An improved kinetic determination for creatinine using the Abbott ABA-100

An improved method for the kinetic determination of creatinine on the Abbott ABA-100 is described. By simplex optimization of the picrate and hydroxide reaction concentration a 35% increase in reaction rate, with respect to the Abbott methodology, was obtained. The new method correlated more closely with the Technician AutoAnalyzer Method (N-30) than did the latter method. The percentage CV for the improved method for high and low quality control sera was 2.4 and 3.6%, respectively. The presence of protein in the standards was found to cause a decrease in the reaction rate with respect to aqueous standards.     

Interesting interferences in a direct serum creatinine reaction

A spectrophotometric study on a direct picric acid reaction for creatinine in severely jaundiced serums is described. A problem appears to be caused by the oxidation of bilirubin which minimizes rising absorbance when using continuous measurement. Simple examples of interferences with the kinetic mode are shown along with the hitherto unreported interference of the drug, Cephalothin, which also undergoes a picric acid reaction. A procedure in which a delta absorbance is obtained after decolorization of the Jaffé complex by acidification is shown as one available means for obviating the bilirubin effect. However, the theory that Jaffé-reactive interferences do not decolorize with the same acid treatment is not totally applicable when the drug, Cephalothin, is present.     

Phototitrimetric determination of sulfhydryl compounds witho-hydroxymercuribenzoic acid using dithiofluorescein as an indicator

Summary A phototitrimetric method for the determination of sulfhydryl compounds in an aqueous medium is described.o-Hydroxymercuribenzoic acid is used as a titrant with dithiofluorescein as an indicator. Disulfide amino acids like cystine and-substituted DL-cystines are reduced with sodium borohydride before titration. All amino acids were determined to the limit of 1.0mol with an error of less than 4%. A selective phototitrimetric method for the determination of-mercaptopyruvate in presence of cysteine and glutathione is described. This method can be extended to the microdetermination of other similar sulfhydryl compounds in biological materials.

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Zusammenfassung Eine phototitrimetrische Bestimmungsmethode für Sulfhydrylverbindungen in wäßriger Phase wurde beschrieben. Als Maßflüssigkeit diento-Hydroxymercuribenzoesäure, als Indikator Dithiofluorescein. Aminosäuredisulfide wie Cystin und-substituierte DL-Cystine werden vor der Titration mit Natriumborhydrid reduziert. Alle Aminosäuren lassen sich bis zur Größenordnung von 1,0Mol mit einem Fehler von weniger als 4% bestimmen. Eine selektive phototitrimetrische Bestimmung von-Merkaptopyruvat in Gegenwart von Cystein und Glutathion wurde angegeben. Das Verfahren läßt sich auch für andere Sulfhydrylverbindungen in biologischem Material anwenden.

     

Study of various substrates for peroxidase-coupled peroxide oxidations

It would appear from this study that the sodium sulfonates of 2, 4-dichloro- and 2, 4-dibromophenol when employed in the presence of 4-AAP produce useful and sensitive, colorimetric, cosubstrate systems. What may prove to be of equal importance is the relatively high initial rate of the coupling reactions, as it has been suggested (29, 30) that this might help alleviate interference by competitive proton donors.As long as the general structure of the phenols is maintained, increased aromaticity tends to add stability to the chromogen. This is, of course, an asset when performing equilibrium analysis.The use of MBTH, in the presence of either aromatic amines or alcohols, appears to have little obvious merit. Apart from the continual increase in absorbance of the blank, that is, a spontaneous coupling of the cosubstrates, as well as the comparatively high blank absorbance, this material does not seem to be as sensitive as some of the other reactions studied.     

Inverse polarographic determination of creatinine with alkaline picrate and 3,5-dinitrosalicylic acid

A selective and highly sensitive inverse polarographic method for the determination of creatinine has been developed, involving the measurement of disappearance of diffusion current for the picric acid wave ($\text{E}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= ?0.501}\text{V}$) when creatinine is added. Creatinine levels of 0–5 μg/ml can be measured accurately.Investigation of 3,5-dinitrobenzoic acid and 3,5-dinitrosalicylic acid revealed the latter to be more reactive and useful for the inverse polarographic determination of creatinine at levels of 0–30 μg/ml.     

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.