Simultaneous Determination of Squalene, α-Tocopherol and β-Carotene in Table Olives by Solid Phase Extraction and High-Performance Liquid Chromatography with Diode Array Detection

Olives, the fruit of the Olea europaea tree, are highly appreciated in olive oil and table olives (20 % of crops) not only for their flavor but also for their nutritional properties, especially for antioxidant compounds such as squalling (SQ), α-tocopherol (TH) and β-carotene (BC). This paper presents a new analytical method for simultaneously determining SQ, TH and BC in table olives by using solid phase extraction (SPE) and high performance-liquid chromatography with diode array detection (HPLC-DAD), avoiding the classic saponification process. The correlation coefficients of calibration curves of the analyzed compounds ranged from 0.998 to 0.999, and the recoveries were in the range of 89.4–99.6 %. The validated method was used to analyze 30 table olive samples from Italy for their content of SQ (537–1,583 mg kg^?1), TH (21–90 mg kg^?1) and BC (0.4–2.6 mg kg^?1). Finally, experiments with HPLC-MS were conducted to compare this novel method with the classic saponification procedure.     

Simple Method for Determination of Fungicides in Citrus Fruits by Liquid Chromatography–Tandem Mass Spectrometry

A simple method for the determination of four fungicides (thiabendazole, imazalil, o-phenylphenol, and diphenyl) in citrus fruits has been developed. After spiking surrogates of these fungicides, the sample was extracted with acetonitrile, salted out, and the water was simultaneously removed using anhydrous magnesium sulfate and sodium chloride. The extract was first purified with a primary secondary amine and octadecylsilane, followed by the addition of dimethyl sulfoxide as a keeper solvent, and subsequently concentrated under a nitrogen stream. The compounds were analyzed by liquid chromatography–tandem mass spectrometry using the atmospheric pressure photoionization interface. The recoveries and relative standard deviations (RSDs) of the fungicides (1 mg kg^?1) were satisfactory (recovery 92–114 %; RSD <10 %). An evaluation with Shewhart QC plots revealed that all data points are within the controlled area, thus confirming the robustness of the method for analyzing the fungicide content of citrus fruits. The sample preparation time for ten samples was approximately 2 h, highlighting the time and labor efficiency of the method.     

Development of a Dispersive Liquid–Liquid Microextraction Method for the Determination of α-Tocopherol in Pigmented Wheat by High-Performance Liquid Chromatography

A dispersive liquid–liquid microextraction (DLLME) method coupled to high-performance liquid chromatography was developed for the analysis of α-tocopherol in grain samples. The DLLME parameters including the type and volume of extractants, the volume of disperser and the addition of salt were examined. The optimized DLLME procedure consisted in the formation of a cloudy solution promoted by the fast addition to the sample (5 mL of saponified sample solution diluted with 5 mL of water) of a mixture of carbon tetrachloride (extraction solvent, 80 μL) and ethanol (dispersive solvent, 200 μL) without the addition of salt, followed by shaking for 5 min and centrifuging for 3 min at 5,000 rpm. Intra- and inter-day repeatability expressed as % RSD were 3.5 and 7.6 %, respectively. The limit of detection and the limit of quantification were 1.9 and 6.3 μg?L^?1. The comparison of this method with the national standardized extraction method, supercritical carbon dioxide extraction, accelerated solvent extraction, and conventional heat-reflux extraction indicates that the DLLME was accurate (no significant differences at the 0.05 % probability level), high efficient, low organic solvent-consuming, and low cost. This procedure was successfully applied to 42 samples of 14 types of purple wheat, for which the content of α-tocopherol exhibited a significantly negative correlation with the pigment content measured by a spectrophotometer. The recovery rates ranged from 90.5 to 103.7 %.     

