A Multi-Wavelength Spectrophotometric Method for the Simultaneous Determination of Five Haemoglobin Derivatives

Abstract

A method is described by which the concentration of deoxyhaemoglobin, oxyhaemoglobin, carboxyhaemoglobin, haemigjobin and sulphaemoglobin in a human blood sample is determined by passing the haemolysate without air contact through a coarse filter and subsequently measuring the absorbance at = 500, 569, 577, 620 and 760 nm. The euouing set of equations is solved by matrix calculation with the aid of a simple computer program. The method has been tested by comparing it with conventional methods for the determination of the various haemoglobin derivatives separately. Eine Viel-Wellenlängen Methode für die simultane spektrophotometrische Bestimmung von fünf HämoglobinderivatenZusammenfassung: Eine Methode, mit der die Konzentration von Desoxyhämoglobin, Oxyhämoglobin, Carboxyhämoglobin, Hämiglobin und Sulfhämogjobin in einer Blutprobe vom Menschen bestimmt werden kann, wird beschrieben. Das Hämolysat durchläuft ohne Luftkontakt ein grobes Filter; die Absorption bei = 500, 569, 577, 620 und 760 nm wird gemessen. Der sich ergebende Satz von Gleichungen wird durch Matrizen-Berechnung mit Hilfe eines einfachen Rechnerprogramms gelöst. Die Methode wurde durch Vergleich mit konventionellen Methoden für die separate Bestimmung der verschiedenen Hämoglobinderivate geprüft.

Further Studies on the Standardization of Neonatal Bilirubin

“…This is the crucial point in our st dy, because by applying formula 4 the absorbance values given in formulae ί and 2 are included. Then we make three errors which compensate each other partly: a) the molar absorbancies of oxyhaemoglobin differ between 455 and 575 nm, A 575 being the higher one (8,9) b) the absorbance value of carotenoids, methaemoglobin and transferrin are higher at 455 nm than at 575 nm (3) c) the oxyhaemoglobin content in neonatal sera can vary considerably and is certainly much higher than in the adult serum sample used for makiiig the Standard (10) and furthermore the carotenoid, methaeinoglobin and transferrin Contents are generally much lower in neonatal sera than in the adult sera used in serum Standards. This may sound very negative.…”

Localized light absorption by hemoglobins of an erythrocyte suspension

“…Variations of their molecular structures causes their absorption coefficients μ i (λ) (i is the hemoglobin form) to assume different properties depending on the wavelength of the light [1-4] although the absolute values are usually much greater than those of other blood chromophores (thrombocytes, plasma) [5, 6]. Therefore, blood absorption spectra μ b (λ) are often used for optical monitoring of its hemoglobin composition and the bulk concentration of derivatives [1,[7][8][9][10][11]. Multi-component analysis [1,3,10,11], least-squares methods, and other well developed mathematical procedures are used to deduce the concentration from the measured function μ b (λ) and known μ i (λ).…”

“…Therefore, blood absorption spectra μ b (λ) are often used for optical monitoring of its hemoglobin composition and the bulk concentration of derivatives [1,[7][8][9][10][11]. Multi-component analysis [1,3,10,11], least-squares methods, and other well developed mathematical procedures are used to deduce the concentration from the measured function μ b (λ) and known μ i (λ). The principal issue with experimental determination of μ b (λ) is the structure of the blood sample, namely, is the hemoglobin in solution after hemolysis of the blood or is it dispersed, concentrating on erythrocytes.…”

Application of CO-oximeter for Forensic Samples

“…The Hb solutions obey the Lambert-Beer Law, and the absorbance measured at selected wavelength is the sum of the absorbance of each Hb derivative [31][32][33][34][35][36]. The wavelength is selected by the combination of absorption maxima and isosbestic points [37].…”