Synthesis of Infant Formula Fat Analogs Enriched with DHA from Extra Virgin Olive Oil and Tripalmitin

Structured lipids (SLs) containing palmitic, oleic, and docosahexaenoic acids for possible use in infant formulas were synthesized by enzymatic acidolysis reactions. The substrates used were tripalmitin, extra virgin olive oil free fatty acids (EVOOFFA), and docosahexaenoic acid single cell oil free fatty acids (DHASCOFFA) in 1:1:1, 1:2:1, 1:3:2, 1:4:2, and 1:5:1 molar ratios. Reactions were carried out at 65 °C for 24 h using Lipozyme^® TL IM lipase. The products were analyzed for total and positional fatty acids by GC-FID, triacylglycerol (TAG) molecular species by HPLC-ELSD, and thermal behavior by DSC. The SLs, SL132, SL142, and SL151 had desirable fatty acid distribution for infant formula use with nearly 60 mol% palmitic acid at the sn-2 position and oleic acid predominantly at the sn-1,3 positions. The total DHA content of SL132, SL142, and SL151 were 7.54, 6.72, and 5.89 mol%, respectively. The major TAG molecular species in the SLs were PPP, OPO, and PPO. The melting completion temperature of SL132 was 37.1, 35.2 °C in SL142, and 32.9 °C in SL151. The SLs synthesized in this study have potential use in infant formulas.     

Biochemical Characterization of Arbequina Extra Virgin Olive Oil Produced in Turkey

Varieties of the olive cultivar Arbequina have recently been cultivated in Turkey. The objective of the study is to characterize and evaluate extra‐virgin olive oils (EVOO) produced from Arbequina grown in the Aegean and Mediterranean regions of Turkey. Major and minor components such as carotenoids, squalene, phenolics and tocopherols were studied to assess their effects on product quality and health benefits. The samples, identified as ArbqI and ArbqA, were from the Izmir and Adana provinces, respectively. Samples were analyzed by GC‐FID to determine fatty acid composition, sterol composition, TAG profile and squalene content. Individual phenolic fractions were analyzed by LC–MS/MS and tocopherol isomers were determined by HPLC. According to the results obtained from this study; Total phenolic content (TPC) of the samples were 454.68 and 50.86 mg Gallic acid/kg oil for ArbqI and ArbqA, respectively. Hydroxytyrosol and tyrosol were determined to be the main phenols. The major tocopherol isomer found in ArbqI and ArbqA was α‐tocopherol with levels of 179.55 and 202.5 mg/kg oil, respectively. β‐Carotene levels in both samples were similar at 0.2 mg/kg. Findings of this study were compared with the literature on Arbequina olive oil produced in different countries. It was determined that Arbequina olive oil of high quality can be produced in Turkey, especially in the Aegean region.     

The Use of FTIR Spectroscopy and Chemometrics for Rapid Authentication of Extra Virgin Olive Oil

The authenticity of high value edible fats and oils including extra virgin olive oil (EVOO) is an emerging issue, currently. The potential employment of Fourier transform infrared (FTIR) spectroscopy in combination with chemometrics of multivariate calibration and discriminant analysis has been exploited for rapid authentication of EVOO from canola oil (Ca‐O). The optimization of two calibration models of partial least square (PLS) and principle component regression was performed in order to quantify the level of Ca‐O in EVOO. The chemometrics of discriminant analysis (DA) was used for making the classification between pure EVOO and EVOO adulterated with Ca‐O. The individual oils and their blends were scanned on good contact with ZnSe crystals in horizontal attenuated total reflectance, as a sampling technique. The wavenumbers of 3,028–2,985 and 1,200–987 cm^?1 were used for quantification and classification of EVOO adulterated with Ca‐O. The results showed that PLS with normal FTIR spectra was well suited for quantitative analysis of Ca‐O with a value of the coefficient of determination (R²) > 0.99. The error, expressed as root mean square error of calibration obtained was relatively low, i.e. 0.108 % (v/v). DA can make the classification between pure EVOO and that adulterated with Ca‐O with one misclassified reported.     

The Effect of Different Cold Storage Conditions on the Compositions of Extra Virgin Olive Oil

Storage conditions can affect the stability and quality of extra virgin olive oil (EVOO). While many studies have reported the influence of high temperature and light exposure during storage, little is known on the influence of the cold storage. The aim of this study was to evaluate the effect of different storage conditions (25, 4.5 and ?27 °C) on the various compositions of EVOO and to determine if cold storage will prolong shelf‐life by retarding hydrolysis and oxidation. The changes of quality indices (FFA, PV, and UV) and natural antioxidants such as α‐tocopherol and phenolic compounds were evaluated periodically during storage. The characterization and quantification of phenolics were achieved by ultra‐performance liquid chromatography—diode array detector (UPLC‐DAD). In addition, 1, 2‐diacylglycerols (DAGs), pyropheophytin A (PPP) were measured to indicate thermal degradation during storage.     

