Chemical Fingerprinting of Olive Oils by Electrospray Ionization-Differential Mobility Analysis-Mass Spectrometry: A New Alternative to Food Authenticity Testing.

Recently, the olive oil industry has been the subject of harsh criticism for false labeling and even adulterating olive oils. This situation in which both the industry and the population are affected leads to an urgent need to increase controls to avoid fraudulent activities around this precious product. The aim of this work is to propose a new analytical platform by coupling electrospray ionization (ESI), differential mobility analysis (DMA), and mass spectrometry (MS) for the analysis of olive oils based on the information obtained from the chemical fingerprint (nontargeted analyses). Regarding the sample preparation, two approaches were proposed: (i) sample dilution and (ii) liquid-liquid extraction (LLE). To demonstrate the feasibility of the method, 30 olive oil samples in 3 different categories were analyzed, using 21 of them to elaborate the classification model and the remaining 9 to test it (blind samples). To develop the prediction model, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) were used. The overall success rate of the classification to differentiate among extra virgin olive oil (EVOO), virgin olive oil (VOO), and lampante olive oil (LOO) was 89% for the LLE samples and 67% for the diluted samples. However, combining both methods, the ability to differentiate EVOO from lower-quality oils (VOO and LOO) and the edible oils (EVOO and VOO) from nonedible oil (LOO) was 100%. The results show that ESI-DMA-MS can become an effective tool for the olive oil sector.

Determination of penicillins in milk of animal origin by capillary electrophoresis: is sample treatment the bottleneck for routine laboratories?

Capillary electrophoresis (CE) is increasingly being used not only for research purposes but also for routine analyses. The latter, however, are especially difficult when the analytes are present at very low concentrations in complex food samples (e.g. penicillins in milk of animal origin). No study of the difficulties encountered in daily practice in sample treatments for the determination of penicillins (PENs) in milk by CE has to our knowledge been reported. Rather than reviewing the main uses of CE for determining PENs in different types of samples, this paper focuses on the weaknesses of available methods for this purpose, which originate in sample treatment rather than in a lack of robustness of the CE technique. Some problems which, based on our own experience, often confront sample treatment and method development in this context are discussed here. Clearly, the greatest source of error in this context is sample processing, which must provide optimal extraction and preconcentration of analytes, and extracts compatible with the separation technique to be used. In this respect, using time-consuming procedures can cause the loss of variable amounts of analytes in different steps. Interestingly, dramatically simplifying the sample preparation process can detract from sensitivity but lead to increased recoveries. As with any methodological development in routine analysis, acceptable results can only be obtained by considering all potentially influential factors.

Easy sample treatment for the determination of enrofloxacin and ciprofloxacin residues in raw bovine milk by capillary electrophoresis.

An easy, selective, and sensitive method has been developed for the determination of enrofloxacin (ENR) and its main active metabolite, ciprofloxacin (CIP), in raw bovine milk using CE with UV detection at 268 nm. Milk samples were prepared by a clean-up/extraction procedure based on protein precipitation with hydrochloride acid followed by being defatted by centrifugation and SPE using a hydrophilic-lipophilic balance cartridge. Optimum separation was obtained using a 50 mM phosphoric acid at pH 8.4 and the total electrophoretic run time was 6 min. Sample preparation by this method yielded clean extracts with quantitative and consistent mean recoveries from 89 to 97% for CIP and from 93 to 98% for ENR. LODs obtained were lower to the maximum residue limits for these fluoroquinolones. The precision of the ensuing method is acceptable; thus, the RSD for peak area and migration time was less than 8.5 and 0.5% for CIP and 9.9 and 0.9% for ENR, respectively. The results showed that the proposed method was efficient showing good recoveries, sensitivity, and precision for the studied compounds and could be satisfactorily applied in routine analysis for the monitoring of ENR and CIP residues in milk, due to its ruggedness and feasibility demonstrated.

Thirty years of capillary electrophoresis in food analysis laboratories: potential applications.

