The Effect of Nitrogen Fertilization on the Yield, Quality and Fatty Acid Composition of Opuntia ficus-indica Seed Oil

Cactus pears are nutritious, drought-tolerant plants that flourish in hot and arid regions. All its plant parts can be consumed by humans and animals. Fruit seed oil production is an important emerging industry in South Africa. As part of an initiative to promote cactus pears as multi-functional crops, dual-purpose cultivars should be identified, and their production increased. The aim of this study was to investigate the role of nitrogen (N) fertilizer on the seed oil yield and quality of Opuntia ficus-indica. The project encompassed a trial using N fertilization from three N sources (limestone ammonium nitrate, ammonium sulfate, urea) and four N application levels (0, 60, 120, 240 kg ha−1). Oil was quantitatively extracted from the seed using the Folch method; fatty acids were quantified using a Varian 430-GC. Seed oil content significantly increased (p = 0.035) with increased N fertilization rates; the oil yield ranged between 7.96 and 9.54%. The composition of the main fatty acids (oleic, palmitic, cis-vaccenic and stearic acid) was significantly influenced; oleic and stearic acid were significantly increased by higher fertilization levels whereas a reducing trend was observed in palmitic and cis-vaccenic acid levels. The highest content fatty acid, linoleic acid, was not significantly influenced.

Improved Folch Method for Liver-Fat Quantification

Fatty liver represents a significant metabolic pathology of excess intrahepatic fat in domestic animals and humans. Quantification of hepatic-fat content is therefore essential for diagnosis and investigation of liver and metabolic disease. However, the reproducibility of hepatic steatosis analysis is often low due to subjective and technical factors. We hypothesized that improvement in tissue-lipids extraction efficiency would contribute to the accuracy and precision of liver-fat determination. To test it, we investigated the effect of standardized tissue sonication on liver-fat quantification by the Folch method in sheep. Liver samples from grownup lambs of lean (n = 16) and fatty (n = 15) livers, and from pregnant ewes (n = 6) who died from pregnancy toxemia (PT), were used for hepatic-fat content determination with or without tissue sonication. In the grown lambs, an average hepatic-fat content of 6.6% was determined in sonicated compared to 5.1% in non-sonicated specimens (P = 0.0002). Similarly, in ewes with PT, an average of 12.5% was determined with sonication compared to 10.8% without it (P = 0.0006), and the reproducibility was higher with sonication (CV of 3.1 vs. 6.1%, respectively). Thus, tissue sonication improved the efficiency of liver-lipids extraction and was significant to the accuracy and precision of hepatic-fat determination. Enzymatic quantification of triglycerides was moderately correlated with the results obtained gravimetrically (r = 0.632, P < 0.005). The reported data provide reliable reference values for pregnancy toxemic sheep. The significant improvement in liver-fat quantification observed with the reported revised protocol is likely applicable to most mammals and humans.

The Effect of the Photoperiod on the Fatty Acid Profile and Weight in Hatchery-Reared Underyearlings and Yearlings of Atlantic Salmon Salmo salar L.

The influence of two light regimes, 16:8 h light/dark (LD 16:8) and 24:0 h light/dark (LD 24:0), in comparison to a usual hatchery light regime (HL), on the fatty acids content and weight gain in hatchery-reared underyearlings (at 0+ age) and yearlings (at 1+ age) of Atlantic salmon in the summer–autumn period was studied. The total lipids were analyzed by Folch method, the lipid classes using HPTLC, and the fatty acids of total lipids using GC. The increase in EPA and DHA observed in October in underyearlings and yearlings salmon (especially under LD 24:0) suggests they were physiologically preparing for overwintering. The changes in fatty acids and their ratios in juvenile Atlantic salmon can be used as biochemical indicators of the degree to which hatchery-reared fish are ready to smoltify. These associated with an increase in marine-type specific DHA and EPA, an increase in the 16:0/18:1(n-9) ratio, in correlation with a reduction in MUFAs (mainly 18:1(n-9)). These biochemical modifications, accompanied by fish weight gain, were more pronounced in October in yearlings exposed to continuous light (LD 24:0). The mortality rate was lower in experimental groups of underyearliings with additional lighting. Exposure to prolonged and continuous light did not affect yearlings mortality rate.