Enzymatic Interesterification Between Pine Seed Oil and a Hydrogenated Fat to Prepare Semi-Solid Fats Rich in Pinolenic Acid and Other Polyunsaturated Fatty Acids

Enzyme catalyzed interesterification (EIE) of pine seed oil (PSO) and a fully hydrogenated soybean oil (FHSBO) were studied in batch reactors in solvent-free media to prepare different semisolid fats rich in polyunsaturated fatty acids (PUFA). Optimal operation conditions found were: 10 % (w/w) enzyme loading, 75 °C and magnetic agitation at 300 rpm. Quasi-equilibrium conditions were reached after 2, 3 and 6 h, when immobilized lipases from Thermomyces lanuginosus (Lipozyme^® TL IM), Candida antarctica B. (Novozym^® 435) and Rhizomucor miehei (Lipozyme^® RM IM) from Novozymes A/S (Bagsvaerd, Denmark) were employed, respectively. Similar distributions of unsaturated to saturated fatty acid (UFA/SFA) residues along the glycerol backbone of the fat products were obtained with both non-selective and sn-1(3) regioselective lipases due to significant spontaneous acyl migration during the reaction. The products had higher UFA/SFA ratios at the sn-2 position (2.4–2.5, 1.4–1.7, and 0.5–0.8 for the trials involving 20, 40 and 70 % FHSBO, w/w, respectively) than the corresponding physical blends (0.8, 0.4 and 0.5, respectively). Fat products containing 3.1–11.6 % (w/w) pinolenic acid (Pn) and 16.1–35.7 % (w/w) linoleic acid (L) at the sn-2 position were prepared. The free acid contents of EIE products prepared with Lipozyme^® TL IM and Novozym^® 435 were 6.1–6.4 and 2.5–2.6, respectively. Residual activities of Lipozyme^® TL IM and Novozym^® 435 diminish by ca. 20 % after 9 reaction cycles.     

Enzymatic interesterification of olive oil with fully hydrogenated palm oil: Characterization of fats

Seven different reaction products were prepared via enzymatic interesterification of extra‐virgin olive oil (EVOO) and fully hydrogenated palm oil (FHPO), by varying the initial weight ratio of EVOO to FHPO from 80 : 20 to 20 : 80. The chemical, physical and functional properties of both the semi‐solid reaction products and the corresponding physical blends of the precursor starting materials were characterized. Fats prepared using large proportions of FHPO contained high levels of TAG species containing only saturated fatty acid residues. By contrast, high levels of TAG species containing both saturated and unsaturated fatty acid residues were found in fat products obtained with the lowest proportions of FHPO. Independently of the initial weight ratio of EVOO to FHPO, the interesterified products were characterized by a higher molar ratio of unsaturated to saturated fatty acid residues at the sn‐2 position, were softer over a wide temperature range, exhibited lower oxidative stabilities and were completely melted at lower temperatures than the corresponding physical blends. Potential applications of the reaction products range from margarines (highest weight ratios of EVOO to FHPO) to frying fats (lowest weight ratios of EVOO to FHPO).     

First Application of Newly Developed FT-NIR Spectroscopic Methodology to Predict Authenticity of Extra Virgin Olive Oil Retail Products in the USA

Economically motivated adulteration (EMA) of extra virgin olive oils (EVOO) has been a worldwide problem and a concern for government regulators for a long time. The US Food and Drug Administration (FDA) is mandated to protect the US public against intentional adulteration of foods and has jurisdiction over deceptive label declarations. To detect EMA of olive oil and address food safety vulnerabilities, we used a previously developed rapid screening methodology to authenticate EVOO. For the first time, a recently developed FT-NIR spectroscopic methodology in conjunction with partial least squares analysis was applied to commercial products labeled EVOO purchased in College Park, MD, USA to rapidly predict whether they are authentic, potentially mixed with refined olive oil (RO) or other vegetable oil(s), or are of lower quality. Of the 88 commercial products labeled EVOO that were assessed according to published specified ranges, 33 (37.5%) satisfied the three published FT-NIR requirements identified for authentic EVOO products which included the purity test. This test was based on limits established for the contents of three potential adulterants, oils high in linoleic acid (OH-LNA), oils high in oleic acid (OH-OLA), palm olein (PO), and/or RO. The remaining 55 samples (62.5%) did not meet one or more of the criteria established for authentic EVOO. The breakdown of the 55 products was EVOO potentially mixed with OH-LNA (25.5%), OH-OLA (10.9%), PO (5.4%), RO (25.5%), or a combination of any of these four (32.7%). If assessments had been based strictly on whether the fatty acid composition was within the established ranges set by the International Olive Council (IOC), less than 10% would have been identified as non-EVOO. These findings are significant not only because they were consistent with previously published data based on the results of two sensory panels that were accredited by IOC but more importantly each measurement/analysis was accomplished in less than 5 min.     

