial least squares analysis was used to develop a calibration equation that could predict the level of adulteration. Cross validation suggested that it was possible to measure the level of adulteration to an accuracy of ±0.9%. External validation of the derived calibation equation gave a standard error of performance of ±2.77%.     

The olive oil oxygen radical absorbance capacity (DPPH assay) as a quality indicator

The antioxidant properties of some single components and the total antioxidant activity of extra‐virgin olive oil have been evaluated by the oxygen radical absorbance capacity (ORAC) method. The total ORAC of the extra‐virgin olive oil was found to be positively correlated with the concentration of total polyphenols, which are important to the shelf life of the product. Among the single phenolic compounds studied, gallic acid showed a higher ORAC than caffeic acid and oleuropein, while among the derivates of oleuropein, hydroxytyrosol was found to be the most active compound among all the phenols studied. The total ORAC of commercial olive oils differed according to the concentration of total polyphenols. The total ORAC of extra‐virgin olive oil was constant during 1 year of storage in rational conditions, whereas it worsened dramatically in olive oil damaged by the lipase‐producing yeast Williopsis californica or by lipase from Pseudomonas spp. The study accomplished on the oily fraction of the fruits before harvesting demonstrated that the total ORAC of the oil of under‐ripe green olives is higher compared to that shown by mature fruits; therefore, through the choice of the harvesting time, it is possible to define also the future content of polyphenols of the oil. The total ORAC test, together with other analyses, can be considered as a qualitative parameter that can contribute to the expression of technological and health virtues of extra‐virgin olive oil.     

Determination of free fatty acids in palm oil by near-infrared reflectance spectroscopy

A near-infrared (NIR) spectroscopy calibration was developed for the determination of free fatty acids (FFA) in crude palm oil and its fractions based on the NIR reflectance approach. A range of FFA concentrations was prepared by hydrolyzing oil with 0.15% (w/w) lipase in an incubator at 60°C (200 rpm). Sample preparation was performed in Dutch cup, and the spectra were measured in duplicate for each sample. The optimized calibration models were constructed with multiple linear regression analysis based on C=O overtone regions from 1850–2050 nm. The best wavelength combinations were 1882, 2010, and 2040 nm. Multiple correlation coefficients squared (R ²) were: 0.994 for crude palm oil, 0.961 for refined-bleached-deodorized (RBD) palm olein, and 0.971 for RBD palm oil. Calibrations were validated with an independent set of 8–10 samples. R ² of validation were 0.997, 0.943, and 0.945, respectively. The developed method was rapid, with a total analysis time of 5 min, and environmentally friendly, and its accuracy was generally good for raw-material quality control.     

Application of a multidisciplinary approach for the evaluation of traceability of extra virgin olive oil

In this work, a multidisciplinary approach for the evaluation of extra virgin olive oil traceability (geographical provenience and botanical differentiation) is presented. Conventional techniques such as major chemical component determination (triacylglycerols, TAG and fatty acids) and other novel approaches as stable isotopic ratio (¹³C/¹²C in combination with ¹⁸O/¹⁶O) and thermal properties obtained from cooling curves and their deconvoluted peaks by means of differential scanning calorimetry were compared. Fifty‐three samples from different Italian regions, diverse cultivars, and two Mediterranean areas (Italy and Croatia) were analyzed with all the three techniques. The oils exhibited different values especially for δ¹⁸O and thermal properties of the deconvoluted peaks of crystallization according to Italian regions and/or cultivars. Data were treated by means of linear discriminant analysis inserting all parameters as predictors in models where the potentiality to discriminate oils was tested. All models revealed a good resolution among categories with selected TAG, δ¹⁸O values, and thermal properties of the deconvoluted peak set at the highest temperature exhibiting the highest weight for the discriminant functions. These findings could give strength to the utilization of new analytical techniques supporting those traditionally employed, also sustained by proper chemometric procedures, as suitable for the resolution of extra virgin olive traceability. Practical applications: Consumers' awareness of extra virgin olive oil traceability has recently increased the interest for new methods that can assess its geographical and botanical origins and new findings in this sector represent a key factor affecting the purchases in non‐producer countries. Multidisciplinary approaches supported by chemometric procedures enable the building of large databases and classification models for the determination of the provenience of extra virgin olive oil.     

Validation of the oxygen radical absorbance capacity (ORAC) parameter as a new index of quality and stability of virgin olive oil

The induction period (IP), determined using accelerated methods such as the Rancimat test, is a parameter that has been used to predict the shelf life of virgin olive oil. The oxygen radical absorbance capacity (ORAC) has recently been proposed as a quality index of virgin olive oil because it measures the efficiency of phenolic compounds in the protection against peroxyl radicals. This study aims to investigate relationships between the ORAC and IP values and proposes ORAC as a new parameter of virgin olive oil stability. The concentrations of phenolics, o-diphenols, tocopherol, β-carotene, lutein, and ORAC and IP values were determined in 33 virgin olive oils. Regression analyses showed that both ORAC and IP values correlate with total phenols and o-diphenols with highly significant indices, whereas the correlations of both ORAC and IP with tocopherols, β-carotene, and lutein were not significant. The ORAC values correlate with the IP values with low but significant indices (R=0.42; P<0.02). The results confirm the key role of phenolic compounds in accounting for the shelf life of virgin olive oil and suggest that the ORAC parameter may be used as a new index of quality and stability.