A simple and sensitive catalytic method for determination of the total iron-binding capacity of blood serum

Summary A catalytic method is proposed for determination of the total iron-binding capacity of blood serum, based on the iron-catalysed oxidation ofp-phenetidine by hydrogen peroxide. Only 0.05 ml of serum is necessary for a single determination. The results obtained by the catalytic method show good correlation with those obtained by a photometric method (with bathophenanthroline) and by atomic-absorption spectrometry.

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Zusammenfassung Eine katalytische Methode zur Bestimmung der Eisenbindungskapazität von Blutserum wurde vorgeschlagen. Sie beruht auf der Fe-katalysierten Oxydation vonp-Phenetidin durch Wasserstoffperoxid. Für eine Bestimmung werden nur 0,05 ml Serum benötigt. Die mit dieser Methode erhaltenen Ergebnisse korrelieren sehr gut mit Werten, die mittels Atomabsorption und Spectralphotometrie (Bathophenanthrolinmethode) erhalten wurden.

     

Microdetermination of serum iron and of iron-binding capacity by precision photometry

Summary Use is made of the high stability of the photometric system of the Enzyrator. This stability, together with the possibility of multiplying the read-out by a factor of 10, make it possible to decrease the sample size by the same factor.

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Zusammenfassung Die besondere Stabilität der photometrischen Anzeige des Enzyrators sowie die Möglichkeit, die Anzeige zu verzehnfachen, bieten die Voraussetzung, die Probengröße um denselben Faktor zu verkleinern.

     

Application of a new ferroin ligand to the automated determination of microgram serum iron and iron binding capacity

Summary Introduction of a new commercially available ferroin ligand by simple modification and substitution into contemporary automated methodology makes possible the rapid, sensitive determination of serum iron and iron binding capacity at theg level. The inclusion of a mildly acidic saline mercaptoacetate diluent before protein dialysis prevents interference from serum copper. Results obtained by the proposed method appear to be almost identical to those obtained by theBabson modified method ofYoung andHicks. The moderate cost of Ferrozine together with the intensity of the ferrous Ferrozine chromophore (M. A. 28000) should make this ligand a popular reagent for automated analysis.

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Zusammenfassung Die Einführung eines neuen handelsüblichen Chelatbildners vom Ferrointyp ermöglicht nach geringfügiger Modifikation der modernen Methodik die rasche und empfindliche Bestimmung des Serum-Eisens und der Eisenbindungskapazität im Mikrogramm-Maßstab.Die Verdünnung der Probe mit schwach saurer, salzhaltiger Mercaptoessigsäure vor der Eiweißdialyse verhindert Störungen durch Serum-Kupfer. Die mit dem vorgeschlagenen Verfahren erzielten Ergebnisse sind mit denen des vonYoung undHicks angegebenen, vonBabson abgeänderten Verfahrens vonYoung undHicks fast identisch. Die geringen Kosten von Ferrozin sowie die Farbintensität von Eisen(II)-ferrozin dürften dieses Reagens für automatische Analysen empfehlen.

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Presented in part at Congressus Biochimiae Clinicae Pragensis, Prague, September 1971.     

A new spectrophotometric method for the determination of total and ferric iron in rain water at the ppb level

A new, simple, selective and sensitive spectrophotometric procedure for the on-site quantification of iron at nano-gram levels in atmospheric precipitations, i.e. rain as sample source is described. It is based on the color reaction of Fe³⁺ with SCN^– ions in the presence of a cationic surfactant, i.e. cetylpyridinium chloride (CPC), in strong HCl solution, and subsequent extraction of the complex with N-octylacetamide into toluene or chloroform. The apparent molar absorptivity of the complex is 2.60 × 10⁵ L mol^–1 cm^–1 at λ_max = 480 nm at an enrichment factor (EF) of 10. The detection limit (causing higher absorbance than the sum of the blank absorbance (0.009) and 3 SD) is 5 ng mL^–1 Fe. Ions commonly associated with iron did not interfere in the present method. The effect of analytical variables, i.e. amount and type of the reagents, acidity, solvent, temperature, dilution, etc., in the determination of iron are discussed. The validity of the present method is checked with GF-AAS. The method has been applied to the determination of iron at the ppb level in rain water samples.     

