Some problems associated with assay of 25-hydroxycalciferol in human serum.

1977 ◽  
Vol 23 (5) ◽  
pp. 806-810 ◽  
Author(s):  
R S Mason ◽  
S Posen
Keyword(s):  
Human Serum ◽  
Silicic Acid ◽  
Rat Kidney ◽  
Extraction Procedure ◽  
Normal Subjects ◽  
Good Alternative ◽  
Alternative Methods ◽  
Control Serum ◽  
Free Material ◽  
Rat Kidney Cytosol

Abstract Methods available for the assay of 25-hydroxycalciferol in human serum are evaluated and compared to one another. Ethanol was chosen for use in the initial extraction procedure and rat-kidney cytosol as the binding protein, although good alternative methods are also available. We used silicic acid for chromatography and found this an essential step. Reproducibility was increased when, after "bound" and "free" material were separated, an aliquot of the supernate was pipetted into the counting vial instead of the entire supernatant fluid being decanted. Beta-lipo-protein added to the assay system was of no advantage; added bovine serum albumin interfered with the assay by giving rise to high blank values. With ethanol extraction, silicic acid chromatography, rat kidney cytosol and separation on dextran-coated charcoal, sera from normal subjects showed a mean 25-hydroxycalciferol concentration of 28.5 microng/liter (range, 13.1 to 43.9) during the fall season. Coefficients of variation for a control serum were 4.9% (intra-assay) and 10.9% (interassay).

DIRECT MEASUREMENT OF ARGININE-VASOPRESSIN IN HUMAN SERUM WITHOUT EXTRACTION PROCEDURE

1975 ◽  
Vol 80 (1_Suppla) ◽  
pp. S130 ◽  
Author(s):  
H. Wagner ◽  
V. Maier ◽  
H.-J. Herrmann ◽  
H. E. Franz
Keyword(s):  
Human Serum ◽  
Direct Measurement ◽  
Arginine Vasopressin ◽  
Extraction Procedure

Microdetermination of Cholesterol and Phospholipid in Cerebrospinal Fluid and Serum by Silicic Acid Column Chromatography

1962 ◽  
Vol 8 (6) ◽  
pp. 598-605 ◽  
Author(s):  
Yung S Shin ◽  
James C Lee
Keyword(s):  
Cerebrospinal Fluid ◽  
Human Serum ◽  
Total Phosphorus ◽  
Silicic Acid ◽  
Column Chromatography ◽  
Silicic Acid Column ◽  
Lipid Phosphorus ◽  
Silicic Acid Column Chromatography ◽  
Phospholipid Ratio

Abstract A method is presented for the determination of cholesterol and phospholipid, which requires 5 µl. of human serum or 1-2 ml. of cerebrospinal fluids. With this method 5-100 µg. of cholesterol and phospholipid can be separated by a modified silicic acid column after elution of the mixture with 1 ml. of chloroform and 3 ml. of methanol. Recovery for 24.6 µg. of cholesterol and 30.5 µg. of phospholipid was 98.4 and 96.7%, respectively. Standard deviations of ± 1.73 and ± 1.24 have been obtained for the reproducibility of cholesterol and phospholipid determinations after chromatography. The method has been applied for the estimation of the cholesterol/phospholipid ratio and of lipid phosphorus in total phosphorus of human cerebrospinal fluids.

Isolation and Measurement of Pancreatic Amylase in Human Serum and Urine

1972 ◽  
Vol 18 (12) ◽  
pp. 1493-1497 ◽  
Author(s):  
L Fridhandler ◽  
J Edward Berk ◽  
M Ueda
Keyword(s):  
Acute Pancreatitis ◽  
Human Serum ◽  
Fallopian Tube ◽  
Normal Subjects ◽  
Pancreatic Amylase ◽  
Salivary Amylase ◽  
Tissue Homogenates ◽  
Quantitative Procedure ◽  
Lines Of Evidence ◽  
Serum And Urine

Abstract We describe a sensitive quantitative procedure for separating isoamylases in human serum, urine, and tissue homogenates. Two components have been discerned with chromatographic characteristics resembling those of pancreatic and salivary amylases, respectively. Several lines of evidence—derived from studies in normal subjects, pancreatectomized patients, and patients with acute pancreatitis—indicate that the pancreas is probably the source of the component in serum and urine that exhibits characteristics of pancreatic amylase. The source of the component resembling salivary amylase has not yet been fully defined. Isoamylase analysis of extracts of fallopian tube and liver revealed two amylase components with chromatographic properties similar to pancreatic and salivary amylases, respectively.

