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PubMed1987

Deproteinizing methods evaluated for determination of uric acid in serum by reversed-phase liquid chromatography with ultraviolet detection.

We evaluated six deproteinizing methods for determination of uric acid in serum by "high-performance" liquid chromatography with ultraviolet detection: those involving zinc hydroxide, sodium tungstate, trichloroacetic acid, perchloric acid, acetonitrile, and centrifugal ultrafiltration (with Amicon MPS-1 devices). We used a Toyosoda ODS-120A reversed-phase column. The mobile phase was sodium phosphate buffer (40 mmol/L, pH 2.2) containing 20 mL of methanol per liter. Absorbance of the eluate was monitored at 284 nm. The precipitation method with perchloric acid gave high recoveries of uric acid and good precision, and results agreed with those by the uricase-catalase method of Kageyama (Clin Chim Acta 1971;31:421-6).

Chemical Precipitation↗
PubMed1996

Production oriented method for the determination of rare earth and other trace elements in rocks by ICP-MS.

The rare earth elements along with Y, Sc, Co, Hf, Nb, Rb, Ta, Th, Ti, U, V and Zr have been determined in ten standard reference materials and ten granitoid samples by inductively coupled plasma mass spectrometry (ICP-MS). Two digestion methods have been used: (a) dissolution with hydrofluoric-perchloric acid followed by fusion of the residue with lithium metaborate, (b) dissolution with hydrofluoric acid-perchloric acid followed by fusion with lithium metaboratesodium perborate and addition of hydrogen peroxide. An evaluation of the two decomposition methods and a comparison of the ICP-MS and X-ray fluorescence results have been carried out. In comparing the two digestion methods, method (b) gave more accurate results for Nb and Ta, and it has been applied to a rock geochemistry project covering whole Finland (about 7000 samples). The statistical stability of the method has been monitored by frequent analyses of reference material SDC-1 (Mica Schist, USGS).

Journal Article↗
PubMed1990

Characterization of a reversed-phase high-performance liquid chromatographic system for the determination of blood amino acids.

High-performance liquid chromatography was used to separate physiological amino acids in perchloric acid supernatants of blood samples. Precolumn derivatization with phenyl isothiocyanate was carried out, starting with 20 microliters of supernatant; 2-10 microliters were injected into a 30-cm Pico Tag column, which was eluted with a gradient of two eluents in 64 min. Stock amino acid solutions prepared in water, hydrochloric acid or perchloric acid showed comparable recoveries on serial dilution (parallelism test). The recovery of crystalline amino acids added to blood in amounts ranging from normal to six times normal was generally satisfactory. The within-assay relative standard deviations were less than 5% for many amino acids. The performance of the system was less than satisfactory for cysteine and methionine. Glutamine and asparagine are interconverted into glutamate and aspartate, respectively, in a time-dependent fashion; a separate measurement of one member of the pair is therefore required in order to assay the other starting from the sum of both chromatographic peaks. The method is suitable for the relatively rapid, sensitive and accurate measurement of blood amino acids in perchloric acid supernatants (in which other relevant metabolites are customarily assayed) over a wide range of physiological concentrations, on very small amounts of sample.

Amino Acids↗
PubMed1999

Analysis of the fatty acid components in a perchloric acid-soluble protein.

