[Development and application of gas chromatographymass spectrometry method for quantitive determination of volatile N-nitrosamines in smoked meat products].

The article presents the results of experimental studies on the development of highly sensitive and selective chromatography-mass spectrometry technique for the determination of 9 N-nitrosamines in food samples (sausage products) using distillation and an automatic solid-phase extraction system on Coconut cartridges for sample preparation. In the elaborated conditions of sample preparation (distillation and solid-phase extraction) and chromatography-mass spectrometric analysis, we achieved a high recovery and efficiency of the separation of nine N-nitrosamines. The quantitation limit was at level of 0.0002 mg/kg with maximum error not exceeding 19%. The complex use of the distillation of N-nitrosamines with the addition of potassium hydroxide in combination with the optimal elution scheme for solid-phase extraction and concentrating the distillate into a Coconut carbon cartridge of 6 ml ensures the recovery of N-nitrosamines from the food product sample (sausage products) up to 93.2-100%. The process of approbation of the chromatography-mass spectrometric method in the samples of food products (sausage products) of various manufacturers revealed the content of N-nitrosamines in the concentration range 0.00029±0.000055÷0.350±0.05 mg/kg. The conducted studies of the content of the sum of highly toxic N-nitrosamines (N-nitrosodimethylamine, N-nitrosodiethylamine) made it possible to disclose that in sample No. 5 the maximum allowable concentration was exceeded by 47 times, in samples No. 2 and 16, to 57.5 and 22.9 times and in sample No. 4 to 88 times, respectively.

[Determination of phthalates in juice product by high-performance liquid chromatography/mass spectrometry].

The paper presents the research results obtained after determination of the persistent organic pollutants of the environment - phthalates in domestic juice products with the use of high-performance liquid chromatography/ mass spectrometry (HPLC/MS/MS) with application of solid-phase extraction as a method of sample preparation. Selective determination of phthalates in fruit and vegetable juices by HPLC/MS/MS method has been provided through parent and child ions' monitoring that are formed during ionization of the molecules of the studied compounds in the electrostatic spray of isomers on a reversed-phase column. The analysis of the samples of juice product revealed the presence of 11 phthalates out of 13 studied in concentration range from 0.4 up to 59.26 mg/dm3. The maximum amount of phthalates was found in juices with pulp 31.9-59.26 mg/dm3 (tetrahedral package), the minimum content of phthalates 0.4 mg/dm3 was detected in flavoured beverages (polyethylene terephthalate packing) and 1 mg/dm3 in the fruit-and-vegetable nectar (glass packing). The most samples contained from 3 to 8 phthalates with total content of 0.4-5.82 mg/dm3. The most prevailing pollutants of the analyzed samples were di-n-octyl phthalate found in 100% of samples, dinonyl phthalate and di (2-ethylhexyl) phthalate, presented in 70-80% of samples. The maximum pollution density of the juice samples was noted for di-n-octyl phthalate, di (2-ethylhexyl) phthalate, diisononyl phthalate and diisobutyl phthalate.

[Determination of N-nitrosodiphenylamine in meat canned food for children by the method of chromato-massspectrometry].

This study demonstrates the results obtained from the GC/MS experimental determination of low concentrations of N-nitrosodiphenylamine in meat canned baby food in the concentration range of 0.027-3.89 mg/kg. The perfect conditions of sample preparation (extraction with organic solvent and solid phase extraction) as well as the application of the chromatography-mass spectrometry allowed us to detect N-nitrosodiphenylamine in samples of the meat canned baby food with high selectivity in concentrations ranged from 0.016 to 5 mg/kg when an error of 23% was assumed. The use of the reaction of transesterification of fatty acids by potassium methylate, the removal of the ester generated from the samples of canned meat by organic solvent (hexane), concentrating of N-nitrosodiphenylamine in the aqueous layer on the cartridges of an automatic solid-phase extraction system provided 99.94% extraction of N-nitrosodiphenylamine from the canned meat samples. The experiment has made evident the presence of N-nitrosodiphenylamine in the samples of canned meat (beef + chicken) with the help of mass-spectrometry method in the mode of full ion scanning using the AMDIS automatic mass-spectral identification system.