CE has generated considerable interest in the research community since instruments were introduced by different trading companies in the 1990s. Nowadays, CE is popular due to its simplicity, speed, highly efficient separations and minimal solvent and reagent consumption; it can also be included as a useful technique in the nanotechnology field and it covers a wide range of specific applications in different fields (chemical, pharmaceutical, genetic, clinical, food and environmental). CE has been very well evaluated in research laboratories for several years, and different new approaches to improve sensitivity (one of the main drawbacks of CE) and robustness have been proposed. However, this technique is still not well accepted in routine laboratories for food analysis. Researching in data bases, it is easy to find several electrophoretic methods to determine different groups of analytes and sometimes they are compared in terms of sensitivity, selectivity, precision and applicability with other separation techniques. Although these papers frequently prove the potential of this methodology in spiked samples, it is not common to find a discussion of the well-known complexity of the matrices to extract analytes from the sample and/or to study the interferences in the target analytes. Summarizing, the majority of CE scientific papers focus primarily on the effects upon the separation of the analytes while ignoring their behavior if these analytes are presented in real samples.

Extracción de Cxitetraciclina en carne de pollo: estudios de rendimiento con aumento de la fase polar del solvente de extracción / Oxytetracycline Extraction in Chicken Meat: Studies on Yield increasing Polar Phase in Extraction Solvent

El consumo de alimentos con residuos de oxitetraciclina (OTC) puede causar diversos efectos tóxicos en el humano. Con la finalidad de extraer y cuantificar dichos residuos en matrices biológicas, como carne de pollo, se han desarrollado diversos métodos. Dentro de los métodos propuestos el más empleado es la extracción líquido-líquido por ser sencillo, rápido y económico. Este tipo de extracción fue aplicada por Furusawa para OTC en pollo, empleando acetonitrilo/hexano en una proporción 5:4 obteniendo una recuperación del 88%. Este trabajo tuvo como objetivo estudiar la recuperación de OTC en carne de pollo ensayada por Furusawa, aumentando la proporción del solvente polar con respecto al hexano (2:1), para su posterior cuantificación mediante cromatografía líquida de alta resolución (HPLC). Para ello se utilizaron 24 porciones de 1 g de tejido muscular perteneciente al muslo de 3 pollos libres de antibióticos, las cuales se fortificaron con soluciones estándares de OTC de 0,1; 0,2; 0,5 y 1 µg/g, obteniendo 6 muestras fortificadas con cada concentración, las cuales fueron almacenadas por 12 horas a 4°C. La extracción del antibiótico se llevó a cabo con acetonitrilo/hexano en proporciones 5:4 y 2:1. En cada caso se evaluó la recuperación, precisión y sensibilidad. Tanto para la proporción 5:4 como 2:1, la concentración de 0,2 µg/g presentó la mayor recuperación, siendo 91,5 y 92,5%, respectivamente; sin embargo, al aumentar la concentración de OTC disminuyó la recuperación. La precisión se incrementó a la concentración de 0,5 µg/g, sin embargo, al duplicar la concentración a 1 µg/g disminuyó dicho parámetro. El límite de detección obtenido para la extracción de OTC con acetonitrilo/hexano en proporción 2:1 fue de 0,09 µg/g. Se recomienda realizar una desproteinización de la muestra previo al proceso de extracción.

