Extraction and Preliminary Purification of Cellulase from Cipangopaludina Cahayensis

ABSTRACT A method is described for the quantitative estimation of C19-Δ16-steroids in human urine. After extraction and preliminary purification by the method of Brooksbank & Haslewood (1961), the steroids were analyzed as chloromethyl dimethylsilyl ethers by gas-liquid chromatography. The specificity, accuracy and precision of the method were found to be satisfactory and comparison with the colorimetric method of Brooksbank & Haslewood (1961) for urinary 3α-hydroxy-5α-androst-16-ene showed a high correlation coefficient, r = 0.97 (P < 0.001). Figures are given for the urinary excretion rate of 3α-hydroxy-5α-androst-16-ene of normal men and women and of some women suffering from hirsutism and from adrenocortical overactivity. The values are given for 3α-hydroxy-5β-androst-16-ene and 3β-hydroxyandrosta-5,16-diene in pools of urine from healthy men and women.

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THE QUANTIFICATION OF SERUM 25-HYDROXYVITAMIN D USING A COMPETITIVE PROTEIN-BINDING ASSAY AFTER PRELIMINARY PURIFICATION

Vitamin D ◽

10.1515/9783110846713.730 ◽

1988 ◽

pp. 730-731

Keyword(s):

Protein Binding ◽

Binding Assay ◽

Hydroxyvitamin D ◽

Protein Binding Assay ◽

Preliminary Purification ◽

Competitive Protein Binding Assay ◽

25 Hydroxyvitamin D ◽

Competitive Protein Binding

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Improved Measurement of Corticosteroids in Plasma and Urine by Competitive Protein-Binding Radioassay

Clinical Chemistry ◽

10.1093/clinchem/19.5.511 ◽

1973 ◽

Vol 19 (5) ◽

pp. 511-515 ◽

Cited By ~ 20

Author(s):

Miguel Ficher ◽

George C Curtis ◽

V K Ganjam ◽

Leon Joshlin ◽

Sarah Perry

Keyword(s):

Protein Binding ◽

Horse Serum ◽

Small Samples ◽

Circadian Cycle ◽

Lower Range ◽

Standard Curve ◽

Preliminary Purification ◽

Corticosteroid Binding Globulin ◽

Competitive Protein Binding

Abstract In the assay of corticosteroids in plasma and urine by competitive protein binding, the use of horse serum or plasma as a source of assay protein gives better sensitivity and somewhat better specificity for cortisol than do previously described procedures. Five to 10 µl of plasma unknown or 50 to 100 µl of urine unknown can be used. Precision for the standard curve is 0.12 ng in the range 0-2.00 ng, and accuracy, measured as the amount of added cortisol recovered from plasma, is about 91%. Because of the improved specificity, the need for preliminary purification by chromatography is decreased for some purposes. The same corticosteroid-binding globulin and standard curve can be used for assaying corticosteroids in urine or plasma. The procedure may be useful for unusually small samples or if discriminations within the lower range of physiological concentrations are needed, as in work with infants and neonates or in the study of small adrenal secretory pulses, such as those occurring at the nadir of the circadian cycle.

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Sewage cleaning by using a phase separator

E3S Web of Conferences ◽

10.1051/e3sconf/202016401020 ◽

2020 ◽

Vol 164 ◽

pp. 01020

Author(s):

Nikolay S. Serpokrylov ◽

Alla S. Smolyanichenko ◽

Elena V. Yakovleva

Keyword(s):

Industrial Wastewater ◽

Local Treatment ◽

Industrial Plant ◽

Urgent Problem ◽

Laboratory Studies ◽

Active Experiment ◽

Preliminary Purification ◽

Treatment Facilities ◽

The Cost ◽

Phase Separator

This article proposes a solution to the urgent problem of treating oil-, fat-containing wastewater. A phase separator of dispersions for preliminary purification of industrial wastewater contaminated with fats is proposed, its effectiveness in the treatment of wastewater at local treatment facilities of an industrial plant for the production of sunflower oil is tested. In order to reduce the cost of acquiring reagents and increase the efficiency of purification of fat-containing wastewater, the use of carbide sludge and sodium hydroaluminate was studied. Laboratory studies conducted by the method of an active experiment.

