Molar absorptivities of human hemoglobin in the visible spectral range

In a recent paper, Burkhard and Barnikol (J. Appl. Physiol.: Respirat. Environ. Exercise Physiol. 52: 124–130, 1982) claimed that the absorption spectra of human (oxy-)hemoglobin are dependent on the total hemoglobin concentration (CHb) and it is suggested that this might also be the case with cyanmethemoglobin (HiCN). Such relationships would invalidate the widely used spectrophotometric methods for the determination of total hemoglobin and the fractions of various hemoglobin derivatives in human blood. Although Burkhard and Barnikol's findings are rather improbable considering earlier data, we measured the millimolar absorptivities of oxyhemoglobin (epsilon HbO2) and cyanmethemoglobin (epsilon HiCN) at various wavelengths over a wide range of concentrations (CHb approximately equal to 0.004–10 mmol x 1(-1)), using two different types of spectrophotometers. epsilon HbO2 and epsilon HiCN proved to be independent of CHb. Moreover the values obtained confirmed those in the earlier literature, whereas those of Burkhard and Barnikol are some 30% higher. Consequently there is no reason to doubt the validity of the generally accepted millimolar absorptivities of human hemoglobin.

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Results of routine determination of clinically significant hemoglobin derivatives by multicomponent analysis.

Clinical Chemistry ◽

10.1093/clinchem/32.6.972 ◽

1986 ◽

Vol 32 (6) ◽

pp. 972-978 ◽

Cited By ~ 24

Author(s):

A Zwart ◽

E J van Kampen ◽

W G Zijlstra

Keyword(s):

Pulmonary Disease ◽

Fetal Hemoglobin ◽

Hemoglobin Concentration ◽

Major Surgery ◽

Chemical Laboratory ◽

Total Hemoglobin ◽

Clinically Significant ◽

Blood Specimens ◽

Total Hemoglobin Concentration

Abstract Our recently developed multiwavelength method for multi-component analysis of hemoglobin (Hb) derivatives (Clin Chem 1984;30:373-379) was adapted for routine use in the clinical chemical laboratory. The method was applied in 4066 determinations on blood specimens from patients awaiting major surgery (n = 3863) or visiting the outpatient department for pulmonary disease (n = 203). Mean total hemoglobin concentration was 141 (SD 14) g/L. The proportion of HbCO was slightly to moderately increased (1.5-10.0%) in 36.5% of all patients; in a few cases it was as high as 15%. Mean methemoglobin was 0.4% (SD 0.2%) in the surgical patients, but 1.5% (SD 0.8%) in the patients with pulmonary disease. In some patients of the latter group the proportion of methemoglobin amounted to 5%. Sulfhemoglobin was found less than 0.4% in all specimens. Interference by paraproteins and by increased concentrations of bilirubin and lipids in plasma was easily detected by means of the performance checks provided by the spectrophotometer (an HP 8451 UV/Vis). The method is equally suitable for measuring blood samples containing fetal hemoglobin.

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Blood replacement in dogs by dextran–hemoglobin

Canadian Journal of Biochemistry ◽

10.1139/o78-153 ◽

1978 ◽

Vol 56 (10) ◽

pp. 981-984 ◽

Cited By ~ 34

Author(s):

Siu-Cheung Tam ◽

Jan Blumenstein ◽

J. Tze-Fei Wong

Keyword(s):

Half Life ◽

Exchange Transfusion ◽

Hemoglobin Concentration ◽

Complete Recovery ◽

Human Hemoglobin ◽

Blood Replacement ◽

Total Hemoglobin ◽

Total Hemoglobin Concentration ◽

Covalent Complex

Exchange transfusions in dogs were performed with a solution of either dextran or a covalent complex between dextran and human hemoglobin. Dogs transfused with dextran alone died when their hematocrit was lowered to 6–10%. Dogs transfused with the dextran–hemoglobin complex, however, survived a reduction of their hematocrit to 2% or below. In the latter animals, the dextran–hemoglobin complex disappeared from the circulation with an average half-life of 2.4 days. Correcting for oxidation of the hemoglobin moiety to methemoglobin, the half-life of functional unoxidized dextran–hemoglobin in the circulation was 1.9 days. In compensation for the loss of dextran–hemoglobin, vigorous erythropoiesis was observed at a rate of close to 5% hematocrit per day over the first 2 days following the exchange transfusion. As a result, the total hemoglobin concentration in blood was maintained at 5–6% during this period, and the animals went on to complete recovery in room air without the need for further transfusion with dextran–hemoglobin.

