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Comprehensive analytical procedures for the determination of volatile and non-volatile, polar and non-polar N-nitroso compounds.

The first comprehensive analytical procedures for the quantitative analysis of N-nitroso compounds are described. The scheme divides N-nitroso compounds into four major, overlapping categories: volatile (Class I), non-volatile, low polarity (Class II), non-volatile, non-ionic, high polarity (Class III) and non-volatile, ionic, high polarity (Class IV). Existing analytical techniques for each class of compound are integrated into an organized and logical sequence of analysis to allow all classes of compounds to be determined. TEA-GC is used for the volatile compounds and TEA-HPLC for the non-volatile. It is emphasized that the coincidence of retention time in either TEA-GC alone or TEA-HPLC alone cannot be taken as sufficient evidence for the identification of N-nitroso compounds, especially for samples from complex matrices. Independent techniques are required to confirm these results. The confirmatory techniques used frequently in our laboratory are: (1) spectroscopic analysis (IR, NMR, UV and MS), (2) formation and identification of derivatives, and (3) parallel TEA-GC/TEA-HPLC techniques. These procedures are now used at Thermo Electron for the comprehensive screening of environmental samples.

Chemical Phenomena

Production of volatiles from decomposing plant tissues and effect of these volatiles on Rhizoctonia solani in culture.

Volatiles, of which NH3 is a major component, were evolved from decomposing immature corn tissue (c:n9) and affected R. solani in culture two ways: they supplied additional nitrogen to the growth medium so that fungal mycelial growth increased; and they raised substrate pH from 5.5 to 8.2 which induced melanization of mycelium. Volatiles increased fungus growth and pigmentation within 2 weeks of amendment addition to soil. Increases were concomitant with NH3 production from corn tissue. More NH3 evolved from decomposing corn tissues of C:N9 and 17 than from those of C-N 33 and 81. More growth and pigmentation occurred in flasks through which volatiles from decomposing corn (C:N9) were passed than in flasks through which volatiles from nonamended soil or decomposing corn (C:N81) were passed. Carbon dioxide from decomposing tissues did not affect growth or pigmentation. Twice as much NH3 evolved from corn tissue (C:N9) which decomposed in saturated soil than from tissue which decomposed in soil at 50% of its water-holding capacity. Pigment production doubled under saturated conditions.

Ammonia

Recent methods for the determination of volatile and non-volatile organic compounds in natural and purified drinking water.

Four analytical protocols for the extraction and preconcentration of organic residues in natural or purified drinking water were investigated and compared: closed loop stripping analysis; simultaneous extraction-distillation; purge and trap analysis; continuous liquid-liquid extraction. Organic extracts were submitted to a variety of separation and identification techniques. Volatiles were determined by conventional capillary column gas chromatography with tandem mass spectrometry, using triple-stage quadrupole instruments. Non-volatile and thermally labile molecules were investigated by several different techniques (high-temperature gas chromatography, capillary column supercritical fluid chromatography, pyrolysis gas chromatography-mass spectrometry, thermospray liquid chromatography with tandem mass spectrometry and conventional fast-atom bombardment with tandem mass spectrometry). Several samples recently examined in the laboratory provide examples of this multitechnique approach for a more complete knowledge of the organic carbon distribution in water-dissolved organic matter, taking into account organic substances with widely different volatilities, polarities and thermal stabilities.

Chlorides

[Effects of processing methods on the amounts of volatile oil of nutmeg and on isolation and characterization of the volatile oil constituents].

In this paper, the authors investigated the effects of various processing methods, i.e., scalding in hot purified talc, simmering wrapped in flour in hot purified talc and stir-frying in smoking wheat bran, on nutmeg (Semen Myristicae) in terms of the quantities of the volatile oil. The experimental results revealed that the amounts of volatile oil contained in nutmeg vary remarkably with the lengths of cooking time and the fluctuation of temperature. Detected by GC-MS-computer, 32 compounds of nutmeg were characterized, and their contents were determined by GC respectively.

Allylbenzene Derivatives

Recent studies in Canada on the analysis and occurrence of volatile and non-volatile N-nitroso compounds in foods.