The effects of vitamins E and C on individual lipides in the liver and skeletal muscles of chicken broilers

The purpose of the work was to find out the changes in the lipid composition of the liver and skeletal muscles of broiler chickens at different stages of growth under the influence of supplements of vitamins E and C in their diet. The studies were conducted on four groups of broiler chickens. The control group was fed a standard compound feed. The first experimental group in addition to the specified compound feed received tocopherol acetate in the amount of 0.1 g/kg, the second — ascorbic acid, 0.25 g/kg. The third experimental group of chickens received tocopherol acetate and ascorbic acid at mentioned doses. The lipids from these tissues were extracted by the Folch method, and the ratio of individual lipid classes was determined by thin layer chromatography on silica gel. The results of the studies indicate the stimulating effect of vitamins E and C on the accumulation of total lipids in the skeletal muscles and liver of broiler chickens. In addition, we can conclude that feeding the broiler chickens of supplements containing vitamins E and C during the period of intensive growth promotes the increase of lipid synthesis in their liver and deposition of synthesized lipids in skeletal muscle. However, these processes are accompanied by a redistribution of the ratio of individual lipid classes in the investigated organs and tissues of chickens

DETERMINATION OF THE QUANTITATIVE CONTENT OF LIPIDS IN A BIOLOGICAL MATERIAL BY THE METHOD OF IR SPECTROSCOPY

A modification of the Folch method has been proposed for the quantitative determination of lipids in biological material, in which, after extraction of lipids with a chloroform / ethanol mixture, lipids are determined using IR spectroscopy. The bands corresponding to the stretching and deformation vibrations of the methyl and methylene groups of lipids and fatty acids were selected as analytical absorption bands: 1396, 1458, 2853, and 2923 cm- 1. These bands do not intersect with the absorption bands of proteins and nucleic acids, thus avoiding the stage of preliminary cleaning of lipids from non-lipid impurities. A multifactor regression model was constructed, which allows experimental data to be described with an error not exceeding 12%.

Polar Lipids in Starch-Rich Commodities to be Analyzed with LC-MS-Based Metabolomics—Optimization of Ionization Parameters and High-Throughput Extraction Protocols

Metabolomics-based approaches are still receiving growing attention with regard to food authenticity testing. Such studies require enormous sample numbers with negligible experimental or analytical variations to obtain statistically reliable results. In this context, an extraction protocol in line with optimized ionization parameters was developed in consideration of potential starch-derived matrix effects focusing on the polar lipids of potatoes. Therefore, well-known extractions (Bligh and Dyer, Folch, Matyash, and a n-hexane-based procedure) were compared in a non-targeted and a targeted approach regarding the extractability of their lipids such as phosphatidylcholines, phosphatidylethanolamines, galacto- and glucocerebrosides, di- and triglycerides, and acylated steryl glucosides. The selected Folch method was also scrutinized in view of its ability to remove the matrix’s starch and consequently improved by substituting trichlormethane with ethyl acetate as a “greener” Folch approach. Moreover, the challenge of starch-derived contamination and imminent ion suppression in the electrospray ionization source (ESI) was addressed by an optimization of ionization parameters varying desolvation settings, removing injection peaks, and increasing the angles and distances of the ESI-device. Long-term stability tests over five days were performed successfully with a combination of appropriate extraction and decreased desolvation settings during ionization. In conclusion, the present methodology provided the basis for on-going large-scale metabolomic studies with respect to the botanical origin of potatoes using UPLC-IMS-QToF (ultra-high performance liquid chromatography ion mobility spectroscopy quadrupole-time of flight mass spectrometer).

