Cholesterol and Its Oxidation Derivatives Content in Market Dairy Products.

Cholesterol oxidation products (COPs) are contaminants of food of animal origin. Increased levels of these compounds in the human body are associated with an increased risk of many non-communicable diseases. Dairy products are mentioned among the main sources of these compounds in the diet. The objective of this study was to evaluate the contents of cholesterol and its oxidized derivatives in eleven groups of dairy products, willingly consumed in European countries. The levels of COPs were determined by applying the GC-TOF/MS method. In the tested products, cholesterol and its oxidation derivatives, such as 7-ketocholesterol, 7α-hydroxycholesterol, 7ß-hydroxycholesterol, 5,6ß-epoxycholesterol and 5,6α-epoxycholesterol, were determined. The studied dairy products differed in their contents and profiles of oxysterols. The highest contents of COPs were found in cheese with internal mold (13.8 ± 2.5 mg kg-1) and Cheddar (11.7 ± 3.5 mg kg-1), while the lowest levels were detected in yoghurt (0.94 ± 0.30 mg kg-1) and kefir (0.57 ± 0.11 mg kg-1). 7-ketocholesterol and 5,6ß-epoxycholesterol were the dominant oxysterols. The ratio of oxidized derivatives to total cholesterol was on average 1.7%. Our results confirmed that dairy products are an important dietary source of COPs. Their levels should be monitored in dairy products to provide the best health quality.

Factors affecting composition of fatty acids in wild-growing forest mushrooms.

The importance of mushrooms as a food source is continually increasing. To investigate how environmental factors affect the nutritional value of mushrooms, we harvested them in eastern Poland, south-central Germany, and northwestern Belgium in plots with similar environmental conditions but varying in tree species composition and richness. We used gas chromatography-mass spectrometry (GC-MS) to analyze the fatty acid (FA) content of the mushrooms. Fungal species identity explained the largest part (40%) of the total variation in FA concentration and composition. Environmental factors accounted for 1-12% of variation. The concentration of FA, especially saturated fatty acids, decreased with increasing understory cover and increasing nitrogen concentration in the topsoil. The effect of tree species richness or tree species identity was negligible. Our results suggest that the nutritional value of mushrooms depends mainly on the species identity of fungi, but that their FA content is slightly higher in forests with less undergrowth and in nitrogen-poor soils.

A Novel Method for the Determination of Squalene, Cholesterol and Their Oxidation Products in Food of Animal Origin by GC-TOF/MS.

Cholesterol present in food of animal origin is a precursor of oxysterols (COPs), whose high intake through diet can be associated with health implications. Evaluation of the content of these contaminants in food is associated with many analytical problems. This work presents a GC-TOF/MS method for the simultaneous determination of squalene, cholesterol and seven COPs (7-ketocholesterol, 7α-hydroxycholesterol, 7ß-hydroxycholesterol, 25-hydroxycholesterol, 5,6α-epoxycholesterol, 5,6ß-epoxycholesterol, cholestanetriol). The sample preparation procedure includes such steps as saponification, extraction and silylation. The method is characterized by high sensitivity (limit of quantification, 0.02-0.25 ng mL-1 for instrument, 30-375 µg kg of sample), repeatability (RSD 2.3-6.2%) and a wide linearity range for each tested compound. The method has been tested on eight different animal-origin products. The COP to cholesterol content ratio in most products is about 1%, but the profile of cholesterol derivatives differs widely (α = 0.01). In all the samples, 7-ketocholesterol is the dominant oxysterol, accounting for 31-67% of the total COPs level. The levels of the other COPs range between 0% and 21%. In none of the examined products are cholestanetriol and 25-hydroxycholesterol present. The amount of squalene, which potentially may inhibit the formation of COPs in food, ranges from 2 to 57 mg kg-1.

Fatty Acid Composition of Pseudocereals and Seeds Used as Functional Food Ingredients.

