Some nitrosoamino acids in bacon adipose tissue and their contribution to the total N-nitroso compound concentration

Zusammenfassung Der Gehalt an Nitrosoaminosäuren und ihr Anteil an den Gesamt-N-Nitrosoverbindungen im Fettgewebe von rohem Schinkenspeck wurde untersucht. In 80% der Proben wurde ein Gehalt an Nitrosoaminosäuren von bis zu 0,2 mg/kg festgestellt. Scheinbare Gesamt-N-Nitrosoverbindungen waren in allen Proben in einer Konzentration von 0,4 bis 3,7 mg (N-NO)/kg vorhanden. Die eigentlichen Nitrosoaminosäuren machen nur etwa 1% der scheinbaren Gesamt-N-Nitrosoverbindungen aus. Die Mehrzahl der scheinbaren Gesamt-N-Nitrosoverbindungen im Fettgewebe findet sich im nichtlöslichen Bindegewebe, wobei der verbleibende Rest chloroformlöslich ist.Summary The levels of nitrosoamino acids and apparent total N-nitroso compounds in the adipose tissue of raw bacon have been examined. Nitrosoamino acids were detected in 80% of the samples in levels of up to 0.2 mg/kg. Apparent total N-nitroso compounds were present in all samples in concentrations of between 0.4 and 3.7 mg (N-NO)/kg. Nitrosoamino acids accounted for approximately only 1 % of the apparent total N-nitroso compounds. The majority of the apparent total N-nitroso compounds in the adipose tissue were associated with the insoluble connective tissue fraction, with the remainder present as chloroform soluble species.

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Einige Nitrosoaminosäuren im fettgewebe von Schinkenspeck und ihr anteil an der Konzentration der Gesamt-N-Nitrosoverbindungen

     

Volatile, non-volatile and total N-nitroso compounds in bacon.

Twenty-five smoked and unsmoked fried bacon samples have been analysed by a group selective procedure to measure the concentration of apparent total N-nitroso compounds (ATNC). The levels of a range of individual N-nitroso compounds, including simple volatile N-nitrosamines, N-nitrosothiazolidines, N-nitrosamino acids and N-nitrosothiazolidine carboxylic acids have also been examined. Concentrations of ATNC varied from 430 to 6800 micrograms(N-NO)/kg with a mean of 2700 micrograms(N-NO)/kg. Protein-bound N-nitrosoproline was the most abundant compound detected in unsmoked bacon, mean 260 micrograms/kg, and on average accounted for 4% of the ATNC concentration. For smoked bacon, bound N-nitrosoproline was detected in levels of up to 890 micrograms/kg and contributed 5% to the ATNC total. The most abundant compound present in smoked bacon was N-nitrosothiazolidine-4-carboxylic acid, mean 660 micrograms/kg, and this accounted for 6% of the ATNC. N-Nitrosothiazolidine, mean 340 micrograms/kg, and 2-(hydroxymethyl)-3-nitrosothiazolidine-4-carboxylic acid, mean 180 micrograms/kg, were the next most prominent compounds detected in smoked bacon. The combined sum of all the individual N-nitroso compounds measured accounted for, on average, 16% of the total ATNC. The identities of the N-nitroso compounds comprising the majority of the ATNC in bacon remain unknown.     

An investigation of the levels of N-nitrosodimethylamine, apparent total N-nitroso compounds and nitrate in beer

Over 170 retail samples of beer have been analysed for N-nitrosodimethylamine (NDMA), apparent total N-nitroso compounds (ATNC) and nitrate. Levels of NDMA ranged from below 0.1 up to 1.2 micrograms/kg with a mean of 0.2 micrograms/kg. ATNC was detected in 42% of the samples in concentrations of up to 569 micrograms (N-NO)/kg. The levels of nitrate ranged from less than 0.2 up to 143 mg/kg with a mean of 16.8 mg/kg. There was no correlation between the amounts of NDMA and ATNC found in the retail beers. Samples taken during the course of fermentation showed that NDMA was unaffected by the bacterial reduction of nitrate which causes ATNC formation. HPLC studies using a photolysis/chemiluminescence detector revealed that the ATNC in beer are highly polar species of as yet unknown identity.     

Characterization studies on insoluble total N-nitroso compounds in bacon adipose connective tissue

Enzymic hydrolysis was employed to solubilize 35% of the total N-nitroso compounds associated with bacon adipose connective tissue. Size exclusion chromatography of the digest showed that 95% of the solubilized N-nitroso compounds had molecular weights equivalent to those of di-, tri- and tetrapeptides. The likely identity of these compounds is discussed in the light of their extractability from acidified solution into ethyl acetate and their thermal and pH stability.     

