Br concentration as an indication of pre-baking bromation of bread products.

Br concentration in bread for baked bread products was shown to be linearly proportional to the amount of Br added per kg of flour used to make the product. Br concentration in bread can be used to help identify those bread products with the greatest likelihood of containing bromate residues. Instrumental neutron activation analysis was used to determine Br in test portions of bread products from commercial bakeries, homemade bread, flour, and unbaked dough. High performance liquid chromatography was used to determine the bromate residue in selected test portions.

Measurement of bromate in bottled water by high-performance liquid chromatography with post-column flow reactor detection.

The objective of this work was to develop a reliable, rugged high-performance liquid chromatographic (HPLC) method for determination of trace levels of bromate (< 10 micrograms/l) in bottled water. HPLC separation was achieved by ion interaction chromatography using a C-18 reversed-phase column and a mobile phase consisting of methanol/water (20:80, v/v) with tetrabutylammonium acetate as the ion interaction reagent. A post-column reaction based on oxidation of o-dianisidine in acidic solution to a product detected at 500 nm provided selective measurement of the oxidants. The limit of detection and the limit of quantitation were 1 and 3 micrograms/l, respectively. Iodate, chlorite, and nitrite were chromatographically separated from bromate and measured by monitoring the post-column reaction. Chloride and chlorate at levels that might be found in bottled water did not interfere with the determination of bromate. Bromate was detected in bottled waters at concentrations up to 40 micrograms/l.

Survey of sulphites determined in a variety of foods by the optimized Monier-Williams method.

A survey of sulphite levels in a variety of foods including dried fruit, dried mashed potatoes, shrimp, canned mushrooms and fruit juices has been conducted. The levels found ranged from 0 ppm in orange juice to 3722 ppm as sulphur dioxide in dried fruit. The foods were analysed by the optimized Monier-Williams procedure.

Determination of free and reversibly bound sulphite in foods by reverse-phase, ion-pairing high-performance liquid chromatography.

The reaction of sulphite with formaldehyde to form hydroxymethylsulphonate (HMS), which is very stable under the controlled conditions of this assay, was used as the first step in an analytical procedure to determine foodborne sulphite. The effect of mobile-phase pH on the stability of HMS during high-performance liquid chromatography was studied. It was found that on-column HMS dissociation to formaldehyde and bisulphite increased with the pH of the mobile phase; therefore the relatively low pH 4.7, at which the dissociation of HMS was approximately 2%, was selected for the analysis. In addition, the release of sulphite from its reversibly bound forms in wine and other foods was examined as a function of the pH of the extraction medium by following the appearance of HMS formed from the reaction of the freed sulphite with formaldehyde. The rate of dissociation of the reversibly bound sulphite was relatively slow at pH 3 but very rapid at pH 7. This difference in kinetics was exploited to develop a procedure to determine free and reversibly bound sulphite in food. The method was challenged by post-reagent spiking studies, i.e. adding the sulphite spike after the food has been blended with the sulphite-protective formaldehyde solution but before proceeding with the remainder of the assay. An average recovery of 100% with a standard deviation of 5.2% (n = 45) was realized at levels of 5, 10 and 20 ppm by weight as sulphur dioxide. Recovery of the sulphite added as the bisulphite addition product of acetaldehyde, a model compound for reversibly bound sulphite, was 95%.

A review of sulphites in foods: analytical methodology and reported findings.

Sulphites in various forms have been added to foods for centuries. Their use became an issue of concern when certain sensitive individuals exhibited adverse reactions to sulphite residues in foods. Analytical methods were developed to monitor these compounds at the regulatory limit of 10 ppm. In this report, analytical methods for determining sulphites in foods are reviewed, along with a critique of their chemistry and procedural schemes. An assessment of the key features of each method category is presented together with some comparative data. The classification scheme used is based upon the fact that determination of the sulphite content of a food is influenced more by the treatment and cleanup of the test solution than by the final determinative step. Based on a 60-year database, the Monier-Williams procedure still remains the method of choice.

Optimized Monier-Williams method for determination of sulfites in foods: collaborative study.

A collaborative study was conducted of the Food and Drug Administration (FDA)-optimized Monier-Williams method for determining sulfites in foods. Twenty-one industry and government laboratories participated in the study, which was jointly sponsored by the National Food Processors Association and FDA. Familiarization samples were shipped to each collaborator. Collaborators were permitted to proceed to the main study only after they demonstrated ability to perform the method to ensure that the study tested the performance of the method itself and not that of the individual laboratories. The study design involved 3 food matrixes (hominy, fruit juice, and protein [seafood]). Each matrix was prepared at 3 sulfite levels--the regulatory level, half the regulatory level, twice the regulatory level--and as a blank. All test samples were analyzed as blind duplicates, which gave each collaborator a total of 24 test portions. Collaborative recoveries gave a reproducibility (among-laboratories) coefficient of variation that ranged from 15.5 to 26.6% for sulfite determined as SO2 by weight in the 3 foods at the 10 ppm level. The optimized Monier-Williams method has been approved interim official first action to replace the AOAC modified Monier-Williams method, 20.123-20.125.