In recent times, the popularity of seeds, other than cereals, in the diet has systematically grown. The fat contained in these products significantly affects their energy value as well as their biological and physicochemical properties, including their susceptibility to oxidation. The objective of this study is to evaluate the fat concentration and fatty acid (FA) composition of popular non-spice seeds used in food as a substitute for cereals or a functional additive. The research material consisted of thirteen groups of seeds derived from the following plants: amaranth, blue poppy, buckwheat, chia, flax, hemp, canihua, milk thistle, pumpkin, plantago, quinoa, sesame, and sunflower. The fat contents and fatty acid profiles differed significantly between the tested products and were dependent on the plant species. In all products, polyunsaturated fatty acids (PUFAs, 40-80% of total FAs) dominated. Linoleic acid was the main FAs in most tested seeds. The exceptions were chia and flax seeds, which were characterized by very high contents of α-linolenic acid, respectively, 62.0 and 51.4% of the total FAs. The share of monounsaturated FAs (mainly oleic acid) in the total FAs content was between 6 and 40%. All tested seeds (especially flax, chia, and hemp) have favorable values for their indexes of atherogenicity and thrombogenicity as well as the hypocholesterolemic/hypercholesterolemic ratio.

The Influence of Supplementation with Zinc in Micro and Nano Forms on the Metabolism of Fatty Acids in Livers of Rats with Breast Cancer.

The aim of this study was to investigate the effect of zinc supplementation (in the form of nano or microparticles) on the profile and metabolism of fatty acids in the liver microsomes of rats with induced breast cancer. The activity of desaturases (Δ5, Δ6, Δ9) and the level of cholesterol and its oxidized derivatives were measured. The aim of this study was also to determine the effect of various forms of zinc supplements on rats that were on 5-, 12- and 15-hydroxyeicosatetraenoic (5-, 12- and 15-HETE) and hydroxyoctadecadienoic (HODE) acids, and the level of prostaglandin E2 (PGE2). Female Spraque-Dawley rats (n = 24) were divided into 2 groups that were supplemented with zinc in the micro form (342 nm) or nano form (99 nm) particles, respectively, and a group with a standard diet (control group). All animals received 7,12-dimethylbenz[a]anthracene twice for the induction of breast cancer. Dietary nano-Zn supplementation increased vaccenic acid content (p = 0.032) and decreased Δ6-desaturase activity (p = 0.006), whereas micro-Zn increased cholesterol (p = 0.006), ∑COPs (total cholesterol-oxidation products) (p = 0.019) and PGE2 (p = 0.028) content. Dietary enrichment with Zn microparticles resulted in lower concentrations of the metabolites 15-, 12- and 5-HETE and HODE. Our study indicates that the effect of zinc supplementation on the metabolism of fatty acids in the liver microsomes under neoplastic conditions depends on the form in which it is administered.

Zinc Affects Cholesterol Oxidation Products and Fatty Acids Composition in Rats' Serum.

The purpose of this work was to evaluate the effect of the nanosized or microsized zinc (Zn) particles on fatty acid profile, enzyme activity and the level of cholesterol, squalene and oxysterols in rats with breast cancer. Rats (female, n = 24) were divided into the following groups: control, and two test groups, whose diets were enriched with either Zn microparticles (342 nm) or Zn nanoparticles (99 nm). All rats were treated twice with the carcinogenic agent; 7,12-dimethylbenz[a]anthracene. In rats whose diet was enriched with zinc (especially in the form of nanoparticles), the number and sizes of tumors were lower. Diet supplementation also significantly reduced the cholesterol (p = 0.027) and COPs (cholesterol oxidation products) levels (p = 0.011) in rats serum. Enriching the diet with Zn microparticles decreased the Δ6-desaturase activity (p < 0.001). Zn influences fatty acids' profile in rats' serum as well as inhibiting desaturating enzymes. A reduced amount of pro-inflammatory arachidonic acid derivatives may be the expected effect.

Determination of Pharmaceuticals, Heavy Metals, and Oxysterols in Fish Muscle.

