A procedure for the determination of lead in green vegetables at concentrations down to 1 microgram/kg.

An accurate and highly sensitive dry ashing/graphite furnace atomic absorption spectrometry (GFAAS) procedure has been developed for the measurement of lead in green vegetables. The accuracy of the method was established by analysis of standard reference materials and confirmed by isotope dilution using an inductively coupled plasma-mass spectrometer (ICP-MS). The limit of detection (wet weight) of the technique is 1-3 micrograms/kg with relative standard deviations ranging from 6% for a sample of kale containing 500 micrograms/kg to 20% for cabbage containing 3.0 micrograms/kg.

An investigation of apparent total N-nitroso compounds in beer.

The concentration of apparent total N-nitroso compounds (ATNC) in beer has been investigated using a group-selective procedure based on chemical denitrosation with hydrogen bromide and chemiluminescence detection of the released nitric oxide. In a survey of samples of 40 brands of beer and lager, detectable levels of ATNC were present in 17 samples at concentrations of 20-100 micrograms N-NO/kg in 11 and 100-500 micrograms N-NO/kg in six. To determine the origin of ATNC in beer the production of a commercial batch was examined in detail. ATNC levels were below the detection limit in the sweet wort (aqueous extract of malt), bitter wort (malt extract boiled with hops) and also at the start of fermentation, but during the course of fermentation the concentration of ATNC increased appreciably and that of inorganic nitrate decreased; detectable, though transitory, levels of inorganic nitrite were observed. None of the brewing ingredients contained sufficiently high enough levels of ATNC to account for the concentration of these compounds present in the beer after fermentation. These findings suggest that the presence of detectable levels of ATNC in some beers is a result of N-nitrosation reactions occurring in the fermenting wort with the nitrosating species derived from reduction of nitrate, due probably to the presence of microbial species with nitrate reductase activity.

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.

N-Nitrosopyrrolidine formation in bacon.

Evidence is presented that the likely major amine precursors to N-nitrosopyrrolidine in cooked bacon are free proline in the adipose tissue and to a lesser extent, collagenous connective tissues. A model system in which animal lipid is replaced with hydrogenated vegetable oil is described and used in investigating the potential role of prolyl derivatives as precursors to N-nitrosopyrrolidine.

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

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.

Comparison of trace element solubility from food items treated separately and in combination.

Solubility of trace elements from various foods (beef, soya, crab, wholemeal and white bread) is assessed with various extractants, at different pH's and after enzyme treatment. Simple extractants do not solubilise trace elements to the same degree as when they are released after digestive enzyme action. Enzymic action can change the nature of the soluble species as well as their amount. Trace element solubility from one food can be affected markedly by the presence of another food, e.g. soya enhances the solubility of iron from beef but suppresses zinc solubility. Trace elements that are bound to the insoluble residue after enzymolysis are soluble in saline solution on subsequent extraction in only isolated instances.

Trace element solubility from food following enzymolysis.

Food items were treated with enzymes simulating gastric and intestinal digestive juices and the amounts of soluble lead, cadmium, zinc, iron, and copper, were determined. Enzyme treatment was conducted in two stages involving (I) pepsin at pH 2,5 followed by (II) pancreatin and amylase at neutral pH. Solubility was determined after each stage and additionally after post enzymolysis acidification. The foods examined comprised wholemeal bread, spinach, canned tomato, ox liver, pig kidney, canned crabmeat, beefburger, and canned corned beef, the last sampled from within the bulk and adjacent to the side seam of the can. Analyte solubility varied with (I) different food items, (II) processing or preparation of foods of similar origin, (III) action of pepsin or pancreatic enzymes and (IV) pH. Reasons for the variations are discussed.

Synthesis of three N-nitroso dipeptides N-terminal in proline and a method for their determination in food.

The terminal N-nitroso derivatives of L-prolyl-L-alanine, L-prolylglycine and L-prolyl-4-hydroxy-L-proline have been synthesized and characterized. A procedure suitable for their analysis in biological samples has been developed and is based on the preparation of an aqueous extract, extraction into an organic solvent, separation by high-performance liquid chromatography and detection by a chemiluminescence method. The average recovery in all cases was above 69% at the level of 200 micrograms/kg using cured meat as a food matrix; the detection limit was 10 micrograms/kg.

Total N-nitroso group analysis of foods. II. Further studies on the precision and sensitivity of the assay.

The total N-nitroso content of foods can be measured by chemical denitrosation with hydrogen bromide and chemiluminescence detection of the cleaved nitric oxide radical. The denitrosation reagent itself causes a significant detector response which has limited the application of the technique to trace analysis. A procedure is described in which the errors associated with this interference are minimized. Application of this method to the trace analysis of aqueous and solid samples is reported together with an investigation of the effects of sample size on the accuracy and sensitivity of the assay as applied to aqueous analytes. The magnitude and significance of the false-positive response from nitrate is discussed in relation to the analysis of cured meats.

Incidence of some non-volatile N-nitroso compounds in cured meats.

Procedures are described for the use of high-performance liquid chromatography and gas chromatography with a chemiluminescence detector in the analysis of N-nitrosoamino acids, N-nitrosothiazolidine-4-carboxylic acid, N-nitroso-oxazolidine-4-carboxylic acids and N-nitroso dipeptides N-terminal in N-nitrosoproline in cured meat products. The detection limit is around 5-10 micrograms/kg. Evidence is presented for the presence of all these species except the N-nitrosated dipeptides.

N-nitroso compounds and polycyclic aromatic hydrocarbons in Icelandic smoked cured mutton.