Simultaneous Determination of 18 Bioactive Compounds in Italian Bitter Liqueurs by Reversed-Phase High-Performance Liquid Chromatography–Diode Array Detection

A simple, fast and accurate method has been developed to simultaneously determine 18 bioactive compounds in Italian bitter liqueurs containing gentian, cinchona, cinnamon, rhubarb, clove, star anise or orange, by reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with diode array detection (DAD). HPLC analysis was performed with a C18 column using methanol and aqueous phosphoric acid (pH 2.5) as mobile phase. Selected wavelengths, i.e. 210, 232, 275, 285, 291, 310 and 368 nm, were used for quantification of compounds. The column temperature was controlled at 30 °C. The correlation coefficients (R ²) of the calibration curves of the analysed compounds were ≥0.9999 in a relatively wide concentration range (0.5–50 μg/ml). The proposed method proved successful in simultaneously analysing 18 bitter liqueurs produced in Italy. The concentration of the most important bitter principles, gentiopicroside, amarogentin, quinine and naringin, ranged as follows: 1.17–299.20, 0.25–32.24, 1.44–6.93 and 0.28–39.99 μg/ml, respectively.     

A New Extraction Method for the Determination of Oxytocin in Milk by Enzyme Immune Assay or High-Performance Liquid Chromatography: Validation by Liquid Chromatography–Mass Spectrometry

There has been a controversy regarding the use of exogenous oxytocin (OT) in milking cattle which may have toxicological consequences during nonphysiological exposure. In the present study, a new sensitive extraction method for OT was developed followed by enzyme immune assay (EIA) or high-performance liquid chromatography (HPLC) analysis. The extraction of OT in milk involves two steps: (1) TCA precipitation of milk proteins and (2) solid-phase extraction (SPE) cleanup process. Without these steps, analysis of OT in milk was not possible. Utilizing EIA as a quantitative tool the limit of detection (LOD) and limit of quantitation (LOQ) were found to be 7.74 and 10.3 pg?ml^?1, precision in terms of intra- and interday coefficient of variation was below 13 % (%RSD, N?=?8), while percent recoveries were between 85 and 92 %. Utilizing UV-HPLC, the LOD, LOQ, precision, and recovery values were found to be 4.1 ng?ml^?1, 9.8 ng?ml^?1, 2–10 %, and 84–91 %, respectively. OT was found to be stable against adverse temperature (up to 100 °C) and pH (2 to 10) and simulated gastric fluid digestibility assay. Four milk samples collected from the market were analyzed, which showed that TCA precipitation and SPE steps are mandatory and the results were validated by LC-MS showing mass ion peak at 1 kD.     

Application of Fast Gas Chromatography–Mass Spectrometry in Combination with the QuEChERS Method for the Determination of Pesticide Residues in Fruits and Vegetables

A fast gas chromatography–mass spectrometry method has been developed for multiresidue determination of up to 56 pesticides in fruits and vegetables in a chromatographic run time of <10 min, using a single quadrupole mass spectrometer operating in selected ion monitoring mode. The well-known acetate-buffering version of the QuEChERS method has been used for sample preparation. Programmable temperature vaporizer injection of 3 μL allowed reaching limits of detection between 0.15 and 15 μg/kg for most compounds in the sample matrices tested. The applicability of the method has been evaluated in apple, orange, carrot, and tomato. Recoveries at three fortification levels (0.01, 0.1 and 0.5 mg/kg) ranged from 70 to 120 % for most compounds, with relative standard deviations below 20 % in all cases. The developed method has been applied to fruit and vegetable samples from different Spanish provinces.     

Multi-Residue Determination of Pesticides in Açai Tropical Fruit (Euterpe oleracea) by Matrix Solid-Phase Dispersion Combined with Liquid Chromatography

An extraction method based on matrix solid phase dispersion was developed to determine bromuconazole, fenbuconazole, parathion-methyl, kresoxim-methyl, and teflubenzuron in açai using liquid chromatography with ultraviolet diode array detector. The best results were obtained using 1.0 g of açai, 3.0 g of neutral alumina as the dispersant sorbent, and cyclohexane/ethyl acetate (1:1, v/v, 20 mL) as an eluting solvent. The method was validated using açai samples fortified with pesticides at three concentration levels (0.25, 0.5, and 1.0 mg kg^?1). Average recoveries (seven replicates) ranged from 66 to 119 %, with relative standard deviations between 1.4 and 20 %. The detection and quantification limits for açai ranged from 0.02 to 0.05 mg kg^?1 and from 0.05 to 0.1 mg kg^?1, respectively.     

