Development and validation of new analytical method for acrolein in air

A new method was developed to determine vapor-phase acrolein in air samples. Air containing vapor-phase acrolein was purged into impingers filled with a dichloromethane solution of N-methylhydrazine. The resulting derivative, 1-methyl-2-pyrazoline, was analyzed by gas chromatography using a nitrogen-phosphorous detector (NPD). The detection limit was 8.9 pg 1-methyl-2-pyrazoline, equivalent to 5.9 pg acrolein. The recovery efficiencies of vapor-phase acrolein were 98.0 +/- 2.9% and 100.3 +/- 3.1% for 150 and 15 micrograms, respectively. This method was satisfactorily applied for determination of acrolein formed from various heated fats. The amounts of acrolein formed in a headspace were 109 micrograms/L from lard, 164 micrograms/L from corn oil, 5.1 micrograms/L from cotton seed oil, and 163 micrograms/L from sunflower oil.     

Determination of aliphatic organic acids by high-performance liquid chromatography with pulsed electrochemical detection.

A new ion exclusion HPLC procedure accomplished with a pulsed electrochemical detection for the determination of several common aliphatic acids is described. A triple-step waveform of the applied potentials, based on the formation/inhibition of PtOH species on the electrode surface, is successfully used for sensitive detection of several aliphatic acids in flowing systems avoiding pre- or postcolumn derivatization and/or cleanup procedures. Under optimal chromatographic conditions (i.e., 50 mM HClO(4)) the proposed method allowed detection limits between 0.5 and 7 microM for all investigated acids, and the dynamic linear range spanned generally over 1 or 2 orders of magnitude. Determination of citric, malic, tartaric, lactic, formic, and acetic acids in several foods and beverages was performed, in approximately 15 min, without the necessity of any sample pretreatment.     

Species-specific identification of seven vegetable oils based on suspension bead array

Species adulteration of vegetable oils has become a main form of adulteration in vegetable oils, severely violating consumer rights and causing disorder in the market. A reliable method of species authentication of vegetable oils is desirable. This paper reports a novel method for identification of seven species of vegetable oils based on suspension bead array. One pair of universal primers and seven species-specific probes were designed targeting rbcl gene of the chloroplast. Each probe was coupled to a unique color-coded microsphere. Biotinylated PCR amplicons of seven oils were hybridized to the complementary probes on microsphere sets. Bound amplicons were detected fluorometrically using a reporter dye, streptavidin-R-phycoeryt hrin (SA-PE). A sample could be analyzed less than 1 h after PCR amplification. With the exception of olive probe, all probes showed no cross-reactivity with other species. Absolute detection limit of the seven probes ranged from 0.01 ng/μL to 0.0001 ng/μL. Detection limit in DNA mixture was from 10% to 5%. Detection of vegetable oils validated the effectiveness of the method. The suspension bead array as a rapid, sensitive, and high-throughput technology has potential to identify more species of vegetable oils with increased species of probes.     

Detection of adulteration of poppy seed oil with sunflower oil based on volatiles and triacylglycerol composition

Although poppy seed oil is an expensive article of trade, no literature about identification methods for adulteration with cheaper vegetable oils, like sunflower oil, has been published. This kind of adulteration is a challenge for routine analytical methods, such as the determination of fatty acid composition, because of almost similar fatty acid ratios. The detection of adulteration of poppy seed oils with sunflower oils at different levels (5-40%, w/w) by using SPME-GC-MS and MALDI-ToF-MS is the subject of our investigation. With the mentioned SPME-GC-MS method, it was possible to detect an admixture of sunflower oils in all relevant (5-40%) amounts by using alpha-pinene as a marker compound. Admixture of sunflower oil with high levels of triolein (high-oleic acid type) could be undoubtedly detected by MALDI-MS down to the 5-10% level. In contrast, adulteration of pure poppy seed oil by "standard" sunflower oils remained indistinguishable using this MALDI-MS.     