[Role of immunological factors in the development of myopic choroidal neovascularization]. / Vliyanie immunologicheskikh faktorov na mekhanizmy formirovaniya miopicheskoi khorioidal'noi neovaskulyarizatsii.

AIM: To study the concentration of cytokines in the aqueous humor of the anterior chamber in patients with myopic choroidal neovascularization (mCNV) and to compare the results to their ophthalmic status. MATERIAL AND METHODS: A total of 19 patients (19 eyes) with mCNV treated with intravitreal ranibizumab were included in the study. The control group consisted of 15 patients (15 eyes) with myopia who had cataract surgery. Age, sex, and refractive error distribution were similar to that in the study group. All patients underwent a detailed ophthalmic examination as well as immunological study of the aqueous humor for cytokines concentrations using flow fluorometry (Bio-Plex Pro Human Cytokine Panel, 27-Plex, Bio-Rad Laboratories, USA). RESULTS: Significant differences in concentrations of 10 cytokines were found between the mCNV and study groups. Vascular endothelial growth factor (VEGF) level was twice as low in patients with mCNV as that in the controls (191.15±142.3 pg/ml and 320.06±170.05 pg/ml, respectively) (p<0.05). The other 9 cytokines were higher in mCNV, namely, platelet-derived growth factor (PDGF-BB), inflammatory and anti-inflammatory cytokines (IL-2, IL-15, IL-17А and IL-5, IL-13, respectively), tumor necrosis factor (TNFα), and chemokines (IL-8, RANTES). The degree of myopia as well as morphological and functional changes in the macular zone were shown to be in close correlation with cytokines involved in inflammation and VEGF. VEGF level appeared to be negatively related to axial eye length, refractive error, and three cytokines: IL-13, INF-γ, and RANTES. At the same time, numerous (6, 8 and more) close correlations were established between inflammatory and anti-inflammatory cytokines, chemokines, and growth factors. CONCLUSION: Patients with mCNV have been found to have higher than usual levels of inflammatory and anti-inflammatory cytokines, chemokines, and growth factors as well as a significantly decreased VEGF concentration. Immunological status of these patients differs from that in other ocular neovascular diseases suggesting possible involvement of alternative pathogenetic mechanisms.

[Toxicological evaluation of nanosized colloidal silver, stabilized with polyvinylpyrrolidone, in 92-day experiment on rats. II. Internal organs morphology].

The aim of the study was to evaluate the safe doses of commercially available nanosized colloidal silver (NCS), stabilized with polyvinilpirrolidone (PVP, food additive E1201) when administered in gastrointestinal tract of rats in the 92-day experiment in terms of the morphological changes in the internals of animals. The sample studied contained non-aggregated nanoparticles (NPs) of silver belonging to size fractions with a diameter of less than 5 nm, 10-20 nm or 50-80 nm. 80% of NPs were inside the range of hydrodynamic diameters 10.6-61.8 nm. The preparation of NCS was administered to growing male Wistar rats. (initial body weight 80 ± 10 g) for 1 month by intragastric gavage and then consumed with food at doses of 0.1, 1.0 and 10 mg/kg of body weight based on silver. The control animals received water or vehicle of nanomaterial--water solution of PVP. After withdrawal of animals from the experiment by exsanguination under ether anesthesia organs (liver, spleen, kidney, ileum) were isolated and their slides were prepared by standard methods following 'by staining with hematoxylin-eosin. Analysis was performed in light optical microscope equipped with a digital camera at a magnification from 1 x 100 to 1 x 1000. It was shown that the experimental animals treated with the NCS developed series of morphological changes in the tissues of the internal organs (liver, spleen and kidney) with the elevation of the range and severity of structural changes with increasing doses of silver. The most sensitive target of NCS action was apparently liver, which has already shown at a dose of 0.1 mg of silver NP/kg of body weight marked eosinophilic infiltration of portal tracts, which was accompanied at doses of 1.0 and 10.0 mg/kg by the emergence of medium and large-drop fat vacuoles in the cytoplasm of hepatocytes, swelling and lympho-macrophage. infiltration of the portal tracts. Detectable changes can be regarded as symptoms of inflammation of hepatocytes, at least, at a dose nanomaterial of 1.0 mg/kg body weight or more. Relative intensity of morphological changes in the internal organs correlated with published data on the biodistribution of silver NP administered to the gastrointestinal tract. It is concluded that the threshold dose corresponding to the minimum adverse effect of NCS is, according to the study of the above, no more than 1.0 mg/kg of body weight based on silver.

