John W Severinghaus And A Freeman Bradley
log pC02 - log pCO 2 pH' - pH When sodium bicarbonate is added, the second term in equation 1 becomes dominant above o.ooim Na, and the equation approximates 2K2 Ah becomes significant at 0.1 m Na Taking two values, as in 2 and computing the sensitivity, S, Thus, the inclusion of bicarbonate ion in the aqueous medium doubles the sensitivity. At higher concentrations, it falls off again as the carbonate ion appeals. At 1.0 m NaHC03, S 0.88 according to equation 1, but this is an approximation...
Determination Of Serum Proteins By Means Of The
By ALLAN G. GORNALL, CHARLES J. BARD A WILL, From the Department of Pathological Chemistry, University of Toronto, Toronto, In the course of an investigation of the biochemical changes following experimental liver injury we felt the need of a simple, rapid, and accurate method for determining the protein fractions in small amounts of serum. Among the simpler procedures known, the biuret reaction seemed to offer Variations and improvements in the application of the biuret reaction to clinical...
Info Iqo
is somewhat better for a 3 1 tartrate ratio, but 0.12 and 0.32 per cent copper sulfate are virtually identical in reagents containing 10 times these amounts of tartrate. Another important property of biuret reagents stabilized with tartrate is the absence of any serious tendency to develop turbidity in the reaction mixture following the addition of clear serum or plasma. Thus one of the main difficulties of Kingsley's reagent, and a concern with Mehl's procedure, is avoided. Clouding will occur...
180 Mg 100 Ml Of Blood Sugar
It is evident that variation in the volumes of solution applied to the paper strips is of no consequence. Generally 100 to 200 fil is applied, the larger volume permitting use of a smaller quantity of tracer beef insulin-I131 for the same counting rate. Assay of insulin in plasma. Mixtures containing unknown samples are prepared at the same time and in the same way as are standard solutions except that the unknown sample is substituted for the human insulin. Plasma insulin is best determined in...
Simple Specific Test for Urine Glucose
Alfred H. Free, Ernest C. Adams, Mary Lou Kercher, Helen M Free, and Marion H. Cook Procedures for the detection of sugar in urine are among the oldest tests known in clinical chemistry. Legends going back to earliest times describe the attraction of insects to urine containing sugar. In many instances it is difficult to separate legend from reality in tracing Among such pioneer American clinical chemists as Stanley Benedict, Otto Folin, Victor Myers, and Donald Van Slyke, there was great...
Commentary to1972 k
Homogeneous Enzyme Immunoassay. A New Immunochemical Technique. Biochemical and Biophysical Research Communications 1972, 47 4 846-851. The enzyme multiplied immunoassay technique EMIT was developed following the introduction of an earlier homogeneous immunoassay technology called the free-radical assay technique FRAT . Both of these homogeneous assay technologies were invented at the Syva Corporation under the research direction of E.F. Ullman. They were initially designed for drugs of abuse...
Biochemical And Biophysical Research Communications
enzyme 2 . Steric inhibition of an enzyme by an antibody might also be envisioned if a hapten were attached to the enzyme near its active site. In a properly selected system binding of a hapten-directed antibody to the hapten might have the same effect as binding of an enzyme-directed antibody directly to the enzyme. In such a system the enzyme activity could be regulated by the amount of antibody available for binding to the enzyme-bound hapten. The amount of free antibody could in turn be...
Results 1
The results on the pure acridine solutions are summarized in Table V and Fig. 1. The data in Table V indicate that at a constant strength of acid various acridines manifest quite different intensities of fluorescence NaOH ADDED NORMALITY OF HCI ADDED NaOH ADDED NORMALITY OF HCI ADDED Fig. 4. Fluorescence characteristics of ethylene dichloride extracts of biological material after these extracts were washed with 2.5 n NaOH, Relative Values for Intensity of Fluorescence Manifested by Six...
