Improved sensitivity in detection of chlormequat by liquid chromatography-mass spectrometry.

Two published separations, using electrospray mass spectrometry (ES-MS), exhibit significant differences in limits of detection (LODs) for chlormequat cation in pear. Separation on ODS1, confirmed to result from ion-exchange, gives shorter analysis times and calibration over a wider concentration range than on an SCX cation-exchange column. The superior LOD using ODS1 (0.04 ng ml(-1) vs. 1.0 ng ml(-1)) results mainly from better chromatographic peak shape. Separation on ODS1 combined with optimised ES-MS detection allows direct quantification of chlormequat on an ion trap instrument at levels lower than those required for residue analysis in foods and also in drinking water.

Determination of the pesticide diflubenzuron in mushrooms by high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry.

A method using high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry (HPLC-APCI-MS) has been developed and validated for the determination of the insecticide diflubenzuron [1-(4-chlorophenyl)-3-(2,6-difluorobenzoyl)urea] in mushrooms. Samples were homogenised with acetone, extracted into dichloromethane-cyclohexane and further cleaned-up by size-exclusion chromatography (SEC). HPLC was performed on an ODS column with methanol-water at 1 ml/min. The limit of detection was 0.02 ng/microl (equivalent to 0.017 mg/kg in the crop). The calibration was linear over the range 0.025-1.0 ng/microl. Recovery of diflubenzuron from spiked mushrooms (0.06-0.58 mg/kg) was 85.5% with a relative standard deviation of 14.5% (n = 56).

Extraction of maleic hydrazide residues from potato crisps and their determination using high-performance liquid chromatography with UV and atmospheric pressure chemical ionisation mass spectrometric detection.

A method was required for the determination of maleic hydrazide residues in potato crisps. A published method for the extraction of the analyte from onions and potatoes was evaluated and found to be inappropriate due to the inability of the extracting solvent to penetrate the oily matrix. A method was developed to overcome this problem; the resulting recovery data (mean = 92.9%, R.S.D. = 8.3%, n = 16) confirmed its efficiency, and was used to analyse 48 retail potato crisp samples. To confirm possible residues identified by screening with HPLC-UV, an HPLC-atmospheric pressure chemical ionization MS method was developed. There was good agreement between the data obtained from the two detection techniques (R2 = 0.978, slope = 1.11).

Studies on the metabolism of deoxynivalenol in the rat.

The metabolism and tissue distribution of [14C]deoxynivalenol have been studied in male PVG rats. Following administration of a single oral 10-mg/kg dose, radioactivity excreted in the urine and faeces accounted, respectively, for 25 and 64% of the administered dose within 96 hr. Less than 0.15% of the dose was detected in the respired air. Very little radioactivity appeared to be retained in any of the tissues examined after 96 hr. HPLC separation of several urinary and faecal metabolites was achieved on a reversed-phase column, using two different elution systems, one at neutral pH and one acidified. Two of the major non-polar HPLC peaks were identified by gas chromatography-mass spectrometry as unchanged deoxynivalenol and 3 alpha,7 alpha,15-trihydroxytrichothec-9,12-dien-8-one.

Levels of selected ortho and non-ortho polychlorinated biphenyls in UK retail milk.

Nineteen pooled samples of retail cow's milk were analysed for ortho (PCBs 28, 52, 101, 118, 138, 153 and 180) and non-ortho (PCBs 77, 126 and 169) polychlorinated biphenyls. Concentrations of individual congeners in whole milk were in the ranges 2 to 95 ng/kg for ortho substituted PCBs (51 to 2440 ng/kg fat) and 0.05 to 0.6 ng/kg for non-ortho substituted PCBs (1.3 to 15.4 ng/kg fat). These values provide a preliminary estimate of background contamination levels for polychlorinated biphenyls in UK milk. Results expressed in TEQs, using the recently proposed WHO/IPCS TEFs, averaged 0.06 ng TEQ/kg whole milk.

Levels of PCDDs, PCDFs and non-ortho PCBs in dietary supplement fish oil obtained in Spain.

Levels of PCDDs, PCDFs and non-ortho PCBs are reported for fish oils available in Spain which are consumed by the population as a dietary supplement. Trace enrichment methodology based on the use of modified silica and activated carbon chromatography was combined with on-column extraction (FMS 100) to separate PCDDs, PCDFs and PCBs from lipid material. The extracts were further cleaned up on disposable Florisil columns and analysed by HRGC/HRMS (PCDD/F and non-ortho PCB fractions). Mean levels found in the samples were 10.50 ppt for total PCDDs and 9.95 ppt for total PCDFs. Calculated PCDD/F I-TEQ values were found to be 2.11 ppt on a lipid weight basis. Total mean levels found for co-planar PCBs (#77, 126 and 169) were 18.26 ppt and the calculated I-TEQ value was 0.31 ppt on a lipid weight basis.