The present paper briefly summarizes the research activities of the Food Research Laboratories, Canadian Health Protection Branch, during the past two years. Data presented indicate that traces of nitrosamines, mainly nitrosodimethylamine and nitrosodiethylamine, can occur in cheese. Several new analytical methods are presented for the determination of non-volatile nitroso compounds, such as nitrosohydroxypyrrolidine, nitrosodiethanolamine, nitrosoproline, nitrosohydroxyproline and nitrosoglyphosphate (a nitrosated herbicide). It is shown that all of the above compounds, even the very polar nitrosoglyphosphate, can be converted to volatile derivatives and then analysed by GLC-MS or by conventional GLC using a specific (such as P-photometric) detector. Traces of nitrosohydroxypyrrolidine have been detected in a few samples of cooked bacon. Contrary to previously reported findings, no significant levels of nitrosoproline were detected in the 6 samples of raw bacon analysed.

Animals

Infrared analysis of volatile anesthetics: impact of monitor agent setting, volatile mixtures, and alcohol.

Infrared analysis can determine exhaled concentrations of the three volatile anesthetics in common use because each absorbs infrared light. Many infrared analyzers use a single source of infrared light at a wavelength of 3.3 microns for measurements of all three agents but cannot identify which agent is in use. Organic gases such as ethanol also absorb infrared light. This study determined the effects on the accuracy of a single-wavelength infrared anesthetic monitor (Capnomac, Puritan-Bennett PB254) of (1) incorrect anesthetic agent setting, (2) mixtures of volatile anesthetics, and (3) ethanol vapor in the analyzed gas. Changing the agent setting on the monitor during steady-state delivery of an agent resulted in readings for the halothane setting five times higher than those for the enflurane setting, and six times higher than those for the isoflurane setting. These ratios reflect the monitor's fixed internal gain for each agent setting. Mixtures of anesthetics had a simple additive effect on the monitor's accuracy. With the monitor set for halothane, 0.2 vol% isoflurane mixed with halothane caused readings 1.2 vol% higher than the true halothane concentration. Conversely, with the monitor set for isoflurane, 1 vol% halothane mixed with isoflurane resulted in readings 0.2 vol% too high. In a model simulating alveolar gas, ethanol vapor corresponding to blood alcohol levels of 0.10, 0.30, and 0.50% had a slight but not clinically significant effect on readings for enflurane and isoflurane but increased readings with the halothane setting 3.5 times the corresponding level of blood alcohol. Clinicians can test for an interfering gas such as ethanol before induction by checking the reading in the halothane setting during preoxygenation.

Anesthesia, Inhalation

Sonochemistry of volatile and non-volatile solutes in aqueous solutions: e.p.r. and spin trapping studies.

Recent spin trapping studies of the free radical intermediates generated by the sonolysis of aqueous solutions are reviewed. Studies of rare gas saturated solutions of volatile solutes (e.g., methanol and ethanol) and of non-volatile solutes (acetate, amino acids, sugars, pyrimidines, nucleotides and surfactants) are consistent with the theory of three reaction zones in aqueous sonochemistry. The very high temperatures and pressures induced by acoustic cavitation in collapsing gas bubbles in aqueous solutions lead to the thermal dissociation of water vapour into hydrogen atoms and hydroxyl radicals. Reactions take place in the gas phase (pyrolysis reactions), in the region of the gas-liquid interface, and in the bulk of the solution at ambient temperature (similar to radiation chemistry reactions). By use of the rare gases with different thermal conductivities, the contributions of individual reaction steps with widely different energies of activation can be evaluated.

Alcohols

Effects of two volatile anesthetics and a volatile convulsant on the excitatory and inhibitory amino acid responses in dissociated CNS neurons of the rat.

1. Effects of two volatile anesthetics [halothane (Hal) and enflurane (Enf)] and a volatile convulsant [hexafluorodiethyl ether (HFE)] on amino acid-induced membrane currents in neurons dissociated from the nucleus tractus solitarius of the rat were examined. The dissociated neurons were voltage clamped in the whole-cell mode of the patch-clamp technique. All drugs were applied with a microperfusion system, termed the "Y-tube" method. 2. The glutamate (Glu)-induced excitatory response was slightly reduced by both the anesthetics. The responses to three agonists at Glu receptor were depressed by Hal (10(-3) M) in the rank order of quisqualate greater than N-methyl-D-aspartate greater than kainate. HFE slightly increased the Glu response at a high concentration of 2 x 10(-3) M. 3. The gamma-aminobutyric acid (GABA)-induced chloride current (ICl) was enhanced by both anesthetics. The dissociation constant (Kd) for the enhancement was 2.3 x 10(-4) M for Hal and 2.1 x 10(-4) M for Enf, and the Hill coefficient was 1.6 for Hal and 1.5 for Enf. HFE depressed the GABA response with a Kd of 8.7 x 10(-5) M and a Hill coefficient of 0.84. 4. Hal (10(-3) M) and Enf (10(-3) M) decreased the Kd of the GABA concentration-response curve from 3.5 x 10(-6) to 10(-6) and 1.9 x 10(-6) M, respectively, without changing the maximum response or the Hill coefficient (1.5). In the presence of HFE (10(-4) M), the Kd was increased to 1.4 x 10(-5) M and the Hill coefficient was slightly changed to 1.2.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids