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The determination of oestradiol and oestrone in the plasma of the domestic fowl by a method involving the use of labelled derivatives

Biochemical Journal ◽

10.1042/bj1060077 ◽

1968 ◽

Vol 106 (1) ◽

pp. 77-86 ◽

Cited By ~ 16

Author(s):

J. E. O'Grady

Keyword(s):

Human Plasma ◽

Paper Chromatography ◽

Domestic Fowl ◽

Specific Activity ◽

High Specific Activity ◽

Double Labelling ◽

Plasma Samples ◽

Labelling Technique ◽

Preliminary Purification

1. A method involving the use of triple-labelled derivatives has been developed for the determination of total oestrone and oestradiol in the plasma of the domestic fowl. The double-labelling technique devised by Svendsen (1960) for the determination of free oestrogens in human plasma was modified to enable the total oestrogen recovery to be determined for each sample. 2. [6,7−3H2]Oestradiol-17β is added to the plasma samples (1–10ml.), which are hydrolysed with acid and the phenolic steroids then extracted and partially purified. The extract is esterified with iodobenzene-p[35S]-sulphonyl chloride of high specific activity. After addition of standard oestrogen [131I]iodobenzene-p-sulphonates the esters are finally purified by paper chromatography. 3. The oestrogens are determined by comparing the 3H/35S and 131I/35S ratios in the purified esters with similar ratios of appropriate standards. 4. With this procedure the recoveries of oestrone and oestradiol after hydrolysis were 70–85% and 72–84% respectively, and after hydrolysis and preliminary purification 38–53% and 39–51% respectively. With this procedure up to 500ng. of oestradiol can be determined. The sensitivity of the technique for oestrone is 3·0ng. and for oestradiol 2·1ng. 5. The ranges of oestradiol and oestrone concentrations found in six plasma samples were 8·3–21·4ng./ml. and 15·2–31·6ng./ml. respectively.

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Further purification and characterization of the acid α-glucosidase

Biochemical Journal ◽

10.1042/bj1080161 ◽

1968 ◽

Vol 108 (2) ◽

pp. 161-167 ◽

Cited By ~ 22

Author(s):

F. Auricchio ◽

C. B. Bruni ◽

V. Sica

Keyword(s):

Rat Liver ◽

Gel Electrophoresis ◽

Gel Filtration ◽

Human Kidney ◽

Purification And Characterization ◽

Preliminary Purification ◽

Kidney Enzyme ◽

Michaelis Constants ◽

Final Degree

1. Centrifugation of rat liver acid glucosidase, which had been purified by adsorption on dextran gel, on a density gradient of sucrose showed the enzyme to be impure. 2. Preliminary purification of the enzyme before the gel filtration improved the final degree of purity of this preparation. Disc gel electrophoresis of this preparation showed a single band of protein. 3. The sedimentation co-efficient and the molecular weight determined on a sucrose gradient were 4·9–5·1s and 76000–83000 respectively for the rat liver enzyme, and 5·6s and 97000 for the acid α-glucosidase purified by means of the same procedure from the human kidney. 4. The Michaelis constants of rat liver and human kidney enzyme were 4·7×10−3m and 13·6×10−3m respectively with maltose as substrate. 5. The enzyme from both tissues was inhibited by tris and by erythritol. The inhibition of the rat liver acid glucosidase by erythritol was competitive.

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Solid-phase enzyme immunoassay of urokinase using monoclonal antibodies

Bioscience Reports ◽

10.1007/bf01122903 ◽

1983 ◽

Vol 3 (4) ◽

pp. 381-388 ◽

Cited By ~ 15

Author(s):

P. Hérion ◽

D. Portetelle ◽

J. -D. Franssen ◽

J. Urbain ◽

A. Bollen

Keyword(s):

Monoclonal Antibodies ◽

Enzyme Immunoassay ◽

Solid Phase ◽

Biological Fluids ◽

Immunosorbant Assay ◽

Rapid Procedure ◽

Preliminary Purification ◽

Enzyme Linked Immunosorbant Assay

Using two monoclonal antibodies directed against urokinase, we have developed a micro enzyme-linked immunosorbant assay (ELISA) to detect and measure urokinase in biological fluids. The system presents the following characteristics: simple and rapid procedure, reproducibility, sensitivity (urokinase levels down to 1 ng/ml) and evaluation of the enzyme in biological fluids such as urine, pleural elfusions, and ascitic fluids without preliminary purification.

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