bluuebunny/crossref_metadata_2025_split huggingface dataset:
Determination of thiamin and thiamin phosphate esters in blood by liquid chromatography with post-column derivatization.
Authors: Kimura M , Itokawa Y 
Published: January 2020

Abstract
We demonstrate a liquid-chromatographic method involving post-column derivatization for determining the concentration of thiamin and its phosphate esters in human blood. Blood, erythrocytes, or plasma is deproteinized and centrifuged. Aliquots of the samples are applied to a mu Bondapak C18 column attached to a "high-performance" liquid chromatograph. Addition of potassium ferricyanide/sodium hydroxide solution to the column effluent with a proportioning pump converts thiamin phosphates into fluorophores, the intensities of which are measured with a spectrofluorophotometer. Thiamin, thiamin monophosphate, thiamin pyrophosphate, and thiamin triphosphate eluted as single peaks; no coeluting substances were detected. Thiamin pyrophosphate was the ester present in greatest concentration, followed by thiamin triphosphate; thiamin monophosphate and thiamin were present in slight amounts. This method allows easy determination of thiamin and its phosphate esters in 0.1 mL of blood.
Kinetic methods that are independent of the rate of reaction.
Authors: Harris R C , Hultman E 
Published: January 2020

Abstract
Kinetic methods for the quantification of substrates offer a number of advantages over equilibrium methods, such as speed and economy. However, in their most commonly practiced form they are liable to error from various sources in addition to those ordinarily encountered in equilibrium methods, owing to variations in enzymic activity between test and standard assays. In certain circumstances the differences in enzymic activity may be such as to completely invalidate the method. Such difficulties may, however, be avoided if three or more measurements are made during the reaction of the sample and are evaluated by one of five means. In each case data from the early course of the reaction are used to predict the total change in sensor signal at infinite time, from which the concentration of the substrate may be calculated as for an equilibrium method.
Discrepancies in measurement of aspartate aminotransferase by continuous-flow analysis.
Authors: West P 
Published: January 2020

Abstract
I compared results for aspartate aminotransferase (EC 2.6.1.1) obtained with a reaction-rate analyzer (LKB 2086 Mark Two), based on IFCC methodology, and a continuous-flow analyzer (the Technicon SMA 2) for 115 patients' sera and seven commercial quality-control sera. The data from the SMA 2 showed a clear positive bias in those sera with activities exceeding 40 U/L (the upper limit of the reference range). Independent data to support the bias of the SMA 2 and other continuous-flow analyzer systems are presented. Application of a correction factor to the SMA 2 data above the upper limit of the range significantly decreased this bias. Failure to apply such a factor to data obtained from continuous-flow analyzers could lead to serious clinical misinterpretation.
Accuracy of lactate dehydrogenase isoenzyme estimations by a thin-layer agarose fluorescent technique: experience with ternary and quaternary mixtures of purified human lactate dehydrogenase isoenzymes.
Authors: Pridgar E M , Leung F Y , Henderson A R 
Published: January 2020

Abstract
We have further assessed the accuracy of the thin-layer agarose fluorescent technique of Elevitch et al. [Am J Clin Pathol 46, 692 (1966)]. Previously, we used semi-purified human lactate dehydrogenase isoenzyme-1 and -5 [Clin Chem 22, 1995 (1976)] and isoenzyme-1 and -2 [Clin Chem 27, 1708 (1981)] to show that this assay accurately measures the proportions of these binary mixtures. In the present study, using ternary and quaternary mixtures of isoenzyme- 1, -2, -3, and -5, we show that the assay gives accurate estimations of all of these isoenzymes, within the errors of the techniques used. We also show that peak area (integration) is more nearly accurate, but less precise, than peak height (amplitude) measurements.
Influence of phospholipid saturation on classical thin-layer chromatographic detection methods and its effect on amniotic fluid lecithin/sphingomyelin ratio determinations.
Authors: Spillman T , Cotton D B , Lynn S C , Bretaudiere J P 
Published: January 2020

Abstract
We compared seven techniques, commonly used for detection of amniotic fluid phospholipids in thin-layer chromatography, with respect to their sensitivity to saturation of the fatty acid carbon-chain of lecithin. The techniques fell into two classes: sensitive and insensitive; those classed as saturation sensitive were less than or equal to 10% as sensitive to fully saturated lecithin as to lecithin with singly unsaturated acid moieties. Color development increased with the number of carbon-carbon double bonds per molecule but required only a single unsaturated acid ester in either the alpha or the beta position. Mixtures of lecithins with defined saturation, when detected by saturation-sensitive methods, mimicked the uneven coloration of amniotic fluid lecithin bands, supporting the hypothesis that this uneven coloration results from natural saturation heterogeneity. Detection techniques representative of these classes are shown to give widely differing values for the lecithin/sphingomyelin ratio for actual specimens of amniotic fluid.
Partial characterization, properties, and clinical significance of a lactate dehydrogenase-immunoglobulin A kappa complex in serum.
Authors: Weijers R N , Mulder J , Kruijswijk H 
Published: January 2020

