Formation of N-nitroso-N-methylurea in various samples of smoked/dried fish, fish sauce, seafoods, and ethnic fermented/pickled vegetables following incubation with nitrite under acidic conditions.

In continuation of our previous studies on N-nitroso-N-methylurea (NMU) formation in cured meats following incubation with nitrite at gastric pH, we extended the investigation to other foods mentioned in the title of this paper. The main objective was to determine whether these foods have the potential to form NMU at pH's that can be found in the human stomach. This was done by nitrosating an aliquot (5 g for fish sauce, 10 g for the others) of each with 7.25 microM to 1.59 mM levels of sodium nitrite for 2 h at room temperature at pH 0.8--1.5 and measuring the amounts of NMU formed. Of the samples tested, fish sauce formed 2--712 ng of NMU, followed in decreasing order by herring (<0.3--688 ng); dried anchovy, shrimp, and other fishes (<0.3--134 ng); crab and lobster paté (<0.3--342 ng); sardines (6--59 ng); oysters and mussels (11--31 ng); dried squid (3--47 ng); kimchi (7--107 ng); and Japanese pickled radish (<0.3--72 ng). Incorporation of 200-2000 ppm of ascorbic acid in the fish sauce and other foods, prior to nitrosation, appreciably inhibited such NMU formation. Although previous researchers in China reported NMU formation in nitrosated samples of fish sauce, this is the first reported formation of NMU upon nitrosation of the other foods mentioned above, and the first reported inhibition of such formation by added ascorbic acid.

Investigation on the possible formation of N-nitroso-N-methylurea by nitrosation of creatinine in model systems and in cured meats at gastric pH.

N-Nitroso-N-methylurea (NMU) is a highly potent direct-acting carcinogen that has been shown to induce cancer in a number of animal species. Although previous research has indicated that nitrosation of creatinine (CRN), a common constituent of meats, dried fish, and seafoods, can form traces of NMU, there is uncertainty as to (1) the yield of NMU and (2) whether detectable amounts of NMU can be formed from cured meats following nitrosation under acidic conditions given the low residual levels of nitrite found in cured meats at the present time. Lack of sensitive and specific analytical methods most likely has hindered progress in research in these areas. An HPLC postcolumn denitrosation-thermal energy analyzer technique and a GC-MS confirmation technique were developed for the determination of NMU in cured meats. Both techniques are highly sensitive (0.5 and 0.03 ppb, respectively) and specific. The optimum pH for NMU formation from CRN ranged between pH 1 and pH 3, and the yields of NMU under variable reactant concentrations ranged between 0.00004 and 0.0046%. When 27 samples of various cured meats (10 g aliquots each) were acidified with HCl (final pH values of 0.8-2.5) and incubated at room temperature for 2 h, without any additional nitrite, 24 gave results below detectable levels but 3 formed 2-26 ng of NMU/10 g of meat. Incubation of the negative meats with additional nitrite (50-500 microg/g of meat) formed 0.6-176 ng of NMU/10 g of sample. Although the amounts of NMU formed were extremely small, this seems to be the first reported formation of NMU from cured meats with and without additional nitrite.

Rapid and sensitive determination of nitrite in foods and biological materials by flow injection or high-performance liquid chromatography with chemiluminescence detection.

A method is described for the determination of nitrite that is based on reduction of nitrite with potassium iodide followed by chemiluminescence detection of the liberated NO using a thermal energy analyzer. The method worked well both in the flow injection mode and upon interfacing with a reversed-phase high-performance liquid chromatography system. Limited data available indicated a good agreement between results obtained by the chemiluminescence method and those obtained by using a well established colorimetric procedure when they were applied for the determination of nitrite in a number of cured meats, human saliva, and baby foods. The chemiluminescence method is, however, much superior to the colorimetric one in its speed, versatility, as well as sensitivity (200 times more), and requires only a minimum of sample preparation. The detection limit of the new method is about 0.1 ng NaNO2 per injection or 5 micrograms/kg in cured meats and other substrates analyzed.

Analytical methods for the determination and mass spectrometric confirmation of 1-methyl-2-nitroso-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid and 2-nitroso-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid in foods.

A method is described for the determination of the two title compounds that is based on: (a) extraction of the acidified sample with methanol, (b) removal of fats and lipids by partitioning of the extract with n-hexane, (c) clean-up on acidic alumina extraction cartridge, and (d) determination by a post-HPLC column chemical denitrosation-thermal energy analyser (TEA) technique or by conventional HPLC-TEA analysis after derivatization of the compounds with diazomethane. Confirmation was carried out by HPLC-mass spectrometry of the free acids and also by gas chromatography-mass spectrometry of the methyl esters. The formation of both 1-methyl-2-nitroso-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid and 2-nitroso-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid in nitrosated samples of several Japanese and Chinese pickled vegetables, one soy sauce, and two cheeses was demonstrated.

The analysis and significance of bound N-nitrosoproline in nitrite-cured meat products.

An alkaline hydrolysis method is described for the release of bound N-nitrosoproline (NPRO) from cured meats that is more efficient than an enzymic method reported previously. The bound NPRO contents of 17 cured meats analysed ranged between non-detectable and 578 micrograms/kg (mean 143 micrograms/kg). Administration of defatted meat powder, prepared from such meats, to rats led to increased excretion of free NPRO in the urine that could not be inhibited by concurrent administration of ascorbic acid. The significance of these findings with regard to the use of such cured meats in in vivo N-nitrosation studies is discussed.