Authenticity Assessment of Extra Virgin Olive Oil: Evaluation of Desmethylsterols and Triterpene Dialcohols

Extra virgin olive oil (EVOO) has a long history of economic adulteration, the detection of which presents significant challenges due to the diverse composition of cultivars grown around the world and the limitations of existing methods for detecting adulteration. In this study, using Method COI/T.20/Doc. No. 30/Rev. 1 of the International Olive Council, the authenticity of 88 market samples of EVOO was evaluated by comparing total sterol contents, desmethylsterol composition, and contents of triterpene dialcohols (erythrodiol and uvaol) with purity criteria specified in the United States Standards for grades of olive oil and olive‐pomace oil. Three of the 88 samples labeled as EVOO failed to meet purity criteria, indicating possible adulteration with commodity oil and/or solvent‐extracted olive oil. Detection of adulteration was also evaluated by spiking an EVOO sample with commodity oil at the 10 % level. As expected, eight of the spiked samples (canola, corn, hazelnut, peanut, safflower, soybean, and sunflower oils, and palm olein) failed to meet purity criteria. Two of the three samples spiked with 10 % hazelnut oil went undetected for adulteration. Overall, a low occurrence rate of adulteration (<5 %), based on purity criteria for desmethylsterols and triterpene dialcohols, was detected for the 88 products labeled as EVOO.     

Effect of Light Exposure on the Quality and Phenol Content of Commercial Extra Virgin Olive Oil during 12-Month Storage

Storage conditions influence the maximum time for which the composition and sensory characteristics of olive oils can be guaranteed. The purpose of this research was to study the quality and phenol content of extra virgin olive oil (EVOO) after storage for 1 year in different types of containers under darkness or light. Three Spanish cultivars with quantitatively different phenol contents were selected for the study. Storage under light conditions impaired the physicochemical and sensorial properties of the three cultivars, and reduced total phenolics, but there was an increase in hydroxytyrosol and tyrosol concentrations. It also markedly decreased their total phenolic content, especially when kept in polyethylene containers exposed to light, with reductions ranging from 4.28% for vanillic acid in Picual oils stored in dark glass containers under dark conditions to 97.82% for ferulic acid in Arbequina oils stored in polyethylene containers under light conditions. There was a reduced concentration of flavonoid and lignan concentrations after 1 year of storage, with the greatest decrease (98.01% of initial content) being observed for in the flavonoid apigenin. These results indicate that EVOO should be stored in dark glass containers under dark conditions for the optimal preservation of its quality and phenol content.     

Novel,Rapid Identification,and Quantification of Adulterants in Extra Virgin Olive Oil Using Near‐Infrared Spectroscopy and Chemometrics

A new, rapid Fourier transform near infrared (FT‐NIR) spectroscopic procedure is described to screen for the authenticity of extra virgin olive oils (EVOO) and to determine the kind and amount of an adulterant in EVOO. To screen EVOO, a partial least squares (PLS1) calibration model was developed to estimate a newly created FT‐NIR index based mainly on the relative intensities of two unique carbonyl overtone absorptions in the FT‐NIR spectra of EVOO and other mixtures attributed to volatile (5280 cm^?1) and non‐volatile (5180 cm^?1) components. Spectra were also used to predict the fatty acid (FA) composition of EVOO or samples spiked with an adulterant using previously developed PLS1 calibration models. Some adulterated mixtures could be identified provided the FA profile was sufficiently different from those of EVOO. To identify the type and determine the quantity of an adulterant, gravimetric mixtures were prepared by spiking EVOO with different concentrations of each adulterant. Based on FT‐NIR spectra, four PLS1 calibration models were developed for four specific groups of adulterants, each with a characteristic FA composition. Using these different PLS1 calibration models for prediction, plots of predicted vs. gravimetric concentrations of an adulterant in EVOO yielded linear regression functions with four unique sets of slopes, one for each group of adulterants. Four corresponding slope rules were defined that allowed for the determination of the nature and concentration of an adulterant in EVOO products by applying these four calibration models. The standard addition technique was used for confirmation.     

