Application of chromatographic and spectroscopic techniques in the evaluation of the lipid fraction of animal products

Lipolysis and oxidation of lipids in foods are the major biochemical and chemical processes that cause food quality deterioration, leading to the characteristic, unpalatable odour and flavour called rancidity. In addition to unpalatability, rancidity may give rise to toxic levels of certain compounds like aldehydes, hydroperoxides, epoxides and cholesterol oxidation products. In this PhD study chromatographic and spectroscopic techniques were employed to determine the degree of rancidity in different animal products and its relationship with technological parameters like feeding fat sources, packaging, processing and storage conditions. To achieve this goal capillary gas chromatography (CGC) was employed not only to determine the fatty acids profile but also, after solid phase extraction, the amount of free fatty acids (FFA), diglycerides (DG), sterols (cholesterol and phytosterols) and cholesterol oxidation products (COPs). To determine hydroperoxides, primary products of oxidation and quantify secondary products UV/VIS absorbance spectroscopy was applied. Most of the foods analysed in this study were meat products. In actual fact, lipid oxidation is a major deterioration reaction in meat and meat products and results in adverse changes in the colour, flavour and texture of meat. The development of rancidity has long recognized as a serious problem during meat handling, storage and processing. On a dairy product, a vegetal cream, a study of lipid fraction and development of rancidity during storage was carried out to evaluate its shelf-life and some nutritional features life saturated/unsaturated fatty acids ratio and phytosterols content. Then, according to the interest that has been growing around functional food in the last years, a new electrophoretic method was optimized and compared with HPLC to check the quality of a beehive product like royal jelly. This manuscript reports the main results obtained in the five activities briefly summarized as follows: 1) comparison between HPLC and a new electrophoretic method in the evaluation of authenticity of royal jelly; 2) study of the lipid fraction of a vegetal cream under different storage conditions; 3) study of lipid oxidation in minced beef during storage under a modified atmosphere packaging, before and after cooking; 4) evaluation of the influence of dietary fat and processing on the lipid fraction of chicken patties; 5) study of the lipid fraction of typical Italian and Spanish pork dry sausages and cured hams

Application of different analytical methods for the determination of phenolics and antioxidant activity in hawthorn (Crataegus spp.) bud and sprout herbal extracts

Hawthorn (Crataegus spp., family: Rosaceae) extracts have been used as pharmaceutical preparations owing to positive effects on cardiovascular system. The AlCl3-based official method employed for the determination of pharmacologically active compounds was compared with other techniques such as Folin-Ciocalteau method and HPLC-DAD. Antioxidant activity was determined by ABTS radical cation assay. Methods were applied on extracts from buds and sprouts collected from common hawthorn (C. monogyna Jacq., C. laevigata (Poir.) DC.) located in Northeastern Italy. Phenolic content determined by AlCl3-based method, Folin-Ciocalteau method, and HPLC-DAD was in the range 23,534-27,728, 75,284-100,616 and 57,317-58,639 mg kg-1 of dry matter (DM), respectively, in buds, and 17,280-19,330, 27,653-38,590, and 30,635-32,185 mg kg-1 DM, respectively, in sprouts. Antioxidant activity ranged from 119,864 to 174,640 and 31,484 to 52,584 mg Trolox eq. kg-1 DM in buds and sprouts, respectively. Phenolic amount and profile were significantly affected by phenological stage and sampling location. Antioxidant activity was related to flavan-3-ol and hydroxycinnamic acid amount, and to non-phenolic substances. AlCl3-based method underestimated total phenolic content owing to lack of selectivity to important phenolic classes whereas Folin-Ciocalteau method was affected by non-phenolic interfering substances. HPLC-DAD proved to be more effective in determining hawthorn phenolics

Composition of phospholipid fraction in raw chicken meat and pre-cooked chicken patties: influence of feeding fat sources and processing technology