Determination of Selected Polychlorinated Biphenyl Residues in Meat Products by QuEChERS Method Coupled with Gas Chromatography–Mass Spectrometry

Polychlorinated biphenyl (PCB) residues in food are an important food safety concern. Simple and sensitive analytical methods are needed to monitor PCB residues and ensure that food is safe for consumption. The aim of this study was to adapt a selective, sensitive, quick, and easy sample treatment for purification of animal fat matrices and to measure the level of PCB28, PCB52, PCB101, PCB118, PCB138, PCB153, and PCB180 in samples of meat products (salami, soudjouk, and sausage) produced in Turkey. The extraction and purification of meat products were performed via the modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) method and PCB levels determined by gas chromatography–mass spectrometry. The linearity was satisfactory for all compounds studied, with correlation coefficients ≥0.99. The limits of determination and the limits of quantification were in the range of 0.144–0.382 and 0.479–1.274 ng g^?1, respectively. Recovery at 3 different spiking concentrations was 95.7–101 % and the relative standard deviations were <3.5 %. This validated method was observed to be more economic and eco-friendly, as it uses a smaller volume of extraction solvents that are also less toxic. The validated method was successfully applied to the analysis of selected PCBs in meat products with satisfactory results. The method’s results indicated the presence of PCBs in some of the meat product samples, although the levels were below the maximum residue limit for food products of animal origin in Turkey (40 ng g^?1 of fat), which is in accordance with EU and Turkish levels.     

Determination of the Nicotine Content in Solanaceae Vegetables by Solid-Phase Extraction Coupled with Ultra High-Performance Liquid Chromatography–Tandem Mass Spectrometry

A validated method based on solid-phase extraction and ultra high-performance liquid chromatography–triple quadrupole tandem mass spectrometry, with electrospray ionization operated in the positive ion mode and multiple reaction monitoring, was developed for the determination of nicotine in Solanaceae vegetables. Sample preparation involved liquid–solid extraction, centrifugation, filtration, and solid-phase extraction. Two kinds of solid-phase extraction adsorbents, based on different retention mechanisms, were investigated. Relatively higher recoveries were obtained with a hydrophilic–lipophilic-balanced cartridge. A deuterated internal standard was used for quantification. The limit of quantification (LOQ) of nicotine in different vegetables was found to be between 0.07 and 0.5 μg/kg. The nicotine levels in the vegetable samples were above the LOQs. The method described here is thus suitable for the analysis of large sample batches, because it provides accurate quantification, high sensitivity and rapid chromatographic separation with facile preparation. The solid-phase extraction cartridges and organic solvents used in this work are easy to obtain, enabling the application of this method in most analytical laboratories.     

A Robust Transferable Method for the Determination of Glyphosate Residue in Liver After Derivatization by Ultra-high Pressure Liquid Chromatography–Tandem Mass Spectrometry

Glyphosate determination in liver is challenging due to this particular molecule/matrix combination. Glyphosate is a very polar molecule and liver composition is highly variable between individuals and species. Since 2014, the Multiannual Control Program (MACP) of the European Union (EU) demands to analyse glyphosate in food of animal origin on a voluntary basis. Moreover, this analysis will be mandatory in 2017. This paper describes a robust and easily transferable method for glyphosate quantification in liver of animal origin by means of liquid chromatography–tandem mass spectrometry (LC-MS/MS). An intensive clean-up was used to eliminate matrix interferences and was combined with a derivatization step which ensures good retention of glyphosate on a conventional reverse-phase LC column. This method allows to meet the MACP requirements without a time-consuming change in the set-up of the routinely used LC-MS/MS system. Furthermore, it allows the use of an LC column and mobile phases often used in multi-residue analysis. The analytical method was validated according to the SANCO/12571/2013 criteria. Isotopic dilution was used to quantify glyphosate, leading to mean apparent recoveries of 115 and 101 % for the low (0.025 mg kg^?1) and the high (0.250 mg kg^?1) fortification levels, respectively. At both levels, the relative standard deviation was below 10 %. The limit of quantification of 0.025 mg kg^?1 was found to be satisfactory as it was below the maximum residue level (MRL) value set at 0.050 mg kg^?1 for glyphosate in liver. It is also the lowest MRL for all commodity types.     