The adsorption of volatile aroma constituents by foods. VIII. Adsorption of volatile carbonyl compounds by amino acids.

The adsorption of some volatile aldehydes and ketones by 22 dry amino-acids was studied. The adsorbed amounts decreased approximately in the order: propanal, biacetyl, hexenal, hexanal, acetone, pentan-3-one, nonan-3-one, undecanal and nonan-5-one, but there were great differences in the adsorption capacity of the amino acids. Lys adsorbed best and often irreversible, arg, his, phe, trp, pro, and val (leu, ile) very good. With cys, thiazolidinecarboxylic acids were formed. In most cases of adsorption exceeding 100 mmol/mol, except for the monoketones, strong discoloration, probably of the maillard type, was observed. Propanal and hexanal were to a great extend converted to aldol addition and condensation products. Tyr and hyp adsorbed less than the corresponding amino-acids without hydroxyl groups. This is attributed to stronger crystal-lattice forces.

Adsorption

Volatile, non-volatile and total N-nitroso compounds in bacon.

Twenty-five smoked and unsmoked fried bacon samples have been analysed by a group selective procedure to measure the concentration of apparent total N-nitroso compounds (ATNC). The levels of a range of individual N-nitroso compounds, including simple volatile N-nitrosamines, N-nitrosothiazolidines, N-nitrosamino acids and N-nitrosothiazolidine carboxylic acids have also been examined. Concentrations of ATNC varied from 430 to 6800 micrograms(N-NO)/kg with a mean of 2700 micrograms(N-NO)/kg. Protein-bound N-nitrosoproline was the most abundant compound detected in unsmoked bacon, mean 260 micrograms/kg, and on average accounted for 4% of the ATNC concentration. For smoked bacon, bound N-nitrosoproline was detected in levels of up to 890 micrograms/kg and contributed 5% to the ATNC total. The most abundant compound present in smoked bacon was N-nitrosothiazolidine-4-carboxylic acid, mean 660 micrograms/kg, and this accounted for 6% of the ATNC. N-Nitrosothiazolidine, mean 340 micrograms/kg, and 2-(hydroxymethyl)-3-nitrosothiazolidine-4-carboxylic acid, mean 180 micrograms/kg, were the next most prominent compounds detected in smoked bacon. The combined sum of all the individual N-nitroso compounds measured accounted for, on average, 16% of the total ATNC. The identities of the N-nitroso compounds comprising the majority of the ATNC in bacon remain unknown.

Animals

Volatile fatty acid metabolism in sheep. 1. Average daily volatile fatty acid production in the rumen of sheep fed lucerne hay.

Changes in the total concentration of the volatile fatty acid (VFA) pool in the rumen were followed over a 24 hour period in 2 groups of sheep, 1 fed at 08h00 and the other twice daily at 08h00 and 20h00. Although similar maximum (143 and 147 meq/1) and average (100, 3 and 102, 1 meq/1) levels were found in the 12 and 24 h groups respectively, the twice daily feeding regimen resulted in a lower variation (S.D.=17, 0 meq/1 and 28, 9 meq/1 respectively). It was concluded from changes in the percentage molar composition of the VFA pool over the same period that the other of VFA absorption from the rumen was propionate greater than acetate greater than butyrate for both groups, but that the differences were less marked for the twice daily fed sheep. Long term infusions of 14C labelled acetic, propionic and butyric acids into the rumen of sheep fed a total of 1 600 g lucerne hay twice daily (08h00 and 20h00), gave an average net total VFA production rate of 4,52+/-1,01 moles/800 g/12 hours irrespective of the acid infused. The net individual turnover rates for acetic (2,81 moles/12 h), propionic acid (0,82 moles/12 h) and butyric acid (0,55 moles/12 h), derived by the subtraction of the inter-conversion factors from the gross production rates of the acids, and expressed as the percentage contribution of each acid to the total net VFA turnover (acetic=62%, propionic=18% and butyric=12%) closely resembled the percentage molar composition of the VFA pool in the rumen (acetic=60%, propionic=23% and butyric=12%). The total net VFA production was found to be directly proportional to the total VFA concentration in the rumen (correlation coefficient=0,83), and the relationships can be described by the equation y=0,034 x + 0,16 where y=VFA production in moles/12 hour and x=VFA concentration in meq/1. A specific VFA production rate of 0,85 moles per 100 g digestible organic matter was calculated from the average daily VFA production rate and the composition of the lucerne hay.