PCDDS and PCDFS in milk from farms in Derbyshire, U.K.

As part of a programme to establish the background levels of PCDD/Fs in milk in the U.K., samples of cows' milk were collected from farms in Derbyshire in 1990 and 1991. The concentrations of PCDD/Fs in the samples from two farms in the Bolsover area of Derbyshire were found to be significantly elevated compared with the normal range for PCDD/Fs in milk in the U.K. Details of the findings and the actions taken as a result are given. The concentrations of PCDD/Fs in milk from those farms previously supplying milk for human consumption are now below the Maximum Tolerable Concentration of 17.5 ng TEQ/kg fat.

Determination of residues of pirimicarb and its desmethyl and desmethylformamido metabolites in fruits and vegetables by liquid chromatography-electrospray/mass spectrometry.

A method was developed for the simultaneous determination of residues of pirimicarb (I) and its desmethylformamido (II) and desmethyl (III) metabolites in plums, peas, green beans, broad beans, carrots, and swedes. The compounds were extracted with ethyl acetate and determined, without cleanup, by reversed-phase liquid chromatography and electrospray mass spectrometry (MS). MS and MS/MS were used concurrently to monitor the protonated molecules and their common collision-induced dissociation product. The limit of detection (signal-to-noise ratio of >3) was 1 ng/mL, corresponding to crop concentrations of <0.0015 mg/kg. All 3 compounds were determined in plums, broad beans, and green beans by MS without interference. Interferences which affected the determination of desmethylformamido-pirimicarb in peas, and to a lesser extent in carrots and swedes, were eliminated by MS/ MS. Recoveries for all 3 compounds, at 0.05 mg/kg for plums and 0.005 mg/kg for other commodities, were in the range 83-124%. No interconversion of I, II and III, occurred during extraction, and the compounds were stable in extracts for > or = 7 days under appropriate conditions.

Contaminants of food : Prioritisation scheme to identify manufactured organic chemicals as potential contaminants of food.

A scheme has been developed to rank 70 industrial organic chemicals in order of their priority for further study as potential contaminants of food. Numerical scales were developed for the following seven key criteria concerning environmental issues, food and toxicity: -Production volume -Pattern of usage -Possible fate in the environment -Likelihood of chemical entering the food chain -Mechanism of entry into the food chain -Persistence and accumulation in the food chain -Toxicity. Each chemical was assigned a score for the above criteria, which were combined to give an overall ranking for the chemicals.This scheme has been endorsed by the MAFF Steering Group on Chemical Aspects of Food Surveillance. It will be used in the assessment of relative priorities for further non-statutory surveillance for these contaminants in the UK food supply.

Automated headspace GC-MS analysis of ethylene dibromide fumigant residues in fresh fruits.

An automated headspace capillary column GC-MS method using selected ion monitoring was developed for the analysis of ethylene dibromide (EDB) residues in fresh fruits. The method was shown to be rapid, to give good agreement with more conventional methods involving distillative extraction, to be free from interferences and to have a limit of detection of 1 microgram/kg. A small survey of a range of fresh fruits including oranges, grapefruit, lemons and mangoes showed no detectable EDB residues in a total of 45 samples obtained from various retail outlets.

Compositional analysis of commercial PVC bottles and studies of aspects of specific and overall migration into foods and stimulants.

Plastics additives and processing aids present severe difficulties for the legislator attempting to establish meaningful controls. In many cases information both on the substances added to the plastic and on the species migrating into food, which may well be different, are lacking. In the studies reported, information has been sought on the actual components employed in two typical commercial plastics. Analysis has been carried out of both the composition and trace constituents of the ingredients of commercial PVC formulations used for manufacturing bottles for orange squash and cooking oil applications. Assessment of maximum leachable material was made by solvent extraction of the individual additives, which was compared with overall migration from the PVC bottles into food stimulants using proposed EEC Statutory Methods. Additionally, specific migration measurements of epoxidized soyabean oil, tris(nonylphenyl) phosphite, nonylphenol, pentaerythritol, and BHT antioxidant were carried out in food stimulants and where possible also in foods.

Determination of pyrrolizidine alkaloids in honey from selected sites by solid phase extraction and HPLC-MS.