Abstract
The existence of a lactate dehydrogenase-immunoglobulin A kappa complex was demonstrated as a marked increase of lactate dehydrogenase activity at the lactate dehydrogenase-3 band in the agar-agarose gel-electrophoresis pattern of sera from seven patients from an unselected group of 21 800 patients. The complex was isolated, in an almost pure form, by gel filtration and affinity chromatography, from the serum of a patient with circulating hepatitis B surface antigen. The complex had a relative molecular mass (Mr) of approximately 445 000 as determined by gel filtration. Electrophoresis in sodium dodecyl sulfate in the absence of reducing agents showed the presence of two subunit bands with Mr 170 000 and 34 000. We therefore propose that the native complex consists of one monomeric immunoglobulin A kappa linked to two tetrameric lactate dehydrogenase molecules. The enzymic activity of the complex is probably from lactate dehydrogenase isoenzyme-5.
Characterization of bacteria by mixed-dye fluorometry.
Authors: Shelly D C , Warner I M , Quarles J M 
Published: January 2020

Abstract
We describe an indirect detection method for bacterial identification and differentiation, based on selective adsorption of several fluorescent dyes. The lipid, protein, and nucleic acid components of fixed whole cells are stained with two mixtures, each containing two fluorochromes. The unadsorbed dyes were measured simultaneously with a video fluorometer [Clin Chem 22: 1483, 1976]. A dye-absorption matrix of the response can be generated, and we did so for each of nine bacteria. These responses were compared to a control or "complete" response matrix, and the response ratios of each bacterial species for each of the four dyes were calculated and plotted to obtain a characteristic pattern. From the response-ratio plots, plus simple pattern-recognition techniques, we could differentiate among all the bacteria. This rapid, sensitive technique is potentially applicable to a wide variety of bacteria.
Controlled dispersion analysis: flow-injection analysis without injection.
Authors: Riley C , Aslett L H , Rocks B F , Sherwood R A , Watson J D , Morgon J 
Published: January 2020

Abstract
Flow-injection analysis is a precise, elegant, and economical technique, but its most troublesome feature is the mode of injection of the sample slug. We describe an alternative approach in which the sample is aspirated by the sample probe. In the simplest version the probe normally rests in reagent; when sampling is to take place, the pump is stopped and the probe is transferred to the sample container. The pump makes a predetermined angular movement, the probe is returned to reagent, and the pump is restarted. In more advanced versions the same approach is combined with the merging zone technique. The system is economical, precise, and capable of full automation in a multichannel discretionary analyzer.
Liquid-chromatographic separation and determination of coproporphyrins I and III in urine.
Authors: Sakai T , Niinuma Y , Yanagihara S , Ushio K 
Published: January 2020

Abstract
We describe a method of determining coproporphyrin I, III, and I plus III in urine by "high-performance" liquid chromatography. Urine is simply injected after dilution with an equal volume of glacial acetic acid. Some urinary coproporphyrin apparently binds zinc without acetic acid treatment. The working linear range of coproporphyrin concentrations is 10 to 2000 micrograms/L of urine. The sensitivity of the method is sufficient to detect as little as 10 micrograms of coproporphyrins per liter of urine. Analytical recoveries for both coproporphyrins were 96.7-106%. Results by the present method and those by an extraction method (Br J Ind Med 31:72-74, 1974) correlate well (r = 0.975). Mean (and range) coproporphyrin I, III, and I plus III concentrations in urine from normal subjects are 33.7 (7-75), 28.6 (0-130), and 62.2 (7-174) micrograms/L, respectively.
Clinical evaluation of immunoinhibition determination of creatine kinase B subunits in coronary care.
Authors: Booth J , McCarthy P J , Walmsley R N 
Published: January 2020