Electron impact mass spectrometry of BHT and its alteration products.

The electron impact (EI) mass spectra of 2,6-di-tert-butyl-4-methylphenol (BHT) and certain of its alteration products are described in detail. Accurate mass measurements confirm the elemental compositions of important fragment ions in the EI spectra. Collisionally activated mass spectra are also used to study fragmentation and suggest common ion structures. The reference spectra provide the basis for identifying various alteration products of BHT by capillary gas chromatography/mass spectrometry (GC/MS) without the necessity of isolating individual components. Application of GC/MS is made to three studies: (i) pyrolysis of hydroperoxy-BHT as a potential pathway to alteration products in food; (ii) GC/MS pyrolysis of hydroperoxy-BHT as a model study; and (iii) alteration of BHT in ethanol/water as a food-simulating solvent.

Sulphite stabilization and high-performance liquid chromatographic determination: a reference method for free and reversibly bound sulphite in food.

A high-performance liquid chromatographic (HPLC) post-column reactor combination has been developed for the determination of sulphite (free and reversibly bound) in 15 different foods. The foods were treated with a pH 5.1 buffered aqueous 2% formaldehyde solution to convert the labile sulphite to the relatively stable hydroxymethylsulphonate. Reverse-phase ion-pairing HPLC with a post-column detector consisting of an initial reaction with KOH followed by colorimetric reaction with Ellman's reagent buffered at pH 6.3 were used for separation and quantitation. The photometric detector at 412 nm quantitated the strongly absorbing 3-carboxy-4-nitrothiophenolate anion, the product of the sulphite reaction with Ellman's reagent. The recovery at levels of 5-100 ppm as sulphur dioxide was greater than 90% by reverse isotope dilution assay. Repetitive analysis of a single grape juice extract containing 20 ppm SO2 showed a relative standard deviation of 2.2%.

Reactions of antioxidants in foods.

The chemical transformations of BHT and BHA in the deep-fat frying of potatoes were studied. Approximately 80% of the BHT and its associated decomposition products was lost from the lard after six batches of french fries. This may have been due to steam distillation caused by the water boiled out of the potatoes. In contrast, 80% of the radiocarbon introduced as radiolabelled BHA was retained by the frying medium even after 12 batches of french fries. The concentration of intact BHA decreased to undetectable levels after four batches. Because of the complexity of the products formed in the heated fats, studies were undertaken on the thermolysis of a hydroperoxy derivative of BHT, 2,6-di-tert-butyl-4-hydroperoxy-4-methylcyclohexa-2,3-dienone++ + (HBHT). The product mixture, which was identified by mass spectrometry and gas chromatography-mass spectrometry, included BHT, 2,6-di-tert-butyl-4-hydroxy-4-methylcyclohexa-2,5-dienone and 3,5-di-tert-butyl-4-hydroxybenzaldehyde. Elemental compositions of other unidentified products were determined by high-resolution mass spectrometry.

Ion chromatographic determination of sulfites in foods.

Ion chromatography (IC) is shown to be a promising technique for the determination of sulfites (SO2, SO2/3-) in foods. Results of a 10 min flash distillation and 10 min IC determination compare favorably with the results from the conventional Monier-Williams method for total sulfite in a variety of food matrices. The IC technique also provides a wealth of additional information, such as (1) sulfite and sulfate (oxidized sulfite) content of the spiking or treatment solution, (2) residual sulfite applied to the food after oxidation losses in the treatment process, (3) free sulfite in foods, and (4) total sulfite in foods. As a further check on the Monier-Williams method, the sulfate content of the trapping solution can be determined by IC. Because the IC technique traps the liberated SO2 in a non-oxidizing rather than an oxidizing medium, it is considered free from interfering sulfides and organic sulfur-containing groups which can give false positives in the Monier-Williams method. IC thus offers a high speed, more sensitive, and cost-effective alternative to conventional techniques for the determination of sulfite in foods.

Reevaluation of Monier-Williams method for determining sulfite in food.

The Monier-Williams distillation procedure has a long history of successful use for determining sulfite in fruit products and wine; however, a systematic evaluation of its accuracy and precision with other food matrices has not been undertaken. We found that the Monier-Williams distillation yielded greater than 90% recovery of sulfite added to foods such as table grapes, hominy, dried mangoes, and lemon juice. Less than 85% recovery was obtained with broccoli, soda crackers, cheese-peanut butter crackers, mushrooms, and potato chips. These results may, in fact, accurately reflect the residual levels of sulfite if a portion of the sulfite undergoes irreversible reaction with some food components. Analysis of commercial food products gave sulfite levels ranging from 1400 ppm in dried apple slices to 25 ppm in cream sherry.