RADIOIMMUNOASSAYS OF 3,5,3′-TRIIODO-L-THYRONINE WITH AND WITHOUT A PRIOR EXTRACTION STEP

1975 ◽  
Vol 80 (1) ◽  
pp. 58-69 ◽  
Author(s):  
A. Burger ◽  
C. Sakoloff ◽  
V. Staeheli ◽  
M. B. Vallotton ◽  
S. H. Ingbar
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Normal Values ◽  
Covalent Binding ◽  
Extraction Procedure ◽  
Mixed Population ◽  
Coupling Agents ◽  
Extraction Step ◽  
The Mean

ABSTRACT Two radioimmunoassays for triiodothyronine (T3) are described, one of which includes an extraction step, while the other does not. To raise antibodies, two carrier proteins and different coupling agents were used, namely haemocyanin and diazotized benzidine or human serum albumin and carbodiimide. In the case of T3 coupled to haemocyanin by diazotized benzidine, evidence of covalent binding of the hapten to the protein was obtained. In the case of T3 coupled to human serum albumin, little evidence of covalent linkage was available. Nevertheless immunization was successful in both cases. The radioimmunoassay in unextracted serum was highly reproducible and precise (intra-assay variability 5.2% inter-assay variability 8.1%). Normal values were determined which clearly indicate a fall in the serum T3 concentration with increasing age. In men the fall occurs in the fifth decade. In women the T3 starts to fall only after 70 years of age. In 31 cases of hyperthyroidism the serum T3 concentration ranged from 2.26 to 10.4 ng T3/ml. In 10 cases of hypothyroidism the values ranged from 0 to 0.8 ng T3/ml. The radioimmunoassay using an extraction procedure was less extensively used since it was found to be less reproducible (intra-assay variability 7.5%, inter-assay 12.25%). The normal values were determined with a mixed population aged 20–50. The mean ± 2 sd was 0.9 ± 0.36 ng T3/ml (n=52). In 17 cases of hypothyroidism the values ranged from 0 to 0.6 ng T3/ml and in 22 cases of hyperthyroidism from 2 to 14.4 ng T3/ml.

Fate and Toxicity of 2-(Fluoromethoxy)-1,1,3,3,3-pentafluoro-1-propene (Compound A)-derived Mercapturates in Male, Fischer 344 Rats 

1998 ◽  
Vol 89 (5) ◽  
pp. 1174-1183 ◽  
Author(s):  
Vinita Uttamsingh ◽  
Iyer A. Ramswamy ◽  
Raymond B. Baggs ◽  
M. W. Anders
Keyword(s):  
Spectroscopic Analysis ◽  
Rat Kidney ◽  
Urinary Metabolites ◽  
Fischer 344 Rats ◽  
Compound A ◽  
Fischer 344 ◽  
Mercapturic Acid Pathway ◽  
Magnetic Resonance Spectroscopic Analysis ◽  
Acid Pathway ◽  
Rat Kidney Cytosol

Background 2-(Fluoromethoxy)-1,1,3,3,3-pentafluoro-1-propene (compound A) is formed in the anesthesia circuit by the degradation of sevoflurane. Compound A is nephrotoxic in rats and undergoes metabolism by the mercapturic acid pathway in rats and humans to yield the mercapturates S-[2-(fluoromethoxy)-1,1,3,3,3-pentafluoropropyl]-N-acetyl-L -cysteine (compound 3) and S-[2(fluoromethoxy)-1,3,3,3-tetrafluoro-1-propenyl]-N-acetyl-L-cys teine (compound 5). These experiments were designed to examine the fate and nephrotoxicity of compound A-derived mercapturates in rats. Methods The deacetylation of compounds 3 and 5 by human and rat kidney cytosol and with purified acylases I and III was measured, and their nephrotoxicity was studied in male Fischer 344 rats. The metabolism of the deuterated analogs of compounds 3 and 5, [acetyl-2H3]S-[2-(fluoromethoxy)-1,1,3,3,3-pentafluoropropyl ]-N-acetyl-L-cysteine (compound 3-d3) and [acetyl-2H3]S-[2-(fluoromethoxy)-1,3,3,3-tetrafluoro-1-propenyl]-N -acetyl-L-cysteine (compound 5-d3), respectively, was measured. Results Compound 5, but not compound 3, was hydrolyzed by human and rat kidney cytosols and by acylases I and III. 19F nuclear magnetic resonance spectroscopic analysis showed no urinary metabolites of compound 3, but unchanged compound 5 and its metabolites 2-(fluoromethoxy)-3,3,3-trifluoropropanoic acid and 2-[1-(fluoromethoxy)-2,2,2-trifluoroethyl]-4,5-dihydro-1,3-thiazol e-4-carboxylic acid were detected in urine. Compound 5 (250 microM/kg) produced clinical chemical and morphologic evidence of renal injury in two of three animals studied. Conclusions Compounds 3 and 5 underwent little metabolism. Compound 5, but not compound 3, was mildly nephrotoxic. These results indicate that compound A-derived mercapturate formation constitutes a detoxication pathway for compound A.