[Control of highly toxic N-nitrosamines (N-nitrosodimethylamine and N-nitrosodiethylamine) content in baby's cereals].

This paper proposes gas chromatography-mass-spectrometry method for determination of N-nitrosamines (N-nitrosodimethylamine and N-nitrosodiethylamine) in dry baby cereals (milk and milk-free). According to the results of the experimental studies, the method of sample preparation has been substantiated. This is the method of distillation with superheated steam, concentration of distillate on cartridges of automatic system of solid phase extraction. Optimal conditions for chromatography-mass spectrometry analysis has been selected (GC/MS). Analysis of the cereals (milk and milkfree) on the content of N-nitrosodimethylamine and N-nitrosodiethylamine in concentration range 0.0055-0.0109 mg/kg allowed to determine high content of the determined components by the sum of N-nitrosamines in cereals' samples of different manufacturers: oatmeal with milk, buckwheat with milk, oatmeal with milk and banana, milk-free maize cereal. In the multigrain milk cereal and buckwheat with milk, peaches and apricots the content N-nitrosamines was not detected (0.0004-0.00066 mg/kg). To confirm the presence of N-nitrosamines identified in a sample of oatmeal with milk, the identification in SCAN mode has been performed. Mass-spectrum of N-nitrosodimethylamine and N-nitrosodiethylamine in examined samples were compared with massspectrums that were included in library bank of mass-spectral data NIST 08.L.

[METHODOLOGICAL APPROACHES TO THE CALCULATION OF ACTUAL AND PREVENTED AS A RESULT OF CONTROL AND SUPERVISORY ACTIVITIES, MEDICAL-DEMOGRAPHIC AND ECONOMIC LOSSES, ASSOCIATED WITH THE NEGATIVE IMPACT OF ENVIRONMENTAL FACTORS].

In the article there are reported the methodological approaches to the calculation of actual and avoided as a result of control and supervisory activities of economic losses caused by mortality, morbidity and disability in the population, associated with the negative impact of environmental factors. There is suggested a consistent solution of the chain of problems: the establishment of cause-and-effect relationships between indices of the health status and indices of the quality of the environment; indices of the quality of the environment and indices of the control and supervision activity of organs and institutions of Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing calculation of the cases of violations of health prevented as a result of the activity ofthe service; evaluation of their economic equivalents. Approbation of approaches on the example of the Russian Federation allowed to establish that as a result of the activity of organs and institutions of Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing in 2013, a positive trend was observed on 51 indices of quality of the habitat environment, there were prevented about 160 thousands of deaths and more than 2 million cases of disease that would be held in conditions of the lack of adequate control and surveillance measures in the field of sanitary and epidemiological welfare of the population. Loss prevention of gross domestic product (GDP) amounted to more than 120 billion rubles, Tax shortfalls in the federal budget--about 25.7 billion rubles. With taking into account the costs of the federal budgetfor the activities of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing in 2013 in part to ensure sanitary epidemiological surveillance in the amount of 11.386 billion rubles there was prevented loss of GDP of 10.56 rubles per 1 rub. of the cost of the federal budget and there was warned the shortfall of taxes to the federal budget of 2.28 rubles per 1 rub. of the cost of the federal budget.

[The elaboration of gas chromatographic method of the determination of N-nitrosamines (N-nitrosodimethylamine, N-nitrosodiethylamine) in biological samples (urine)].