A Rapid and Sensitive Method for Determination of Melamine in Fish,Shrimp, Clam,and Winkle by Gas Chromatography–Mass Spectrometry with Microwave-Assisted Derivatization

A method for rapid and sensitive determination of melamine in aquatic products by gas chromatography–mass spectrometry with microwave-assisted derivatization was proposed in this paper. Melamine was extracted from aquatic product samples using methanol, and the extract was cleaned with a mixed-mode cationic exchange solid phase extraction column. After elution with 5 % ammonia–methanol solution and drying with nitrogen, the residue was derivatized using N,O-bis(trimethylsilyl)trifluoroacetamide containing 1 % trimethylchlorosilane under microwave irradiation for 1 min with a power of 420 W, then detected with gas chromatography–mass spectrometry, and quantified by the external standard method. Some important parameters such as extraction solvent, microwave irradiation power and time, and derivatization reagent volume were investigated and optimized. The results showed that methanol could effectively extracted melamine from aquatic products as well as precipitated the protein in samples. Under the optimum conditions, the detection limit for melamine was as low as 0.006 mg/kg, and the linear range was from 0.02 to 50 mg/kg with a correlation coefficient of 0.9997. The proposed method was applied to the analysis of melamine in aquatic products (fish, shrimp, clam, and winkle), and the recovery for melamine was 89.65–105.16 % with relative standard deviation of 3.0–6.0 %.     

Inhibition of Lipid Oxidation in Chicken by Carnosine and Dietary α-Tocopherol Supplementation and its Determination by Derivative Spectrophotometry

The antioxidant activity of carnosine in chicken meat, and its relationship to dietary α-tocopherol supplementation, was examined. Broiler chickens were fed diets containing 30 (basal) or 200 (supplemental) mg α-tocopherol acetate kg⁻¹ feed for 6 weeks. Raw and cooked thigh meat patties containing carnosine (0–1·5%) were prepared. Lipid oxidation, during refrigerated storage under fluorescent light, was assessed by monitoring malonaldehyde formation, using the TBA assay and single wavelength (conventional) or first derivative spectrophotometry. In raw patties, added carnosine improved oxidative stability for up to 10 days of refrigerated storage. In cooked patties, the 1·5% carnosine level provided the best antioxidant protection during 7 days of storage. Carnosine (1·5%) was at least as effective as supplemental α-tocopherol in improving the oxidative stability of raw and cooked patties. The presence of both antioxidants had an additive effect on oxidative stability. Overall, the use of derivative spectrophotometry improved the specificity of the TBA assay for monitoring MDA formation in refrigerated meats.     

A Rapid Determination of Ergosterol in Grains Using Gas Chromatography–Mass Spectrometry Method Without Derivatization

Food Analytical Methods - A simple, rapid and sensitive GC-MS method using on-column injection was developed and validated to determine ergosterol (ERG) levels in maize and wheat. In this method,...     

Determination of free α-lipoic acid in foodstuffs by HPLC coupled with CEAD and ESI-MS

A simple and rapid method for determination of free α-lipoic acid in different food matrices has been developed. It consists of extraction of α-lipoic acid with 0.5% glacial acetic acid in methanol by sonication, quantitative analysis of the extract by isocratic RP-HPLC (acetonitrile/methanol/50 mM potassium dihydrogen phosphate buffer adjusted to pH 3 with phosphoric acid: 350/65/585, v:v:v) at a flow rate of 0.45 ml/min coupled with coulometric electrode array detection at potentials between +300 and +700 mV and qualitative analysis by LC–ESI-MS in the negative ion mode for confirmation. Egg, dried egg powder, mayonnaise, fine peas and potatoes were analysed and free α-lipoic acid contents ranged from 0.1 to 4.2 μg/g with recoveries between 70% and 94%. Limits of quantitation were between 0.1 and 0.3 μg/g. This newly developed method can be used to establish a database for the content of free α-lipoic acid in different foodstuffs.     