Info Gze
Values for 1 1 0.00384 T and a' may be found in reference 18, Values for 1 1 0.00384 T and a' may be found in reference 18, Observing the above-mentioned precautions, analysis for CO2 is made as follows. Draw up 0.03 ml of serum to the mark. Plug with mercury and follow with 0.03 ml. of lactic acid. Follow with 0.01 ml. of caprylic alcohol. Draw up 0.1 ml. of water and follow with sufficient mercury to bring all the reaction mixture beyond the stopcock. Close the stopcock which is now sealed...
Molecular Diagnostics
37. Pauling, L., Itano, H. A., Singer, S. J. and Wells., I. C. 1949 Sickle cell anemia, a molecular disease. 38. Hyypia, T., Jalava, A., Larsen, S. H., Terho, P. and Hukkanen, V. 1985 Detection of Chlamydia trachomatis in clinical specimens by nucleic acid spot hybridization. 39. Maskos, U. and Southern, E. M. 1992 Oligonucleotide hybridisations on glass supports a novel linker for oligonucleotide synthesis and hybridisation properties of oligonucleotides synthesised in situ. This page is...
Chemical And Diagnostic Specificity Of Laboratory Tests
Effect of Hemolysis, Lipemia, Anticoagulants, Medications, Contaminants, and Other Variables WENDELL T. CARAWAY, PH.D. McLaren General Hospital, St. Joseph Hospital, and the Flint Medical Laboratory, Flint, Michigan It is desirable that clinical laboratory procedures have a high degree of diagnostic specificity. Chemical analysis of biologic material is based upon the presence of functional groups in organic compounds or upon the separation and measurement of ions. Such analyses are inherently...
Summary And Conclusion
1. An immunoassay for plasma insulin in man is presented, based on the reaction of human insulin, competing with beef insulin-I131, with insulin-binding antibodies in the sera of guinea pigs immunized with beef insulin. The method is sensitive to less than 1 U of insulin, permitting measurement of insulin concentrations in 10 to 20 xl of plasma. 2. Human insulin added in vitro to plasma is recovered quantitatively, and measured endogenous insulin concentrations decrease proportionately on...
Specific Analytes
Colorimetric Analysis with the Duboscq 14. Folin, O. and Wu, H. 1919 15. Van Slyke, D. D. and Neill, J. M. 1924 The determination of gases in blood and other solutions by vacuum extraction and manometric measurement. I. Bilirubin With the Filter Photometer 16. Malloy, H. T. and Evelyn, K. A. 1937 The determination of bilirubin with the photoelectric colorimeter. 17. Gornall, A. G., Bardawill, C. J. and David, M. M. 1949 Determination of serum proteins by means of the biuret reaction. Methods...
References 1
COMMENTARY TO Separation of Tissue and Serum Creatine Kinase Isoenzymes by Ion Exchange Column Chromatography. Clinical Chemistry 20 1 36-40. The measurement of isoenzymes in serum in order to improve the specificity of an enzyme assay for disease detection began with the work of Oscar Bodansky in 1937. He used taurocholate inhibition to differentiate bone, kidney and intestinal alkaline phosphatase ALP, EC 3.1.3.1 in normal serum and in two patients with Paget's disease 1 . Zone...
A Correction Of Comparator Standard To Eliminate Error Of Serum Adsorption
Our previous experiments showed that serum adsorbed a certain amount of bromides this amount increasing proportionally to the concentration of the bromides, and that the amount adsorbed is practically independent of the particular blood serum between 1 and 4.1 per cent . This adsorption loss is indicated in chart 11 by the shaded portion, which represents the difference in the amount of bromides estimated from watery solution and from serum solutions of known quantities of bromides. The...
Info Ggy
Determination of Urea by Urease Decomposition and Distillation. Transfer 5 cc. of the tungstic acid blood filtrate to a clean and dry Pyrex ignition tube capacity about 75 cc. . The graduated Pyrex tubes recommended for the non-protein nitrogen determination should never be used for urea determinations, because they have contained Nessler solutions and Nessler solutions leave behind films of mercury compounds which destroy the urease. If those tubes must be used, they should first be washed...