Determination of nonprotein amino acids and betaines in vegetable oils by flow injection triple-quadrupole tandem mass spectrometry: a screening method for the detection of adulterations of olive oils

A novel screening method using an automated flow injection electrospray ionization tandem mass spectrometry system is proposed for the simultaneous determination of five nonprotein amino acids (β-alanine, alloisoleucine, ornithine, citrulline, pyroglutamic acid) and three betaines (glycine betaine, trigonelline, proline betaine) after derivatization with butanolic HCl. MS/MS experiments were carried out in a triple-quadrupole instrument using multiple reaction monitoring mode in <2 min. The proposed method provided high fingerprinting power to identify the presence of five of the studied compounds in different types of vegetable oils (soybean, sunflower, corn, olive) with LODs at parts per billion levels. The method was validated, and different mixtures of extra virgin olive oil with seed oils were analyzed, achieving the typification for the detection of adulterations in extra virgin olive oils up to 2% w/w. The nonprotein amino acid ornithine was confirmed as a marker for adulteration in the olive oils analyzed.     

Capillary electrophoresis of free fatty acids by indirect ultraviolet detection: application to the classification of vegetable oils according to their botanical origin

A method for the determination of fatty acids in vegetable oils by capillary electrophoresis with indirect UV-vis detection has been developed. The separation of fatty acids was optimized in terms of Brij surfactant nature and concentration and organic modifier (2-propanol) percentage. The optimal background electrolyte consisted of 10 mM p-hydroxybenzoate, 5 mM Tris at pH 8.8, 80 mM Brij 98, 40% acetonitrile, and 10% 2-propanol. Under these conditions, vegetable oils from five botanical origins (avocado, corn, extra virgin olive, hazelnut, and soybean) were analyzed and the fatty acid contents established. Linear discriminant analysis (LDA) models were constructed using fatty acid peak areas as predictors. An excellent resolution among all category pairs was obtained, and all samples were correctly classified with assignment probabilities of >95%.     

Determination of free proline and monosaccharides in wine samples by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD)

A sensitive and selective analytical method for the simultaneous separation and quantitative determination of proline and free monosaccharides in wine samples by high-performance anion-exchange chromatography coupled with pulsed amperometric detection is described. Under optimized experimental conditions, a complete separation was obtained in less than 30 min, using an isocratic elution with 10 mM NaOH and 1 mM Ba(OAc)(2). No postcolumn addition of strong bases to the eluent for enhancing detection sensitivity was needed. Upon 25-fold sample dilution and purification to avoid interference of tannins, pigments, and phenolic compounds, the fingerprinting of common monosaccharides (i.e., arabinose, glucose, fructose, galactose, and xylose) and proline in wines, musts, and vinegars can be easily accomplished. The method allows high recovery and satisfies the necessary requirements for accuracy, repeatability, and sensitivity. Values obtained for proline content ranged from 470 to 1190 mg/L in "Aglianico" red wines (mean value, 870 +/- 192 mg/L, n = 21) and from 168 to 286 mg/L in white wines (mean value, 208 +/- 32 mg/L, n = 11). Lower levels were found in musts of red and white grapes, 550 and 87 mg/L, respectively. The lowest content of proline, ca. 10 mg/L, was found both in white and red vinegars.     

Determination of aflatoxins in vegetable oils

A simple method is proposed for determination of aflatoxins in vegetable oils. The method was successfully applied to both crude and degummed oils. The oil sample, dissolved in hexane, was applied to a silica column and washed with ether, toluene, and chloroform; aflatoxins were eluted from the column with chloroform-methanol (97 + 3). As quantitated by thin layer chromatography and liquid chromatography, the oils analyzed contained aflatoxin B1 at levels of 5-200 micrograms/kg. Recoveries of aflatoxin B1 standards added to aflatoxin-free oils were between 89.5 and 93.5%, with coefficients of variation of 6.3-8.0%.     