Modified QuEChERS Combination with Magnetic Solid-Phase Extraction for the Determination of 16 Preservatives by Gas Chromatography–Mass Spectrometry

In this paper, based on the mechanism of the quick, easy, cheap, effective, rugged and safe (QuEChERS) method, a novel graphene grafted silica-coated Fe₃O₄ (Fe₃O₄@SiO₂@G) was synthesized and applied as the efficient magnetic solid-phase extraction (MSPE) adsorbent for rapid cleanup of vegetable samples prior to analyzing 16 preservative residues by gas chromatography–mass spectrometry (GC-MS). The method, which took advantages of the novel nanoparticle adsorbent and an external magnetic field separation targets from samples, not only could avoid the time consuming of the traditional solid-phase extraction, but also could be developed for simultaneous determination of 16 preservative residues in vegetables. Various experimental parameters that could affect the extraction efficiencies have been investigated. Under the optimum conditions, 16 preservatives showed good linearity over the range of 0.02–2.00 mg/L and correlation coefficients (R ²) of 0.9946–0.9998. The limits of detections (LODs) were in the range of 0.21–11.50 μg/kg. The recoveries of 16 preservatives ranged from 78.3 to 116.7 %, and the relative standard deviations (RSDs) ranged from 1.4 to 11.9 %.     

Headspace Liquid-Phase Microextraction Followed by Gas Chromatography–Mass Spectrometry for Determination of Furanic Compounds in Baby Foods and Method Optimization Using Response Surface Methodology

In the present study, a sensitive, rapid, and simple method for determination of furanic compounds in baby foods has been developed. Headspace liquid-phase microextraction (HS-LPME) coupled with gas chromatography–mass spectrometry was used to extract and measure furan, 2-methylfuran, and 2,5-dimethylfuran in baby foods. Effective parameters such as salt amount (NaCl), stirring rate, temperature, and time of extraction were optimized using response surface methodology based on a central composite design to obtain the best conditions for extracting furanic compounds. The optimum parameter values were 1 g NaCl, 700 rpm stirring rate, 40 °C extraction temperature, and 15 min extraction time. The calibration curves were linear over the range of 0.2–200 ng?mL^?1 (R ² ?>?0.99) for all compounds, and the repeatability of the method, described as relative standard deviation, ranged between 3.84 and 7.06 % (n?=?6). The recovery of spiked baby food sample after extraction ranged between 89.33 and 103.64 %, and the best enrichment factor was achieved about 972-fold for furan. The limits of detection and quantitation ranged between 0.021 and 0.038 ng?g^?1 and 0.069 and 0.126 ng?g^?1, respectively. The merit figures of the HS-LPME/GC-MS method showed that it can be considered as a new, fast, and effective alternative method for investigating furanic compounds in baby foods.     

Validation of an Electrochemical Detection–High-Performance Liquid Chromatography Method for Simultaneous Determination of Lignans in Flaxseed (Linum usitatissimum L.)

Flaxseed is a major source of lignans, which are important bioactive compounds. The aims of this work were to validate a liquid chromatographic method for the rapid and simultaneous determination of the main lignans in flaxseed using high-performance liquid chromatography coupled with electrochemical detection and to analyze the composition of commercial samples of flaxseed. The performance criteria of the proposed method demonstrate that the method can be used for the analysis of secoisolariciresinol diglucoside (SDG), secoisolariciresinol (SECO), matairesinol (MATA), pinoresinol (PINO), lariciresinol (LARI), hydroxymatairesinol (HYDROXY), and isolariciresinol (ISOLARI) in flaxseeds at suitable levels. Calibration curves were determined for six different concentrations of standard solutions injected in triplicate. The sensitivity of the calibration curve was evaluated considering the confidence intervals of the intercept and slope. The limit of detection and limit of quantification were 7.4 and 10.9 μg/l, respectively, for LARI and 17.7 and 37.5 μg/l, respectively, for MATA. The relative standard deviation of repeatability values were lower than 2.59 %, which are acceptable because the Horwitz ratio values were 0.1 for all of the lignans. The recoveries of lignans were in the range of 74–100 % of SECO, which are consistent with the literature. The precision of the proposed method was determined by analyzing four flaxseed samples of different years and varieties. SDG was the main lignan present in all the samples, followed by ISOLARI and HYDROXY.