绵羊、牛和猪脂肪组织中挥发性脂肪酸的组成分析

为了研究不同畜种之间脂肪组织中挥发性脂肪酸的组成成分,利用同时蒸馏萃取技术和气相色谱技术对猪、牛和羊脂肪组织中的挥发性脂肪酸进行了测定.结果表明,绵羊、牛和猪脂肪组织中总挥发性脂肪酸分别为600.512、456.029、202.371 mg/kg.在猪脂肪组织中没有检测到支链脂肪酸;在牛脂肪组织中检测到了4-甲基辛酸,质量比为7.508 mg/kg,远小于羊脂肪组织中4-甲基辛酸的含量,其质量比为28.119 mg/kg;在羊脂肪组织中,检测到种类较为丰富的挥发性脂肪酸,其中对羊肉膻味起主要作用的4-甲基辛酸,在羊脂肪组织中有较高的含量,这一特征与猪和牛的挥发性脂肪酸形成显著的区别.     

The application of a chemical denitrosation and chemiluminescence detection procedure for estimation of the apparent concentration of total N-nitroso compounds in foods and beverages

The apparent total N-nitroso content of foods can be measured by a procedure based on chemical denitrosation and chemiluminescent detection of the eliminated nitric oxide. Procedures have been established which substantially reduce the 'apparatus blank' response to the denitrosating agent and allow total nitroso contents down to 10 micrograms (N-NO)/kg to be measured reproducibly on a 1-g sample. Typically, duplicate analyses of samples containing 10-1000 micrograms (N-NO)/kg differ by less than 15% of their mean. Potentially the method can be subject to some interference from compounds other than N-nitroso compounds, but at least in some commodities these interfering compounds do not exist in measurable amounts.     

Examination of some fermented foods for the presence of apparent total N-nitroso compounds

The fermented foods yoghurt, bread and cheese were analysed for the presence of apparent total N-nitroso compounds (ATNC) by a group-selective procedure involving direct chemical denitrosation and chemiluminescence detection of the released nitric oxide. The levels of ATNC were below the 20 micrograms(N-NO)/kg detection limit in all 20 yoghurts, 23 of the 24 bread samples and 28 out of 31 different varieties of cheese analysed. ATNC were detected in most of those cheese samples manufactured with added nitrate, including Edam, Gouda and Havarti, in concentrations ranging from 30 to 210 micrograms(N-NO)/kg.     

AN INVESTIGATION INTO THE APPARENT TOTAL N-NITROSO COMPOUNDS IN MALTS

The levels of apparent total N-nitroso compounds in a range of UK malts have been investigated, and the effects of various kilning procedures are reported. Concentrations of N-nitrosodimethylamine and N-nitrosoproline have been determined for selected malts kilned with and without sulphuring. Low or non-detectable levels of apparent total N-nitroso compounds are consistently found only in malts kilned under low NOX conditions. Sulphuring appreciably reduces N-nitrosodimethylamine but has little effect on production of the majority of non-volatile N-nitroso compounds.     

The effect of castration on the headspace aroma components of cooked lamb

Minced lamb samples (lean meat or adipose tissue) from 12-week-old ram lambs and wethers were fried in the absence of added fat. The headspace aroma components were isolated onto Tenax GC prior to thermal desorption. A total of 132 compounds were identified and a further 31 were partially characterised by GC-MS, 49 of which are reported here for the first time as components of sheep meat aroma. One of these, 4,6-dimethyl-1,3-oxathiane, which has not previously been reported in any meat, was associated with a ‘stale/wet animal’ odour. All aroma isolates were dominated by products of lipid degradation with aliphatic aldehydes present in the highest relative amounts. Certain branched-chain fatty acids have been implicated in the characteristic aroma of sheep meat, and 4-methyloctanoic and 4-methylnonanoic acids were identified at higher levels in the adipose tissue from entires than from wethers.     

Surveillance of preservatives and their interactions in foodstuffs.

The organization of food surveillance in the UK is described, in particular as it has been applied to preservatives and their interaction products in foods. Applications of nitrates and nitrites as preservatives are discussed, together with the consequential exposure of consumers to these anions and their reaction products. Analytical methods for the determination of volatile and non-volatile N-nitroso compounds are referred to in relation to the results in food surveillance studies. Concentrations of Apparent Total N-nitroso Compounds (ATNC) averaged 2900 micrograms(N-NO)/kg in fried smoked bacon compared with 2400 micrograms/kg in fried unsmoked bacon; of this, known volatile and non-volatile N-nitroso compounds accounted for only 10-20%. ATNC were not detected in cheeses except those manufactured with added nitrate when ATNC levels up to 210 micrograms(N-NO)/kg were detected. Further studies are needed to determine the identity and toxicological properties of the non-volatile N-nitroso compounds.     

