A sensitive method for the determination of chlorine-36 in foods using accelerator mass spectrometry.

A method using accelerator mass spectrometry (AMS) has been developed to offer a more sensitive alternative to scintillation techniques for the determination of chlorine-36 ((36)Cl) in foods. The main problem in method development was the potential interference of the sulfur-36 ((36)S) isobar. This was overcome by reducing the sulfur level of acid digests of food by precipitation of chloride as silver chloride, then purification by washing, dissolution and reprecipitation to present silver chloride as the AMS target. The limit of detection was around 0.1 Bq kg(-1) and the limit of quantitation was around 0.2 Bq kg(-1). The AMS method was only semi-quantitative at the lowest levels of interest. To test the method a few samples of milk (five) and blackberries (three) collected from near two nuclear power stations as potential sources of contamination were analysed. Blackberries spiked at 0.2 Bq kg(-1) and milk spiked at 0.1 Bq kg(-1) could be distinguished from method blanks. There was no (36)Cl detectable in the unspiked samples.

Bioavailability of selenium from fish, yeast and selenate: a comparative study in humans using stable isotopes.

OBJECTIVE: To measure the bioavailability of selenium from cooked and raw fish in humans by estimating and comparing apparent absorption and retention of selenium in biosynthetically labelled fish with labelled selenate and biosynthetically labelled selenium in brewers yeast. DESIGN: The intervention study was a parallel, randomised, reference substance controlled design carried out at two different centres in Europe. SETTING: The human study was carried out at the Institute of Food Research, Norwich, UK and at TNO Nutrition and Food Research, Zeist, The Netherlands. SUBJECTS: In all, 35 male volunteers aged 18-50 y were recruited; 17 subjects were studied in Norwich (UK) and 18 in Zeist (Netherlands). All of the recruited subjects completed the study. INTERVENTIONS: Biosynthetically labelled trout fish (processed by two different methods), biosynthetically labelled brewers yeast and isotopically labelled selenate were used to estimate selenium apparent absorption and retention by quantitative analysis of stable isotope labels recovered in faeces and urine. Subjects consumed the labelled foods in four meals over two consecutive days and absorption was measured by the luminal disappearance method over 10 days. Urinary clearance of isotopic labels was measured over 7 days to enable retention to be calculated. RESULTS: Apparent absorption of selenium from fish was similar to selenate and there was no difference between the two processing methods used. However, retention of fish selenium was significantly higher than selenate (P<0.001). Apparent absorption and retention of yeast selenium was significantly different (P<0.001) from both fish selenium and selenate. CONCLUSION: Fish selenium is a highly bioavailable source of dietary selenium. Cooking did not affect selenium apparent absorption or retention from fish. Selenium from yeast is less bioavailable.

Urinary biomarkers for assessing dietary exposure to caffeine.

The feasibility of using metabolites specific to caffeine as urinary biomarkers to be employed in the estimation of dietary caffeine intake is reported. The influence of inter-individual differences in the metabolism of caffeine and the effect of volunteer phenotype on the interpretation of potential biomarkers has been investigated using urinary caffeine metabolite data. This method of phenotype determination accurately reflected the rate constant for the cytochrome P4501A2 (CYP1A2)-catalysed 3-demethylation of caffeine in vivo. Three studies with up to 20 human volunteers demonstrated that a 24-h urine collection after a caffeine dose allows quantification of the metabolites excreted; that the ratios of selected metabolites used to classify the volunteers into fast, intermediate or slow caffeine metabolizers by CYP1A2 phenotype gave a similar result (2:7:3, slow:intermediate:fast) to that found in the general population (1:7:2); and that three metabolites, 1,7-dimethylxanthine, 1,7-dimethyluric acid and 1-methylxanthine, could be studied further as potential biomarkers for caffeine dietary intake.

Use of the stable isotope (106)Cd for studying dietary cadmium absorption in humans.