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.     

Physical,Chemical, and Biological Characterization of Veiled Extra Virgin Olive Oil Turbidity for Degradation Risk Assessment

Six different 300 kg batches of olive fruits are processed and the resulting six 20 kg batches of oil are collected at the end of the “decanter.” These batches of oil are subjected to four different water and solid particle separation treatments so as to obtain the following oil samples: veiled oil, filtered oil, “solid particle‐only” oil, and “water‐only” oil. The applied separation treatments show that water content has an important role in the degree of turbidity. High water content values (>0.2% w/w) are related to water activity values of >0.6 which are suitable for chemical and enzymatic reactions. The veiled oil samples are contaminated by microorganisms, but non‐proportional behavior occurs between the microbial cell count and the water and solid particle contents. Practical Applications: The results of this study recommend a multi‐approach method to characterize turbidity, based on control markers such as the degree of turbidity, water content and water activity, solid particle content, microbial contamination, and phenolic compound content. In this way, each degree of turbidity can be associated with a different level of risk of veiled extra virgin olive oil degradation during shelf life.     

Optimization of Reaction Conditions in the Enzymatic Interesterification of Soybean Oil and Fully Hydrogenated Soybean Oil to Produce Plastic Fats

Semisolid fats obtained from oils and fats through enzymatic interesterification have interesting applications. The effect of certain reaction parameters (enzyme concentration, moisture content, reaction time, substrate ratio, temperature, and agitation level) over the enzymatic interesterification of fully hydrogenated soybean oil (FHSO) and refined soybean oil (SO) using two immobilized enzyme types (Lipozyme RM IM and Lipozyme TL IM), was studied with a fractional factorial design (FFD). The reaction products were analyzed with respect to melting point (mp), by-products content and triacylglycerols (TAG) composition. It was found that substrate ratio, reaction time, and their interaction presented the most significant contributions to mp, varying this from 43.4 to 61.5 °C. The highest contributions to by-product content were presented by time and its interaction with the amount of molecular sieves, mainly for Lipozyme TL IM. Through the models obtained, theoretical conditions to achieve minimal by-product generation and mp were found, being 5.0 % (w/w_subst.) of any of both lipases, 24 h, 70:30 (oil:fat,  % w/w), 65 °C, 230 rpm, and absence of molecular sieves. Regression models for TAG groups as a function of significant factors and interactions were constructed, offering useful information to establish the reaction conditions for obtaining a product with a target mp or chemical composition.     

Oleocanthal,a Phenolic Derived from Virgin Olive Oil: A Review of the Beneficial Effects on Inflammatory Disease

Virgin olive oil (VOO) is credited as being one of many healthful components of the Mediterranean diet. Mediterranean populations experience reduced incidence of chronic inflammatory disease states and VOO is readily consumed as part of an everyday dietary pattern. A phenolic compound contained in VOO, named oleocanthal, shares unique perceptual and anti-inflammatory characteristics with Ibuprofen. Over recent years oleocanthal has become a compound of interest in the search for naturally occurring compounds with pharmacological qualities. Subsequent to its discovery and identification, oleocanthal has been reported to exhibit various modes of action in reducing inflammatory related disease, including joint-degenerative disease, neuro-degenerative disease and specific cancers. Therefore, it is postulated that long term consumption of VOO containing oleocanthal may contribute to the health benefits associated with the Mediterranean dietary pattern. The following paper summarizes the current literature on oleocanthal, in terms of its sensory and pharmacological properties, and also discusses the beneficial, health promoting activities of oleocanthal, in the context of the molecular mechanisms within various models of disease.     

A Comparative Study of O/W Nanoemulsions Using Extra Virgin Olive or Olive‐Pomace Oil: Impacts on Formation and Stability