The issues of the elaboration of a method for the determination of N-nitrosamines (N-nitrosodimethylamine, N-nitrosodiethylamine) in urine by means of the method of capillary gas chromatography with the use of a thermionic detector are considered. There were performed investigations on the study of the efficacy of the extraction of N-nitrosamines from the urine by steam distillation and gas chromatographic detection of headspace. With the aim of the maximal recovery of N-nitrosamines from the urine and setting parameters of the extraction two method were used to prepare the bioassay for the analysis the alkalization with potassium hydroxide and the addition of salting out reagent--neutral salts of alkali and alkaline earth metals. During the process of performed studies there was found that the greatest degree of extraction of N-nitrosamines from the urine by the method of headspace analysis is achieved if using the salting-out agent in an amount of 16 g of sodium sulfate and for N-nitrosodimethylamine is 99%, for N-nitrosodiethylamine--100%.

[Characteristics of regulatory system in children exposed to the environmental chemical factors].

In children residing in areas with a high content of a number of aromatic hydrocarbons in ambient air and organochlorine compounds in drinking water there were studied the blood levels of these compounds, as well as the assessment of the indices of the immune and neuroendocrine systems was performed. The higher blood content of phenol and formaldehyde has been established and there was identified an array of organochlorine and aromatic compounds not detected in the control group children. In the blood of the children of a study group there was found an imbalance of indices of cellular components of innate and adaptive immunity, pro- and anti-inflammatory cytokines, as well as increased concentrations of free thyroxine and serotonin in the blood serum, which indicates to a change in the functions of regulatory systems in children exposed to organochlorine and aromatic compounds.

[Experience of justification of hygienic standards of food safety with the use of criteria for the risk population health].

In the article there is presented the experience of justification of hygienic standards of food safety with the use of criteria for the risk for population health. Health risk assessment under the impact of tetracyclines with food showed that the content of residual amounts of these antibiotics at the level of 10 mg/kg (permissible residual tetracycline accepted in Customs Union Member Countries (CUMC) will not increase the risk to public health, including the most sensitive groups of the population. The assessment ofthe health risk associated with the receipt of ractopamine with food, showed that eating foods containing ractopamine at ADI level (0-1 mg/kg body weight), and even at the limit of quantification levels in meat products, is inadmissible because of unacceptable risk of functional disorders and diseases of the cardiovascular system. The results of the substantiation of the permissible levels of nitrates content in crop production showed that at the level of exposure according to hygienic standards established in the CUMC as at the recommended and actual consumption levels of products ofplant origin, the health risk as carcinogenic and non-carcinogenic, does not exceed acceptable levels. The results of the assessment of the risk associated with the permissible levels of L. monocytogenes in certain food groups showed that an exposure level of hygienic standards established in the CUMC, standards of Codex Alimentarius Commission and EU documents (before release to the market by the manufacturer) the health risk does not exceed the maximum permissible level of the appearance of serious diseases. Adoption of standards of Codex Alimentarius Commission and the EU (for handling products in the market) is not acceptable because it can lead to an unacceptable risk of listeriosis for the population of the Russian Federation as a whole, and for the most sensitive groups.

[Methodical ware for the hygienic risk assessment of vanadium exposure to the children's health].

In the article there is considered the complex of methodological approaches for the detection of vanadium in the air and biological substrates of the population for the practical use in the frameworks of public health monitoring in areas with localization of steel industry facilities. The developed complex of methods on the base of mass spectrometry with inductively coupled plasma (ICP-MS) allowed to perform the hygienic assessment of the quality of objects of the environment in the territory located in the zone of the impact of emissions of ferrovanadium production (the city of Tchusovoy of the Perm Krai). From the results of the study there was established the significant excess of the vanadium content in the ambient air of the studied area in relation to the control territory and to the reference concentration for chronic inhalation exposure (RfCxp) to 6.0 times. There was revealed a significant excess of vanadium content in the blood of children residing in the study area, with respect to the regional background levels (0.0001-0.00016 mg/dm3). Complex clinical laboratory and chemical-analytical studies of biosubstrates of the children population allowed to substantiate the marker of the inhalation exposure (the vanadium content in the blood) and its reference level (0.0023 dm3).