Hydroponically grown wheat was intrinsically labelled with the stable isotope 106cadmium (106Cd) and the flour made into a porridge. The abundance of the isotope in the porridge was approximately 30 times the natural abundance, but the total level of Cd in the porridge was 0.03 mg/kg fresh weight, which was the same as expected in a normal diet. Cadmium measurements were made using inductively coupled plasma-mass spectrometry (ICP-MS). The porridge was eaten at breakfast by adult and infant volunteers. Bulked faecal collections were analysed for unabsorbed Cd. Initial results suggest that the apparent absorption of Cd may be higher than 5% as commonly quoted, but longer faecal collection times may be necessary to confirm this.

Lack of effect of dietary chromium supplementation on glucose tolerance, plasma insulin and lipoprotein levels in patients with type 2 diabetes.

Chromium is essential for the regulation of insulin action, thereby influencing carbohydrate and lipid metabolism. An uncontrolled pilot study was designed to measure the habitual daily intake of chromium in a group of healthy individuals with type 2 diabetes and to monitor the effect of daily supplementation with high chromium yeast on glucose tolerance, plasma insulin and lipoproteins. Twelve free-living adults with type 2 diabetes underwent a glucose tolerance test (GTT) on recruitment, at 4 weeks (after a 7-d duplicate diet collection) and at 12 weeks (following 8 weeks daily supplementation with 100 micrograms of chromium). Urine samples were collected on the day before and the day of each GTT. Blood samples were taken at half hourly intervals for 3 hours during the GTT and the plasma glucose, cholesterol, triglyceride, HDL, LDL and insulin concentration measured. The chromium content of diets and urine samples was determined. Fasting glucose concentrations and glucose area under the curve profiles did not alter significantly post supplementation with the chromium rich yeast. No significant changes in insulin and lipoprotein concentrations were observed. The results of this study do not support the hypothesis that individuals with type 2 diabetes benefit from yeast-based chromium supplements (100 micrograms/day).

Urinary monitoring of saccharin and acesulfame-K as biomarkers of exposure to these additives.

A method was developed to determine the levels of two intense sweeteners, saccharin and acesulfame-K, in human urine. Measurement of these two analytes in urine allowed an assessment to be made of dietary exposure to the sweeteners using intake/excretion curves. This paper describes an intake/excretion study in which 22 volunteers consumed known amounts of saccharin and acesulfame-K at five different levels ranging between 1.4 and 70 mg of saccharin (0.7-30% of the ADI for a 60 kg person) and 2.2-102 mg of acesulfame-K (0.4-19% of the ADI for a 60 kg person). Urine collections were then carried out by the volunteers for 24 h after each dose. The data obtained from this study were used to construct intake/excretion curves for both sweeteners. To test the methodology developed, 188 volunteers aged between 3 and 74 years were asked to carry out a 24-h urine collection whilst keeping a 48-h food diary. Comparison of the intake data obtained using the biomarker approach with the questionnaire-based results was generally good, although discrepancies due to the format and assessment of the questionnaire data were noticed.

The analysis of cyhexatin residues in apples, pears and kiwi fruit using inductively coupled plasma mass spectrometry as an initial screen for total tin, with confirmation by gas chromatography-mass spectrometry.

Inductively coupled plasma-mass spectrometry (ICP-MS) has been used to determine concentrations of total tin in samples of apples, pears and kiwi fruit, following acid digestion. It was found that the background levels of tin in these fruits were normally below 0.06 mg/kg, which was equivalent to the target reporting limit for cyhexatin for this work of 0.2 mg/kg. The procedure was used to screen a total of 72 retail samples. Three apple samples contained tin at concentrations greater than 0.06 mg/kg. These three samples were re-analysed by a second confirmatory method using gas chromatography-mass spectrometry (GC-MS) as the determinative step. Results obtained by this method demonstrated that in fact, none of the samples contained cyhexatin residues at or above a reporting limit of 0.1 mg/kg. The use of ICP-MS as a preliminary screen substantially reduced the number of samples requiring analysis by GC-MS, thus reducing the cost of the survey (in terms of staff hours) by approximately 30%. The screening method could potentially be applied to other organometallic pesticide residues.