Five samples of Icelandic smoked cured mutton were analysed for volatile N-nitrosamines, N- nitrosoamino acids, total N-nitroso content and several polycyclic aromatic hydrocarbons. Levels varied considerably from sample to sample but generally the levels of polycyclic aromatic hydrocarbons and N-nitrosamines (as judged by N- nitrosoproline , N- nitrosohydroxyproline and total N-nitroso content) varied in the same way. It is suggested that in this product the smoking procedure is responsible for both groups of contaminants and that the curing process is relatively unimportant.

Comparison of a capillary gas chromatographic and a high-performance liquid chromatographic method of analysis for polycyclic aromatic hydrocarbons in food.

Five food samples have been analysed for a number of polycyclic aromatic hydrocarbons (PAHs) by capillary gas chromatography with flame ionization detection (GC-FID) and high-performance liquid chromatography (HPLC) with fluorescence detection. The methods give similar results and there is no significant difference in their repeatability. The capillary GC method is favoured where it is desirable to analyse for a large number of PAHs, whereas the HPLC method is preferred for the individual analysis of a smaller number of PAH isomers.

Estimation of nitropolycyclic aromatic hydrocarbons in foods.

A method is described for the sample clean-up and estimation of nitropolycyclic aromatic hydrocarbons (nitro-PAH) in foods. The analysis involves the novel use of a coupled capillary gas chromatograph/thermal energy analyser and provides a detection limit for 1-nitropyrene of 12 pg (equivalent to 0.02 micrograms/kg for a 50-g sample). Only 3 out of 24 samples contained detectable quantities of nitro-PAHs; these were in the range 0.2-2.0 micrograms/kg. In contrast to the widespread occurrence of the parent PAHs in foodstuffs, the presence of nitro-derivatives appears to be minimal.

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.

N-nitrosamine analysis in foods: N-nitrosoamino acids by high-performance liquid chromatography/thermal energy analysis and total N-nitroso compounds by chemical denitrosation/thermal energy analysis.

The total N-nitroso content of foods can be measured by chemical denitrosation and chemiluminescent detection of the eliminated nitric oxide. Appropriate procedures substantially reduce the 'system response' to the denitrosating agent, so that N-nitroso group contents down to 10 micrograms/kg can be measured on a one-gram sample. Using N-nitrosamine standards added to beer, the coefficients of variation are approximately 10% and 5% at N-nitroso contents of 19 and 94 micrograms/kg, respectively. In cured meats, the coefficient of variation for unidentified N-nitroso compounds is 26% for a 0.3-g sample containing 600 micrograms/kg. Some interference from non-nitroso compounds is possible, but, in some commodities at least, these interfering compounds are not detectable. Conditions have been established that allow measurement of N-nitrosoamino acids in foods using a high-pressure liquid chromatograph interfaced to a Thermal Energy Analyzer, without the need for prior derivatization. After extraction of lipids with hexane, nitrosoamino acids are extracted with ethyl acetate and subjected to appropriate clean-up stages prior to high pressure liquid chromatography on Microbondapak CN with a hexane:ethanol:acetic acid mobile phase and Thermal Energy Analyzer detection. Recoveries from cured meat are in the 55-75% range for N-nitrososarcosine, N-nitrosoproline and N-nitrosohydroxyproline; elution is complete within seven minutes.

Analysis of polycyclic aromatic hydrocarbons in UK total diets.

Analysis of UK total-diet samples for polycyclic aromatic hydrocarbons was carried out using a simplified sample clean-up and a high-performance liquid chromatography dual fluorescence detector system. The results indicate that cereals and oils/fats contribute the major part (approximately one third each) of the polycyclic aromatic hydrocarbons in these total diets. Fruit, sugars and vegetables provide much of the remainder (approximately one quarter) while meat, fish, milk and beverages make relatively minor contributions. These results are compared with others in the current literature on polycyclic aromatic hydrocarbons in foods. The levels in the UK diet seem to be at least as low as those found elsewhere.

Dietary surveys on a population at Shipham, Somerset, United Kingdom.

The results are reported of three dietary surveys and crop sampling surveys carried out at Shipham in 1979. Concentrations of cadmium, lead and zinc in crops were higher than would normally be expected. Copper concentrations in crops were normal, and the results for mercury showed that mercury translocation from soil to crops was very low. Although cadmium dietary intakes were found to be greater than those expected from a normal UK diet, dietary lead and zinc intakes were normal. Dietary copper intakes at Shipham were a little lower than national average intakes. Four participants in the duplicate diet study, or 6% of the study population, had cadmium intakes higher than 0.4 mg per week.

The effect of oxygen on nitrosamine formation in bacon.

Bacon has been cooked in air and in nitrogen atmospheres and the effect on the formation of N-nitrosodimethylamine and N-nitrosopyrrolidine has been studied. Under nitrogen, reductions of 50%-90% were obtained in the concentrations of both nitrosamines in the cooking vapour. The results are discussed in terms of the likely role of nitric oxide in nitrosamine formation in cooking bacon.

Gas chromatographic determination of vinylidene chloride monomer in packaging films and in foods.

Headspace electron-capture-gas chromatographic methods are described for the quantification of vinylidene chloride monomer in poly(vinylidene chloride) containing films and in film-packaged foodstuffs. Quantification of vinylidene chloride was possible at levels down to 0.001 mg/m2 in the films and 0.005 ppm in the foodstuffs. The results of a small survey of the amounts of this compound in packaging films and foodstuffs purchased from retail outlets are reported.