Abstract
One hundred patients with chest pain of cardiac origin were evaluated on the basis of clinical findings, electrocardiograph results, and total creatine kinase (CK) and creatine kinase B-subunit (CK-B) activity (as determined by immunoinhibition with the Boehringer CK-MB kit) in serum. All patients diagnosed as having had an acute myocardial infarction had increased values for both CK-B and total CK. In no case was normal total CK activity associated with an increased CK-B, nor was normal CK-B associated with an increased total CK. During collection of data for reference ranges, we found 10 patients who had no evidence of cardiac disease but had various other diseases, who exhibited high values for CK-B in serum; four of these had normal values for total CK. We conclude that estimations of CK-B in serum by this method added no more diagnostic information than did data on total CK in the evaluation of chest pain.
Fluorometric procedure for determining "lamellar body" phospholipids in the particulate fraction of amniotic fluid after filtration.
Authors: Egberts J , Soederhuizen W , Gebhardt D O 
Published: January 2020

Abstract
A rapid (30-min) semiautomated continuous-flow procedure is described for use in assessing the phospholipids of the particulate ("lamellar body") fraction of human amniotic fluid. The method is based on measuring the difference in fluorescence of 1,6,-diphenyl-1,3,5-hexatriene added to amniotic fluid before and after micropore filtration. The filtration step removes "lamellar body" particles, which are considered to contain the fetal lung surfactant. The phospholipid values for the filtered particles are independent of background fluorescence, which increases when amniotic fluid is contaminated by bilirubin pigments or blood components. Over a wide range (3-150 mumols/L) the fluorescence increases linearly with the phospholipid concentration of the amniotic fluid. There is a good agreement between the value for particulate "lamellar body" phospholipid, the ratio of the "lamellar body" phospholipids to total amniotic fluid phospholipids, and the lecithin/sphingomyelin ratio.
Radioimmunoassay for bupropion in human plasma: comparison of tritiated and iodinated radioligands.
Authors: Butz R F , Smith P G , Schroeder D H , Findlay J W 
Published: January 2020

Abstract
We evaluated the potential usefulness of 125I-labeled p-hydroxybupropion in a direct radioimmunoassay for bupropion in human plasma as compared with a currently used [3H]bupropion dextran-coated charcoal method. In both radioimmunoassay methods succinoylpropylbupropion antiserum was used that was highly specific for unchanged drug, cross reactivities with known bupropion metabolites being less than 0.3%. However, the use of 125I-labeled p-hydroxybupropion afforded greater sensitivity (0.3 microgram/L vs 0.6 microgram/L with [3H]bupropion) and was readily adaptable to the more convenient polyethylene glycol separation method. Between-assay CVs were 3.8 to 12.2% (mean 7.6%) with the 125I-based radioimmunoassay and 5.1 to 11.5% (mean 7.5%) with the 3H-based assay. Agreement between the two radioimmunoassay determinations of buproprion in human plasma samples collected over a 60-h period after oral drug administration was excellent (slope = 1.086, r = 0.989). We find the 125I-based assay a convenient and suitable alternative to the [3H]bupropion assay in pharmacokinetic studies in humans.
Quantification of lipoprotein cholesterol in serum from children with different lipoprotein profiles: heparin-calcium precipitation and ultracentrifugation compared.
Authors: Srinivasan S R , Webber L S , Whitaker C F , Berenson G S 
Published: January 2020

Abstract
We compared the serum lipoprotein cholesterol concentrations in subgroups of children (n = 360), ages 5-17 years, as measured by the heparin-Ca2+ and preparative ultracentrifugation methods. Children were grouped from the total population on the basis of their previous results for serum beta- and pre-beta-lipoprotein cholesterol (Group I: low beta- and low pre-beta-; Group II: high beta- and low pre-beta; Group III: high beta- and high pre-beta-; Group IV: low beta- and high pre-beta-). The values for very-low-density (VLDL) cholesterol by ultracentrifugation method were 44, 53, 15, and 10 mg/L greater than the values for pre-beta-lipoprotein cholesterol by the heparin-Ca2+ method in Groups I, II, III, and IV, respectively; the differences were not significant in Group IV. The values of low-density (LDL) cholesterol were 64, 137, 144, and 73 mg/L less than the values for beta-lipoprotein cholesterol in Groups I, II, III, and IV, respectively (p less than 0.005). On the other hand, high-density (HDL) cholesterol concentrations in the respective four groups were 10, 37, 93, and 52 mg/L greater than alpha-lipoprotein cholesterol concentrations; the differences were significant for Groups II, III, and IV (p less than 0.005). Overall, the values for LDL-cholesterol correlated highly with beta-lipoprotein cholesterol (r = 0.94), whereas correlations for VLDL- and HDL-cholesterol values with pre-beta-lipoprotein cholesterol (r = 0.76) and alpha-lipoprotein cholesterol (r = 0.77) were somewhat lower. The differences between these two methods may result from their different operational definitions for measuring serum lipoproteins and the possibility that without appropriate corrections the values obtained by preparative ultracentrifugation do not serve as reference values.
Ultrafiltrable calcium and magnesium in ultrafiltrates of serum prepared with the Amicon MPS-1 system.
Authors: D'Costa M , Cheng P T 
Published: January 2020