Info Yzp
This page is intentionally left blank page 279 20. Wide, L. and Gemzell, C. A. I960 COMMENTARY TO An Immunological Pregnancy Test. Acta Endocrinologica Copenhagen In the 1951 movie People Will Talk, Cary Grant played a physician who in an early scene has just told his patient, played by Jeanne Crain, that she is pregnant. She slumps into the office chair, her face buried in her hands. She asks if he is certain, the test was only done this morning she says. Grant assures her that the results are...
Info Qhh
sequence 3'-AGGTCGCCGCCC was dipped into 30 d of a solution containing the complementary oligonucleotide 5'-TCC-AGCGGCGGG 30,000 c.p.m., 80 fmol in 30 d of 0.1 M NaCl, 37 C . After hybridisation for various times and at different temperatures the stick was removed from the tube, rinsed, and the bound material eluted by dipping the stick into 0.1 M NaCl at 50 C. Typical hybridisation yields were 4 at 37 C for 30 min 5.3 at 30 C for 55 min 13 at 30 C for 16 hrs. Nonspecific binding was between...
Lecturer In Physiology
From the Physiology Department, St. Mary's Hospital Medical School, London 1,t is well known that the estimation of blood-glucose by reducing methods lack specificity and give values which are too high. Enzymes, however, enable a greater selectivity of the reducing substances to be made. In the presence of glucose oxidase, which ia highly specific for glucose, glucose is oxidised to gluconic acid and hydrogen peroxide Koilin and Hartroe 1948 . The hydrogen peroxide can, in the presence of...
Info Jnw
Oligonucleotide syntheses were performed with approx. 4 mg CPG or 120 mg ballotini before and after ammonia treatment of the support and the detritylation effluent collected. Since cleavage is expected to produce silanol groups which will also take part in a coupling reaction coupling yields measured in this way are the sum of the reaction of these silanol and of linker hydroxy alkyl groups, given by the formula ODm 1 - x ODa xODs with x amount of cleavage ODm optical density measured at 498 nm...
Addendum
page 343 27. Friedman, S. M. and Nakashima, M. 1961 COMMENTARY TO Single Sample Analysis With the Sodium Electrode. Analytical Eisenman, Rudin and Casby in 1957 described the chemical formula for the composition of cation sensitive glass that was selective for sodium or for other ions such as potassium with a change in the formulation 1 . With one of the formulas of glass they constructed an ion selective electrode ISE for measuring sodium in aqueous solutions in the presence of potassium and...
Caraway Mow
dine. The latter may be a somewhat more specific measure of circulating thyroxine in hypothyroidism. In the same report, 15 cases of hyperthyroidism are described with normal serum protein-bound iodine. It is suggested that this may reflect either random variation in the hyperthyroid group or a diminution in thyroxine-binding protein with concurrent elevation in metabolically active free thyroxine. Bence Jones proteins in urine have the common property of precipitating when heated to 40 to 60...
Info Cxp
Zone Electrophoresis in Open-Tubular Glass Capillaries James W. Jorgenson and Krynn DeArman Lukacs Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina 27514 A system for performing zone electrophoresis In open-tubular glass capillaries of 75 im inside diameter and with applied voltages up to 30 kV is described. The small inside diameter of these capillaries allows efficient dissipation of the heat generated by the application of such high voltages. However, the...
Literature Cited
1 Wleme, R. J. In Chromatography A Laboratory Handbook of Chromatographic and Electrophoretic Methods, 3rd ed., Heftmann, E., Ed. Van Nostrand Relnhold New York, 1975 Chapter 10. 2 Mikkers, F. E. P. Everaerts, F. M. Verheggen, Th. P. E. M. J. Chro-matogr. 1979, 169, 11-20. 3 GkJdings, J. C. Sep. Sei. 1969, 4, 181-189. 4 Hinckley, J. O. N. J. Chromatogr. 1975, 109, 209-217. 5 Brown, J. F. Hinckley, J. O. N. J. Chromatogr. 1975, 109, 218-224. 6 Pretorius, V. Hopkins, B. J. Schleke, J. D. J....