Nanoemulsions are considered an innovative approach for industrial food applications. The present study explored the potential use of olive‐pomace oil (OPO) for oil‐in‐water (o/w) nanoemulsion preparations and compared the effectiveness of extra virgin olive oil (EVOO) and OPO at nanoemulsion formulations. The ternary‐phase diagrams were constructed and the o/w nanoemulsions properties were evaluated in relation to their composition. The results showed that it is possible to form OPO nanoemulsions using Polysorbate 20 or Polysorbate 40. Nanoemulsions with EVOO and OPO presented desirable properties, in terms of kinetic stability (emulsion stability index % [ESI%]), mean droplet diameter (MDD), polydispersity index (PDI), ζ‐potential, viscosity, and turbidity. EVOO exhibited lower surface and interfacial tension forming nanoemulsions with a high ESI% and a low MDD. However, OPO led to nanoemulsions with a high ESI% but with a higher MDD. It was observed that by increasing the emulsifier concentration the MDD decreased, while increasing the dispersed phase concentration led to a higher MDD and a lower ESI%. Finally, nanoemulsions with the smallest MDD (99.26 ± 4.20 nm) and PDI (0.236 ± 0.010) were formed using Polysorbate 40, which presented lower surface and interfacial tension. Specifically, the nanoemulsion with 6 wt% EVOO and 6 wt% Polysorbate 40 demonstrated an interfacial tension of 51.014 ± 0.919 mN m^?1 and an MDD of 99.26 ± 4.20 nm. However, the nanoemulsion with 6 wt% OPO and 8 wt% Polysorbate 20 presented an interfacial tension of 54.308 ± 0.089 mN m^?1 and an MDD of 340.5 ± 7.1 nm.     

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.     

Influence of the Atmosphere Composition during Malaxation and Storage on the Shelf Life of an Unfiltered Extra Virgin Olive Oil: Preliminary Results

The nutraceutical and organoleptic quality of the extra virgin olive oil (EVOO) depends on a process that begins with the olive ripening and ends with the packaging and storage. As the EVOO quality decay is mainly induced by the oxidation process, the gaseous composition of the atmosphere during both malaxation and storage can play a key role. Because of that, the aim of this research work is to determine the influence of the gaseous composition of the atmosphere used during malaxation and storage on an unfiltered EVOO shelf life, and to individuate the best “malaxation atmosphere × storage atmosphere” combination to adopt to more effectively slow down the oil quality decay rate during storage. The combined experimental conditions used during malaxation and storage significantly affect the quality decay rate of the EVOO over one year of storage for all the evaluated parameters. In conclusion, the gaseous composition of the atmosphere used during both malaxation and storage deeply affects the shelf life of the unfiltered EVOO as regards its chemical and sensory features, especially when air is used. On the contrary, the best results are obtained when an inert gas (nitrogen or argon) is used during both phases. Practical Applications : The regular consumption of EVOO represents one of the main cornerstones of the Mediterranean diet. According to the FDA (Food and Drug Administration), this product can be considered as a healthy food because of its extraordinary nutraceutical and organoleptic properties, so that if consumers consume half a tablespoon of olive oil per day—without increasing their daily calorie intake—they can significantly reduce the incidence of several degenerative diseases as well as the risks associated with them. The knowledge of the impact of different “malaxation atmosphere × storage atmosphere” combinations on the quality decay rate during storage of an unfiltered EVOO can allow to identify the best operating conditions to be used during its production and storage, in order to preserve the nutraceutical and organoleptic properties of this product over time, thus extending its commercial life in terms of both quality parameters and healthy properties.     

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.     

Enzymatic Production of Fatty Acid Methyl Esters by Hydrolysis of Acid Oil Followed by Esterification

Acid oil, a by-product of vegetable oil refining, was enzymatically converted to fatty acid methyl esters (FAME). Acid oil contained free fatty acids (FFA), acylglycerols, and lipophilic compounds. First, acylglycerols (11 wt%) were hydrolyzed at 30 °C by 20 units Candida rugosa lipase/g-mixture with 40 wt% water. The resulting oil layer containing 92 wt% FFA was used for the next reaction, methyl esterification of FFA to FAME by immobilized Candida antarctica lipase. A mixture of 66 wt% oil layer and 34 wt% methanol (5 mol for FFA) were shaken at 30 °C with 1.0 wt% lipase. The degree of esterification reached 96% after 24 h. The resulting reaction mixture was then dehydrated and subjected to the second esterification that was conducted with 2.2 wt% methanol (5 mol for residual FFA) and 1.0 wt% immobilized lipase. The degree of esterification of residual FFA reached 44%. The degree increased successfully to 72% (total degree of esterification 99%) by conducting the reaction in the presence of 10 wt% glycerol, because water in the oil layer was attracted to the glycerol layer. Over 98% of total esterification was maintained, even though the first and the second esterification reactions were repeated every 24 h for 40 days. The enzymatic process comprising hydrolysis and methyl esterification produced an oil containing 91 wt% FAME, 1 wt% FFA, 1 wt% acylglycerols, and 7 wt% lipophilic compounds.