A method was developed for the determination in honey of the Ragwort (Senecio jacobaea) derived pyrrolizidine alkaloids jacoline, jacozine, jacobine, seneciphylline and senecionine, combining solid-phase extraction with high performance liquid chromatography and atmospheric pressure chemical ionization mass spectrometric detection. The method allowed determination of individual alkaloids and offered a considerable improvement in terms of speed, sensitivity and specificity over previous approaches, but was not suitable for determination of jaconine, a minor alkaloid in Ragwort. Instrument calibrations were linear over the range 0.005 to 100 micrograms/ml, equivalent to approximately 0.001 to 2.0 mg/kg in honey with the extraction method used and allowing for observed recoveries. Detection limits in honey were 0.002 mg/kg. Recoveries for most of the alkaloids were between 57 and 70%. The alkaloids have been determined in a number of samples of honey selected after pollen identification and counting. The alkaloids were not detectable in samples containing two grains or less of Ragwort pollen per gram of honey. Samples collected in late July and August contained Ragwort pollen at 15-21 grains/g and total alkaloid concentrations of 0.011-0.056 mg/kg. Similar contributions to the total were made by jacozine, seneciophylline and senecionine, with jacobine making a larger and jacoline a smaller contribution. Two samples of honey containing Ragwort pollen at 24 and 16 grains/g had total alkaloid concentrations of 0.42 and 1.48 mg/kg respectively (not corrected for recovery). The alkaloid profile in these samples was dominated by seneciphylline and senecionine which together comprised 90-95% of the total. Alkaloids were not detected in retail honeys.

Gas chromatographic determination of 1,1,1-trichloroethane in vinyl chloride polymers and in foods.

A gas chromatographic method is described for the quantification of levels of 1,1,1-trichloroethane in vinyl chloride polymer resins and in poly(vinyl chloride) (PVC) bottles used in the packaging of foods. By application of electron-capture detection the method can be extended to foods which have been stored in PVC bottles and 1,1,1-trichloroethane can be estimated with a limit of detection in the foodstuff of between 0.002 and 0.01 ppm depending on the food. The results of a small survey of the amounts of this compound in PVC bottles and foodstuffs purchased from retail outlets are reported.

Analysis of sulphonamides using supercritical fluid chromatography and supercritical fluid chromatography-mass spectrometry.

Packed-column supercritical fluid chromatography has been used for the separation of mixtures of sulphonamides on silica and amino-bonded stationary phases utilizing carbon dioxide with methanol modifier as the mobile phase. The effect of modifier concentration, column pressure and modifier identity on retention was also studied. Packed-column supercritical fluid chromatography-mass spectrometry (SFC-MS) of these mixtures utilizing both moving-belt and modified thermospray interfaces was also studied. The identification of sulphamethazine in a spiked porcine kidney extract was performed by SFC-MS using the moving-belt interface.

Migration from plasticized films into foods. 3. Migration of phthalate, sebacate, citrate and phosphate esters from films used for retail food packaging.

A UK survey of plasticizer levels in retail foods (73 samples) wrapped in plasticized films or materials with plasticized coatings has been carried out. A wide range of different food-types packaged in vinylidene chloride copolymers (PVDC), nitrocellulose-coated regenerated cellulose film (RCF) and cellulose acetate were purchased from retail and 'take-away' outlets. Plasticizers found in these films were dibutyl sebacate (DBS) and acetyl tributyl citrate (ATBC) in PVDC, dibutyl phthalate (DBP), dicyclohexyl phthalate (DCHP), butylbenzyl phthalate (BBP), and diphenyl 2-ethylhexyl phosphate (DPOP) in RCF coatings, and diethyl phthlate (DEP) in cellulose acetate. Foodstuffs analysed included cheese, pate, chocolate and confectionery products, meat pies, cake, quiches and sandwiches. Analysis was by stable isotope dilution GC/MS for DBP, DCHP and DEP, GC/MS (selected ion monitoring) for BBP and DPOP, and GC with flame ionization detection for DBS and ATBC, but with mass spectrometric confirmation. Levels of plasticizers found in foods were in the following ranges: ATBC in cheese, 2-8 mg/kg; DBS in processed cheese and cooked meats, 76-137 mg/kg; 76-137 mg/kg; DBP, DCHP, BBP, and DPOP found individually or in combination in confectionery, meat pies, cake and sandwiches, total levels from 0.5 to 53 mg/kg; and DEP in quiches, 2-4 mg/kg.

Migration from plasticized films into foods. 2. Migration of di-(2-ethylhexyl)adipate from PVC films used for retail food packaging.