Standards in the Clinical Chemistry Laboratory
There has been no coordinated effort among various groups to bring about specifications and definitions for various standards and reference samples for the clinical chemistry laboratory. Suggested definitions and a review of the standards available to the clinical chemist will be presented in this discussion. In the reference frame of usual analytical laboratory practice the primary standard is defined as a pure chemical substance that is used for the purpose of assaying a volumetric solution...
D D Van Slyke and J M Neill 525
of magnitude. Thereby the percentage error in gas measurement becomes the minimum attainable with given absolute errors in pressure and volume. The apparatus consists of a short pipette with the upper stem closed by a stop-cock, the lower connected with a glass tube. The Fig. 2. Extracting chamber, showing mercury-sealed rubber joint at bottom and attachment of Stadie shaker at top. Gas is at 2 cc. volume for pressure reading. Fig. 2. Extracting chamber, showing mercury-sealed rubber joint at...
Summary 1
Among the methods of preparing stable, one piece biuret reagents, the use of sodium potassium tartrate appears most promising. The tartrate-stabilized reagent devised by Weichselbaum has been investigated in some detail and has been modified to contain less copper, more alkali, and less or no potassium iodide. A simple procedure for the determination of serum total protein, albumin, and globulins is described. 766 serum protein determination The biuret reaction is approved as a simple, rapid,...
Lipemia
Serum appears lipemic when the content of neutral fat is elevated. Although normally less than 200 mg. per 100 ml., the neutral fat may reach values of the order of 10,000 mg. per 100 ml. of serum. Colorimetric analytic methods, in which serum is still present in the final test mixture, but not in the blank, result in elevated optical densities in the presence of lipemia. Depending on the method used, falsely elevated values for total protein, thymol turbidity, amylase, transaminase, and so on,...
Info Syg
thyroxine 2 , Whitehead and Beale obtained low values in 14 euthyroid subjects, viz., a mean of 4.6 fig thyroxine 100 ml, corresponding to a thyroxine iodine of 3.1. Comparing this value with the mean protein-bound iodine value corrected for recovery in 100 euthyroid subjects obtained by Bodansky et al., viz., a mean of 7.1 1.5, our own combined values for thyroxine iodine in 20 healthy subjects and in 20 euthyroid patients, viz., 6.6 1.3 fig 100 ml, are intermediate between the two but much...
Info Lyq
of the urine samples were 1 or more with Benedict's test but were negative with the glucose oxidase test. Two of these urines contained over 0.3 ascorbic acid which was a sufficient quantity to account for the positive test. In addition, among these urines were samples containing galactose, lactose, and fructose. There were 36 urine samples that gave positive reactions with the glucose oxidase test and negative reactions with Benedict's test. In these cases the amount of glucose present was...
A
Fig. 5. Construction of overflow-type pipet. A. 0.5-mm. bore capillary heated and blown. B. Pipet drawn from A at both ends. C. Pipet holder. D Assembled pipet B and C joined with rubber-collar cut from soft rubber tubing. Fig. 5. Construction of overflow-type pipet. A. 0.5-mm. bore capillary heated and blown. B. Pipet drawn from A at both ends. C. Pipet holder. D Assembled pipet B and C joined with rubber-collar cut from soft rubber tubing. bore tubing by means of a one-holed rubber stopper....
Info Nfm
Fig. 7. Typical course of change in total CK and MB activities after myocardial infarction MB found in cardiac tissue. Correlation between MB yields and total CK was not observed. A typical time course for post-infarct activities of total CK and MB is shown in Figure 7. These activities paralleled one another until 36 h after infarct, when MB could no longer be detected. Column reproducibility was evaluated by repeated analysis of a serum pool fortified with MB isoenzyme from an extract of...
Info Fpa
similarly except that cysteine was omitted. Although incubation and dialysis alone led to a 26 to 36 per cent loss in endogenous insulin concentration in the control samples, cysteine was almost completely effective in destroying the endogenous insulin Table I . In simultaneous experiments insulin-I131 was found to be virtually completely destroyed under these conditions as determined by paper chromato-electrophoresis. To evaluate cellulose adsorption of endogenous insulin, a minute amount of...
