Evaluation of a nephelometric assay for haptoglobin and its clinical usefulness.

1979 ◽  
Vol 25 (12) ◽  
pp. 2007-2010 ◽  
Author(s):  
F Van Lente ◽  
A Marchand ◽  
R S Galen
Keyword(s):  
Correlation Coefficient ◽  
Coefficient Of Variation ◽  
Binding Capacity ◽  
Clinical Correlation ◽  
Intravascular Hemolysis ◽  
Hemolytic Disease ◽  
Serum Haptoglobin ◽  
Hemoglobin Binding ◽  
Significant Interference

Abstract Serum haptoglobin has been advocated as an indicator of intravascular hemolysis. We have evaluated a nephelometric determination of serum haptoglobin. The assay is sensitive and exhibits within-run precision in the range of 2.5-7.4% coefficient of variation (CV) and between-run precision of 7.0% (CV). In addition, when haptoglobin values determined with the nephelometric assay were compared with hemoglobin-binding capacity determined by electrophoresis, the correlation coefficient was 0.968. The assay is essentially independent of phenotype and free of significant interference by hemolysis. The clinical correlation of haptoglobin values obtained for 100 selected patients with the nephelometric technique correlated well, if less than 250 mg/L, with the presence of hemolytic disease.

Manual Salicylate-Hypochlorite Procedure for Determination of Ammonia in Kjeldahl Digests

1982 ◽  
Vol 65 (5) ◽  
pp. 1076-1079
Author(s):  
Helen E Fukumoto ◽  
George W Chang
Keyword(s):  
Correlation Coefficient ◽  
Spectrophotometric Method ◽  
Coefficient Of Variation ◽  
Precise Timing ◽  
Coefficients Of Variation ◽  
Moderate Volume ◽  
Kjeldahl Method

Abstract The determination of nitrogen in Kjeldahl digests of urine and feces has been simplified by using a manual spectrophotometric method with salicylate and hypochlorite. It neither involves the hazards of the phenol-hypochlorite method, nor requires an automated analyzer. We determined the conditions which minimize the need for precise timing and neutralization of acidic digests and obtained coefficients of variation of about 0.8%. Agreement between the spectrophotometric method and the conventional micro-Kjeldahl method was excellent; a correlation coefficient of 0.9992 and a coefficient of variation of the estimate of 2.1% were obtained. This method is well suited for a laboratory with a moderate volume of samples. We could process at least 200 digests per day, in contrast with the 60 per day which we had previously done by the Kjeldahl distillation and titration.

Determination of Serum-Hemoglobin Binding Capacity and Haptoglobin-Type by Acrylamide Gel Electrophoresis

1966 ◽  
Vol 46 (3_ts) ◽  
pp. 385-389 ◽  
Author(s):  
Thomas G. Ferris ◽  
Robert E. Easterling ◽  
Kenneth J. Nelson ◽  
Richard E. Budd
Keyword(s):  
Gel Electrophoresis ◽  
Binding Capacity ◽  
Hemoglobin Binding

Radioimmunoassay for pteroylglutamic acid.

1981 ◽  
Vol 27 (5) ◽  
pp. 701-703 ◽  
Author(s):  
J Hendel
Keyword(s):  
Detection Limit ◽  
Bovine Serum Albumin ◽  
Coefficient Of Variation ◽  
Clinical Applicability ◽  
Specific Determination ◽  
Pteroylglutamic Acid ◽  
Tetrahydrofolic Acid ◽  
Oral Doses ◽  
Significant Interference

Abstract I describe a radioimmunoassay for specific determination of pteroylglutamate in plasma, erythrocytes, and urine. Antiserum was raised in rabbits immunized with a conjugate of pteroylglutamic acid and methylated bovine serum albumin. The lowest detection limit of the assay was 0.1 microgram/L; the coefficient of variation ranged from 4.01 to 14.63%. No significant interference was demonstrated from tetrahydrofolic acid, 5-methyl-tetrahydrofolic acid, or methotrexate. Under normal sampling and storing conditions dihydrofolic acid was oxidized to pteroylglutamic acid and estimated as such in the assay. The clinical applicability of the assay is demonstrated by measurements of pteroylglutamate absorption rates after oral doses of pteroylglutamic acid or pteroyltriglutamic acid and by the estimation of the accumulation of pteroylglutamate and dihydrofolate in plasma during therapy with methotrexate.

Kinetic determination of serum haptoblobin with a centrifugal analyzer.

1976 ◽  
Vol 22 (12) ◽  
pp. 1962-1967 ◽  
Author(s):  
T S Kickler ◽  
P F Gong ◽  
G F Johnson ◽  
H M Solomon
Keyword(s):  
Binding Capacity ◽  
Regression Line ◽  
Normal Serum ◽  
Least Squares Regression ◽  
Kinetic Determination ◽  
Automated Method ◽  
Hemoglobin Binding ◽  
Total Peroxidase Activity ◽  
Assay Conditions

Abstract We describe a method for determining haptoglobin with a centrifugal analyzer that is based on haptoglobin combining stoichiometrically with hemoglobin to form a complex that has peroxidase-like activity proportional to the quantity of haptoglobin present. Under assay conditions, unbound hemoglobin exhibits only a small fraction of the total peroxidase activity. Activity is measured colorimetrically at 405 nm after reaction with o-dianisdine and ethyl hydrogen peroxide. The procedure is standardized by saturating aknown amount of hemoglobin with a serum whose hemoglobin binding capacity exceeds the amount of hemoglobin in the assay system. The mean and mean within-run precision of our method, determined by performing 17 replicate assays of both a pooled normal serum and a 10-fold dilution of the serum, was 1.13 g/liter (CV, 2.9%), and 106 mg/liter (cv, 5.8%), respectively. The 95 percentile estimate of the normal range by our method is 0.45-1.85 g/liter hemoglobin binding capacity. When results by our automated method were compared to those by a manual method [Scand. J. Clin. Lab. 2nvest. 18, 80 (1965)], the slope of the unweighted linear least-squares regression line was .970 the y-intercept 26 mg/liter, and the correlation coefficient .995.