A UK survey of di-(2-ethylhexyl)adipate (DEHA) levels in retail foods (83 samples) wrapped in plasticized PVC film has been carried out, examining a wide range of different food types obtained from retail and take-away outlets. Foodstuffs analysed included fresh meat and poultry, ready-cooked poultry, cheese, fruit, vegetables and baked goods such as cakes, bread rolls and sandwiches. Analysis by stable isotope dilution GC/MS showed DEHA levels ranging from 1.0 to 72.8 mg/kg in uncooked meat and poultry, 9.4 to 48.6 mg/kg in cooked chicken portions, 27.8 to 135.0 mg/kg in cheese, less than 2.0 mg/kg in fruit and vegetables and 11 to 212 mg/kg in baked goods and sandwiches. The level of DEHA migration correlated with the extent of contact between the film and exposed fatty portions of the food, whether that was the mayonnaise filling of a sandwich or the surface fat from a joint of uncooked meat. The level of DEHA in meat exposed to plasticized film was not reduced significantly by volatilization or chemical transformation on subsequent cooking by grilling or frying.

Screening, confirmation, and quantification of sulphonamide residues in pig kidney by tandem mass spectrometry of crude extracts.

Collisionally activated dissociation mass spectra, observed with a hybrid tandem instrument, of the chemical ionization protonated molecular ions of sulphonamide drugs have been used as the basis of a rapid screening procedure for these drugs in crude extracts of pig's kidney by scanning to detect the parents of a characteristic daughter fragment. Extracts were introduced without chromatography by a moving belt interface. Detection limits of 0.1 mg/kg were achieved. Confirmation was made by obtaining daughter ion spectra of the protonated molecular ions. Multiple reaction monitoring with a stable isotope analogue as internal standard permitted the quantification of targeted compounds with high sensitivity and precision.

Detection of residues of chloramphenicol in crude extracts of fish and milk by tandem mass spectrometry.

The collision-induced dissociation mass spectrum, observed with a hybrid tandem instrument, of the ammonia chemical ionization protonated molecular ion of chloramphenicol was used for the detection of residues of the drug in biological samples. The extracted oil from fish was subjected to a rapid clean-up on a pre-packed silica gel cartridge prior to non-chromatographic tandem mass spectral analysis. Fat extracted from milk was analysed directly by on-line combined high-performance liquid chromatography/tandem mass spectrometry with rapid elution of chloramphenicol. Identification was on the basis of agreement of the daughter ion spectra obtained from sample extracts with that of the chloramphenicol standard. Detection was unambiguous at 0.5 mg kg-1. The sensitivity advantage normally expected with multiple reaction monitoring was not achieved owing to the effect of neutral noise phenomena.

Determination of residues of propamocarb in wine by liquid chromatography-electrospray mass spectrometry with direct injection.

A simple, sensitive and reliable liquid chromatography-mass spectrometry method with direct injection of diluted samples is reported for the determination of propamocarb residues in wine. Red and white wines were diluted 40- and 20-fold, respectively, using water. Liquid chromatography was performed with a mobile-phase gradient and detection was by electrospray mass spectrometry in a positive ionization mode. Propamocarb was detected as the protonated molecular species at m/z 189. Using matrix-matched calibrant solutions, a calibrated range equivalent to 0.05-2.0 mg kg(-1) in red and white wines and limits of detection of 0.025 mg kg(-1) for white wine and of 0.05 mg kg(-1) for red wine (0.00125 microg ml(-1) of sample solution injected) were readily achievable. Recovery of propamocarb hydrochloride from wine spiked before dilution was in the range 91-115%. The chromatograms were free of isobaric interferences. In a small wine survey (72 samples), no residues of propamocarb were detected above 0.1 mg kg(-1).

Formation of gas-phase clusters monitored during electrospray mass spectrometry: a study of quaternary ammonium pesticides.

Gas-phase cluster formation between the quaternary ammonium pesticides paraquat, diquat, difenzoquat, chlormequat and mepiquat, and chloride and acetate anions present in a liquid chromatography (LC) mobile phase, has been studied using electrospray mass spectrometry. The clusters of paraquat, mepiquat and chlormequat were revealed over the entire m/z range of the mass spectrometer, and their formation is dependent on the concentrations of both the cationic and the anionic species. Mepiquat and chlormequat form clusters of the type [2M(q)(+) + A(-)](+), where M(q)(+) is the quaternary ammonium cation and A(-) is the anion. Paraquat forms a cluster species with ammonia and also an ion-pair complex with chloride anions. Diquat and difenzoquat did not form observable ion-pair complexes or clusters with any of the anions studied. Competitive binding of acetate and chloride anions reflects the higher charge density of chloride, which forms the dominant clusters with mepiquat and chlormequat. The formation of cluster species has implications for the quantification of quaternary ammonium pesticides and may have an influence on the linearity of calibrations.