Polifenoles Totales, Antocianinas y Capacidad Antioxidante (DPPH y ABTS) durante el procesamiento del licor y polvo de cacao

Polifenoles Totales, Antocianinas y Capacidad Antioxidante (DPPH y ABTS) durante el procesamiento del licor y polvo de cacao Total polyphenols, anthocyanins and antioxidant capacity (DPPH and ABTS) during processing of cocoa liquor and cocoa powder Chávez Rivera Ronald Eduardo y Ordoñez Gómez, Elizabeth Susana, Universidad Nacional Agraria de la Selva, Apartado 156 DOI: https://doi.org/10.33017/RevECIPeru2013.0006/ Resumen El presente trabajo de investigación se desarrolló en los laboratorios del (CIPNA – CIDBAM) – UNAS. Los objetivos fueron: cuantificar el contenido de polifenoles totales, antocianinas y determinar la capacidad antioxidante por su capacidad de inhibir radicales libres 1,1-diphenyl-2-picrilhydrazyl (DPPH) y ácido 2,2-azinobis(3etilbenzotiazoline)-6-acidosulfonico (ABTS)durante el proceso del licor de cacao y polvo de cacao. Las muestras fueron molidas y desgrasadas por el método Folch. Se preparó extractos hidroalcohólicos que consistió en pesar 10 g de cacao en 100 mL (agua/etanol 50:50v/v), macerado por 24 h, filtrado y centrifugado a 10000 rpm/10min/4ºC. Los resultados fueron analizados mediante el diseño completo al azar (DCA) y el diseño de bloques completo al azar (DBCA), en los tratamientos donde hubo diferencia estadística se aplicó la prueba de Tukey (p<0,05), utilizando el programa SAS versión 9.0. Para analizar todos los tratamientos de manera conjunta se utilizó el análisis multivariado con componentes principales, utilizando el programa estadístico InfoStat versión 2011. El mayor contenido de polifenoles totales durante el procesamiento del licor de cacao y polvo de cacao fue en el grano 6,394±0,095 g EAG/100 g y el menor fue para el grano molido 4,036±0,105 g EAG/100 g. El mayor contenido de antocianinas fue en el grano 1,490±0,043 mg cianidina-3-glucósido/g y el menor fue para el grano molido 0,074±0,003 mg cianidina-3-glucósido/g. y la mayor capacidad de inhibición del radical DPPH fue IC50=51,158±0,117 μg/mL y la mejor capacidad de inhibir del radical ABTS fue IC50=34,918±0,054 μg/mL. Descriptores: cacao, polifenoles, antocianinas, antioxidante, DPPH, ABTS 236 Abstract This research was developed in the laboratories of (CIPNA – CIDBAM) - UNAS. The objectives were to quantify the content of total polyphenols, anthocyanins and to determine their antioxidant capacity by their ability to inhibit free radicals 1,1-diphenyl-2-picrilhydrazyl (DPPH) and 2,2 - azinobis (3etilbenzotiazoline)-6-acidosulfonic (ABTS) during the process of cocoa liquor and cocoa powder. The samples were ground and defatted by the Folch method. Hydroalcoholic extracts were prepared by weighing 10 g of cocoa in 100 mL (water / ethanol 50:50 v / v), macerating it for 24 h, filtering and centrifuging at 10000 rpm/10min/4ºC. The results were analyzed using a complete randomized design (CRD) and a complete randomized block design (CRBD), and in the treatments with statistical differences a Tukey test (p <0.05) was used using SAS version 9.0. To analyze all treatments in conjunction a principal component multivariate analysis was used using the statistical software InfoStat version 2011. The highest content of total polyphenols in the process of making cocoa liquor and cocoa powder was found in the grain with 6.394 g ± 0.095 g EAG/100 g and the lowest was found in the grounded grain with 4.036 ± 0.105 g EAG/100 g. The highest content of anthocyanins was in the grain with 1.490 ± 0.043 mg cyanidin-3-glucoside/g and the lowest was in the grounded grain with 0.074 ± 0.003 mg cyanidin-3-glucoside/g and the greatest inhibitory capacity for the DPPH radical was IC50 = 51.158 ± 0.117 µg/mL and the greatest capacity to inhibit the ABTS radical was IC50 = 34.918 ± 0.054 µg/mL. Keywords: Cocoa, poly-phenols, anthocyanins, antioxidant, DPPH, ABTS 235

Application of FTIR Spectroscopy and Chemometrics for Halal Authentication of Beef Meatball Adulterated with Dog Meat

