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Methods of Biochemical Analysis, Volume 11

Biochemical analysis - ACS Publications. Reagentov Anal. A,, Izd. Tulyupa, F.

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Keagenlov Anal. Leningrad, USSli. Comenianae, Geol. Electro- anal. Interfacial Electrochem. Acta, 51, A,, Zh. A,, Klyueva, Nauk Uzb. SSR, 27 8 , 37 A,, Kapitsa, N. SSR, 27 lo , 42 Leningrad , 44, Tashkent, Gos. SSR, 26 l , 29 A , , Lenchenko, T. Leningrad , 44, 4. Warsaw , 15, Biochemical Analysis Morton K. It would be presumptuous for any reviewer to claim that he intends to completely cover all of the publications in this dynamic area.

get link As pointed out by the last reviewer in this series I n , the large volume of pertinent reports encompasses journals in chemistry, biology, physics, electronics, and engineering. Today journals in medicine and computer and nuclear science, as well as in instrumentation must be included. There are literally thousands of references that have appeared during the four-year period of this report.

The enormity of the assignment is empha- sized by the fact that each of the bimonthly Biochemical Methods Sections o j Chemical Abstracts contains over entries and the Journal of Lipid Research, which maintains a cumulative monthly listing of pertinent references to lipid methods, had accumulated references in this limited area through the first eleven months of 5 A. It is mandatory that this review be selective rather than comprehensive and precludes incorporation of routine applications or minor modifications of accepted techniques or methods. This reviewer will attempt to cover the subgroupings included in previous reviews of this same title and to emphasize new developments in biochemical analysis with par- ticular reference to those areas of interest to the writer.

Limited reference will be made to the areas of biochemical methodology considered in other reviews in this series. There have been numerous volumes published in continuing series, each devoted to specific areas of biochemistry. The volume included chapters devoted to oxygen electrode measurements, separation and determination of bile pigments liquid scintillation counting, gas chromatography, fluorometric assay of enzymes, assay of blood phenylalanine and tyrosine, determination of urea, ammonia, and urease, and separation, identification, and estimation of prostaglandins 56B.

The topics in included gel filtration, free zone electrophoresis, optical rotary dispersion and circular dichroism, automated peptide chromatography, use of the dansyl reaction, and steroid analysis by gas chromatography 57B. There was also a supplemental volume devoted entirely to the analysis of biogenic amines and their related enzymes 60B. The Methods in Medical Research series also contains reviews of interest to the biochemist.

Original Research ARTICLE

For example, the latest issue 10SB contains sections devoted to adsorption chromatography, gas-liquid chromatography, ion-exchange chromatography, electrochromatography, and the use of the techniques in the isolation and characteristics of a variety of metabolites in man. Colowick and Nathan 0. Kaplan as Editors-in-Chief, has continued to present reviews and step procedures for a variety of biochemical methods.

Reviews in other specialized fields have been considered in volumes of annual series. The Essays in Biochemistry series has continued to present reviews aimed a t overall views of biochemical subjects 29BB. A new series of Critical Reviews has appeared during the period of this report with the stated objective of providing reviews that organize, evaluate, and present what is known about a subject.

The first volume in biochemistry has considered interferon, blood coagulation, steroid receptors, chymotrypsin, structural predicting from amino acid sequence, insulin, and asparaginase 68B. The Automation in Analytical Chemistry series, presents a compilation of papers describing automated biochemical analysis by continuous flow technique 4 l B , 4ZB, B.


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Each of these volumes contains approximately papers of automation methodology in the areas of biochemistry, industrial chemistry, and other fields of biology. Numerous papers related to biochemical methodology are collected in the Protides of Biological Fluids series. Each volume contains papers from three different areas of protein chemistry.

During the period of this report the following have been considered: genetics and antibody response, molecular variation, and electrofocusing l l I B , conformation and structure of protein molecules, proteins of bodily fluids llOB , membranes, complement activity, and separation methods B.

Many texts have appeared during the four years of this report. There have also been books that have considered techniques in protein chemistry dB, 81B, 98B , isoenzymes SI? New journals appearing during this review period have been devoted to chemical instrumentation I l B , chromatography OB ,steroids 89B ,computers 8SB, B , bioenergics 12B , chemico-biological reactions 1S5B and general biochemistry 72B.

Other new journals have been concerned with clinical investigation 96B , cellular immunology 80B , and general topics in the field 72B. There have been several reviews of ultracentrifugal techniques and sedimentation theory , 2lC, 26C, 44c, 7 w. Homogenizers have been described for large scale production of brain homogenate 60C and for the continuous isolation of cell constituents SSC.

Ehrlich ascites tumor cells have been totally disintegrated in 6 seconds in a disintegrator which uses glass beads IC. A zonal centrifuge rotor has been developed to permit gradient recovery from the rotor center light end of the gradient or from the rotor edge heavy end of the gradient SC. Another rotor and centrifuge permits continuous sample flow for large scale virus isolation 7C, 9C. The rotor can hold a ml gradient and has a ml stream volume. It is capable of a centrifugal force of 83, X g. A most useful application of ultracentrifugation has been the development of a microanalytical system.

Sample and reagents are brought together in cuvettes in the rotor by centrifugal force. The rotor containing the many cuvettes spins past a beam of light and the signal is displayed on an oscilloscope. A peak is observed continuously for each sample 6C.

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In an early version of the system, 15 analyses for biuret protein determinations are completed in as little as 30 seconds after the rotor is started 6C. The cuvettes are emptied and rinsed during rotation allowing time between runs of less than two minutes. Methods are being developed to permit analysis of simple substances, active groups, or enzyme activity 8'2. Along similar lines a path rotor has been described for the precipitation of proteins and removal of the pellets. The samples and precipitating reagents are in separate chambers and are brought together in a sedimentation chamber by centrifugal force during rotation.

The supernatant then decants itself when the rotor comes to rest 6C. A method has been described for studying sedimentation in angle-head centrifuges using a titanium rotor, polycarbonate tubes and an electronic integrator A device has been developed to prepare reproducible density gradients with a Technicon proportioning pump 6 I C. Sucrose gradients have been stored at "C and used after a minute thaw a t room temperature 64C. Devices and techniques for the control, accurate collection, fractionation, and analysis of cesium chloride or sucrose gradients have been described 16C, 3OC, 43C.

The usefulness of isokinetic gradients and their preparation for zonal rate centrifugation has been described 4OC, 6OC, SSC , as have flow through cuvettes for the evaluation of density gradients 34C, 66C. Solid cesium chloride has been used to create an internal standard for the determination of buoyant density R4C. It was shown that the stability of brain fractions was improved by substituting in the gradient iso-osmotic ficoll-sucrose for hyperosmotic sucrose 26C. Immunoprecipitation has been applied to identification of antigens following centrifugation in an acrylamidesucrose linear gradient and photopolymerization 46C.