scholarly journals Studies on the Serum Haptoglobin Level in Hemoglobinemia and Its Influence on Renal Excretion of Hemoglobin

Blood ◽  
1957 ◽  
Vol 12 (6) ◽  
pp. 493-506 ◽  
Author(s):  
CARL-BERTIL LAURELL ◽  
MARGARETA NYMAN
Keyword(s):  
Intravenous Injection ◽  
Threshold Value ◽  
Free Hemoglobin ◽  
Electrophoretic Method ◽  
Serum Haptoglobin ◽  
Hemoglobin Binding ◽  
Constant Rate ◽  
Haptoglobin Level ◽  
Binding Power

Abstract A short survey is given of the literature on haptoglobin, the hemoglobin-binding serum protein, its properties and biologic variations. The principles of an electrophoretic method for quantitative determination of the serum haptoglobin are described. Electrophoretic studies showed that haptoglobin has a high affinity for hemoglobin at physiologic pH and that every haptoglobin molecule can bind at least 2 hemoglobin molecules. Observations made following the intravenous injection of hemoglobin showed: that hemoglobin administered intravenously is bound by the haptoglobin; that free hemoglobin is not demonstrable until more hemoglobin has been injected than can be bound by the haptoglobin; that the complex hemoglobin-haptoglobin is eliminated from the plasma after intravascular hemolysis or intravenous administration of hemoglobin without being excreted in the urine; that the hemoglobin-haptoglobin complex is removed from the plasma at a constant rate during the major part of the elimination period; that the haptoglobin level will fall to nil within 24 hours, if the amount of hemoglobin injected is sufficient to bind all the haptoglobin available. During the following days the rate of formation of haptoglobin can be studied. From the data available it can be concluded that hemoglobinuria cannot appear until the amount of hemoglobin administered intravenously or the amount liberated intravascularly exceeds the binding power of the haptoglobin and the reabsorption capacity of the tubules. The variation observed by earlier authors in the so-called renal threshold for hemoglobin on intravenous injection of hemoglobin can be explained among other things by the variation in the haptoglobin content in one and the same subject, i.e., if the haptoglobin level is low, the threshold value will also be low, and vice versa.

Spectrophotometric Difference Method for Determination of Capsaicin

1979 ◽  
Vol 62 (5) ◽  
pp. 998-1000
Author(s):  
Joseph J Dicecco
Keyword(s):  
Correlation Coefficient ◽  
Difference Method ◽  
Coefficient Of Variation ◽  
Activated Alumina ◽  
Alumina Column

Abstract Capsaicin is extracted from capsicum spice with acetone. The extract is cleaned up on an activated alumina column and assayed by using a spectrophotometric difference method (SDM). The coefficient of variation of replicate samples for 2 pungency levels was 0.36%. The correlation coefficient between the organoleptic assay and SDM was 0.95.

scholarly journals Effects of the Drug Disulphine Blue on Routine Biochemical Investigations

1983 ◽  
Vol 20 (5) ◽  
pp. 317-320 ◽  
Author(s):  
Stephen P Halloran ◽  
David J Torrens
Keyword(s):  
Binding Capacity ◽  
Total Iron ◽  
Blue Dye ◽  
Colorimetric Determination ◽  
Reaction Conditions ◽  
Successive Sampling ◽  
Absorbance Peak ◽  
Iron Binding Capacity ◽  
Significant Interference

Major interference by Disulphine Blue in the colorimetric determination of amylase, albumin, protein, iron and total iron-binding capacity is described. The drug, an intense blue dye, was administered intravenously to a patient before surgery to allow demarcation of devitalised bone. Successive sampling showed the drug to have a half-life of 30 hours, to remain visible for 150 hours in both urine and blood, and to cause significant interference with the five analytes for up to 2 1/2 days. The 640 nm absorbance peak of Disulphine Blue was shown to be pH-dependent, and therefore the reaction conditions of individual methods may influence the degree of interference.

scholarly journals Serum Protein Binding of Myoglobin, Hemoglobin and Hematin

Blood ◽  
1960 ◽  
Vol 16 (5) ◽  
pp. 1579-1585 ◽  
Author(s):  
MUNSEY S. WHEBY ◽  
O'NEILL BARRETT ◽  
WILLIAM H. CROSBY
Keyword(s):  
Serum Protein ◽  
Binding Capacity ◽  
Normal Serum ◽  
Starch Gel Electrophoresis ◽  
Serum Protein Binding ◽  
Hemolytic Disease ◽  
Total Hemoglobin ◽  
Hemoglobin Binding ◽  
Human Serum Protein ◽  
Starch Gel

Abstract Using paper and starch-gel electrophoresis we have demonstrated binding of hemoglobin, myoglobin and hematin by human serum protein which is not a haptoglobin. This protein was present in all sera tested except that from two patients with hemolytic anemia and was also present in serum genetically deficient in haptoglobin. The binding capacity of this protein is low and contributes only slightly to the total hemoglobin binding capacity of normal serum. However, it may represent most or all of the binding in sera from patients with hemolytic disease.

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