The effect of dietary lipid sources (animal fat, AF vs. vegetable oil, VO) was investigated on the phospholipid (PL) fraction of raw chicken meat and precooked chicken patties. PL classes were determined on extracted lipids by high-performance liquid chromatography (HPLC) connected with an evaporative light-scattering detector (ELSD). The HPLC-ELSD method showed good intraday repeatability (less than 5% for different PL) and limits of detection ranging from 0.006 mg/mL (phosphatidylethanolamine) to 0.045 mg/mL (sphingomyelin). Diet did not consistently influence PL in chicken breast. A significant difference was assessed in meat batter where total PL were at levels of 349.0 and 388.2 mg/100 g of product in batter from broilers fed AF and VO diet, respectively. Cooking oil absorption caused an important decrease in PL (lower than 300 mg/100 g of product in pre-fried patties). Diet was an effective means to control the PL fatty acid profile in chicken, particularly the levels of polyunsaturated fatty acids (PUFA). In minced breast and thigh and meat batter from birds fed a VO-supplemented diet, the relative amount of PUFA was 46, 81 and 28% higher than that determined in the corresponding samples obtained from broilers fed an AF-based diet

Retention of alkylresorcinols, antioxidant activity and fatty acids following traditional hulled wheat processing

This study compared alkylresorcinols, fatty acids and antioxidant activity retention following hulled wheat processing by traditional and modern plants. Glume removal and kernel crushing were considered as characterising steps. Samples were collected from two traditional einkorn wheat bulgur processing plants in Turkey, one emmer wheat processing plant in Armenia and a modern two stage processing plant in Italy, also including pearling. Whole kernels showed higher alkylresorcinol contents (759\u20131037 mg kg 121dm) and antioxidant activity with respect to the correspondent crushed products. Following crushing a decrease of total alkylresorcinol content was observed, with lowest amounts detected in the semolina-like fractions. Pearling did not determine a significant decrease of alkylresorcinols of the pearled output. However the waste obtained from pearling, mainly composed of bran parts, showed the highest alkylresorcinol amount (1493 mg kg 121dm) and antioxidant activity. The waste from pearling resulted also the richest fraction of unsaturated alkylresorcinol homologues and unsaturated fatty acids. Among fatty acids, C18:2, C18:1 and C16 resulted the most abundant compounds in all fractions. All the plants showed similar performance, with higher alkylresorcinol decreases due to the loss of specific parts of kernel, such as germ and bran

Assessing the effect of traditional hulled wheat processing on bioactive compounds retention

Glume removal is the preliminary operation to make hulled wheats suitable to human consumption. Traditionally glumes are separated from kernels by means of stone mills, simultaneously causing kernel crushing, a loss of kernel parts and associated phytochemicals. The aim of this study was to compare bioactive compound retention of hulled wheat products obtained by traditional processing still used in Turkey and Armenia, with a more update plant located in Italy. On-plant samplings were carried out in two einkorn wheat bulgur processing plants in Turkey, and one emmer wheat processing plant, in Armenia and Italy. Whole and crushed kernels sampled at different processing stages were analysed for their phytochemical compound content. For the Italian plant, also the pearling process was considered. Whole kernels showed higher contents of lipid and phytochemical compounds than the correspondent processed fractions, with special respect for tocopherols and phytosterols. Carotenoid content was mainly affected by genotype. Pearling had a lower effect on the content of phytochemicals; however the correspondent waste fraction showed very high amounts of lipids and other compounds. All the plants showed similar retention of phytochemicals upon processing, with higher losses occurring in smaller kernel crushed fractions, mainly because of the loss of the germ

Alkylresorcinols and fatty acids in primitive wheat populations of Italian and Black sea region countries origin

Alkylresorcinols, fatty acid content, and antioxidant activity were analysed in Triticum monococcum, T. dicoccum, T. timopheevi, T. palaeo-colchicum and T. macha accessions from Turkey, Georgia, Armenia, Bulgaria and Italy, also including modern durum wheat (T. durum) and bread wheat (T. aestivum) varieties as controls, grown in a common environment. Total alkylresorcinol content ranged between 478.3 and 1153.9 mg kg 121 dm, with the highest contents detected in T. aestivum, T. macha and some T. monococcum accessions. All alkylresorcinol homologues effectively contributed at genotype discrimination. Hexaploid wheats showed higher A19 (23.3\u201327.5%) and lower A23 (14.0\u201315.9%) and A25 (4.3\u20138.1%) relative contents, with respect to other species. C18:2 (50.4\u201362.3%), C18:1 (14.5\u201325.8%) and C16 (13.3\u201316.7%) were the most abundant fatty acid compounds in all accessions. PUFA/MUFA, C18:1/C16 and C18:1/C18:2 ratios significantly differed among genotypes, with special respect to T. monococcum, T. aestivum and T. macha. Significant differences were also observed for protein and total phenolic content, and antioxidant activity. The data indicate that some genotypes could represent interesting sources of specific compounds, with a potential use either as promising material for the selection of improved varieties or raw materials to manufacture foods with naturally enhanced content of health promoting compounds