Nitrate, nitrite, and N-nitroso compounds in Finnish foods and the estimation of the dietary intakes

An estimate of the dietary intakes of nitrite, nitrate and N-nitroso compounds is presented, based on the analytical data supplied by the Finnish Food Quality Control. Figures on the food consumption of the Finnish population, taken from a national dietary survey, and food consumption of 1768 children and adolescents over a 48-h recall period were used. The mean daily dietary nitrate intakes were estimated to be about 55 mg for the total population and for children and adolescents. The mean nitrite intakes were 1.88 mg for the total population and 1.07 mg for children and adolescents. The intake of N-nitroso compounds (NDMA) was estimated to be 0.08 micrograms for the total population and 0.02 micrograms for children. Nitrates were found to originate mainly from vegetables (80%), nitrites from meat products (97%) and nitrosamines mostly from fish products and beer. A comparison of the estimate of dietary intake of with ADI values indicated that the nitrite intake of the total population was 23% and by children 39% of ADI. The average weight was approximately 60 kg for adults and 20 kg for children. When measured average weight (39 kg) was used, and the nitrite intake was found to be 28% of ADI. Nitrate intakes from food additives were 2.5% and 1.4% of the ADI value, respectively. When the total nitrate intake from various food sources was compared with the ADI (which is given only for food additives), the estimated nitrate intake of the total population was 25% and that of children 28% of the ADI value.     

Composition of subcutaneous adipose tissue of dry-cured pork forelegs as affected by desalting and boiling: The effects of vacuum-packaging

The effects of desalting and boiling, with or without vacuum-packaging, on the composition of the subcutaneous adipose tissue of dry-cured pork forelegs were investigated. The adipose tissue contained 7.1% water and 91.0% lipids. The main fatty acids in glycerides (about 89% of total lipids) were oleic (39.6%), palmitic (23.9%), linoleic (13.7%) and stearic (12.6%) acids. The main fatty acids in free fatty acids (about 11% of total lipids) were oleic (36.0%), linoleic (28.4%) and palmitic (13.1%). The culinary treatment (desalting and boiling) caused significant decreases in dry matter and lipid contents. These decreases were not as great in vacuum-packaged samples because the lipid losses were prevented during the boiling stage. No significant changes in lipid fraction proportions (glycerides and free fatty acids) were observed. The samples boiled with vacuum-packaging showed higher contents of polyunsaturated fatty acids than did the samples boiled without vacuum-packaging.     

Distribution of volatile branched-chain fatty acids in various lamb tissues

Volatile fatty acids (C4–C11) including even-, odd-, and branched-chain members in lamb tissues were quantitatively analyzed. Volatile branched-chain fatty acids (BCFA) were more concentrated in subcutaneous adipose tissue samples (rump, shoulder, breast) than in perinepheric adipose or muscle tissues. Perinepheric adipose tissue contained relatively high quantities of n-chain, even-numbered fatty acids and very low levels of BCFA. Greater variation existed in fatty acid profiles among similar subcutaneous adipose tissues from different lambs than between samples of adipose tissue from different carcass sites from a given lamb sample. 4-Methyl- and 4-ethyloctanoic acids were present at concentrations greatly above threshold levels in all lamb fats tested, and thus upon hydrolysis would contribute species-related flavors to lamb. 4-Methylnonanoic concentrations in lamb fats ranged from nondetectable to greater than the threshold level, and therefore this compound would not always contribute to the species-related flavors of lamb. Lean meat samples contained very low concentrations of 4-methyl- and 4-ethyloctanoic acids.     

Review of methodologies for the determination of nonvolatile N-nitroso compounds in foods

An attempt has been made to briefly review methods available for the determination of total N-nitroso compounds, N-nitrosamides, N-nitrosamino acids, and miscellaneous other nonvolatile N-nitroso compounds in foods and beverages, giving special emphasis to the progress made during the past five years. It appears that a wide variety of acceptable methods are available for N-nitrosamino acids and simple hydroxylated N-nitrosamines, but none of them has yet been adequately validated. Only limited progress has been made for the analysis of N-nitrosamides, N-nitroso sugar amino acids, and other N-nitroso compounds. Various mass spectrometric methods available for the determination and confirmation of these compounds are also discussed.     

A method for the investigation of free and protein-bound N-nitrosoproline in beer

Hydrolysis of beer peptides and proteins by both mild enzymatic and vigorous alkaline conditions has established that peptide-bound N-nitrosoproline residues are not present in beer in significant quantities. Free N-nitrosoproline remains the single most abundant identified N-nitroso compound, and this accounts for up to 10% of the apparent total N-nitroso compounds (ATNC). Both hydrolysis and extractability studies strongly indicate that the majority of the ATNC are not associated with beer peptides, but rather are likely to be low molecular weight possessing in some cases an acidic functional group.     