Consumption of food that contains oxytetracycline (OTC) residues may produce several toxic effects in human beings. In order to extract and quantify such residues in biological matrices, like chicken meat, several methods have been developed. Among these methods, liquid-liquid extraction is the mostly used, because is quick, simple and inexpensive. This extraction method was applied by Furusawa for OTC in chicken, using acetonitrile/hexanes in a 5:4 proportion, obtaining an 88% recovery. The objective of this research was to study extraction of OTC in chicken meat assayed by Furusawa, raising polar solvent proportion in relation to hexanes (2:1), and further quantifying by means of high performance liquid chromatography (HPLC). Twenty four portions of 1 g from 3 OTC free chickens were fortified with OTC standard solutions: 0.1, 0.2, 0.5 and 1 µg/g, obtaining 6 fortified samples for each concentration, stored for 12 hours at 4°C. Antibiotic extraction was performed using acetonitrile/hexanes in 5:4 and 2:1 proportions. Recovery, precision and sensitivity were analyzed in all samples. Either 5:4 or 2:1 proportions, 0.2 µg/g concentration obtained the higher recovery, 91.5 and 92.5%, respectively; however when OTC concentrations raised, recovery became lower. Precision increased at 0.5 µg/g concentration, but, fell down when concentration duplicated: 1 µg/mL. Detection limit obtained for OTC extraction using acetonitrile/hexanes in 2:1 proportion were 0.09 µg/g. Deproteinization is recommended previously to extraction process.

Aminas biógenas y bacterias en salchichón tipo milano: Efecto del tiempo de almacenamiento / Bacterial and Biogenic Amines in Milano Type Sausage: Storage Time Effect

La presencia de aminoácidos libres, bacterias descarboxilantes, temperatura y tiempo de almacenamiento son factores que favorecen la formación de aminas biógenas. El objetivo de este trabajo fue determinar el efecto del tiempo de almacenamiento sobre la presencia de aminas biógenas y bacterias en salchichones tipo milano. Se utilizaron 9 salchichones enteros de tres marcas comerciales (tres unidades por marca), con fecha de vencimiento de dos meses. Las muestras se analizaron 60, 30 y 0 días antes del vencimiento. Se determinaron aminas biógenas por cromatografía líquida de alta resolución (HPLC), recuento de bacterias ácido lácticas (BAL), enterococos (ENTC) y enterobacterias (ENTB) según ICMSF. En la marca 1 se observó una disminución significativa (P < 0,05) del número de BAL entre los muestreos, la marca 2 se mantuvo constante, mientras que la marca 3 aumentó significativamente (P < 0,05) entre el primer y segundo muestreo, para mantenerse constante hasta el vencimiento. No hubo diferencias significativas en los ENTC de las marcas 1 y 3, mientras que en la marca 2 se encontraron diferencias significativas (P < 0,05) entre la primera fecha y las dos últimas, en las que se incrementó el número de UFC. Los factores marca, tiempo de almacenamiento e interacción de ambos tuvieron un efecto significativo (P < 0,05) sobre el contenido de aminas biógenas, ENTC y BAL. Durante el almacenamiento BAL predominó sobre ENTC. No se encontraron ENTB. Se detectó la presencia de tiramina, histamina, cadaverina y putrescina; las concentraciones de tiramina e histamina encontradas podrían producir intoxicación alimentaria en individuos susceptibles.

Free aminoacids presence, decarboxilating bacteria, temperature and storage time may induce biogenic amine formation. The objective of this research was to determine storage time effect on biogenic amine and bacteria presence in milano type sausage. Nine (9) complete sausages of three brands (3 units per brand) were used, two months from expiration date. Samples were analyzed 60, 30 and 0 days before expiration. Biogenic amines were determined by means of liquid chromatography (HPLC), lactic acid bacteria (LAB), enterococci (ENTC) and enterobacteria (ENTB) according to ICMSF. In brand 1 a significant diminishment (P < 0.05) in LAB number, brand 2 remain stable, meanwhile brand 3 raised significantly (P < 0.05) between first and second sampling, and kept this values until expiration date. There were no significant differences in ENTC of brands 1 and 3, but in brand 2, the number raised, and significant differences (P < 0.05) between first and later dates were found. Brand and storage time and interaction between them had a significant effect (P < 0.05) on biogenic amine content, ENTC and LAB. LAB was higher than ENTC during storage. ENTB were not found. Tiramine, histamine, cadaverin and putrescine were detected; tiramine and histamine concentrations could produce a food poisoning in susceptible individuals.