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PubMedDate not supplied

Determination of inorganic arsenic in marine food samples by hydrochloric acid distillation and flow-injection hydride-generation atomic absorption spectrometry.

A simple, rapid, and reliable method was developed for determination of inorganic As in biological samples such as fish fillet. Inorganic AS was distilled from the sample as AsCl3 with HCl. The separated inorganic AS was determined by flow-injection hydride-generation atomic absorption spectrometry after prereduction with KI and HCl. The influences of various concentrations of KI, ascorbic acid, and HCl in the prereduction stage; NaBH4 as the reductant; and HCl as the carrier solution on analytical results were studied. Digestion was performed in a Kjeldahl digestion system for 75 min with 4 mL nitric acid and 1 mL sulfuric acid at 380 degrees C. The concentrations of inorganic As in samples were less than 0.1 mg/kg dry weight for fish fillet and somewhat higher for crustaceans and bivalve molluscs. The total and inorganic As contents of various marine biological samples and certified reference materials were determined.

Animals↗

PubMedDate not supplied

Optimized determination of calcium in grape juice, wines, and other alcoholic beverages by atomic absorption spectrometry.

This paper describes a study of the different methods of sample preparation for the determination of calcium in grape juice, wines, and other alcoholic beverages by flame atomic absorption spectrometry; results are also reported for the practical application of these methods to the analysis of commercial samples produced in Spain. The methods tested included dealcoholization, dry mineralization, and wet mineralization with heating by using different acids and/or mixtures of acids. The sensitivity, detection limit, accuracy, precision, and selectiviy of each method were established. Such research is necessary because of the better analytical indexes obtained after acid digestion of the sample, as recommended by the European Union, which advocates the direct method. In addition, although high-temperature mineralization with an HNO3-HCIO4 mixture gave the best analytical results, mineralization with nitric acid at 80 degrees C for 15 min gave the most satisfactory results in all cases, including those for wines with high levels of sugar and beverages with high alcoholic content. The results for table wines subjected to the latter treatment had an accuracy of 98.70-99.90%, a relative standard deviation of 2.46%, a detection limit of 19.0 microg/L, and a determination limit of 31.7 microg/L. The method was found to be sufficiently sensitive and selective. It was applied to the determination of Ca in grape juice, different types of wines, and beverages with high alcoholic content, all of which are produced and widely consumed in Spain. The values obtained for Ca were 90.00 +/- 20.40 mg/L in the grape juices, 82.30 +/- 23.80 mg/L in the white wines, 85.00 +/- 30.25 mg/L in the sweet wines, 84.92 +/- 23.11 mg/L in the red wines, 85.75 +/- 27.65 mg/L in the rosé wines, 9.51 +/- 6.65 mg/L in the brandies, 11.53 +/- 6.55 mg/L in the gin, 7.3 +/- 6.32 mg/L in the pacharán, and 8.41 +/- 4.85 mg/L in the anisettes. The method is therefore useful for routine analysis in the quality control of these beverages.

Alcoholic Beverages↗

PubMed2005

Lipoic acid decreases exhaled nitric oxide concentrations in anesthetized endotoxemic rats.

We recently demonstrated that lipoic acid suppresses endotoxin-stimulated expression of inducible nitric oxide synthase and nitric oxide production in mouse macrophages. In this study, we tested whether lipoic acid suppresses these inflammatory mediators in the lungs of rats. Rats were assigned to receive either no special treatment, endotoxin alone, or pretreatment with lipoic acid followed by endotoxin. After anesthetizing the rats and injecting them intraperitoneally with lipoic acid (100 mg/kg) at 4 h and again at 1 h before treatment, the rats then received either endotoxin (0.01 mg/kg) or its vehicle solution. Exhaled gas was sampled every 15 min and concentrations of nitric oxide in the samples were measured using a chemiluminescence analyzer. After 150 min of exposure to endotoxin, the lungs were harvested and snap-frozen in liquid nitrogen for subsequent analysis. Lipoic acid attenuated endotoxin-induced increases in exhaled nitric oxide concentrations (P<0.001) and iNOS (P<0.05). These findings support the hypothesis that lipoic acid inhibits endotoxin-stimulated formation of intrapulmonary nitric oxide.

Anesthesia↗

PubMedDate not supplied

Determination of arsenic, cadmium, cobalt, chromium, lead, molybdenum, nickel, and selenium in fertilizers by microwave digestion and inductively coupled plasma-optical emission spectrometry detection: collaborative study.