Direct transesterification of total fatty acids of adipose tissue, and of freeze-dried muscle and liver with boron-trifluoride in methanol

The purpose of this study was to demonstrate direct transesterification of fatty acids of tissue total lipids with boron-trifluoride in methanol. Adipose tissue, muscle, and liver samples obtained from market-weight lambs, were transesterified with 14% boron-trifluoride in methanol at 80 °C. Total lipids of additional samples were first solvent-extracted and then transesterified with 14% boron-trifluoride in methanol. Thin layer Chromatographic evaluation revealed quantitative conversion of triacylglycerol- and polar lipid-fatty acids to fatty acid methyl esters using direct transesterification. No differences in fatty acid composition were observed for adipose tissue (p > 0.05); however, for muscle, 20:4 was greater (p = 0.01) with direct transesterification. For liver, greater concentrations of fatty acids were observed for the direct transesterification method (p < 0.02). Weight percentages of adipose and muscle fatty acids were unchanged from 15 to 180min of direct transesterification; however, 120 and 90 min of direct transesterification were required for adipose tissue and muscle, respectively, for complete transesterification. It was concluded that for analysis of total fatty acids, one-step direct transesterification with boron-trifluoride in methanol is a useful and acceptable procedure.     

DEVELOPMENT OF METHODS FOR THE CHARACTERISATION OF NON-VOLATILE N-NITROSO COMPOUNDS IN MALT

The effect of pH on extractability and the chromatographic separation of non-volatile N-nitroso compounds in aqueous wort extracts of malt have been studied. The apparent total N-nitroso compounds (ATNC) contain acidic functional groups and consist of a large number of minor components.     

HPLC-DAD Analysis to Identify the Phenolic Profile of Rhododendron Honeys Collected from Different Regions in Turkey

In this study, phenolic compounds of Rhododendron honey (also known as mad honey) samples collected from the Black Sea Region were identified using high performance liquid chromatography-diode array detector system. The major phenolic substances in Rhododendron honeys were found to be chlorogenic and coumaric acids with the amounts of 0.11–191.54 mg/kg and 0–82.83 mg/kg, respectively. Gallic and ferulic acids were detected in the most honey samples. Additionally, significant correlations were determined between the phenolic substances. The present study showed that Rhododendron honeys contained higher quantities of phenolic acids than flavonoids. Chlorogenic and coumaric acids were the dominant phenolic substances detected in honey samples.     

Dietary risk assessment of organophosphorus and dithiocarbamate pesticides in a total diet study at a Brazilian university restaurant

In this study ready-to-eat food samples were collected in the production line of the university restaurant of the University of Brasilia, Brazil, which serves non-vegetarian and vegetarian meals daily. Samples were analysed for the presence of ten organophosphorus insecticides (OPs) by GC/FPD, after extraction with ethyl acetate and anhydrous sodium sulfate (LOQ = 0.002 mg kg(-1)), and for dithiocarbamate fungicides, as CS(2), using the spectrophotometric method (LOQ = 0.05 mg kg(-1)). About 43% of the 175 samples analysed contained at least one OP compound at levels up to 1.83 mg kg(-1). Methamidophos was the compound most detected (37.7%), present in most of the soup, soybean and salad samples. No OP residues were found in fruit juice, beans and bran rice samples. The cumulative acute intake of OPs was estimated using methamidophos and acephate as index compounds (IC). The total cumulative intake represented 9.1% and 47.7% of the methamidophos ARfD for the non-vegetarian and vegetarian diets, respectively. When acephate was used as IC, the total intakes represented 20.7% and 116% of the ARfD for the non-vegetarian and vegetarian diets, respectively. Dithiocarbamates were detected in 70% of the 177 samples analysed, at levels up to 0.51 mg kg(-1) CS(2); all salad samples were positive and no residues were found in fruit juice. The chronic intake of dithiocarbamates represented 8.6 and 8.9% of the ADI (mancozeb) for the vegetarian and non vegetarian diets, respectively.     

Subcellular Distribution and Composition of Lipids in Muscle and Adipose Tissues

Subcellular distribution of fatty acids in the membranes and cytoplasm of muscle and adipose tissues from USDA Select beef carcasses was determined. Cholesterol in subcellular fractions of intramuscular adipose tissue also was quantified. There was a greater percentage of saturated fatty acids in cytoplasmic, than in membrane, fractions of muscle. With the exception of intramuscular adipose tissue, monounsaturated fatty acids constituted a greater percentage of the total lipid in the cytoplasm than in membranes. There was a greater percentage of polyunsaturated fatty acids in the membrane, than in the cytoplasmic, fraction. Cholesterol averaged 118 mg per 100 g intramuscular adipose tissue with 54% in the cytoplasm and 46% in the membranes.