Inclusion of sardine oil (SO) in diets for laying hens significantly increases the n-3 polyunsaturated fatty acids (PUFAs) in the egg, but these are more sensitive to oxidation, so the storage time and temperature can cause a decrease in their concentration. Therefore, the objective of this study was to determine the effect of algae Macrocystis pyrifera, Enteromorpha spp., and Sargassum sinicola on n-3 PUFA contents in eggs from laying hens fed diets supplemented with sardine oil and stored for different times (0, 15, and 30 days) and temperatures (20°C and 4°C), for 8 weeks. One hundred and twenty hens were divided into four treatments: T1 (commercial diet), T2 (2% SO+10% M. pyrifera), T3 (2% SO+10% Enteromorpha), and T4 (2% SO+10% S. sinicola). At the end, 50 eggs per treatment were collected to quantify total lipids and egg n-3 PUFAs at different times (0, 15, and 30 days) and temperatures (20°C and 4°C) of storage. The results were analyzed using a 3×3×2 factorial design, and Tukey test to compare means (P<0.05). The results show that M. pyrifera and S. sinicola had a better effect on eicosapentaenoic acid, while Enteromorpha was better for docosahexaenoic acid. In relation to time and temperature, the content of the fractions analyzed in the three treatments at 15 days/4°C had a lower loss compared with eggs analyzed at day 0/20°C.

Abstract. Curcumin has gained great attention in the last decades due to its fascinating properties for humans, such as anti-inflammatory or as cytotoxic against cancer. These effects are also claimed for pets such as cats and dogs, where curcumin administration is a daily practice routine. However, curcumin presents poor oral bioavailability, driving scientists to look for new delivery systems. In the last decades, several analytical methods for the quantification of curcumin in plasma have been published. To our knowledge there are no published reports on the effect of the level of hemolysis in the determination of this compound. In the present paper, a highly specific, sensitive and selective method is presented using Molecular Reaction Monitoring (SRM) using positive ionization (ESI+) mode. Curcumin and clopidogrel bisulfate – used as internal standard (IS) – were separated on an Acquity UPLC BECH Shield RP 18 column (1.7µm, 2.1 X 100mm) with 0.1% formic acid in acetonitrile and water in proportion of 60:40 (v/v). The analyte transitions were 369.3→177.06 m/z for curcumin and 322→212.05 m/z for IS. The method was fully validated and showed good linearity (R2 ≥ 0.999) over the range of 3-160 ng/mL. The Relative Standard Deviation (RSD) were less than 6% for intra-and inter-day analysis and recovery spanned 85-95%. We proved that the degree of hemolysis impaired curcumin quantitation. This method was applied to test curcumin bioavailability in both a mucoadhesive nanocapsule formulation and traditional capsules in dogs that attended routine veterinary consultation.Resumen. La curcumina ha ganado gran atención en las últimas décadas debido a sus propiedades terapéuticas para los humanos, como antiinflamatorio o citotóxico contra el cáncer. Estos efectos también se observan en pequeñas especies como gatos y perros, donde la administración de curcumina se ha vuelto una alternativa. Sin embargo, la curcumina presenta una baja biodisponibilidad oral, lo que impulsa a los científicos a buscar nuevos sistemas de administración. En las últimas décadas, se han publicado varios métodos analíticos para la cuantificación de curcumina en plasma. Actualmente, no hay informes publicados sobre el efecto del grado de hemólisis en la determinación de este compuesto. En este trabajo se desarrolló un método específico, sensible y selectivo utilizando el Monitoreo de reacción seleccionado (SRM) en modo de ionización positiva (ESI +). La curcumina y el bisulfato de clopidogrel, utilizado como patrón interno (IS), se separaron en una columna Acquity UPLC BECH Shield RP 18 (1,7 μm, 2,1 X 100 mm) con ácido fórmico al 0,1% en acetonitrilo y agua a una proporción de 60:40 (v/v). Las transiciones de los analitos fueron 369.3 → 177.06 m/z para curcumina y 322 → 212.05 m/z para IS. El método fue validado y demostró ser lineal (r2 ≥ 0.999) en el rango de 3-160 ng/mL. La desviación relativa estándar (RSD) fue inferior al 6% para el análisis intra e interdía y el porcentaje de recuperación fue 85-95%. Se descubrió que el grado de hemólisis afecta la cuantificación de curcumina. El método desarrollado se aplicó para evaluar la biodisponibilidad de curcumina tanto en una formulación de nanocápsulas mucoadhesivas como en cápsulas tradicionales en perros que asistieron a consultas veterinarias de rutina.