There is increasing regulatory interest in the non-nutritive metals content of fertilizer materials, but at present there is no consensus analytical method for acid digestion and instrument detection of those elements in fertilizer matrixes. This lack of method standardization has resulted in unacceptable variability of results between fertilizer laboratories performing metals analysis. A method has been developed using microwave digestion with nitric acid at 200 degrees C, followed by inductively coupled plasma-optical emission spectrometry instrument detection, for the elements arsenic, cadmium, cobalt, chromium, molybdenum, nickel, lead, and selenium. The method has been collaboratively studied, and statistical results are here reported. Fourteen collaborators were sent 62 sample materials in a blind duplicate design. Materials represented a broad cross section of fertilizer types, including phosphate ore, manufactured phosphate products, N-P-K blends, organic fertilizers, and micro-nutrient materials. As much as possible within the limit of the number of samples, materials were selected from different regions of the United States and the world. Limit of detection (LOD) was determined using synthetic fertilizers consisting of reagent grade chemicals with near zero levels of the non-nutritive elements, analyzed blindly. Samples with high iron content caused the most variability between laboratories. Most samples reasonably above LOD gave HorRat values within the range 0.5 to 2.0, indicating acceptable method performance according to AOAC guidelines for analyses in the mg/kg range. The method is recommended for AOAC Official First Action status.

Fertilizers↗

PubMed1999

Presence and distribution of arsenical species in beers.

The total content of arsenic and of its inorganic (As(III) and As(V)) and organic (monomethylarsonic acid, MMAA, and dimethylarsinic acid, DMAA) species were determined in a set of 21 alcoholic and alcohol-free beer samples using the technique of Hydride Generation Atomic Absorption Spectrometry. For total arsenic analysis, beer samples were dried and then microwave digested with nitric acid in polytetrafluoroethylene containers. For the speciation analysis, beers were previously subjected to ion exchange chromatography to elute the mentioned inorganic and organic arsenical species. Both microwave digestion and chromatographic separation methods were validated from certified reference materials and prepared standard solutions, respectively. The results obtained are presented in terms of the distribution and occurrence of arsenical species in the samples. The As levels of the beer samples were in the range of 1.5-12.4 micrograms/l. The influence of the production process for the alcohol-free beers in the speciation of arsenic is discussed. In alcoholic beers MMAA was the most abundant species, and for non-alcoholic beers inorganic As(III) was similar to the organic species. An estimated intake of total As of 0.47 microgram/person/day and 11.4 micrograms/person/day was obtained for average consumers and for heavy drinkers, respectively.

Alcohol Drinking↗

PubMed1977

Histochemical methods for proteins in tissues submitted to decalcification. A histophotometric study.

4 histochemical methods for demonstration of protein acid groups: Nihydrin-Schiff, Millon, DDD and DMAB-nitrite were applied in sections of rat pancreas fixed in the following fixatives: 10% formalin, 10% formalin in saline, ZENKER and GENDRE, and treated with the following decalcifying solutions: 5% nitric acid, 5% trichloroacetic acid, 33% formic acid, 5% EDTA pH=7.0 JENKINS, GREEP and citrate buffer pH=4.5. The colour intensity of each reaction was measured photometrically over the zymogen granules region of the acinar cells, using a Zeiss histophotometer, and compared with controls, i.e., fixed tissue which have not been treated with decalcifying solutions. The results have shown that for each method there is more than one combination fixative-decalcifying solution which gives results similar to those of the control.

Animals↗

PubMed2006

On-line preconcentration system using mini-column packed with a chelating resin for the characterization of seasonal variations of trace elements in seawater by ICP-MS and ICP-AES.

An on-line column preconcentration technique coupled with inductively coupled plasma-mass spectrometry (ICP-MS) and -atomic emission spectrometry (ICP-AES) was developed using a mini-column (ca. 3 mm i.d., 40 mm length), that was packed with chelating resin (0.2 g) of iminodiacetic acid groups, Muromac A-1. After the preconcentration step, the column was washed with ammonium acetate buffer (pH 5.5) and water to remove major elements, such as Ca and Mg, and then eluted with 4 ml of 2 mol l(-1) nitric acid. Eleven trace elements (Al, V, Fe, Co, Ni, Cu, Zn, Cd, Pb, Th and U) in seawater were determined by ICP-MS/AES. Recoveries for most of the elements tested were over 90%, although those for Al, V and Th were around 70%. The accuracy of the proposed method was evaluated by analyzing a standard reference material of seawater (NASS-4, NRC Canada). The values of Fe, Co, Ni, Cu, Zn, Cd and Pb obtained with the present method showed good agreement with the certified values as judged from the standard deviation. The method was successfully applied to characterize seasonal variations of trace elements in deep seawater (DSW) and surface seawater (SSW). In addition, no serious decrease in analytical performance of the present column system was observed during the experimental period of about 1 year.

