[Antitumor enzyme L-lyzine-alpha-oxidase from Trichoderma harzianum Rifai F-180 and investigation of its action on L-lysine oxidation by capillar electrophoresis].

Trichoderma harzianum Rifai F-180, an organism producing the antitumor enzyme L-lysine-alpha-oxidase was cultivated and the enzyme was isolated and purified under the manufacturing conditions. The effect of L-lysine-alpha-oxidase on oxidation of L-lysine was investigated for the first time by capillary electrophoresis and the procedure conditions were developed. The reaction of L-lysine oxidative deamination is described and location of the reaction components picks on the elecrophoregrams was identified. The average rate of the catalytic reaction of L-lysine oxidation equal to 0.46 RU/min (7.7 x 10(-3) RU/sec) was determined. The use of the antitumor enzyme L-lysine-alpha-oxidase is recommended as a drug for the treatment of superficial tumors and tissue relative oxygen excess.

Molecular absorption spectrometry (MAS) by electrothermal evaporation in a graphite furnace-XIII Determination of traces of fluoride by mas of AlF after liquid-liquid extraction of fluoride with triphenylantimony(V) dihydroxide.

The determination of traces of fluoride by means of the molecular absorption of AlF volatilized in graphite cuvettes is described. An extraction method for separation and preconcentration of the fluoride has been developed, to avoid matrix effects. The fluoride is extracted with 10(-3)M triphenylantimony(V) dihydroxide in M1BK, and stripped with 0.025M barium hydroxide. The method is sensitive and specific for fluoride. The detection limit is about 0.3 ng of fluoride, and the fluoride content of 6 ml of 10(-7)M solution can be determined. The determination is possible in presence of various ions, but it is estimated that usage of trace matrix separation leads to a considerable improvement in the relative detection limits (by 2-3 orders of magnitude), levels as low as 0.01 ppm being detectable in some matrices.

Membrane filtration studies of aquatic humic substances and their metal species: a concise overview. Part 2. Evaluation of conditional stability constants by using ultrafiltration.

The assessment of conditional stability constants of aquatic humic substance (HS) metal complexes is overviewed with special emphasis on the application of ultrafiltration methods. Fundamentals and limitations of stability functions in the case of macromolecular and polydisperse metal-HS species in aquatic environments are critically discussed. The review summarizes the advantages and application of ultrafiltration for metal-HS complexation studies, discusses the comparibility and reliability of stability constants. The potential of ultrafiltration procedures for characterizing the lability of metal-HS species is also stressed.

Determination of trace heavy metals in waters by atomic-absorption spectrometry after preconcentration by liquid-phase polymer-based retention.

A new method for the determination of metals in waters by flame atomic-absorption spectrometry is described. The metals are retained by water-soluble polymers in a membrane filtration cell and the factors which influence their determination are discussed. The method has been applied to the determination of Ni, Cu, Zn, Hg and Cd in drinking and river water with poly(ethyleneimine) and its thiourea derivative as complexing polymers. The metals were determined in the aqueous concentrate after a 250-fold preconcentration by 2% polymer solution at pH 7. The metal recoveries were at least 92%, and the limits of detection (ng/mg) were 0.012 for Cu, 0.006 for Zn, 0.03 for Ni, 0.004 for Cd and 0.0001 for Hg (cold vapour method). When a new type of membrane filtration cell is used even higher preconcentration factors can be achieved and lower concentrations can be determined.

Molecular absorption spectrometry (MAS) by electrothermal evaporation in a graphite furnace-IX Determination of traces of bromide by mas of AlBr after liquid-liquid extraction of bromide with triphenyltin hydroxide.

The determination of traces of bromide by molecular absorption spectrometry (MAS) of AlBr (with electrothermal volatilization) is described. It is possible to determine 25 ng of bromide. Many problems are caused by various matrices, so an extraction method for separation (and also preconcentration) was developed. The combination of bromide extraction with triphenyltin hydroxide, stripping with 0.025M barium hydroxide and determination of the extracted bromide by MAS of AlBr (after addition of aluminium ions) gives a very sensitive and selective method for determination of traces of bromide in micro or macro samples, in the presence of large amounts of other species, including chloride and iodide.