Acetates

Gas chromatographic determination of volatile anaesthetic agents in blood. Part 1. Preparation of standard gas mixtures of volatile anaesthetic agents.

A method for preparing standard gas mixtures of the volatile anaesthetics halothane, enflurane and isoflurane is described. Static mixtures of gases of known concentration can be prepared manometrically by measuring the required pressure of anaesthetic gas into a bulb and diluting to atmospheric pressure with air. Standard gas mixtures in the concentration range 0-4% V/V can be prepared with an accuracy of +/- 0.01% V/V, and the relative standard error of measurements of a single standard concentration is less than 0.8%. Significant adsorptive losses in the gas sampling valve were observed for gas standards prepared in the absence of any diluent gas. These losses were not detected for measurements of standards made up to atmospheric pressure in air. A comparison with calibration procedures currently in practice is presented.

Anesthetics

[Rapid preliminary diagnosis of anaerobic bacterial infections. II. Investigation of the presence of volatile and non-volatile fatty acids (C1-C6) in pus, purulent fluids and dressing by application of gas chromatography].

Studies were performed in the years 1986-1990 and dealing with 405 clinical materials originating from infections within the abdominal cavity, maxilla, reproductive organ, post-operative wounds as well as dressings, extremities, blood, bile and other types of material and usefulness of analysis by gas chromatography of presence of lower fatty acids in materials for rapid and preliminary diagnostics with strictly anaerobic bacteria was confirmed. Presence in chromatograms of isobutyric, butyric isovaleric, valeric, isocaproic and caproic acid, and partly of propionic acid, is a good indication of infection with strictly anaerobic bacteria. Moreover, presence of succinic acid may suggest presence of anaerobic infection, similarly as presence in chromatograms of three or more various lower fatty acids. Rapid, approximative diagnosis of anaerobic bacteria by application of gas chromatography is worth of broader application. Investigation of clinical material by this method lasts only 1-2 hours and obtained results my present a basis for preliminary diagnosis of the etiological factor and for direction of chemotherapy, which is very important in most early period of treatment of infections.

Bacteria, Anaerobic

[Volatile ashes and their biological effect. 2. Fibrogenic effect of volatile ashes].

In experiments on white Wistar rats fibrogenic effects of 6 samples of fly-ashes collected from electric precipitators in power engineering plants have been evaluated. The coal came from different national deposits. All the ashes have been found to contain: quartz and mullite, 3 ashes contained additionally orthoclase, whereas 1, apart from quartz and mullite, contained kaolinite; naturally radioactive elements (Ra226, K40, Th228) and trace elements (As, Ba, Be, Cd, Ce, Cu, Fe, Pa, Mo, Ni, Pb, Se, U Zu). Experimental pneumoconiosis was induced through intratracheal administration of single doses of 50 mg of dust; the experiment was carried out at 3 time intervals of 3, 6 and 9 months. The fibrogenic activity was evaluated both qualitatively (histopathological methods) and quantitatively (lung weight, hydroxyproline content in lungs, dust elimination from lungs); control groups consisted of animals which obtained NaCl solution and quartz sands. Fly-ashes were found to exhibit different fibrogenic effects, yet, their fibrogenic activity was weaker, compared to quartz sands. No clear correlation was found between fibrogenic effects of ashes and test physico-chemical properties, such as the content of SiO2, trace elements or naturally radioactive elements. Analysis of occupational diseases (for the period section): (1979-1983) demonstrated occupational diseases of dust-related aetiology among power engineering workers, pneumoconioses, constituting 7.8% of 127 cases of occupational diseases.

Animals