Liquid chromatographic determination of aspartame in dry beverage bases and sweetener tablets with confirmation by thin layer chromatography.

A liquid chromatographic method is described for the determination of aspartame in dry beverage bases and sweetener tablets. The sample was mixed with the mobile phase, the pH was adjusted to within +/- 0.1 pH unit of the mobile phase, and the sample was diluted to volume with the mobile phase. The solution was filtered and a 10 microL aliquot was injected onto a C18 reverse phase column. Aspartame was quantitated with an ultraviolet detector. Recoveries of aspartame ranged from 94 to 111%. The dry beverage bases contained 5-13% aspartame and the sweetener tablets contained 19% aspartame. The presence of aspartame was confirmed by using thin layer chromatography.

Gas-liquid chromatographic determination of dehydroacetic acid in squash and wine.

A gas-liquid chromatographic procedure has been developed for the determination of the preservative dehydroacetic acid (DHA) in frozen cut squash and white wine. The cleanup procedure uses the acidic character of DHA to enhance separation from interfering substances. Recoveries were 93-104% from squash fortified at 30, 65, and 130 ppm and 96-99% from wine fortified at 50, 100, and 200 ppm. The method can be used to establish that the amount of DHA used in squash does not exceed the permissible level of 65 ppm.

Gas chromatographic determination of sorbitol, mannitol, and xylitol in chewing gum and sorbitol in mints.

A method has been developed for determination of sorbitol, mannitol, and xylitol in chewing gum and sorbitol in mints. Chewing gum is partitioned between methylene chloride and water; the mint is simply dissolved in water. The aqueous extract is dried and the residue is derivatized with pyridine-acetic anhydride to form the corresponding peracetates. The derivatives are quantitated by gas chromatography using a 9 ft x 2 mm column packed with 10% Silar 10C on Chromosorb W/AW. Average recoveries of these sugar alcohols ranged from 96 to 102%.

Time required to achieve homogeneity in swine feed mixtures.

FD&C Red No. 3 was mixed with 20 kg pig feed to give a concentration of 0.1%. A mixing time of 30 min was sufficient to achieve homogeneity for this mixture. For larger amounts or more flocculent types of additives, a longer time may be required. Ammoniated glycyrrhizin was mixed with 8 separate batches of pig feed at a concentration of 1%; 1 h was sufficient mixing time.

High pressure liquid chromatographic determination of ethoxyquin in paprika and chili powder.

A method is described for the determination of ethoxyquin (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline) in paprika and chili powder. Ethoxyquin is extracted from the spice with hexane and partitioned into 0.3N HCl. After adjusting the solution to pH 13-14, ethoxyquin is extracted into hexane, and the hexane layer is evaporated to dryness. An acetonitrile solution of the residue is then analyzed by reverse phase high pressure liquid chromatography with detection at 254 nm. The mobile phase is water-acetonitrile with ammonium acetate buffer. Recoveries from samples fortified at 50, 100, and 200 ppm averaged 92% with a coefficient of variation of 2.3%. The method was applied to a number of commercial samples of paprika and chili powder. Ethoxyquin was found in paprika samples at levels up to 63 ppm and in chili powder samples at levels up to 20 ppm.

Reverse phase ion pair high pressure liquid chromatographic determination of ethylenediaminetetraacetic acid in crabmeat and mayonnaise.

A method is described for the determination of ethylenediaminetetraacetic acid (EDTA) in crabmeat and mayonnaise. EDTA is extracted from the food sample with water and converted to its copper chelate, which is then quantitated by reverse phase ion pair high pressure liquid chromatography with ultraviolet detection. Maximum sensitivity is obtained with detection at about 254 nm; higher wavelengths may be used for enhanced specificity. Cleanup procedures for crabmeat and mayonnaise were improved by using a radiotracer method. Analyses of crabmeat and mayonnaise samples spiked at 3 different levels showed greater than 90% recovery of EDTA.

Quantitative determination of isosorbide dinitrate and two metabolites in plasma.

A GLC method for the determination of plasma isosorbide dinitrate and two metabolites, isosorbide 2-mononitrate and isosorbide 5-mononitrate, is described. The three substances are extracted from alkalinized plasma with ether. Quantitation is effected by electron-capture detection after GLC separation with a 30% SE-30 column. The unusually heavy liquid phase loading is necessary to eliminate irreversible adsorption on the solid support. The electron-capture detector provides excellent sensitivity and specificity because of the electronegative nature of the nitric ester. The method was used to study the blood levels of isosorbide dinitrate and two metabolites in four beagle dogs after single oral doses of 40 mg of isosorbide dinitrate and in two human volunteers after a sublingual dose of 10 mg of isosorbide dinitrate.