Molecular absorption spectrometry by electrothermal evaporation in the graphite furnace-X Determination of chloride traces by AlCl ma in graphite cuvettes after liquid-liquid extraction of chloride with triphenyltin hydroxide.

The determination of traces of chloride by means of the molecular absorption of AlCl in graphite cuvettes is described. An extraction method for separation and preconcentration of the chloride has been developed, to avoid matrix effects. Extraction was done with 0.03M triphenyltin hydroxide in o-xylene, and stripping with 0.025M barium hydroxide. The method is sensitive and specific for chloride. The detection limit is about 1.5 ng of chloride, and the chloride content of 20 ml of 10(-7)M solution can be determined. The determination is possible in presence of excess of bromide and iodide.

Determination of ortho- and pyrophosphates in waters by extraction chromatography and flow-injection analysis.

Extraction-chromatographic separation of ortho- and pyrophosphate anions on an inert support modified with an organotin extractant was studied and used for their subsequent determination in a flow-injection system. The proposed FIA manifold includes an extraction-chromatographic mini-column, on which the phosphate anions are separated and preconcentrated, and a post-column spectrophotometric detector. For the determination of orthophosphate, the absorbance of the reduced 12-molybdophosphoric acid is monitored at 660 nm. The sum of ortho- and pyrophosphate can be determined after preliminary hydrolysis of pyrophosphate to orthophosphate in neutral solution at 50 degrees by use of inorganic pyrophosphatase. For a sample volume of 6 ml, the calibration graph is linear within a range of 5.0-100.0 ng/ml P. The limit of detection is 0.3 ng/ml P. The recovery of the ions to be determined is not less than 96%, the relative error is not worse than 4%. The proposed method was used for the analysis of river water samples.

Ultrafiltration and determination of Zn- and Cu-humic substances complexes stability constants.

This study exhibits that size fractionation of humic substances (HS) and their metal complexes by ultrafiltration is an efficient procedure for simultaneous determination of stability constants. Using sequential-stage ultrafiltration and a radiotracer technique the HS-Cu and HS-Zn complexes studied can gently be size-fractionated and their free metal fractions simply be discriminated. The conditional stability constants Ki obtained for size fractions of these HS metal complexes exhibit a clear molecular size dependence. Accordingly, the highest Ki values (6.6 for Zn and 6.4 for Cu) are found in the HS fractions of >105 kDa. Moreover, the overall stability constants K found for Cu (log K=5.5) and Zn complexes (log K=4.5) of the aquatic HS complexes studied are quite comparable to those reported in the literature.

Water-soluble polymers for spectrophotometric and flow injection determination of cobalt with nitroso-R-salt.

The water-soluble polymers poly(ethylenimine), quaternized poly(ethylenimine), and poly-4-vinyl-(N-benzyltrimethyl) ammonium chloride were found to be able to change the kinetics of the reaction of cobalt with nitroso-R-salt and the optical density of the cobalt complex formed. The optimum pH range for the complex formation was a wide range from 1 to 10 and at pH 2 the reaction developed instantly. The calibration graph was linear in the range 0.005-2 mug ml(-1). The effective molar absorptivity coefficient of the complex was equal to (2.8+/-0.08)10(3). A spectrophotometric determination method for cobalt with nitroso-R-salt in the presence of water-soluble polymers (before and after membrane preconcentration) and a colorimetric flow injection method were developed. For the flow injection-based spectrophotometric determination, the calibration graph was linear in the concentration range of 0-4.0 mug ml(-1) cobalt with a regression coefficient of 0.9992. The relative standard deviation (R.S.D.) for the determination of 1.0 mug ml(-1) cobalt was 0.9% (ten replicate injections), and at all concentration measured, the R.S.D. of the data was below 5.0%. The proposed FI procedure was applied to river water samples after membrane preconcentration of cobalt. The limit of detection was 4 ng ml(-1).