Beef meatball is one of the favorite meat-based food products among Indonesian community. Currently, beef is very expensive in Indonesian market compared to other common meat types such as chicken and lamb. This situation has intrigued some unethical meatball producers to replace or adulterate beef with lower priced-meat like dog meat. The objective of this study was to evaluate the capability of FTIR spectroscopy combined with chemometrics for identification and quantification of dog meat (DM) in beef meatball (BM). Meatball samples were prepared by adding DM into BM ingredients in the range of 0–100% wt/wt and were subjected to extraction using Folch method. Lipid extracts obtained from the samples were scanned using FTIR spectrophotometer at 4000–650 cm-1. Partial least square (PLS) calibration was used to quantify DM in the meatball. The results showed that combined frequency regions of 1782–1623 cm-1 and 1485-659 cm-1 using detrending treatment gave optimum prediction of DM in BM. Coefficient of determination (R2) for correlation between the actual value of DM and FTIR predicted value was 0.993 in calibration model and 0.995 in validation model. The root mean square error of calibration (RMSEC) and standard error of cross validation (SECV) were 1.63% and 2.68%, respectively. FTIR spectroscopy combined with multivariate analysis can serve as an accurate and reliable method for analysis of DM in meatball.

Abstract T P86: Lipid Profiling Reveals Galatosylceramide Increasing in Plasma of Atherosclerotic Stroke

The development and innovation of new technique in metabonomics research and its application in medicine provides important resolution in screening biomarkers and targets for further investigation. The metabonomics research has unique advantages of facility, objection and reliability compared with other -omics studies. Lipidomics research was development based on metabonomics and has supreme merits in study of lipids related disease. Method: The carotid arthrosclerosis patients with stroke suffered before were recruited in this research. The items of lipid related laboratory examinations of these patients are normal because of well controlling under drug-taken. The peripheral blood of these patients was drawn and the plasma was isolated after centrifugation. The lipids from plasma were extracted by the standard Folch method. Then the lipids samples were injected for lipid profile analysis by 2D QoF LC/MS. The qualitative differentiations of these lipids were based on their m/z value, characteristic of the compounds fragmentation, direct and relative retention time of each lipid class. On the establishment of the lipids identification, we validated this newly 2D QoF LC/MS method following these measurements including its calibration equations, low of the detection and relative standard deviation. Result: Compared with native subjects, 518 different types of lipids molecular from 17 classes were identified from plasma of atherosclerosis patients by the method of 2D QoF LC/MS; The galactosylceramide and glucoceramide were identified separately although they were same of molecular weight; We proved that the levels of galactosylceramide, rather than glucoceramide were increasing in atherosclerotic stroke patients. Conclusion: the increasing levels of galactosylceramide in plasma of atherosclerosis patients hint that the immune effects induced by lipids play an important role in atherosclerosis.

Enhancement of Lipid Extraction from Marine Microalga,ScenedesmusAssociated with High-Pressure Homogenization Process

Marine microalga,Scenedesmussp., which is known to be suitable for biodiesel production because of its high lipid content, was subjected to the conventional Folch method of lipid extraction combined with high-pressure homogenization pretreatment process at 1200 psi and 35°C. Algal lipid yield was about 24.9% through this process, whereas only 19.8% lipid can be obtained by following a conventional lipid extraction procedure using the solvent, chloroform : methanol (2 : 1, v/v). Present approach requires 30 min process time and a moderate working temperature of 35°C as compared to the conventional extraction method which usually requires >5 hrs and 65°C temperature. It was found that this combined extraction process followed second-order reaction kinetics, which means most of the cellular lipids were extracted during initial periods of extraction, mostly within 30 min. In contrast, during the conventional extraction process, the cellular lipids were slowly and continuously extracted for >5 hrs by following first-order kinetics. Confocal and scanning electron microscopy revealed altered texture of algal biomass pretreated with high-pressure homogenization. These results clearly demonstrate that the Folch method coupled with high-pressure homogenization pretreatment can easily destruct the rigid cell walls of microalgae and release the intact lipids, with minimized extraction time and temperature, both of which are essential for maintaining good quality of the lipids for biodiesel production.