The Determination Of Bilirubin With The Photoelectric Colorimeter
By HELGA TAIT MALLOY and KENNETH A. EVELYN From the Department of Medicine, McGill University Clinic, Royal Victoria The main source of error in the determination of bilirubin in serum by the diazo reaction has been the loss caused by adsorption on the protein precipitate. In addition, the lack of an accurately matching artificial standard and the sensitivity of the azobilirubin color to changes in pH have made accurate colorimetric determinations impossible, without the aid of some type of...
Sulfanilamide Determination Ytj
what slight modification of the method, if any, is necessary to determine it accurately. However, we must caution investigators against accepting results with a new compound until control recoveries from blood and urine have been made. In the determination of sulfanilamide by diazotization and coupling in acid solution, the use of N- l-naphthyl ethylenediam-ine dihydrochloride offers the following advantages over N,N-dimethyl-l-naphthylamine dimethyl-a-naphthylamine 1 reproducibility and...
Reagents And Preparations
Antigen. Ampuls containing 1500 IU of a commercial preparation Pregnyl, Organon of human chorionic gonadotropin HCG was used. Adjuvant. Ramon's adjuvant Ramon ct ttl. 1 lt W gt was prepared by mixing 48 ml of mineral oil, 24 ml of 0.9 per cent saline and 1 gt g f lanoline. The mixture was autoclaved and homogenized. Antisera. Normal white adult male and female rabbits were used for immunization. Rabbit anti-HCG-sera was prepared by subcutaneous injection of 12 000 IU of HCG, suspended in 2 ml...
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index of reliability than methods used routinely in clinical laboratories. In performing a number of analyses for urea in blood, a technician must perform many consecutive steps, each with care, in order to arrive at a correct result. The number of operations that must be performed provides ample opportunity for error. As is frequently the situation, the technician is required to perform several other kinds of analyses as well, and the opportunity for error becomes greatly magnified. By...
Caraway Kfi
ation of voltage as a source of error in coulometric titration. Am. J. Clin. Path., 34 195, 1960. 187. Woodard, H. Q. The clinical significance of serum acid phosphatase. Am. J. Med., 27 902-910, 1959. 188. Wootton, I. D. P. Standardization in clinical chemistry. Clin. Chem., 3 401405, 1957. 189. Wr6blewski, F. The clinical significance of transaminase activities of serum. Am. J. Med., 27 911-923, 1959. 190. Wr6blewski, F., and LaDue, J. S. Lactic dehydrogenase activity in blood. Proc. Soc....
Predictive Value Of A Single Diagnostic Test In Unselected Populations
WHEN a new test for a disease is being evaluated it is customary to perform the test in two selected groups of subjects those with an indisputable diagnosis of the disease by other criteria and those from the normal population who have no evidence of the disease and in whom all the factors known to result in a higher than normal risk of the disease can be excluded.13 The test results may be expressed dichotomously as positive or negative, or by some numerical units along a scale, usually with...
The New England Journal Of Medicine 1
sence of unlabeled drug, and the mixture incubated at 25 C for 15 minutes. As shown in Figure 1, competition between labeled and unlabeled digoxin for antibody binding sites determines the amount of labeled digoxin-antibody complex present at equilibrium. Separation of bound from free labeled digoxin Fig. 2 was achieved by the dextran-eoated charcoal technic of Herbert et al.,11 resulting in selective binding of free digoxin to the coated charcoal, which was then separated by centrifugation....
Info Kng
5. Annino, J. S., and Relman, A. S. The effect of eating on some of the clinically important chemical constituents of the blood. Am. J. Clin. Path., 31 155-159,1959. 6. Arst, H. E., ManninGj R. T., and Delp, M. Serum leucine ammopeptidase activity findings in carcinoma of the pancreas, pregnancy and other disorders. Am. J. M. Sc., 238 598-609, 1959. 7. Babson, A. L. Opportunities for application of the scientific method in the routine clinical laboratory. Am. J. M. Technol., 26 379-385, 1960....