Headspace solid-phase microextraction use for the characterization of volatile compounds in vegetable oils of different sensory quality

Headspace solid-phase microextraction (HS-SPME) was used to isolate the volatile compounds, which are formed during peroxidation of fatty acids in vegetable oils. Isolated compounds were characterized by GC-MS and quantified using GC with FID detection. Four fibers for HS-SPME method development were tested, and the divinylbenzene/carboxene/PDMS fiber was selected as providing the best detection of analyzed compounds. Extraction curves, limits of detection, repeatability, and linearity were investigated for 14 aldehydes, ketones, hydrocarbons, and alcohols being products of fatty acids autoxidation. Limits of detection for 11 of these were below 1 microg/L. For quantitative purposes, to minimize the influence of temperature on hydroperoxide formation and the changes in the volatiles profile of the extracts, sampling was performed at 20 degrees C. For compound characterization by GC-MS, sampling temperature of 50 degrees C was applied. The developed method was applied to the analysis of refined and cold-pressed rapeseed oil stored at 60 degrees C for 10 days, and for 10 different vegetable oils of various degree of peroxidation. All samples were subjected to sensory analysis. The results of PCA sensory analysis were related to the amount of volatile compounds isolated by SPME method. In cases where the amount of compounds was highest, the samples were perceived as the worst, whereas those with low levels of volatile compounds were the most desired ones according to sensory evaluation. The relation was observed for both total volatiles, quantified C5-C9 aldehydes, and 14 compounds selected in method development. SPME revealed to be a rapid and sensitive method for the extraction and quantitation of trace volatile compounds from plant oils even at ambient temperature.     

Development of two stable isotope dilution assays for the quantitation of acrolein in heat-processed fats

Two stable isotope dilution assays were developed for the quantitation of acrolein in fats and oils using [(13)C(3)]-acrolein as the internal standard. First, a direct GC-MS headspace method, followed by an indirect GC-MS method using derivatization with pentafluorophenyl hydrazine, was established. Analysis of six different types of oils varying in their pattern of fatty acids showed significant differences in the amounts of acrolein formed after heating at various temperatures and for various times. For example, after 24 h at 140 °C, coconut oil contained 6.7 mg/kg, whereas linseed oil was highest with 242.3 mg/kg. A comparison of the results showed that the extent of acrolein formation seemed to be correlated with the amount of linolenic acid in the oils. Although the acrolein concentrations were lowered in all six oils after frying of potato crisps, linseed and rapeseed oil still contained the highest amounts of acrolein after frying. By applying both methods on different thermally treated fats and oils, nearly identical quantitative data were obtained.     

NIR spectroscopy and partial least-squares regression for determination of natural alpha-tocopherol in vegetable oils

Near-infrared (NIR) spectroscopy and partial least-square regression were used for determination of alpha-tocopherol in edible oils after extraction with ethanol. The standard error of calibration and the standard error of prediction were calculated for evaluation of the calibration models. The chemometric calibration model was prepared in spectral region 6500-4500 cm(-1) for standard alpha-tocopherol solutions (0.54-53.54 mg/mL). Obtained mean concentrations of natural alpha-tocopherol in different types of oils varied from 17.53 to 57.10 mg/100 g. Net analyte signal calculation was used to estimate detection limit (DL = 0.12 mg/mL), quantification limit (QL = 0.40 mg/mL), sensitivity (SEN = 0.045 mg/mL), and selectivity (SEL ranged between 0.24 and 0.54% of the measured reflectance signal) of the proposed NIR method. The comparable precision (RSD = 0.68-2.80% and 0.79-3.06%) and accuracy (recovery, 97.2-102.4% and 96.8-103.2%) for the proposed NIR and standard HPLC methods, demonstrate the benefit of the NIR method in the routine analysis of alpha-tocopherol in vegetable oils.     

Mass spectrometry and identification of sterols in vegetable oils as butyryl esters and relative quantitation by gas chromatography with flame ionization detection

Electron ionization mass spectrometry (MS) of sterol butyrates is described. Fragmentation of common sterol butyrates is related to structure and is discussed in relation to the fragmentation of free sterols and of commonly used sterol derivatives. Derivatized samples of vegetable oils are introduced using a 10 m capillary gas chromatographic (GC) column for complete separation of the sterol butyrates. Quantitation of sterol butyrates in vegetable oils by packed column GC/flame ionization detection is based on percent relative area of peaks identified by MS. Results of analyses of sunflower, castor, rapeseed, and virgin olive oils, and other oils are presented. These techniques have been applied to the rapid screening of marketed olive oils for possible adulteration.     