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An oximeter for measuring hemoglobin concentration and oxygen content

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1989.257.5.h1705 ◽

1989 ◽

Vol 257 (5) ◽

pp. H1705-H1711 ◽

Cited By ~ 1

Author(s):

A. P. Shepherd ◽

V. T. Randal ◽

J. M. Steinke ◽

J. L. Schmalzel

Keyword(s):

Whole Blood ◽

Capillary Tube ◽

Nonlinear Function ◽

Hemoglobin Concentration ◽

Infrared Light ◽

Oxyhemoglobin Saturation ◽

Light Emitting ◽

Total Hemoglobin ◽

Wide Range ◽

Total Hemoglobin Concentration

We have developed an oximeter that measures both the total hemoglobin concentration in whole blood and the percentage of the hemoglobin saturated with oxygen. The oximeter uses red and infrared light-emitting diodes to illuminate a capillary tube filled with a sample of whole blood. Light scattered by the blood travels a short distance down the length of the capillary tube and reaches a photodetector, the output of which is amplified, digitized, and fed into a microprocessor. The microprocessor computes the total hemoglobin concentration as a nonlinear function of the infrared light intensity. Oxyhemoglobin saturation is computed from the ratio of the logarithms of the intensities of red and infrared light. Our instrument has the following advantages over existing oximeters: 1) it provides a measurement of total hemoglobin concentration, 2) it is immune to the calibration shifts that fluctuations in total hemoglobin concentration cause in other oximeters, 3) it is accurate over a wide range of oxygen saturation, and 4) the blood samples are not diluted and can thus be preserved for further analysis. A detailed parts list and circuit diagram are presented, and sources of error are discussed.

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Photometric determination of glycosylation of hemoglobin in diabetes mellitus.

Clinical Chemistry ◽

10.1093/clinchem/26.12.1683 ◽

1980 ◽

Vol 26 (12) ◽

pp. 1683-1687 ◽

Cited By ~ 25

Author(s):

C V Subramaniam ◽

B Radhakrishnamurthy ◽

G S Berenson

Keyword(s):

Blood Glucose ◽

Blood Glucose Concentration ◽

Blood Glucose Control ◽

Hemoglobin Concentration ◽

Thiobarbituric Acid ◽

Photometric Determination ◽

Diabetic Patients ◽

Wide Range ◽

Average Blood Glucose

Abstract We evaluated glycosylation of hemoglobin (HbA + HbA1) in 25 control subjects and in 133 diabetic patients who were in various stages of blood glucose control, by measuring ketoamine-linked hexoses in hemoglobin. These hexoses were converted by digestion with 10 mol/L acetic acid for 16 h at 100 +/- 5 degrees C to 5-hydroxymethylfurfuraldehyde, which was quantitated by reaction with 2-thiobarbituric acid. Glycosylation of hemoglobin was expressed as micromoles of hydroxymethylfurfuraldehyde per gram of globin protein (the "HMF index"). A mean HMF index of 1.67 (SD = 0.23) was obtained for controls; that for diabetic patients was 2.93 (SD 0.95). The index correlated well (r = 0.83, p < 0.001) with average blood glucose concentration as measured during the preceding 16 weeks, over a wide range of glucose values (1 to 6 g/L). It correlated even better (r = 0.92, p < 0.001) when corrected for variations in hemoglobin concentration. Thus measurement of ketoamine-linked hexoses of hemoglobin or HMF index provides an independent and useful alternative to the currently used methods that measure only HbA1 or HbA1c.

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Oxygen affinity of blood in altitude Sherpas

Journal of Applied Physiology ◽

10.1152/jappl.1979.47.2.337 ◽

1979 ◽

Vol 47 (2) ◽

pp. 337-341 ◽

Cited By ~ 44

Author(s):

M. Samaja ◽

A. Veicsteinas ◽

P. Cerretelli

Keyword(s):

Sea Level ◽

Concentration Ratio ◽

Oxygen Affinity ◽

Hemoglobin Concentration ◽

Cell Counts ◽

Blood Cell Counts ◽

Total Hemoglobin ◽

The Mean ◽

Total Hemoglobin Concentration ◽

2,3 Dpg

Oxygen equilibrium curves on blood within 6 h from sampling have been estimated from polarographic measurements of oxyhemoglobin concentration, in 13 male 14- to 50-yr old Sherpas residing at 3,850 m above sea level (Kumjung, Nepal). In samples with red blood cell counts = 4.7 +/- 0.8 (SD) x 10(6)/mm3, total hemoglobin concentration [Hb] = 17.0 +/- 1.9 g/dl, and hematocrit = 53.3 +/- 5.0, the mean oxygen half-saturation of hemoglobin (P50) (pH = 7.4 and PCO2 = 40 Torr) was 27.3 +/- 1.8 Torr. The P50 of altitude Sherpas was not significantly different from that of acclimatized lowlanders (28.2 +/- 1.3; n = 7), sea-level Caucasian residents (26.5 +/- 1.0; n = 17), and Sherpas at sea level (27.1; n = 3). The 2,3-diphosphoglyceric acid-to-hemoglobin concentration ratio ([2,3-DPG]/[Hb]) in altitude Sherpas was 1.22 +/- 0.03, the same as that of acclimatized Caucasians (1.22 +/- 0.10). The Bohr effect measured for the blood of one altitude Sherpas by the ratio deltalog P50/deltapH was -0.32 and -0.45 at PCO2 levels of 40 and 20 Torr, respectively. These values are not significantly different from those found in Caucasians at sea level where deltalog P50/deltalpH was -0.35 and -0.42, respectively. It is concluded that the P50 in native highlanders is not significantly different from that observed in sea-level dwellers. [2,3-DPG]/[Hb] at altitude, both in natives and in newcomers, is 20% higher than in sea-level residents.

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