Icteric Serum Research Articles

A modification of the ceric oxidation method of butanol-extractable iodine using a new extraction filter

A modification of the BEI method previously introduced by the author is described. The main feature of this modification is the utilization of a new extraction-filter the use of which shortens considerably the time required for serum extraction with no loss of over-all methodological precision. Average time required for analyzing 6 samples is 3.5 hr, and 24 samples can be run in 7 hr. With this modified procedure, quantitative recoveries of thyroxine iodine or free iodide added to sera were effected. A normal range for 250 patients was established between 3.0 and 7.0 μg per 100 ml serum with a mean ± S.D. of 4.5 ± 1.1 μg per 100 ml. Bilirubin was found not to interfere with the analysis although in highly icteric serum (total bilirubin 16.3 mg per 100 ml) recovery of added thyroxine iodine was slightly lower than in normal serum.

Conjugated and Unconjugated Bilirubin Determined in Icteric Sera by Direct Spectrophotometry

The difference between the absorption curves of conjugated and unconjugated bilirubin is used for determining their ratio. The error due to oxyhemoglobin is eliminated.Icteric serum is diluted at pH 8.3 and absorbances are read at three wavelengths: 460, 440, and 415 mμ. Per cent unconjugated bilirubin is calculated according to equations given in the text.The difference between the total amount of bilirubin determined as azopigments and the fraction of unconjugated bilirubin expresses the amount of bilirubin-conjugates in icteric sera where absorbance due to nonbilirubin yellow pigments is negligible.Values obtained by this method, by the Fog 1964 method, and by chloroform extraction agree within narrow limits.

僻地病院の輸血後肝炎

It is very difficult to collect accurate statistios on post transfusion hepatitis at university hospitals of other large medical centers. Because of its long incubation period, and because many of the patients at these hospitals come from a distance. Therefore, by the time the symptoms of serum hepatitis appear, the patients have returned to their homes—many to remote rural areas.Therefore, the authors chose the Yosanoumi Hospital in the northern part of Kyoto Prefecture as suitable for a more accurate statistical study of post transfusion hepatitis. Liver function tests were performed periodically for 6 months in all patients receiving transfusions at this hospital.1) Between August, 1963 and December, 1964, 51 patients received transfusions at this hospital. Serum hepatitis developed in a total of 43% of these cases, 17.6% with and without 25.5% jaundice.2) Amount tuberculous patients the incidence of icteric serum hepatitis was lower, probably because they were kept quiet and lived a better regulated life for more than 6 months after operation.3) The incubation period renged from 37 to 112 days. In 86% of the cases liver dysfunction appeared less than three months after transfusion.4) The author made it a routine to wash syringes and needles immediately after injection or transfusion and to sterilize them in an autoclave. Therefore, non-transfusion serum-hepatitis could be ruled out in this study.

Studies on the Determination of Bile Pigments

Abstract Tests for total bilirubin on 10 icteric sera by 5 diazo procedures and a spectrophotometric method showed fair agreement with most samples but an occasional grossly discrepant result. Results for free and conjugated bilirubin by diazo methods, acetone precipitation, and extraction of serum with ethyl acetate and chloroform were widely discrepant. Problems in documentation of accuracy of such procedures are discussed. It is concluded that unavailability of reference standards of bilirubin conjugates poses a serious obstacle to solution of the analytical problems.

Comparison of Technics for Estimation of Albumin by Determination of Its Dye-Binding Capacity and by Paper Electrophoresis

Abstract Values for serum albumin obtained by the methyl orange and 2-(4’-hydroxyazobenzene) benzoic acid (HABA) dye-absorption methods have been compared with values obtained by Spinco Procedure B paper-electrophoresis system. A good correlation of albumin values has been found by the three methods in serums with albumin/globulin ratios ranging from 0.19 to 3.00. Modifications of the dye-absorption procedures are presented which permit use of reagents ordinarily employed for automatic analyses on small numbers of specimens. A decrease in ionic strength of the methyl orange buffer tends to increase the sensitivity of this procedure. Temperature affects the HABA dye/albumin reaction. Bilirubin, free hemoglobin, and lipid interfere with both dye procedures. Means for eliminating most of these interferences have been presented. The methyl orange procedure appears to be superior to the 2-(4 -hydroxyazobenzene) benzoic acid method for estimating albumin in icteric serum and serum containing macroglobulins. Albumin values of serum containing abnormal proteins were compared with values obtained by paper electrophoresis and both dye procedures. Estimation of albumin by the two dye procedures appears to be simple, precise, and fairly specific.

Direct Determination of Calcium in Icteric Serum

Abstract A rapid method suitable for the direct titrimetric determination of calcium in icteric sera is presented. The major portion of serum chromogens are oxidized to colorless products by either a mixture of hydrogen peroxide and potassium ferricyanide or by sodium hypochlorite. Ethylenediamine tetraacetic acid (EDTA) is the titrant used and either calcein or Cal-Red is suitable as the specific calcium indicator. Results compare favorably with those obtained by the method of Clark and Collip.

A Mercurimetric Chloride Determination Suitable for Icteric Sera

Abstract A modification of the direct mercurimetric determination of serum chloride suitable for icteric sera is presented. The method is simple, rapid, and accurate. No deproteinization of the sample is required. The results agree well with the standard protein-free filtrate procedure of Schales and Schales.

