Multicomponent analysis of hemoglobin derivatives with reversed-optics spectrophotometer.

We describe a method for the simultaneous determination of the five clinically relevant hemoglobin derivatives (Hb, HbO2, HbCO, Hi, SHb) in a blood sample by means of a reversed-optics spectrophotometer (Hewlett-Packard HP8450 A UV/Vis). A built-in computer program is used for multicomponent analysis in an overdetermined system, i.e., a system in which the number of independent equations used exceeds the number of unknowns to be determined. First, the spectra of the five hemoglobin derivatives are measured in a series of different human blood samples. Thereafter, the multicomponent method for the simultaneous determination of the five hemoglobin derivatives is tested by comparison with conventional methods for the separate determination of oxygen saturation, HbCO, Hi, and SHb fractions. The multicomponent (multiwavelength) method is sufficiently reliable, accurate, and easy to justify its use in physiological chemical research as well as its routine application in the clinical chemical laboratory.

[1]  Y. Chang,et al.  The structure of the chromophore of sulphymyoglobin , 1967 .

[2]  W. Zijlstra,et al.  Sulfhaemoglobin. Absorption spectrum, millimolar extinction coefficient at lambda = 620 nm, and interference with the determination of haemoglobin and of haemiglobincy anide. , 1977, Clinica chimica acta; international journal of clinical chemistry.

[3]  W. Barnikol,et al.  Molar absorptivities of human hemoglobin in the visible spectral range. , 1985, Journal of applied physiology.

[4]  D. Drabkin,et al.  Spectrophotometric studies; the crystallographic and optical properties of the hemoglobin of man in comparison with those of other species. , 1946, The Journal of biological chemistry.

[5]  W. Zijlstra,et al.  Carboxyhaemoglobin. Spectrophotometric determination tested and calibrated using a new reference method for measuring carbon monoxide in blood. , 1977, Clinica chimica acta; international journal of clinical chemistry.

[6]  R. Benesch,et al.  Equations for the spectrophotometric analysis of hemoglobin mixtures. , 1973, Analytical biochemistry.

[7]  Zijlstra Wg,et al.  Haemoglobinometry: challenges and pitfalls. , 1970 .

[8]  W G Zijlstra,et al.  Molar absorptivities of human hemoglobin in the visible spectral range. , 1983, Journal of applied physiology: respiratory, environmental and exercise physiology.

[9]  O. W. van Assendelft,et al.  Spectrophotometry of haemoglobin derivatives , 1970 .

[10]  W G Zijlstra,et al.  A Multi-Wavelength Spectrophotometric Method for the Simultaneous Determination of Five Haemoglobin Derivatives , 1981, Journal of clinical chemistry and clinical biochemistry. Zeitschrift fur klinische Chemie und klinische Biochemie.

[11]  G. Kwant,et al.  A new reference method for the determination of the oxygen content of blood. , 1976, Clinica chimica acta; international journal of clinical chemistry.

[12]  Leslie J. Brown,et al.  A New Instrument for the Simultaneous Measurement of Total Hemoglobin, % Oxyhemoglobin, % Carboxyhemoglobin, % Methemoglobin, and Oxygen Content in Whole Blood , 1980, IEEE Transactions on Biomedical Engineering.

[13]  O. Siggaard‐Andersen.  Experiences with a new direct-reading oxygen saturation photometer using ultrasound for hemolyzing the blood. , 1977, Scandinavian journal of clinical and laboratory investigation. Supplementum.

[14]  Bradley E. Copeland,et al.  Spectrophotometry of Haemoglobin Derivatives , 1971 .

[15]  K. Evelyn,et al.  MICRODETERMINATION OF OXYHEMOGLOBIN, METHEMOGLOBIN, AND SULFHEMOGLOBIN IN A SINGLE SAMPLE OF BLOOD , 1938 .

[16]  F. Rodkey,et al.  Effects of carboxyhemoglobin on the determination of methemoglobin in blood. , 1974, Biochemical medicine.

[17]  J. Miller,et al.  A SOURCE OF ERROR IN THE CYANMETHEMOGLOBIN METHOD OF DETERMINATION OF HEMOGLOBIN CONCENTRATION IN BLOOD CONTAINING CARBON MONOXIDE. , 1965, American journal of clinical pathology.

[18]  W. Zijlstra,et al.  OXYGEN BINDING-CAPACITY OF HUMAN HEMOGLOBIN - HUFNERS FACTOR REDETERMINED , 1977 .

[19]  B. No̸rgaard-Pedersen,et al.  Hemoglobin pigments. Spectrophotometric determination of oxy-, carboxy-, met-, and sulfhemoglobin in capillary blood. , 1972, Clinica chimica acta; international journal of clinical chemistry.

[20]  J. Peisach,et al.  The preparation and some physical properties of sulfhemoglobin. , 1978, The Journal of biological chemistry.

[21]  G. Kwant,et al.  Oxygen dissociation curves for whole blood, recorded with an instrument that continuously measures pO2 and sO2 independently at constant t, pCO2, and pH. , 1982, Clinical chemistry.

[22]  W K Barnikol,et al.  Dependence of visible spectrum [epsilon (lambda)] of fully oxygenated hemoglobin on concentration of hemoglobin. , 1982, Journal of applied physiology: respiratory, environmental and exercise physiology.

[23]  J. Berzofsky,et al.  Sulfheme proteins. I. Optical and magnetic properties of sulfmyoglobin and its derivatives. , 1971, The Journal of biological chemistry.

[24]  W. Zijlstra,et al.  Determination of hemoglobin derivatives with IL 282 CO-oximeter as compared with a manual spectrophotometric five-wavelength method. , 1981, Clinical chemistry.