Adsorption↗

PubMedDate not supplied

Colorimetric determination of total iodine in foods by iodide-catalyzed reduction of Ce+4.

An earlier acid digestion determination of iodine in foods was modified to provide an improved detection limit and to allow for the analysis of a greater variety and larger amounts of foods. The organic material in the sample was oxidized overnight by concentrated nitric acid, followed by digestion in a mixture of concentrated sulfuric and 70% perchloric acid. The iodine was determined by an automated colorimetric method based on the iodide-catalyzed reduction of Ce+4 by As+3. The method had an average relative standard deviation of 3.1% for the samples analyzed, and a detection limit of 0.1 ng/mL in the digested solution and 5 ng/g in a 2 g sample prior to digestion. The recovery of added iodine ranged from 90.3 to 101.3%, using external standards. Samples analyzed included NBS Standard Reference Material 1549, and composites of a variety of dairy products, meat, eggs and fish, cereals, and potatoes. The iodine detected in these samples ranged from 9 ng/g for the potato group to 3360 ng/g for the standard reference material.

Autoanalysis↗

PubMed1996

Synthesis of indole-N-propionitrile, N-propionic acid and N-propionamide derivatives of potential biological activity.

Cyanoethylation of a selected group from indole derivatives 1a-i using acrylonitrile in pyridine and potassium hydroxide afforded the corresponding indole-1-propionitrile 2a-h derivatives which readily hydrolysed in basic medium to the corresponding indole-1-propionic acid derivatives 4a-c. A several attempts to cyclize the above products-to the corresponding dihydropyrroloquinolin-4-one derivatives. The indole-1-propionamide derivatives reacted with diethyl carbonate and ethyl acetate under Claisen condensation also, with diazonium salts and nitric acid.

Amides↗

PubMed1985

Germinability of coat-lacking spores of Bacillus megaterium.

Upon treatment with acid, the germinability of both intact and coat-lacking spores of Bacillus megaterium ATCC 19213 exhibited similar features. Namely, when the spores previously germinated by alanine in the presence of phosphate buffer were converted to H-spores by treatment with nitric acid, germination proceeded at a very low speed in a same germination medium. When H-spores converted to Ca-spores by treatment with calcium acetate and subsequently germinated, germination proceeded at a speed higher than that of native spores and occurred even in the absence of buffer. These results suggest that the site of exchangeable cations concerned with germinability must not exist in the coat.

Alanine↗

PubMed2002

Effect of surface chemistries and characteristics of Ti6Al4V on the Ca and P adsorption and ion dissolution in Hank's ethylene diamine tetra-acetic acid solution.

This study examined the influence of chemistries and surface characteristics of Ti6Al4V on the adsorption of Ca and P species and ion dissolution behavior of the material exposed in Hank's solution with 8.0 mM ethylene diamine tetra-acetic acid at 37 degrees C. The variation of chemistries of the alloy and nano-surface characteristics (chemistries of nano-surface oxides, amphoteric OH group adsorbed on oxides, and oxide thickness) was effected by surface modification and three passivation methods (34% nitric acid passivation. 400 degrees C heated in air, and aged in 100 degrees C water). X-ray photoelectron spectroscopy and Auger electron spectroscopy were used for surface analyses. The chemistries of nano-surface oxides in a range studied should not change the capability of Ca and P adsorption. Nor is the capability affected significantly by amphoteric OH group and oxide thickness. However, passivations influence the surface oxide thickness and the early stage ion dissolution rate of the alloy. The rate-limiting step of the rate can be best explained by metal-ion transport through the oxide film, rather than hydrolysis of the film. Variation of the chemistries of titanium alloy alters the electromotive force potential of the metal, thereby affecting the corrosion and ion dissolution rate.

Adsorption↗

PubMed2003

Factors affecting gallbladder motility: drugs.

Various drugs and medications that inhibit or stimulate gallbladder contraction and basal tone in humans are described. Active gallbladder contraction may be achieved using synthetic hormones such as cholecystokinin, caerulein and motilin, cholinomimetic drugs such as bethanecol, prostigmine, and erythromycin due to its motilin-like effect. Furthermore, cisapride and cholestyramine, may have some excitatory activity on the gallbladder muscle. Intravenous amino acids also induce gallbladder contraction through the release of cholecystokinin. Inhibition of gallbladder contraction induced by a meal, or reduction of the basal fasting tone may be achieved by using atropine and other cholinergics, and by inhibitory hormones such as somatostatin, the nitric acid releaser arginine, the calcium channel antagonist nifedipine, and progesterone. Other drugs such as trimebutine, loperamide and ondansetron may negatively affect gallbladder contraction.