Removal of Bilirubin by Albumin Binding during Intermittent Peritoneal Dialysis

Since the introduction of intermittent peritoneal dialysis by Grollman et al.1 in 1951 a number of reports have appeared attesting to its efficacy in the removal of dialyzable substances in acute renal failure,2 3 4 5 hypercalcemia,4 barbiturate intoxication,4 salt poisoning,6 salicylism7 and hepatic coma.8 The toxic substances involved in these conditions are water soluble and freely dialyzable. Bilirubin, however, is poorly soluble in water and is almost entirely bound to protein in icteric serum.9 The present report of a case of an unusual combination of acute hepatitis, drug-induced hemolytic anemia and renal failure describes the accelerated removal of bilirubin induced by the . . .

A rapid micro-method for determining serum calcium

1. 1. A micro method for the determination of calcium in serum by direct titration under ultra-violet light with EDTA, using the fluorescent indicator calcein, is described. Good recoveries of calcium from serum, both directly and after ashing, have been obtained. 2. 2. The effect of possible interfering substances present in serum has been investigated. Magnesium and phosphate do not interfere in the concentrations present in serum and the determination can be made on haemolysed and icteric sera. 3. 3. Thirty sera have been analysed by fluorescent titration, murexide titration and by the Kramer-Tisdall technique. No significant difference has been found between the results given by the three methods. 4. 4. A convenient apparatus for carrying out titrations under ultra-violet light has been described.

Serum bilirubin and the yellow color of icteric serum.

Serum bilirubin and the yellow color of icteric serum.

Influence of the surface tension in serum and urine on the flamephotometric analysis.

AbstractThe experiments show that the low surface tension that may occur in diluted icteric sera (probably also in specimens of urine) have no effect on the flame photometric analysis, whether or not this is carried out with internal standard. As pointed out before (Ryssing, 1952), for other reasons it is of advantage to use internal standard.

Photometric studies on the diazo reaction for serum bilirubin.

Photometric studies on the Malloy-Evelyn procedure for the determination of serum bilirubin have shown that the azobilirubin color intensity obtained when the diazo reagent acts for 30 min. upon a known amount of bilirubin added to serum in the form of the sodium salt is less than when the reaction takes place with the same amount of bilirubin in chloroform-alcohol solution. This suggests that the usual calibration curve prepared from bilirubin in chloroform-alcohol solution, when used as standard for serum bilirubin determinations, may give values which are about 10% less than the true bilirubin content of the serum. It has also been shown that with the 1 : 10 dilution of the Malloy-Evelyn procedure the azobilirubin from icteric sera does not obey Beer’s law beyond a serum bilirubin content of 15 mgm. per 100 ml. Evidence is submitted that the rate of azobilirubin color formation can be markedly accelerated by increasing the strength of the diazo reagent, and a new reagent is proposed by the use of which the time of color development can be reduced from 30 min. to 5 min.

SERUM IRON AND IRON-BINDING CAPACITY OF THE SERUM IN CHILDREN WITH SEVERE MEDITERRANEAN (COOLEY'S) ANEMIA

The blood of children with severe Mediterranean anemia was found to have a high serum iron and an absence of a latent or unbound iron capacity of the serum. Whereas serum iron levels comprise approximately one third of the total iron-binding capacity in normal children and adults, the two values are equal in the severe form of Mediterranean anemia. The iron-binding capacity of the serum in this condition is therefore fully saturated and quantitatively lower than in normal children. The absence of a measurable latent iron-binding capacity does not depend entirely on the liberation of iron from exogenously administered blood. Comparable changes were also noted in two nonanemic adult carriers possessing the trait of the disease, in two children with the mild form not requiring transfusions, and in one severely anemic child with Mediterranean anemia who had never been transfused. Similar changes occurred in three patients with spherocytic anemia and in five children with sickle cell anemia, one of whom had never been transfused. In every instance therefore of the advanced form of Mediterranean anemia and in some of the nonanemic and mildly anemic forms of this disease, a fully saturated metal-binding protein is present in total amounts usually less than those found in normal children. In contrast, children with iron deficiency anemia manifest a reduction in percentage saturation of iron-binding protein, an increased level of unbound latent iron-binding capacity and a markedly elevated total iron-binding capacity. The failure to demonstrate a latent iron-binding capacity of the serum in asymptomatic persons adds another criterion to those already established for the identification of carriers of the trait of Mediterranean anemia who constitute the source of its transmission. The presence of bilirubin in increased concentrations in the serum of patients with severe Mediterranean anemia does not interfere with its capacity to combine with iron. The capacity for binding iron was restored when Fraction IV-7 was added to samples of icteric sera of patients in whom no capacity had previously been demonstrated. The high serum iron and absence of latent iron-binding capacity observed in children with severe Mediterranean anemia receiving multiple transfusions correspond to the changes regarded as diagnostic of transfusion hemosiderosis and hemochromatosis. While these changes remain unaltered in Mediterranean anemia, their reversibility has been demonstrated in two children with hemophilia who received weekly transfusions of small amounts of blood. Following cessation of this form of therapy a decrease of serum iron and a gradual restoration of iron-binding capacity were noted.

A SIMPLE METHOD FOR THE DETERMINATION OF PHENOLTETRACHLORPHTHALEIN IN BLOOD SERUM

The clinical determination of hepatic function by means of phenoltetrachlorphthalein was first proposed by Rowntree, Hurwitz and Bloomfield. Rosenthal¹improved the technic of the test by injecting the dye intravenously and examining the blood serum after from fifteen to sixty minutes for its dye content. The technic he employed entailed the addition of several drops of 5 per cent. sodium hydroxid to the serum, and the comparison of the test specimen with known concentrations of the dye. He found retention of phenoltetrachlorphthalein in the blood serum of both patients and experimental animals in which there was severe damage to the liver. We have made use of Rosenthal's technic in a clinical and experimental study of liver function. These results are to be published elsewhere. In carrying out the test, not infrequently specimens were found in which the presence of slight amounts of hemolysis, lipemia and icteric serum greatly interfered