A new spectrophotometric method for the determination of total and ferric iron in rain water at the ppb level

A new, simple, selective and sensitive spectrophotometric procedure for the on-site quantification of iron at nano-gram levels in atmospheric precipitations, i.e. rain as sample source is described. It is based on the color reaction of Fe3+ with SCN- ions in the presence of a cationic surfactant, i.e. cetylpyridinium chloride (CPC), in strong HCl solution, and subsequent extraction of the complex with N-octylacetamide into toluene or chloroform. The apparent molar absorptivity of the complex is 2.60 x 10(5) L mol(-1) cm(-1) at lambdamax = 480 nm at an enrichment factor (EF) of 10. The detection limit (causing higher absorbance than the sum of the blank absorbance (0.009) and 3 SD) is 5 ng mL(-1) Fe. Ions commonly associated with iron did not interfere in the present method. The effect of analytical variables, i.e. amount and type of the reagents, acidity, solvent, temperature, dilution, etc., in the determination of iron are discussed. The validity of the present method is checked with GF-AAS. The method has been applied to the determination of iron at the ppb level in rain water samples.     

Determination of total iron binding capacity of serum by capillary electrophoresis

Summary A capillary electrophoresis (CE) technique for determining total iron binding capacity (TIBC) of serum has been developed. The optimum serum pretreatment involves the following major steps: at first, saturate serum transferrin with Fe⁺³; then, dissociate them completely after removing excess unbound Fe. Finally, complex the released iron with phenanthroline, a chromophore, to make suitable for the CE analysis. Ammonium acetate (pH=5.0) was used as CE background electrolyte solution. In this system, a good linear correlation coefficient was maintained over the range 0.5≈10 μM (r=0.9979,n=12). Seven adult serum samples were studied and the TIBC parameters measured. In the present system, 10≈30 μL serum is sufficient for determination. The study shows that the CE technique described is a powerful method for rapid, efficient, sensitive and reliable analysis and hence particularly suitable for clinical application.     

Comparison of modifiers for determination of arsenic, antimony and selenium by atomic absorption spectrometry with atomization in graphite tube or hydride generation and in-situ preconcentration in graphite tube

The study was performed to compare the effect of magnesium modifier (magnesium nitrate) with that of other modifiers (palladium nitrate and nickel nitrate) in determination of arsenic, antimony and selenium by atomic absorption spectroscopy with atomization in a graphite tube, with generation of hydrides and in situ preconcentration in a graphite tube. The assumed criterion of a modifier performance was the magnitude of the analytical signal. It was found that in determinations with atomization in a graphite furnace the effects of all these modifiers were comparable, while in those with hydride generation and in situ preconcentration in a graphite tube the magnesium modifier showed poorer performance (25% decrease of the analytical signal). In determinations of arsenic and selenium the analytical signal obtained with magnesium salt as a modifier was comparable with those obtained in the presence of all other modifiers.     

Direct determination of iron in urine and serum using graphite furnace atomic absorption spectrometry

A simple, rapid and low-cost method for the routine determination of iron in urine and serum using graphite furnace atomic absorption spectrometry is described which may provide an alternative to the more widespread automated spectrophotometric methods. The urine and serum samples were simply diluted with water prior to analysis. Matrix modification was found to be redundant. The standard additions technique or the use of matrix matched standards (addition calibration) was found to be unnecessary and, therefore, the calibration was performed using aqueous standards. For serum analysis the degree of dilution could be reduced by using the less sensitive 302.0-nm resonance line, yielding more precise determinations, and for urine analysis, interferences were eliminated by means of a L'vov platform. The interferences that exist in the presence of nitric acid are also discussed. Finally, the presence of background absorption was investigated by means of Zeeman effect atomic absorption.     

3, 6-pyridazinediol as a specific spectrophotometric reagent for the determination of iron

Iron(III) reacts with 3,6-pyridazinediol in presence of nitric acid to give pink color having maxima at 480 mμ. The color reaction which is specific for iron(III), has 1 μg/ml as visual limit of identification and provides the basis of a new spectrophotometric method for the determination of iron. The quantitative assessment of the tolerable amount of different ions which do not interfere with the determination are reported. The mechanism of the color reaction is not clear.     

Spectrophotometric evaluation of interferences in three iron reactions for the determination of serum total cholesterol