The present study aimed to assess the levels of 98 multi-class pharmaceuticals including cardiovascular drugs, antidepressants, hypnotics, antibiotics, and sulfonamides occurring in the muscle tissue of fish caught in the Baltic Sea. The following fish species were collected: perch (Perca fluviatilis); flounder (Platichthys flesus); turbot (Scophthalmus maximus); plaice (Pleuronectes platessa); cod (Gadus morhua callarias); bream (Abramis brama); crucian (Carassius carassius). Additionally, in the examined fish muscle the levels of heavy metals and trace elements were determined (As; Ag; Au; Ba; Cd; Co; Cr; Cu; Hg; Li; Mo; Ni; Pb; Sb; Se; Sn; Tl; V) as well as the levels of cholesterol and its 5 derivatives (7-ketocholesterol; 7α-hydroxycholesterol; 7ß-hydroxycholesterol; 5ß,6ß-epoxy-cholesterol; 5α,6α-epoxycholesterol). In the performed studies 11 out of 98 examined pharmaceuticals were detected in fish muscle. The levels of pharmaceuticals in fish muscle varied depending on the species. In the tissues of bream and crucian, no pharmaceuticals were found. Mercury, lead and arsenic were detected in the muscles of all examined fish. Based on the hazard factor for Hg, Pb, Cd, Ni (target hazard quotient, THQ < 1), it was found that the consumption of the studied fish does not constitute a health risk. However, the THQ for As remained >1 indicated possible risk from those metals. In the examined fish muscle the total cholesterol oxidation products (COPs) level of oxysterols were, respectively: 6.90 (cod) µg/g-4.18 µg/g (perch), which corresponded to 0.7-1.5% of cholesterol. The main COPs evaluated were 7-ketocholesterol (0.78 ± 0.14-1.79 ± 0.06 µg/g), 7ß-hydroxycholesterol (0.50 ± 0.04-3.20 ± 2.95 µg/g) and 5ß,6ß-epoxycholesterol (0.66 ± 0.03-1.53 ± 0.66 µg/g). The assessment of health hazards due to contaminations is necessary, which may help to introduce national legislation and global standards aimed at reducing or even eliminating the exposure to contaminants.

Giblets and abdominal fat of pomegranate seed oil fed chickens as a source of bioactive fatty acids.

The aim of present study was to determine whether chickens' (broilers Ross 308, n = 180, sex ratio 1:1) diet modification with different doses of grape or pomegranate seed oil will favourable change fatty acids and cholesterol content in selected giblets (liver and heart) or wastes (adipose tissue). It was also verify whether generated changes would make the giblets and wastes more valuable as dietary components or by-products for food industry. From 22 to 42 day of life, five diets were administered to chickens. Control grower diet (CON) contained 5% of soy oil, whereas in the experimental grower diets part of soy oil (1.5% or 2%) was replaced with specific amount of grape or pomegranate seed oil (GRAP 1.5; GRAP 2.0; POM 1.5; POM 2.0 respectively). Fatty acids and cholesterol content were determined with gas chromatography with flame-ionization detection. Pomegranate seed oil improved fatty acids profile more favourably than grape seed oil, which makes it a valuable additive in chickens' feeding. Abdominal fat of pomegranate seed oil supplemented chickens appeared to be the richest sources of rumenic acid and n3 polyunsaturated fatty acids, which allows to suggest its use in manufacturing of meat products to obtain foodstuffs rich in those essential nutrients. In principal component analysis (PCA), two principal components: PC1 and PC2, which were enough to explain 29.91% of variance of initial variables, allowed to a good separation of chickens fed with both doses of pomegranate seed oil from animals from control and grape seed oil fed groups. Because poultry addresses all nutritional, institutional and consumer requirements, enrichment of giblets in rumenic acid by pomegranate seed oil incorporation into chickens' diet may provide a valuable dietary source of bioactive fatty acids for consumers, especially of low-income countries.

Evaluation of 5-hydroxymethylfurfural content in market milk products.

5-Hydroxymethylfurfural (5-HMF) is a cyclic aldehyde formed during non-enzymatic browning reactions. In milk, it is, on the one hand, a contaminant, on the other, a quality indicator. The objective of this study was to evaluate the content and selected factors affecting the concentration of 5-HMF in cows' milk and first infant milk. The content of 5-HMF in market milk varied widely, the average was 54.8 µg L-1 (3.3-136.8). There was no significant difference in the concentration of this contaminant between UHT, HTST pasteurised and micro-filtered milk. It was also shown that the content of lipid components did not affect the concentration and kinetics of the 5-HMF formation in milk subjected to thermal treatment. Lactose-free milk was characterised by a level of this compound much higher than regular products. The 5-HMF average contents in powder cows' milk and first infant milk were respectively 642 and 2315 µg kg-1.

Chemometric Analysis of Fatty Acids Profile of Ripening Chesses.