We had previously found that a perchloric acid-soluble protein (PSP1) occurs in rat liver, and that this novel protein inhibits protein synthesis in a rabbit reticulocyte lysate system (T. Oka, H. Tsuji, C. Noda, K. Sakai, Y.-H. Hong, I. Suzuki, S. Muñoz, Y. Natori, J. Biol. Chem. 270 (1995) 30060-30067). In the present study, we analyzed lipid components bound to PSP1. Native PSP1 was purified from rat liver using Sephadex G-75, DE-52 cellulose and IgGPSP-affinity chromatography, and the lipid components were extracted. The components obtained from the purified PSP1 were shown to be free fatty acids by thin-layer chromatography. By GC-MS, six major fatty acids were identified as 14:0, 16:0, 18:0, 18:1, 18:2 and 20:4. 1 mol of PSP1 contained 1.26 mol of total fatty acid components. The fatty acid-binding assay of PSP1 showed that the Bmax was 1.25 mol fatty acid/mol PSP1 and the Kd value for palmitic acid was 6.03 microM. The concentration of PSP1 mRNA in rat liver increased 2.3-fold by the administration of peroxisome proliferator, bezafibrate. These findings show that PSP1 is a fatty acid-binding protein-like protein, which is involved in the intracellular metabolism of fatty acid and is quite different from the known fatty acid-binding proteins.

Animals↗
PubMed1990

Mechanism of acid-induced folding of proteins.

We have previously shown [Goto, Y., Calciano, L. J., & Fink, A. L. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 573-577] that beta-lactamase, cytochrome c, and apomyoglobin are maximally unfolded at pH 2 under conditions of low ionic strength, but a further decrease in pH, by increasing the concentration of HCl, refolds the proteins to the A state with properties similar to those of a molten globule state. To understand the mechanism of acid-induced refolding of protein structure, we studied the effects of various strong acids and their neutral salts on the acid-unfolded states of ferricytochrome c and apomyoglobin. The conformational transition of cytochrome c was monitored at 20 degrees C by using changes in the far-UV CD and in the Soret absorption at 394 nm, and that of apomyoglobin was monitored by changes in the far-UV CD. Various strong acids (i.e., sulfuric acid, perchloric acid, nitric acid, trichloroacetic acid, and trifluoroacetic acid) refolded the acid-unfolded cytochrome c and apomyoglobin to the A states as was the case with HCl. For both proteins neutral salts of these acids caused similar conformational transitions, confirming that the anions are responsible for bringing about the transition. The order of effectiveness of anions was shown to be ferricyanide greater than ferrocyanide greater than sulfate greater than thiocyanate greater than perchlorate greater than iodide greater than nitrate greater than trifluoroacetate greater than bromide greater than chloride.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Observations on the quantitation of the phosphate content of peptides by fast-atom bombardment mass spectrometry.

Equimolar mixtures of the phosphorylated and dephosphorylated forms of several peptides have been subjected to fast-atom bombardment mass spectrometry (FABMS), to investigate whether the stoichiometry of phosphorylation can be determined from the relative molecular-ion abundances of the phospho and dephospho derivatives. It is concluded that quantitation can be achieved for peptides with large positive or negative hydrophobicity/hydrophilicity indices (delta F values) where addition of a phosphate group does not alter the distribution of the peptide within the matrix significantly. For peptides with small positive or negative delta F values, phosphopeptides tend to be partially suppressed by their dephosphorylated counterparts. Suppression can be partially or totally overcome by conversion of the peptide to a hydrophobic derivative, and by the selection of an appropriate matrix. Alternatively, addition of a very strong acid, perchloric acid, can even reverse the original suppression effect. This last effect is believed to be due to the increased ionic strength in the matrix, which forces a relatively hydrophilic analyte to the matrix surface; and the ability of such a phosphorylated analyte to form a more stable gas-phase cation.

Chemical Phenomena↗
PubMed1985

Simple fluorimetric determination of selenium in food and biological materials. A validation study.

A method for the determination of selenium in food and biological materials has been developed. The method involves sample decomposition in a closed system at 150 degrees C with nitric acid, perchloric acid treatment of the digest, reduction of selenium to the tetravalent state with hydrochloric acid followed by Se-2,3-diaminonaphthalene (DAN) complex formation and extraction in a single-step operation. Performance characteristics of the method (accuracy and repeatability) were extensively tested for eleven reference materials and ten practice samples covering both a wide variety of matrices and a wide range of selenium content. Almost all results for reference materials were within the 95% confidence range of the certified value or the best estimate thereof. The coefficient of variation for all samples ranged from 2.3 to 10.1%, average value 4.7%. Average recovery for spiking experiments-levels 0.060, 0.075 and 0.150 micrograms/g was 102%; range 84-114%.