Rare earth elements as nonabsorbable fecal markers in studies of iron absorption.

The use of rare earth elements as nonabsorbable fecal markers for studies of iron absorption from sources labeled extrinsically with stable isotopes was evaluated. On 3 successive days 13 healthy fasting adults were given different stable isotopes of iron with samarium, ytterbium, or dysprosium. On day 1, three meals were given with 57Fe (1 mg per meal) plus samarium (0.33 mg per meal); on day 2, identical meals (taken with a calcium supplement to reduce iron bioavailability) were given with equivalent amounts of 58Fe-labeled iron and ytterbium; on day 3, a well-absorbed reference dose of 54Fe (3 mg) was given with 1 mg Dy. A complete fecal collection was carried out for 5-9 d and each stool was analyzed for rare earth elements by inductively coupled plasma-mass spectrometry and iron isotopes by thermal ionization quadrupole mass spectrometry. Mean recovery of rare earth elements was 101%, indicating that they are totally unabsorbed. The excretory pattern of the iron isotopes and the rare earth elements was very similar; the correlation coefficients between samarium and 57Fe, ytterbium and 58Fe, and dysprosium and 54Fe were 0.992, 0.989, and 0.988, respectively (P < 0.001). Iron absorption was calculated as the difference between isotope dose and fecal excretion. Mean (+/-SEM) iron absorption was 16.7 +/- 2.4%, 4.3 +/- 1.6%, and 40.3 +/- 3.1% on days 1-3, respectively. Predicted values estimated from the first 4 d of pooled feces, using the rare earth element recovery data to produce corrected figures for unabsorbed isotope, were in close agreement: 19.1 +/- 2.1%, 4.6 +/- 1.7%, and 40.8 +/- 3.1%, respectively (P < 0.001). With the diet of medium iron bioavailability and with the highly bioavailable reference dose it was possible to predict iron absorption accurately from only one or two stools, provided that they were sufficiently enriched with isotope and a rare earth element.

Determination of lead in wine by graphite furnace atomic absorption spectrophotometry: interlaboratory study.

An interlaboratory study of a graphite furnace atomic absorption spectrophotometry (GFAAS) method for the determination of lead in wine was conducted. Seventeen laboratories from France, United States, and the United Kingdom, using a variety of GFAAS instruments, took part in the study. The method incorporated a novel matrix-matching procedure to minimize matrix effects between standards and samples. Six wine test materials were prepared and sent to participants as 12 blind duplicate or split level samples. There was good agreement between results obtained from participants and target values (24-279 micrograms/L) obtained with an inductively coupled plasma-mass spectrometry method. The precision of the GFAAS method was well within the range predicted by the Horwitz equation for the 6 test materials analyzed. Repeatability standard deviations ranged from 3 to 17%. Reproducibility standard deviations were in the range of 10 to 30%. The method is recommended for use for official purposes.

Element residues in food contact plastics and their migration into food simulants, measured by inductively-coupled plasma-mass spectrometry.

Polymers intended for food contact use have been analysed for inorganic residues which can be attributed to a range of substances employed as polymerization aids (e.g. catalysts), or to additives incorporated into the polymer to fulfil a specific task (e.g. lubricants). The migration of these residues into food simulants was studied. Residues were determined by using the multi-element capability of Inductively-Coupled Plasma-Mass Spectrometry (ICP-MS). Semi-quantitative analysis was carried out on acid digests of polymer materials, obtained by microwave heating in sealed Teflon containers. Limits of detection in the polymer were generally less than 1 mg/kg. Migration experiments were carried out with three food simulants and under two sets of conditions. Analysis for element residues was performed directly or, in the case of olive oil, on an emulsion of the simulant. Migration of certain elements into aqueous simulants was observed: Zr from polystyrene (650 micrograms/kg), Sb from polyethylene terephthalate (4 micrograms/kg) and Mg from acrylonitrile/butadiene/styrene copolymer (50 micrograms/kg). In all cases, where limits of detection permit, the levels which migrated from polymer to stimulant were less than proposed limits on migration.