Determination of the enantiomeric composition of gamma-lactones in edible oils by on-line coupled high performance liquid chromatography and gas chromatography

A new method is proposed for the determination of the enantiomeric composition of gamma-lactones in different vegetable edible oils (i. e., olive oil, almond oil, hazelnut oil, peanut oil, and walnut oil), and its potential for authenticity control is underlined for a limited number of samples. The method is based on the direct injection (i.e., without requiring a sample pretreatment step) in on-line coupled reversed phase liquid chromatography to gas chromatography (RPLC-GC) using a chiral stationary phase in the GC-step. Different experimental values for both speed of sample introduction into GC and volume of the transferred fraction are considered to improve the recoveries obtained. Relative standard deviations lower than 10% and detection limits ranging from 0.06 to 0.22 mg/L were achieved for the investigated gamma-lactones.     

Detection of tricresyl phosphates and determination of tri-o-cresyl phosphate in edible oils

Tricresyl phosphate (TCP) in contaminated edible oils was extracted using acetonitrile and detected by thin layer chromatography as well as gas chromatography (GC). The chromatoplate was developed with isooctane-ethyl acetate (90 + 10) and visualized by spraying with 2,6-dichloroquinone chloroimide. TCP gives a characteristic blue-violet spot when heated at 100 degrees C for 15 min. The method is direct and sensitive and can be used to detect as low as 2.5 micrograms TCP or TOCP (tri-o-cresyl phosphate). GC was carried out using 10% OV-101 as the stationary phase and flame ionization detection for confirmation and quantitation of TOCP in oils.     

Quantitative determination of hydroxy pentacyclic triterpene acids in vegetable oils

A simple and precise analytical method for the determination of hydroxy pentacyclic triterpene acids (HPTAs) in vegetable oils was developed. The acidic fraction was isolated by solid-phase extraction using bonded aminopropyl cartridges, and the extract was silylated and analyzed by gas chromatography. Repeatability and recovery of the method were determined. In virgin olive oils, similar amounts of oleanolic (3beta-hydroxyolean-12-en-28-oic) and maslinic (2alpha,3beta-dihydroxyolean-12-ene-28oic) acids and traces of ursolic (3beta-hydroxyurs-12-en-28-oic) acid were found. The main factor affecting HPTA concentration was the oil quality since that increases as the quality decreases, while olive variety, olive ripeness, and oil extraction system had less influence. In crude olive pomace oils, the concentrations were very much higher than in virgin olive oils. During refining processes, total or significant losses of HPTAs were observed. Esterified derivatives of HPTAs were not found.     

Solid-phase microextraction for studies on the enantiomeric composition of filbertone in hazelnut oils

The enantiomeric distribution of filbertone was determined in unroasted and roasted hazelnut oils of different geographical origins by using solid-phase microextraction (SPME) and capillary gas chromatography. An optimization procedure including SPME fiber, extraction time, exposure temperature, and sample volume enabled the best conditions to be selected. Under the optimized conditions, detection limits were in the micrograms per liter level for both enantiomers of filbertone with relative standard deviation values of 7.1 and 4.9% for R-filbertone and S-filbertone, respectively. The proposed approach allowed the rapid determination of the enantiomeric composition of filbertone and demonstrated that its variability is an inherent property of the natural compound. Analysis of two batches of hazelnut oils obtained from either unroasted or roasted hazelnuts showed, in general, significantly higher amounts of filbertone in roasted hazelnut oils.     

Non-detectable levels of trans-fatty acids in peanut butter

The fatty acid composition of 11 brands of peanut butter and paste freshly prepared from roasted peanuts was analyzed with emphasis on isomeric trans-fatty acids. No trans-fatty acids were detected in any of the samples in an analytical system with a detection threshold of 0.01% of the sample weight. Hydrogenated vegetable oils are added to peanut butters at levels of 1--2% to prevent oil separation. Some hydrogenated vegetable oils are known to be sources of trans-fatty acids in the human diet. The addition of these products was not found to result in measurable amounts of trans-fatty acids in the peanut butters analyzed.     