Animals↗
PubMed2005

Suitability of a malachite green procedure to detect minimal amounts of phosphate dissolved in acidic solutions.

The study describes the suitability of a colorimetric method (malachite green procedure) for detection of minimal amounts of phosphate (7.3-29.1 micromol/L) in different acidic solutions (hydrochloric acid, oxalic acid, maleic acid, perchloric acid, tartaric acid, citric acid, lactic acid and acetic acid) adjusted to pH 2.0. A mixture of the respective phosphate concentrations with distilled water served as control. The experiments were run with ten repeats in series. Assessment of intra- and interassay coefficient of variation and lower limit of quantification revealed that depending on the acid used, the applied method is a reliable and suitable tool to detect and quantify minimal phosphate contents in small samples of acidic solutions that have the potential to cause erosive dental lesions.

Acids↗
PubMed2006

Comparative metal distribution in hair of Pakistani and Libyan population and source identification by multivariate analysis.

Using nitric acid-perchloric acid wet digestion based FAAS method ten selected metals (Cd, Co, Cr, Fe, K, Mn, Na, Ni, Pb and Zn) were determined in the scalp hair of male Pakistani and Libyan donors (n = 62), between 3-54 years age and residing in typical urban areas. The study was taken up to identify sources of metal distribution in two diverse population segments exposed to different environments. Sodium emerged with the highest mean hair concentration of 511.0 microg/g and 496.9 mug/g for Libyan and Pakistani donors respectively, followed by K, at 266.2 microg/g and 242.4 microg/g for the two donor categories. Cadmium showed the lowest mean hair concentration for both Pakistani (0.380 microg/g) and Libyan (0.530 microg/g) donors. Levels of Na, K, Pb, Cr and Cd were higher in hair of Libyan donors compared with Pakistani counterparts which showed higher comparative levels of Zn, Fe, Co, Ni and Mn. Strong metal-to-metal correlations were found between Na-K (r = 0.638) and Cd-Co (r = 0.574) for Pakistani donors, while for Libyan donors, Na-K (r = 0.680) and Fe-Mn (r = 0.624) correlations emerged as strongly significant. Only K was found to have a strong positive correlation (r = 0.611) with age for Pakistani donors while this correlation was significantly negative (r = -0.500) for Libyan donors, probably arising from individual food habits. Other metals showed no viable relationship with age. Principal Component Analysis (PCA) and Cluster Analysis (CA) of the data evidenced food, industrial emissions and automobile emissions as possible sources of metal distribution. The results of the present study are compared with those reported for subjects from other regions of the world.

Adolescent↗
PubMed1984

Optimal extraction conditions for high-performance liquid chromatographic determination of nucleotides in yeast.

Different extraction methods of nucleotides from the yeast Saccharomyces cerevisiae were compared. A new extraction solution--formic acid saturated with 1-butanol--was found to be more effective than the commonly used solutions of trichloroacetic acid, perchloric acid, or formic acid alone. Using this solution the optimal extraction conditions were established. Nucleotide recovery was evaluated by adding standard nucleotides to the extraction medium and carrying them together with the cells through the whole extraction procedure. Nucleotides were separated and quantitated by high-performance liquid chromatography on an anion-exchange column.

1-Butanol↗
PubMed1990

Determination of tin in biological samples using gaseous hydride generation-inductively coupled plasma-atomic emission spectrometry.

A highly sensitive method determining for sub-microgram/gram levels of tin in biological samples is described. Tin hydride reduced by sodium borohydride and trichloroacetic acid solution was introduced into inductively coupled plasma after separation of liquid and excess hydrogen by an improved gas/liquid separator, and emission intensity was measured at a wavelength of 189.989 nm. Samples were decomposed by a nitric acid-perchloric acid mixture and analyzed after dilution by a standard addition technique. The relative standard deviation was 1.2% for a 10 ng/ml tin standard solution with a detection limit of 30 pg/ml.