Determination of copper, zinc and aluminium from dietary sources in the femur, brain and kidney of guinea pigs and a study of some elements in in vivo intestinal digesta by size-exclusion chromatography inductively coupled plasma mass spectrometry.

The multi-element facility of an inductively coupled plasma mass spectrometer was used to monitor other elements during a project to investigate aluminium uptake from some foods by guinea pigs and to characterize aluminium in the intestinal digesta by size exclusion chromatography. Copper, zinc and aluminium were measured in the femur, brain and kidney. Aluminium, copper, zinc and manganese were measured in the solid digesta and soluble fraction of the digesta. The soluble fraction was separated by size exclusion chromatography and the behaviour of aluminium, copper, zinc, manganese, strontium and rubidium was monitored. The effect of citrate on each of these elements was assessed in aqueous standards and the complex matrix of the digesta.

Mass spectrometric methods for studying nutrient mineral and trace element absorption and metabolism in humans using stable isotopes. A review.

Mass spectrometric methods for determining stable isotopes of nutrient minerals and trace elements in human metabolic studies are described and discussed. The advantages and disadvantages of the techniques of electron ionization, fast atom bombardment, thermal ionization, and inductively coupled plasma and gas chromatography mass spectrometry are evaluated with reference to their accuracy, precision, sensitivity, and convenience, and the demands of human nutrition research. Examples of specific applications are described and the significance of current developments in mass spectrometry are discussed with reference to present and probable future research needs.

Aluminium uptake from some foods by guinea pigs and the characterization of aluminium in in vivo intestinal digesta by SEC-ICP-MS.

The uptake of ingested aluminium (Al) from food items commonly consumed in a normal human diet was investigated by feeding five test diets to guinea pigs. Al concentrations were measured in the femur, brain, kidney and upper intestinal contents. Consumption of these diets did not lead to elevated Al levels in brain. Levels of Al in the bone were elevated in animals fed sponge cake with a permitted Al-containing additive, and the presence of citrate as orange juice enhanced bone deposition and increased kidney Al levels. Less than 1% of Al in the upper intestinal contents was found in the soluble fraction, and characterization by SEC-ICP-MS indicated that this Al was not present as Al-citrate.

Lead in feed incident--multi-element analysis of cattle feed and tissues by inductively coupled plasma-mass spectrometry and co-operative quality assurance scheme for lead analysis of milk.

Contaminated cattle feed was imported into the UK in 1989 and resulted in lead toxicity in some animals. Rapid analyses for lead and several other possible contaminating elements were required for feed and cattle tissues. Microwave dissolution of samples with measurement by ICP-MS was used for multi-element determinations. Lead was found to be the major contaminant. Lead levels in milk samples were measured by several laboratories during the crisis and an analytical quality assurance scheme was devised to monitor the quality of the data. The scheme allowed any poorly performing laboratories to be rapidly identified and excluded from the survey.

Lead contamination during domestic preparation and cooking of potatoes and leaching of bone-derived lead on roasting, marinading and boiling beef.