Development of a stable lipoprotein diluent for use in reconstituting lyophilized human serum for the preparation of clear, hyperlipidemic quality-control materials.

1979 ◽  
Vol 25 (8) ◽  
pp. 1377-1380 ◽  
Author(s):  
G J Proksch ◽  
D P Bonderman
Keyword(s):  
Quality Control ◽  
Sodium Bicarbonate ◽  
Human Serum ◽  
Simple System ◽  
Lipoprotein Concentration ◽  
Control Serum ◽  
Quality Control Materials

Abstract We describe a simple system for the preparation of a hyperlipidemic control serum. Beta- and pre-beta-lipoproteins are removed from the serum and prepared as a stable diluent; the extracted serum is then lyophilized. Upon addition of the lipoprotein diluent, which was observed to contain only liproprotein and sodium bicarbonate, the serum reconstitutes rapidly, usually within 5 min. By a suitable adjustment of the lipoprotein concentration in the diluent, a hyperlipidemic control serum may be produced with desired concentrations of lipids. Because the lipoproteins are not included in the destructive lyophilization step, the resulting product has remarkable clarity, precision, and stability.

NORMAL IN VIVO KINETICS OF FACTOR VIII (F VIII) AND FACTOR IX (F IX) TREATED WITH TRI (N-BUIYL) PHOSPHATE (TNBP) AND TWEEN 80 FOR INACTIVATION OF VIRUSES

1987 ◽  
Author(s):  
C Baumgartner ◽  
B A Perret ◽  
E Meili ◽  
M Furlan ◽  
H Friedli ◽  
...  
Keyword(s):  
Body Weight ◽  
Solvent Extraction ◽  
Extraction Procedure ◽  
Coagulation Factor ◽  
Factor Ix ◽  
Alternative Methods ◽  
Maximum Increase ◽  
In Vivo Kinetics ◽  
Lipid Solvent

Heat treatment has been cxxnmonly used for the sterilisation of coagulation factor concentrates. This causes, however, considerable loss of coagulation factor activity? therefore alternative methods have been developed. Two new virus-inactivated coagulation factor preparations were recently introduced by our institution. Their manufacturing procedure includes a lipid solvent extraction step: The cryoprecipitate (F VIII preparation) or the first DEAE eluate (F IX preparation) is incubated with 0.3 % TNBP and 1 % Tveen 80 at 24°C for at least 12 hours. (Horowitz, Transfusion 25 : 516-522, 1985). Single doses of these preparations (F VIII: median 20.5, range 6-33 U/kg, F IX: median 25, range 9-46 U/kg body weight) were given intravenously to 28 hemophilic patients with minor or no bleeding. F VIII or F IX levels in plasma were determined before and at intervals up to 48 h after injection. The recovery was calculated from the maximum increase of activity and an plasma volume of 41 ml/kg body weight. The plasma half life was calculated according to the procedures described by Morfini (Thrcmb. Res. 42: I-III, 1986). Results are shown in the table below.No side effects were recorded. Hemostasis was satisfactory in all patients with bleedings (n = 13). These results are within the range expected for conventional and heat treated F VIII and F IX preparations. We conclude that the lipid solvent extraction procedure as used here does not influence the in vivo kinetics and the function of F VIII and F IX and does not induce any acute toxicity.

Improved assessment of plasma lipoprotein patterns. IV. Simple preparation of a lyophilized control serum containing intact human plasma lipoproteins.