Animals↗
PubMed2000

Assays for total homocysteine and other thiols by capillary electrophoresis-laser-induced fluorescence detection. I. Preanalytical condition studies.

In recent papers, we presented a new analytical method for thiol quantification in serum. It is based on the use of capillary electrophoresis and laser-induced fluorescence to analyze thiol 6-iodoacetamidofluoresceine (IAF) derivatives. Quantitative results of homocysteine, glutathione, cysteine-glycin, and cysteine were shown (Clin. Chem. 45 (1999) 412). A comprehensive comparison of the quantitation of homocysteine in serum, using high-performance liquid chromatography/conventional fluorescence detection and fluorescence polarization immunoassay was also used (E. Caussé et al., Electrophoresis 21 (2000) 2074). Sample preparation prior to derivatization with IAF had never been investigated. In this work we present the results of quantitation of thiols in serum and plasma with three different anticoagulants widely used: ethylenediaminetetraacetic acid (EDTA), heparin, and sodium citrate. We show that serum and EDTA plasma gave the same results. Then serum protein precipitations by acetonitrile, acetone, sulfosalicylic acid, perchloric acid and trichloracetic acid, prior to derivatization by IAF, were also investigated. Their influence on the concentrations of the thiols were determined. Sulfosalicylic acid and acetonitrile precipitations are well adapted, whereas acetone cannot be used.

Adult↗
PubMed2003

The influence of short strong hydrogen bonding on the structure and the physicochemical properties of alkyl-N-iminodiacetic acids in solid state and aqueous systems.

Alkyl-N-iminodiacetic acids with varying alkyl chain lengths have been prepared and characterized with respect to structure, acidic properties, and ability to form aggregates in water. The alkyl-N-iminodiacetic acids are the group of ligands with the lowest molecule weight which can be characterized as chelating surfactants, compounds with surface chemical properties which at the same time have a high ability to bind metal ions. The solid alkyl-N-iminodiacetic acids have a unique structure with neutral zwitterionic units linked together to polymer chains through a short strong hydrogen bond, d(O(-H)...O) approximately 2.46 A, and where the nu(O-H) stretching vibration at ca. 720 cm(-1) supports the presence of such a hydrogen bond. The polymer chains are cross-linked together to bilayers through relatively strong hydrogen bonds between ammonium and carboxylate groups, and where the parallel alkyl groups are interdigitating each other; the bilayer surface consists of hydrophilic iminodiacetic acid groups. The acidic properties of monomeric alkyl-N-iminodiacetic acids in water are in the expected ranges with pK(a) values of about 1.7, 2.3, and 10.3. n-Octadecyl-N-iminodiacetic acid, present as aggregates in water, displays very acidic properties of the first proton, and a substantially weakened acidity of the second proton, pK(a2) = 5.5-7.5, depending on ionic strength, and pK(a3) = 9.5-10.5. This pattern of the acidic constants strongly indicates that the polymer structure with short strong hydrogen bonds is maintained in the aggregates and that such bonds can exist in aqueous systems if they are supported by a strong and rigid backbone structure, as the bilayers of well-organized long interdigitating alkyl chains in the studied systems. Hydrogenbis(methyl-N-iminodiacetic acid) perchlorate precipitates from perchloric acidic solutions of methyl-N-iminodiacetic acid. The structure is built up of dimers of zwitterionic methyl-N-iminodiacetic acid units linked together by an extra proton in a short strong hydrogen bond, d(O(-H)...O) approximately 2.456(6) A, and nu(O-H) = 789 cm(-1).

Journal Article↗
PubMed1984

Phosphorylation of membrane-located proteins of soybean in vitro and response to auxin.