Info Rlj
The copper solution is adjusted to give proportionate reductions with 0.12 to 0.4 mg. of dextrose. This covers the range of hypoglycemic and hyperglycemic bloods. But in extreme cases it is better to use 3 or 1 cc. of the filtrate, instead of 2 cc., adding water to the standard or to the unknown so as to equalize the concen- page 199 15. Van Slyke, D. D. and Neill, J. M. 1924 COMMENTARY TO The Determination of Gases in Blood and Other Solutions by Vacuum Extraction and Manometric Measurement....
Results And Discussion
1300 ANALYTICAL CHEMISTRY, VOL. 53, NO. 8, JULY 1981 Figure 1. Zone electrophoretic separation of dansyl amino acids A unknown impurity, B c-labeled lysine, C dllabeled lysine, D asparagine, E isoleucine, F methionine, G serine, H alanine, I glycine, J and K unknown impurities, L dllabeled cystine, M glutamic acid, N aspartlc acid, O cysteic acid. The concentration of each derivative is approximately 5 x 1CT4 M, dissolved in operating buffer. Figure 1. Zone electrophoretic separation of dansyl...
Normality Of
NaOH added normality of HCI added Fig.3. Fluorescence characteristics of the fluorescent material contained in ethylene dichloride extracts of biological material in the absence of an alkaline wash. The dog and human from which the samples were obtained had received atabrine for several days previously. chloride was returned to an aqueous phase by extraction with 0.1 n hydrochloric acid. Ethylene dichloride extracts of human and dog plasma, whole blood, and urine Fig. 4 prepared as in the...
References Vjd
1. Wichers, E., Standards for reagent chemicals. Anal. Chem. 23, 1537 1951 . 2. American Chemical Society, Reagent Chemicals American Chemical Society Specifications, 3. Standard Materials, National Bureau of Standards Miscellaneous Publication 241. Issued 4. Lashof, T. W., and Macurdy, C. B., Precision Laboratory Standards of Mass and Laboratory Weights, National Bureau of Standards Circular 547. Section I, Issued 5. Hughes, J. C., Testing of Glass Volumetric Apparatus, National Bureau of...
Info Yxb
a Only people with plasma triglycerides less than 400 mg 100 ml are included. b LDLf Cldl calculated with the preparative ultracentrifuge see text . LDLjs Cldl calculated by the estimation procedure see text . ' jLDLc - LDU the absolute value of LDLcH-DU . X db -value .025, n -1 SD n. X tolerance-value .95, .95, n SD. a Only people with plasma triglycerides less than 400 mg 100 ml are included. b LDLf Cldl calculated with the preparative ultracentrifuge see text . LDLjs Cldl calculated by the...
Chemometrics
32. Levey, S. and Jennings, E. R. 1950 The use of control charts in the clinical laboratory. Interferences in Analytical Accuracy Chemical and diagnostic specificity of laboratory tests. Predictive value of a single diagnostic test in unselected populations. 36. Barnett, R. N., Cash, A. D. and Junghans, S. P. 1968 Performance of kits used for clinical chemical analysis of cholesterol. This page is intentionally left blank page 403 32. Levey, S. and Jennings, E. R. 1950 COMMENTARY TO The Use of...
Gemini Centrifugal Analyzer
Fia. 4. Electrode analysis of sodium concentration in whole blood after addition of weighed amounts of NaCl. The actual plasma Na concentration achieved was measured by flame photometry. Fia. 4. Electrode analysis of sodium concentration in whole blood after addition of weighed amounts of NaCl. The actual plasma Na concentration achieved was measured by flame photometry. a small constant. This strongly suggests that some proportion of the sodium in the plasma is inaccessible to the electrode...
Determination of Bilirubin 1
which protein precipitation and consequent loss of bilirubin by 2. The interfering effect of yellow serum pigments in the color determination has been overcome by the use of a specially selected light filter, which also eliminates the necessity for artificial color 3. It has been shown that all the bilirubin in serum will react with diazo reagent even in the presence of serum proteins, provided a sufficiently high concentration of alcohol 50 per cent is present. 4. The new method has been found...