Gallbladder↗

PubMed1991

Free radical-induced carbonyl content in protein of estrogen-treated hamsters assayed by sodium boro[3H]hydride reduction.

Oxidative damage to proteins is known to occur via conversion of side chain amino groups to corresponding carbonyl derivatives. Such damage to enzymes and purified proteins has been quantified previously by reduction with sodium boro[3H]hydride and subsequent measurement of the incorporation of 3H into amino acid fractions. In this study, the NaB3H4 reduction assay was modified to permit the quantitation of free radical-mediated oxidative damage to proteins obtained from animals. Modifications included additional extractions of protein isolates with organic solvents to remove lipids and with nitric acid to remove metal ions. The modified assay has first been validated in vitro by measuring changes in levels of oxidative damage to bovine serum albumin exposed to xanthine plus xanthine oxidase (2-fold increase), to hydrogen peroxide and iron(II) sulfate (5-fold increase), or to gamma radiation (30-fold increase over controls, respectively). gamma radiation of isolated hamster kidney protein also raised the carbonyl content in a dose-dependent manner. The modified assay has then been validated in vivo by measuring the changes in oxidative damage to lung tissue in animals exposed to approximately 85% oxygen (2-fold increase) or to different doses of paraquat (5-fold increase with the high dose over controls, respectively). The assay was then used to examine free radical-mediated oxidation introduced by acute or chronic treatment of hamsters with estrogens, since both synthetic and natural estrogens induce kidney tumors in this species. Priming of hamsters for 3 days with 20 mg/kg/day diethylstilbestrol and treatment with 100 mg/kg of this drug on the 4th day resulted in a 160% increase in free radical modification of renal proteins. Oxidative damage to kidney proteins was also assayed in hamsters treated with estradiol implants for up to 7 months, a regimen known to induce kidney tumors. Significant increases in covalent oxidative modification to renal proteins over values in age-matched controls were detected after 1, 2, and 7 months of continuous estradiol exposure. It is concluded that the modification of the NaB3H4 reduction assay is a useful postlabeling method for monitoring free radical action in vivo. Furthermore, it is postulated that free radical damage in estrogen-treated hamster kidney plays a role in estrogen-induced carcinogenesis.

Animals↗

PubMed2001

Molybdenum determination in iron matrices by ICP-AES after separation and preconcentration using polyurethane foam.

A procedure is proposed for the separation and determination of molybdenum in iron matrices by a batch process. It is based on the solid-phase extraction of the molybdenum(V) ion as thiocyanate complex on polyurethane (PU) foam. The extraction parameters were optimized. Using 0.20 mol L-1 hydrochloric acid, a thiocyanate concentration of 0.10 mol L-1, 100 mg of polyurethane foam and shaking time of 10 min, molybdenum (5-400 micrograms) can be separated and preconcentrated from large amounts of iron (10 mg). Desorption was carried out instantaneously by conc. nitric acid or acetone. Distribution coefficients, sorption capacity of the PU foam and coefficients of variation were also evaluated. The effect of some ions on the separation procedure was assessed. Iron(III) should be reduced to iron(II). The proposed procedure was used to determine molybdenum in standard iron matrices such as steel and pure iron. The achieved results did not show significant differences with certified values.

Journal Article↗

PubMed2005

Suppression of arachidonic acid metabolism and nitric oxide formation by kudzu isoflavones in murine macrophages.

Inhibitory effect of kudzu isoflavones on arachidonic acid metabolism and nitric oxide (NO) production in lipopolysaccharide activated RAW 264.7 macrophages were investigated. Isoflavone aglycones, such as daidzein, genistein, biochanin A, and formononetin significantly suppressed arachidonic acid release (50 microM). Biochanin A, which displayed the most active inhibition on arachidonic acid release in HT-29 human colon cancer cells, exhibited its most potent suppression in RAW 264.7 cell (by 86%) without showing cytotoxicity. However, isoflavone glucosides, puerarin and daidzin, showed lower inhibitory activities on the release of arachidonic acid and its metabolites. In NO formation, biochanin A showed marked inhibition, by 62% (50 microM), followed by genistein, daidzein, formononetin, and daidzin, 56, 39, 33, and 8%, respectively. 5,7-Dihydroxyl group in the A-ring of isoflavones could be a key functional group responsible for the strong inhibitory activity of biochanin A and genistein on NO production. These activities may contribute to the antiinflammatory and anticarcinogenic properties of kudzu isoflavones.

Animals↗

PubMed2005

15-hydroxyeicosatetraenoic acid depressed endothelial nitric oxide synthase activity in pulmonary artery.