Lead concentrations were measured in boiled, mashed potatoes and in baked potatoes that had been prepared and cooked in domestic kitchens. Levels of lead in the boiled, mashed potatoes ranged from below the 1 microgram/kg limit of detection up to 18 micrograms/kg with a mean of 6 micrograms/kg (wet weight). In the large majority of cases the lead in the tap water was the predominant source of the metal. Higher amounts of lead (range 11 micrograms/kg to 56 micrograms/kg, mean 27 micrograms/kg) were present in baked potatoes and this was attributed to soil adhering to the potato skin. The extent of leaching of lead from bone during cooking has also been investigated. For beef stocks there was little evidence to suggest that significant migration of bone lead occurred. For beef casseroles, marinaded in red wine, some leaching did occur from beef joints containing elevated amounts of bone lead; however the levels were all below 350 micrograms/kg and, on average, less than double that found in casseroles prepared from normal joints where the bone lead levels were an order of magnitude less.

Preliminary study of metals in proteins by high-performance liquid chromatography-inductively coupled plasma mass spectrometry using multi-element time-resolved analysis.

The feasibility of multi-element and multi-isotope determinations during the on-line chromatographic separation of protein matrices was investigated using time-resolved software with inductively coupled plasma mass spectrometry. Size-exclusion chromatography was used to separate a mixture of known proteins and the behaviour of associated elements was monitored. It was possible to scan either the entire mass range or a selected number of isotopes. Reversed-phase chromatography was used to separate Zn-containing species in chicken meat that had been intrinsically and extrinsically labelled with stable isotopes of Zn. The ratio obtained for the intrinsic label agreed with that obtained by other workers using thermal ionization mass spectrometry. Time-resolved analysis permitted rapid data acquisition, thus reducing the number of chromatographic runs required. However, at present, data handling can be time consuming. Further software developments will add to the advantages that this technique offers.

Apparent zinc absorption by rats from foods labelled intrinsically and extrinsically with 67Zn.

A variety of foods (peas (Pisum sativum), chicken meat, eggs, goat's milk, human milk) enriched with the stable isotope 67Zn were prepared by means of intrinsic- and extrinsic-labelling procedures. They were fed to rats and apparent absorption of 67Zn determined from faecal excretion measurements using thermal ionization mass spectrometry. There were significant differences in the absorption of the extrinsic and intrinsic label which differed in magnitude between the foods tested. The extrinsic 67Zn was less well absorbed in peas, chicken meat, eggs, and human milk than intrinsic 67Zn, but in goat's milk the extrinsic 67Zn was better absorbed than the intrinsic label. These results demonstrate that extrinsically-added stable Zn isotopes do not fully exchange with endogenous Zn in many foods, and illustrate the need for caution when using extrinsic labels for Zn bioavailability studies.

Application of high-performance liquid chromatography--inductively coupled plasma mass spectrometry to the investigation of cadmium speciation in pig kidney following cooking and in vitro gastro-intestinal digestion.

The speciation of cadmium in retail pig kidney has been examined by size-exclusion chromatography (SEC) coupled directly to inductively coupled plasma mass spectrometry (ICP-MS). Approximately 35% of the cadmium from uncooked kidney was soluble after aqueous extraction at pH 8 and SEC - ICP-MS revealed three discrete peaks whose retention times corresponded to estimated relative molecular masses of 1.2 x 10(6), 7.0 x 10(4) and 6 x 10(3)-9 x 10(3). In the cooked kidney, 35% of the Cd was soluble and was all associated with a peak of a relative molecular mass (Mr) of 6 x 10(3)-9 x 10(3). After simulated gastric digestion of cooked pig kidney at pH 2.5, 60% of the cadmium was solubilised and associated with a species of Mr less than 1 x 10(3). When the digest was also subjected to simulated intestinal digestion at pH 6.8, a single peak, which corresponded to 20% of the original cadmium, was eluted. This peak co-eluted with the single peak extracted at pH 8.0 from the cooked kidney. It was also of similar estimated Mr to the single broad peak observed after simulated gastro-intestinal digestion of equine renal metallothionein (Mr = 1.1 x 10(4]. The results suggest that the majority of soluble cadmium in retail pig kidney is associated with a metallothionein-like protein that survives both cooking and simulated in vitro gastro-intestinal digestion.

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.