A spectrophotometric and chemical evaluation of reported interferences for three iron reactions for the determinations of serum cholesterol has been presented. It has been shown that all three reactions are affected by various interfering substances, such as 2-thiouracil, nitrate, azide, bromide, diethylstilbesterol, and steroids. Spectral differences between the reactions are probably due to solvent and anion effects.The incorporation of uranyl acetate as a precipitating agent into the ferric acetate-uranyl acetate procedure did not make the results obtained comparable with the Abell-type extract of a very icteric serum. Incorporation of ferrous sulfate does not noticeably affect the intensity or stability of color development with standards.As proposed, the ferric acetate-uranyl acetate procedure for the determination of cholesterol represents a modified iron reagent, but the reaction mechanism and the procedure described for it is neither new nor direct. The use of the ferric acetate reagent for serum cholesterol as opposed to ferric chloride, ferric ammonium chloride, or ferric perchlorate appears to have no real advantages as a color reaction. In fact the reaction is less sensitive while reagent preparation is more tedious, time consuming, and expensive than the ferric chloride procedure. Although no quantitative studies were performed, the only effects that chloride had on the reactions were in the region of 400 nm, a wavelength sufficiently far enough away from the 560-nm peak that it did not affect determinations. The concept that the ferric acetate-uranyl acetate reagent contains only acetate and sulfate anions is nullified as soon as one adds serum to the reagents.     

Direct spectrophotometric determination of thiocyanate in serum and urine with a continuous-flow analyzer

A spectrophotometric method for the rapid measurement of thiocyanate in serum and urine without separation from interfering substances is described. Thiocyanate reacts immediately with chloramine-T in presence of iron(III) chloride catalyst to give cyanogen chloride, which reacts with a mixture of γ-picoline (4-methylpyridine) and barbituric acid to form a soluble violet—blue product, which is measured at 605 nm. Other components of physiological fluids react more slowly and do not interfere if the reaction time of the chlorinating step is kept very short. The proposed procedure is compared with a highly selective method, based on the oxidation of thiocyanate to cyanide, and good agreement was obtained for both serum and urine. The method is readily adapted to a continuous-flow procedure with a Technicon AutoAnalyzer.     

Selective spectrophotometric determination of iron(III) with EDTA

A spectrophotometric determination of iron as its iron (III)-EDTA-H₂O₂-NH₃ complex is described; up to 10-fold amounts of metals that form EDTA complexes absorbing at the same wavelength do not interfere because hydrogen peroxide reacts with thciron(IIl)-EDTA complex but does not affect the EDTA complexes of coppcr(II), nickel(II), cobalt(ltl) and chromium(Ilt).     

Spectrophotometric determination of serum iron with a new reagent

The use of 2,2′-dipyridyl-2-pyridylhydrazone (DPPH) as a new “ferroin” type color reagent for the spectrophotometric determination of serum iron is presented. The determination is performed on protein-free filtrates at pH 1.6 to 3.0. The reagent has been found to be specific for serum iron, under the proposed conditions. The color of the water-soluble iron-DPPH complex is considerably stable.     

The determination of germanium by atomic absorption spectrometry with a graphite tube atomizer

A carbon filament atom reservoir and a graphite tube atomizer are compared as methods for the determination of germanium by atomic absorption spectrometry. It is shown that the filament technique is not applicable, probably because of loss of the sample as a volatile oxide species which results in inefficient atomization. The design of a small graphite tube atomizer is described; with this, a limit of detection for germanium of 3·10^-10 g was found. For a 20-μl sample, this corresponds to a concentration of 0.015 p.p.m. The interfering effect of 13 anions and cations is studied.     

A new catalytic kinetic spectrophotometric method for determination of iron

The catalytic effect of iron(III) on the oxidation of reduced Rhodamine B with hydrogen peroxide in acetic acid medium has been studied. The reaction is highly accelerated by potassium thiocyanate. A new catalytic kinetic spectrophotometric method for the determination of iron has been developed. Iron(III) can be determined by the fixed time method with a detection limit of 4 x 10(-11) g/ml.     

2-Oximinodimedone monoguanylhydrazone as spectrophotometric reagent for determination of iron

The synthesis, properties and analytical possibilities of 2-oximinodimedone monoguanylhydrazone as spectrophotometric reagent have been examined. A new method for the determination of iron has been developed in a concentration range varying from 0·2–5·0 ppm of iron; the molar absorptivity is 1·1×10⁴ litres mol⁻¹ cm⁻¹ at 600 nm and the relative error is ±0·3%. The method has been applied satisfactorily to the determination of iron in different inorganic samples.     

Flow-injection analysis with the iron-induced perbromate-iodide reaction: spectrophotometric determination of iron

Total iron is determined by a flow-injection spectrophotometric technique. The production of I(-)(3) from the iron(II)- or iron(III)-induced perbromate-iodide reaction is monitored at 353 nm. Calibration graphs are linear from 10 to 100 ng/ml with correlations up to 0.9998 and can be extended up to 10 microg/ml by appropriate adjustment of conditions. The average sampling rate is 30 samples/hr. Detection limits and relative standard deviations compare well with those of other FIA methods.