Comparison of two extraction methods (high pressure extraction vs. maceration) for the total and relative amount of hydrophilic and lipophilic organosulfur compounds in garlic cloves and stems. An application to the Italian ecotype “Aglio Rosso di Sulmona” (Sulmona Red Garlic)

Garlic-based extracts have been surveyed as healthy promoting supplements in relation to their content of organosulfur compounds. The present study investigated the effect of high pressure extraction and maceration, and four extraction solvents (three hydroalcoholic mixtures and sunflower oil) on the total and relative amounts of the main organosulfur compounds of clove and stem extracts of the Italian ecotype \u201cAglio Rosso di Sulmona\u201d (Sulmona Red Garlic). Organosulfur compounds were more abundant in cloves than in stem extracts. High pressure extraction led to higher recoveries of hydrophilic compounds than maceration whereas the highest amounts of lipophilic compounds were detected in macerates. A higher alliin-to-allicin conversion ratio was detected in clove, in comparison to stem extracts. Sunflower oil fostered allicin decay to compounds such as ajoenes, vinyldithiins and sulfides. The use of specific extraction methods may lead to standardised grade extracts, both from native raw materials and by-products

The impact of sesquiterpene lactones and phenolics on sensory attributes: An investigation of a curly endive and escarole germplasm collection

In the present study, curly endive (Cichorium endivia L. var. crispum) and escarole (Cichorium endivia L. var. latifolium) accessions were investigated for their sensory characters (bitterness, astringency and herbaceous flavour) and acceptance in relation to sesquiterpene lactone and phenolic content. Different facets of the perception of these sensory traits in relation to lactones and phenolics were brought out. Lactucopicrin and kaempferol malonyl glucoside were consistently related to bitterness, astringency and herbaceous flavour perceptions. Overall acceptance was significantly and inversely related mainly to bitterness. The generic statement that sesquiterpene lactones and phenolic compounds are determinants of bitterness and other related sensory characters does not seem to be fully consistent with our data, that indicated how the balance of different compounds affects these traits individually, in a rather complex manner, with a prevailing negative impact of phenolics. Bitter, astringent, and herbaceous perceptions were significantly affected by variety, with curly endive showing on average higher scores in comparison to escarole, with particular respect to bitterness

Variation of sesquiterpene lactones and phenolics in chicory and endive germplasm

Fresh edible parts of 32 endive (var. crispum and latifolium) and 64 chicory accessions (Head radicchio, Sugarloaf, Leafy radicchio, and Witloof types) were analyzed for their sesquiterpene lactone (SL) and phenolic concentrations, and HPLC profile. Six SL and 20 phenolic compounds, belonging to hydroxycinnamic acids, flavonoids, and anthocyanins, were quantified. Total SL were in the range of 128\u20132045 and 383\u20132497 mg kg 121 d.m. for endive and chicory, respectively, whereas total phenolics ranged from 2207 to 15,235 and from 1356 to 77,907 mg kg 121 d.m. in the two species. An ample variability and significant differences were detected between endive and chicory and among variety and type, within species. On average, the highest SL concentration was verified in Sugarloaf chicory, whereas red Head radicchio chicories showed the highest phenolic concentration. The two species were clearly separated on the basis of discriminant analysis, with Witloof chicory showing some similar features to endive

Donation After Circulatory Death: When Withdrawing Life-Sustaining Treatments Is Ethically Acceptable

The possibility to determine death based on cardiocirculatory criteria in controlled cases, namely when there is a request to withhold treatment\u2014or, more frequently, withdraw it\u2014specifically recalls the recent Italian law on advance treatment directives and leaves the following question unanswered: Under what conditions is the patient's request legally and ethically acceptable? We present three ethical proportionality criteria for supporting physicians\u2019 decision-making facing patients\u2019 requests of treatment withdrawal, namely: 1. irreversible pathology with an ominous and worsening prognosis; 2. within an evaluation considering both clinical data and the patient's history; and 3. facing burdens that are no longer bearable. We finally argue that reflection over controlled donor may be a model for giving medicine the chance to responsibly deal with broader end-of-life issues