1982 ◽  
Vol 28 (6) ◽  
pp. 1335-1337 ◽  
Author(s):  
H Wieland ◽  
D Seidel
Keyword(s):  
Human Serum ◽  
Human Plasma ◽  
Clinical Chemistry ◽  
Plasma Lipoprotein ◽  
Plasma Lipoproteins ◽  
Simple Preparation ◽  
Control Serum

Abstract Addition of sucrose (413-825 mmol/L) to human serum allows plasma lipoproteins to be lyophilized without denaturation. A control serum so prepared is especially suited for use in monitoring determinations of apolipoproteins and quantitative lipoprotein electrophoresis. It is clear upon reconstitution; hence, the described procedure may also be useful for preparation of control sera for general clinical chemistry without de-lipoproteinization.

scholarly journals Characterization of glutathione S-transferases in rat kidney. Alteration of composition by cis-platinum

1988 ◽  
Vol 252 (1) ◽  
pp. 127-136 ◽  
Author(s):  
G M Trakshel ◽  
M D Maines
Keyword(s):  
Specific Activity ◽  
Rat Kidney ◽  
Male Rats ◽  
Glutathione S Transferase ◽  
Aberrant Form ◽  
Glutathione S Transferases ◽  
Exchange Matrix ◽  
Rat Kidney Cytosol

We have developed chromatographic and mathematical protocols that allowed the high resolution of glutathione S-transferase (GST) subunits, and the identification of a previously unresolved GST monomer in rat kidney cytosol; the monomer was identified tentatively as subunit 6. Also, an aberrant form of GST 7-7 dimer appeared to be present in the kidney. This development was utilized to illustrate the response of rat kidney GST following cis-platinum treatment in vivo. Rat kidney cytosol was separated into three ‘affinity families’ of GST activity after elution from a GSH-agarose matrix. The affinity peaks were characterized by quantitative differences in their subunit and dimeric compositions as determined by subsequent chromatography on a cation-exchange matrix and specific activity towards substrates. By use of these criteria, the major GST dimers of affinity peaks were tentatively identified. The major GST dimers in peak I were GST 1-1 and 1-2, in affinity peak II it was GST 2-2, and in peak III they were GST 3-3 and 7-7. GST 3-6 and/or 4-6, which have not been previously resolved in kidney cytosol, were also present in peak II. Alterations in the kidney cytosolic GST composition of male rats were detected subsequent to the administration of cis-platinum (7.0 mg/kg subcutaneously, 6 days). This treatment caused a pronounced alteration in the GST profile, and the pattern of alteration was markedly different from that reported for other chemicals in the kidney or in the liver. In general, the cellular contents of the GSTs of the Alpha and the Mu classes decreased and increased respectively. It is postulated that the decrease in the Alpha class of GSTs by cis-platinum treatment may be related to renal cortical damage and the loss of GSTs in the urine. The increase in the Mu class of GSTs could potentially stem from a lowered serum concentration of testosterone; the latter is a known effect of cis-platinum treatment.

Concanavalin A-bound selenoprotein in human serum analyzed by graphite furnace atomic absorption spectrometry

1994 ◽  
Vol 40 (1) ◽  
pp. 62-70 ◽  
Author(s):  
R Daher ◽  
F Van Lente
Keyword(s):  
Human Serum ◽  
Atomic Absorption ◽  
Concanavalin A ◽  
Graphite Furnace ◽  
Direct Determination ◽  
Absorption Spectrometry ◽  
Normal Subjects ◽  
Acute Phase Reactant ◽  
Single Chain ◽  
Total Selenium

Abstract We developed an assay for the direct determination of selenium in serum with a Perkin-Elmer Model 4100ZL Zeeman atomic absorption spectrometer and Ag-Cu-Mg modifier. We used this assay to analyze concanavalin A-bound selenoprotein (CABSP) in human serum after concanavalin A (ConA) affinity chromatography. The CABSP was identified as a single-chain glycoprotein of 57.3 kDa. Carbohydrate units were N- and O-linked to the protein. The selenium moiety was selenocysteine. Total selenium, glutathione peroxidase (GPX; EC 1.11.1.9), ConA-bound selenium (CABS), and alpha 1-acid glycoprotein (AAG) were determined in normal subjects and patients with various pathological conditions. CABS accounted for 44.1% +/- 6.3% of total selenium in sera from normal subjects and 46.5% +/- 3.9% to 55.1% +/- 8.1% in sera from patients with a variety of diseases. Total selenium in serum was well correlated with serum CABS (r = 0.860), but not with serum GPX activity (r = 0.117), for all patients studied. Serum CABS increased in normal subjects after selenium supplementation. Serum CABSP did not behave as an acute-phase reactant, compared with AAG.

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