The number of different types of cheese worldwide exceeds 4000 and dairy fat, composed of about 400 fatty acids (FA), is one of the most complex dietary fats. Cheeses are valuable sources of different bioactive FA, i.e., conjugated FA (CFA). The aim of present study was to determine FA profile of commercially available ripening cheeses, with the special emphasis on CFA profile. Multivariate analyses (cluster analysis (CA), principal component Analysis (PCA), and linear discriminant analysis (LDA)) of chromatographic data have been proposed as an objective approach for evaluation and data interpretation. CA enabled the differentiation of ripening cheeses from fresh cheeses and processed cheeses. PCA allowed to differentiate some types of ripening cheese whereas proposed LDA model, based on 22 analyzed FA, enabled assessing cheeses type with average predictive sensitivities of 86.5%. Results of present study clearly demonstrated that FA and CFA content may not only contribute to overall nutritional characteristics of cheese but also, when coupled with chemometric techniques, may be used as chemical biomarkers for assessing the origin and/or the type of ripening cheeses and the confirmation of their authenticity, which is of utmost importance for consumers.

The Effect of Diet Supplementation with Pomegranate and Bitter Melon on Lipidomic Profile of Serum and Cancerous Tissues of Rats with Mammary Tumours.

The aim of this study was to present overall lipid profile of organisms with ongoing neoplastic process and applied diet supplementation with pomegranate seed oil (PSO) and bitter melon extract (BME). The following were quantified in serum and cancerous tissues of rats suffering from mammary tumours: fatty acids, conjugated fatty acids and sterols, their oxidised metabolites (malondialdehyde and oxysterols) and lipoxygenase (LOX) metabolites of polyunsaturated fatty acids. The obtained results indicate that abnormalities in lipid metabolism accompany neoplastic process. These differences concern all classes of lipids and most pathways of their transformation, with the special emphasis on lipid peroxidation and LOX-mediated metabolism. Cancer process appears to be so detrimental that it may conceal positive influence of dietary modifications. The lack of anticarcinogenic properties of PSO and BME in this model may be due to their antioxidant properties or elevated levels of conjugated linoleic acids (CLA), which change CLA isomer activity from anti- to pro-tumorigenic. As CLA are the product of conjugated linolenic acids (CLnA) endogenous metabolism, high CLA levels may be explained by applied diet enrichment.

Iron in red meat-friend or foe.

In developed countries, due to high content and bioavailability, red and processed meats are the main sources of iron in the diet. Adequate intake of this nutrient is essential for the proper development and functioning of the human body, and its deficiencies are associated mainly with the occurrence of anemia, which is one of the most widespread nutritional problems in the world. However, excessive intake of iron can be detrimental to health. Studies have shown that high consumption of red meat and its products, and thereby iron, particularly in the form of heme, increases the risk of non-communicable diseases, including cancers, type II diabetes and cardiovascular disease. Due to the high nutritional value, the presence of red meat in the diet is preferable, but according to World Cancer Research Fund International its consumption should not exceed 500g per week. Furthermore, there are several potential ways to suppress the toxic effects of heme iron in the diet.

INFLUENCE OF POMEGRANATE SEED OIL AND GRAPE SEED OIL ON CHOLESTEROL CONTENT AND FATTY ACIDS PROFILE IN LIVERS OF CHICKENS.

Oils are important food ingredients, mainly as a source of unsaturated fatty acids. The offer of novel edible oils from herbs, spices and fruit seeds has grown and many of them are used as functional food and dietary supplements but also as feed additives in animal feeding. Poultry meat is recommended.in properly balanced diet and its consumption in Poland has been growing. The objective of present study was to verify if the supplementation of chickens' diet with grape seed oil or pomegranate seed oil influences cholesterol content and fatty acids (FA) profile in their livers. Ross 308 chickens (n = 24) were fed with fodder enriched with grape seed oil (G group) or pomegranate seed oil (P group). Diet of control group (C group) was based on soybean oil. FA analysis in livers as well as cholesterol content was made with gas chromatography. We observed significant increase in fat content when part of soybean oil was replaced by grape seed oil (p = 0.0002). Its highest amount was detected in G group (4.44 ± 1.53%) whereas the lowest in C group (1.73 ± 0.53%). Applied supplementation did not change total cholesterol content. Its content ranged from 233.0 ± 12.2 mg/100 g in G group to 234.6 ± 29.7 mg1100 g in C group. However, chickens' diet modification with grape seed oil and pomegranate seed oil influenced the FA profile in livers. We detected the presence of punicic acid (cis-9, trans-11, cis-13 C18:3, PA) in livers of chicken fed with pomegranate seed oil. Pomegranate seed oil is one of natural sources of conjugated linolenic acids (CLnA), which predominate in this oil (PA >70% of all FA). However, in livers PA constituted only 0.90 ± 0.10% of all fatty acids. Furthermore, we detected substantial amounts of rumenic acid (cis-9, trans-11 C18:2, RA) - the major isomer of conjugated linoleic acids (CLA). Its natural sources in diet are meat and milk of ruminants, but incorporation of pomegranate seed oil into chickens' diet caused a significant increase of its share in fatty acids pool in their livers (3.73 ? 0.79% in P group in relation to 0.08 ± 0.03% in G group and 0.02 ± 0.00% in C group, p < 0.0001). It proves that PA is effectively converted into RA in chickens organisms. Pomegranate seed oil seems to be an interesting feed additive in chicken feeding which can improve FA profile of poultry meat.