Isolated membranes of soybean incorporate (32)P from gamma-[(32)P]ATP in vitro. The incorporation was rapid and did not require added calcium. When displayed on 10% sodium dodecyl sulfate-polyacrylamide gels, several protein bands were revealed. An apparent auxin (2,4-dichlorophenoxyacetic acid) stimulation of (32)P incorporation into material from membrane vesicles insoluble in trichloroacetic acid-perchloric acid may be reflected partly in enhanced incorporation into protein bands with apparent molecular weights of 45,000 and 50,000. Additionally, a low molecular weight component was sometimes observed where incorporation was stimulated 2- to 3-fold by auxin. However, protein-bound radioactivity represented only a small fraction of the total radioactivity of the acid-insoluble material. Other labeled constituents, not retained on the gels, may contribute to the apparent, rapid (10 s or less) auxin response of the isolated membranes. Stimulation of incorporation into the low molecular weight component was given by diglyceride plus calcium, constituents known to augment protein kinase activities in other systems.

Journal Article↗
PubMed1984

The application of scanning transmission electron microscopy (STEM) to the study of thin anodic films on nickel and nickel-molybdenum alloys.

Anodized thin-film samples of nickel, molybdenum and a nickel-13 w/o molybdenum alloy have been analysed by scanning transmission electron microscopy, using selected area diffraction, microdiffraction and X-ray micro-analysis. Thin-film samples were obtained by ion thinning with argon and by electropolishing in acetic acid-perchloric acid. Electropolishing is the preferred technique for these studies, producing a surface with roughness on a scale of 1 nm. The anodized films on nickel and nickel 13 w/o molybdenum alloy exhibit a rugosity with a 'particle' size of 3 nm. The crystal structure of the films is similar to f.c.c. nickel oxide. Molybdenum enrichment was detected in the anodized alloy surfaces but there is no electron diffraction evidence for a crystalline molybdenum oxide. The electron diffraction pattern from anodized pure molybdenum suggests that the surface film is amorphous.

Alloys↗
PubMed2006

Multivariate analysis of the selected metals in the hair of cerebral palsy patients versus controls.

Seventeen metals were measured in scalp hair samples from cerebral palsy patients (CPPs) and controls. Samples were collected from 95 CPPs and 93 controls. The nitric acid-perchloric acid wet digestion procedure was used for quantification of the selected metals by flame atomic absorption spectrophotometry. The concentrations of Ag, Ca, Cd, Co, Cr, Li, and Mg were significantly higher and those of Cu, Fe, K, Mn, Na, Ni, Pb, and Sb were lower in the hair of CPPs compared with controls. Astrong positive correlation was found between Ca and Mg in the hair of controls but not in that of CPPs. Antimony was found significantly negative in terms of its correlation with Co and Cu in CPPs group but not in the controls. Principal component analysis (PCA) of the data extracted seven factors for CPPs and six factors for controls. Cluster analysis (CA) was also used to support the PCA results. The study evidenced some specific source of Mg and Sb in the hair of CPPs.

Adolescent↗
PubMed2004

Urinary excretion of nickel after dermal application of Nigerian light crude oil.

We examined the pattern of excretion of crude oil nickel (CON) in urine and feces. Adult male guinea pigs were treated with a single dose of 5 mL/kg b.w. Nigerian Bonny light crude oil applied on the skin. Untreated guinea pigs served as controls. The CON content was measured spectrophotometrically by the dimethylglyoxime reaction following nitric acid:perchloric acid: sulfuric acid (3:1:1) digestion of blood, urine, and feces samples at 2, 4, 8, and 16 days after the crude oil application. We found a peak of CON excretion in the urine at 2 days and a return to the control level at 16 days. There was no change in the CON content in the feces during the observation time. We found a peak of CON content in the blood at 8 days and a return to control values at 16 days. The implication of these excretion and transport patterns of CON is discussed vis-á-vis the potential application of the urinary nickel content as a valid biochemical indicator of human exposure to environmentally spilled crude oil.

Administration, Cutaneous↗