15-hydroxyeicosatetraenoic acid (15-HETE) plays an important role in hypoxia-induced pulmonary vasoconstriction. Release of nitric oxide (NO) is apparently decreased and activity of endothelial nitric oxide synthase (eNOS) is impaired in chronic hypoxia. However, little is known whether 15-HETE contributes to eNOS/NO pathway in the constriction induced by 15-HETE. We examined the response of rat pulmonary artery (PA) rings to 15-HETE, the production of NO, total eNOS expression and the phosphorylation of eNOS in bovine pulmonary artery endothelial cells (BPAECs) stimulated by 15-HETE. Rat PA rings were divided into three groups: endothelium intact group, endothelium denuded group, and nitro-L-arginine methyl ester (L-NAME, 0.1 mmol/L, an inhibitor of eNOS) group. Constrictions to 15-HETE were significantly enhanced in endothelium denuded group and L-NAME group (both P< 0.05 vs endothelium intact group, n= 9); BPAECs were incubated in different conditions to test nitrite production by Greiss method. Nitrite production was significantly reduced by 1 mumol/L 15-HETE (P<0.05), and increased by the lipoxygenase inhibitors, 10 mumol/L cinnamyl 3,4- dihydroxy-[alpha] -cyanocinnamate (CDC, P< 0.05) and 0.1 mmol/L nordihydroguiairetic acid (NDGA, P< 0.01 ); Western blot analysis of extracts from BPAECs incubated with 15-HETE in different time was carried out to test total eNOS expression, and the expression was changed unobviously. Immunoprecipitation (IP) and Western blot analysis of cell extracts from BPAECs treated with 2 mumol/L 15-HETE in different length of time were accomplished, using phospo-eNOS-threonine 495 (Thr495, an inhibitory site) antibody for IP, and eNOS or 15-lipoxygenase (15-LO) antibodies for Western blot. 15-HETE depressed eNOS activity by increasing the levels of phospho-eNOS-Thr 495. The data suggest that eNOS/NO pathway is involved in PA constrictions induced by 15-HETE and that 15-HETE depresses eNOS activity by phosphorylation in Thr495 site. The protein interaction between phospho-eNOS (Thr495) and 15-LO is discovered for the first time.

Animals↗

PubMed2001

Ion exchange chromatographic determination of olpadronate, phosphate, phosphite, chloride and methanesulfonic acid.

A method based on single column ion chromatography with UV detection was developed for purity testing and assay of monosodium olpadronate. The analyte aqueous solution is precipitated with methanol to enhance the impurities/olpadronate molar ratio, thus improving purity determination at trace levels. The resulting solution is injected into a standard chromatographic system with UV detector in indirect mode with a Waters IC Pak HR column using diluted nitric acid as the mobile phase. The method was fully validated according to ICH guidelines for the determination of phosphite, phosphate, chloride and methanesulfonic acid in olpadronate being suitable for purity testing and assay.

Chlorides↗

PubMed1994

Identification of covalently bound amino-terminal myristic acid in endothelial nitric oxide synthase.

Endothelial nitric oxide synthase (eNOS) is unique among the nitric oxide synthase family of proteins due to the presence of an N-myristoylation consensus sequence elucidated from the cloning of its cDNA. Although eNOS was metabolically labeled with [3H]myristic acid and mutation of glycine 2 in the N-myristoylation consensus sequence changed the particulate localization of the enzyme to a cytosolic form, the definitive characterization of eNOS as an N-myristoylprotein has not been demonstrated. Therefore, the purpose of the present study was to determine the nature of the fatty acid incorporated into eNOS. Wild-type or G2A mutant (mutation of glycine 2, the myristic acid acceptor site, to alanine) eNOS-transfected COS cells and bovine aortic endothelial cells (BAEC) were metabolically labeled with [3H]myristic acid for 5 h. The radiolabel was primarily incorporated into membrane-associated eNOS from wild-type transfected COS cells and cultured BAEC but not into the mutant eNOS from G2A-transfected COS cells. Qualitatively similar amounts of immunoreactive protein were found in wild-type and G2A-transfected cells. In addition, linkage of the radiolabel to eNOS was insensitive to hydroxylamine treatment, and incorporation of the radiolabel into eNOS was abolished by cyclo-heximide. Chemical analysis of the fatty acid released by acid methanolysis of labeled eNOS verified the 3H-labeled fatty acid as protein-bound myristic acid. These results unequivocally demonstrate that eNOS incorporates myristic acid via an amide linkage with the amino-terminal glycine of the enzyme as a co-translational modification.

Alanine↗