The effect of meat cuts and thermal processing on selected mineral concentration in beef from Holstein-Friesian bulls.

The impact of meat cuts (nine muscles and liver) and thermal processing on selected mineral (potassium, sodium, phosphorus, magnesium, zinc, iron, including heme form) concentration in beef from Holstein-Friesian bulls was evaluated in the present study. The mineral's content widely varied depending on the tissue type (skeletal muscles/liver, except zinc) and between the different bovine muscles. The greatest diversity between the muscles demonstrated was zinc (3.5-6.9 mg 100 g(-1)f/w) and iron (1.7-2.3 mg 100 g(-1)f/w), however, there were no significant differences in heme iron to total iron ratio (average 74%). Thermal processes conducted on longissimus dorsi muscles also significantly affected mineral concentration. Grilled, roasted and fried bovine meat was characterised by a higher content (by 6-26%) of most studied minerals (except sodium) as compared to raw meat. Sodium levels in processed meat were 16-33% lower than in raw samples.

Vitamin B12 content in raw and cooked beef.

The aim of this work was to evaluate the vitamin B12 content and profile of cobalamin bioactive forms in raw and cooked beef. The study showed that vitamin B12 distribution is unequal among beef muscles. The content of this nutrient ranges from 0.7 to 1.5 µg/100 g of tissue. In beef, three biologically active forms of vitamin B12 were determined, i.e.: methylcobalamin, adenosylcobalamin and hydroxycobalamin; the dominating form of vitamin B12 was adenosylcobalamin (68% of total cobalamins). The process of roasting and grilling had little effect on the vitamin B12 content in the final product as compared to the raw meat. The fried product was characterised by about a 32% lower content of cobalamins than in raw meat.

Application of the SPE reversed phase HPLC/MS technique to determine vitamin B12 bio-active forms in beef.

Vitamin B12 is an animal origin nutrient of a substantial importance in human diet. Its concentration in foodstuffs is low and its chemical forms are diverse, which significantly hampers its precise determination. The determination method of choice is HPLC (high performance liquid chromatography) coupled with inductively-coupled-plasma mass spectrometry (ICP-MS). The main disadvantage of this method is high instrumentation cost and complexity of handling. The aim of this work was to develop a novel approach for determination of vitamin B12 bio-active forms in beef and beef liver. The proposed method comprises the following steps: (i) vitamin B12 is cleaved off from peptides using thermal denaturation in a weakly acidic environment; (ii) sample is cleaned-up using liquid-liquid extraction and reversed phase solid phase extraction; and finally (iii) vitamin B12 is determined using HPLC and single-quadrupole mass spectrometer with ESI source. Vitamin B12 concentrations in various beef meats were in the 2.84-3.95 µg 100g(-1) range. Average B12 concentration in beef liver was 153,60 µg 100g(-1) (n = 15). Major forms of B12 present in beef meat include adenosine cobalamin (AdoCbl) and in smaller quantities hydroxycobalamin (OHCbl). Major forms of vitamin B12 present in beef liver include OHCbl (48.2%), AdoCbl (33.8%), methylocobalamin (MeCbl, 16.3%), and cyanocobalamin (CNCbl, 1.7%). Thermal treatment noticeably decreases B12 the content in meat. Depending on conditions of treatment, B12 concentrations in the 1.04-2.20 µg 100g(-1) range were found in processed meats.