Direct liquid chromatography method for the simultaneous quantification of hydroxytyrosol and tyrosol in red wines

A validated HPLC method with fluorescence detection for the simultaneous quantification of hydroxytyrosol and tyrosol in red wines is described. Detection conditions for both compounds were optimized (excitation at 279 and 278 and emission at 631 and 598 nm for hydroxytyrosol and tyrosol, respectively). The validation of the analytical method was based on selectivity, linearity, robustness, detection and quantification limits, repeatability, and recovery. The detection and quantification limits in red wines were set at 0.023 and 0.076 mg L(-1) for hydroxytyrosol and at 0.007 and 0.024 mg L(-1) for tyrosol determination, respectively. Precision values, both within-day and between-day (n = 5), remained below 3% for both compounds. In addition, a fractional factorial experimental design was developed to analyze the influence of six different conditions on analysis. The final optimized HPLC-fluorescence method allowed the analysis of 30 nonpretreated Spanish red wines to evaluate their hydroxytyrosol and tyrosol contents.     

Development of multiresidue analysis for twenty phthalate esters in edible vegetable oils by microwave-assisted extraction-gel permeation chromatography-solid phase extraction-gas chromatography-tandem mass spectrometry

A novel multiresidue analysis method is developed for the determination of twenty phthalate esters at the μg/kg level in edible vegetable oils by microwave-assisted extraction-gel permeation chromatography-solid phase extraction-high resolution gas chromatography-tandem mass spectrometry (MAE-GPC-SPE-HRGC-MS/MS). The samples were extracted with methanol under microwave incubation. Cleanup was carried out with GPC followed by a further C18 SPE column and then separated by the HP-5MS capillary column under a temperature program. The eluents were qualitatively and quantitatively determined by tandem mass analyzer with selected reaction monitoring (SRM) type and positive ion mode. The calibration curves showed good linearity in the range 5 μg/kg to 2.50 mg/kg with correlation coefficients larger than 0.999. Low detection limits (LODs) of 0.218-1.367 μg/kg and quantification limits (LOQ) of 0.72-4.51 μg/kg were achieved. The mean recoveries were in the range from 93.04% to 104.6% at 5, 15, and 40 μg/kg spiked levels, and the relative standard deviations (RSDs) were in the range of 1.01% and 5.26% (n = 7). This method could potentially overcome the interference from large amounts of lipids and pigment. The real sample test showed this method can be used for sensitive and accurate determination and confirmation of phthalate ester residues in high-fat and complex samples.     

Influence of time, surface-to-volume ratio, and heating process (continuous or intermittent) on the emission rates of selected carbonyl compounds during thermal oxidation of palm and soybean oils

The aim of this work was to compare the emission rates of selected carbonyl compounds (CC) produced by palm and soybean oils when heated at 180 degrees C in the presence of air, through different time intervals and at different surface-to-volume ratios ( S/ V), in continuous and intermittent processes. The CC were collected and derivatized onto silica C18 cartridges impregnated with an acid 2,4-dinitrophenylhidrazine solution, followed by extraction with acetonitrile and analysis by HPLC-UV and, in some cases, HPLC-MS with electrospray ionization. Among the CC quantified, namely, acetaldehyde, acrolein, propanal, butanal, hexanal, 2-heptenal, and 2-octenal, acrolein was the main emission in both oils and all S/ V ratios, followed by hexanal and 2-heptenal. The soybean oil has presented greater emission rates of acrolein than palm oil. When different S/ V ratios used during the heating process of the oil were compared, the emission rates, in general, were directly related to them, although saturated and nonsaturated CC have had different behaviors toward oxidation reactions. During intermittent heating, there was a trend of increasing emission rates of saturated aldehydes, whereas the opposite was observed with unsaturated aldehydes, probably due to the reactivity of the double bond present in these compounds.