Abstract
We evaluated the Amicon micropartition system (MPS-1) for preparing ultrafiltrates of serum for use in evaluating ultrafiltrable Ca and Mg. We found no adsorption of either to the filter and 99.6% retention of serum proteins on the membrane. Ultrafiltrate volumes recovered (100-450 microL) varied with centrifugation time (10-30 min) and temperature. Centrifugation time did not affect the measured concentration of ultrafiltrable calcium and magnesium, and pH change in the 1-mL serum specimen during a 30-min centrifugation at room temperature was negligible. There was an inverse relationship between temperature and ultrafiltrable Ca and Mg concentrations. The precision (CV) between filters ranged from 1.2 to 5.1% for ultrafiltrable Ca and 1.5 to 2.7% for ultrafiltrable Mg. The correlation between ultrafiltrable Ca (y) and ionized Ca (x) in samples from 115 patients with calcium-related metabolic disorders was good (y = 1.04x + 0.18; r = 0.9128). We find the MPS-1 to be a simple and convenient tool for the rapid production of serum ultrafiltrates.
Development and clinical evaluation of a microcentrifugal analyzer method for determining creatine kinase MB isoenzyme.
Authors: Massey T H , Butts W C 
Published: January 2020

Abstract
We have adapted to a microcentrifugal analyzer an immunoinhibition assay for measuring the activity of creatine kinase MB by using an inhibitory antibody for the M monomer. The method actually measures half the MB activity, but results are not multiplied by two because atypical isoenzymes of creatine kinase, including BB, IgG-BB, and the isoenzyme derived from mitochondria, are also detected, if they are present. Results correlated well with an electrophoresis method for 36 serum samples. Myocardial infarction was assessed in 175 patients admitted to our coronary-care unit, with respect to sensitivity (100%) and specificity (98%) when a decision point of 100 U/L (30 degrees C) was chosen for total creatine kinase activity (dithiothreitol-activated) and 6 U/L (30 degrees C) for the isoenzyme (by immunoinhibition). Atypical isoenzymes are easily recognized and confirmed by electrophoresis when the MB activity (by immunoinhibition) exceeds 6 U/L and 20% of the total creatine kinase activity.
Interference of fetal hemoglobin and labile glycosylated hemoglobin with measurements of glycosylated hemoglobin.
Authors: Yatscoff R W , Tevaarwerk G J , Clarson C L , Warnock L M 
Published: January 2020

Abstract
We examined the effect of fetal hemoglobin and labile glycosylated hemoglobin on a number of diverse methods used to measure glycosylated hemoglobin. Samples were supplemented with various amounts of cord blood to give proportions of fetal hemoglobin ranging from 1 to 20% of total hemoglobin concentration. Procedures in which the separation of hemoglobin A1 from the major hemoglobin A fraction is based on differences in ionic properties (cation-exchange chromatography and electrophoresis) are subject to interference by fetal hemoglobin, whereas procedures that base the quantitation on other properties (colorimetry and affinity column chromatography) are not. The same procedures that are affected by the presence of fetal hemoglobin are also subject to interference by labile glycosylated hemoglobin. We conclude that the affinity chromatographic and colorimetric methods may give a more nearly accurate determination of glycosylated hemoglobin.
Automated rate-immunonephelometric determination of serum prealbumin (transthyretin).
Authors: Jacob R A , Gorman N 
Published: January 2020

Abstract
We describe an automated determination of serum prealbumin (transthyretin), based on the maximum rate of light scatter of an antigen-antibody precipitate. Optimal dilutions of antibody reagent and of 100-microL serum samples are given. Within- and between-assay CVs are less than 5%. Results by this method are nearly identical with those obtained by a radial-immunodiffusion technique. Measuring the rate of light scatter provides a considerably faster test than equilibrium scatter, electrophoretic, or radial-immunodiffusion methods. Treatment before analysis to clear lipemic sera obviates falsely high results.
Optimization of enzyme-based assays in coagulation testing.
Authors: Coleman P L , Perry J F , Wehrly J A 
Published: January 2020

Abstract
Optimized assays for antithrombin III and plasminogen have been developed based on a study of the kinetic parameters Km and Kcat for four commercially available substrates: the p-nitroanilide derivatives of D-Phe-pipecolyl-Arg (S-2238), and toluenesulfonyl-Gly-Pro-Arg (Chromozym TH), which are thrombin substrates; D-Val-Leu-Lys (S-2251), a plasminogen/streptokinase substrate; and alpha-N-carbobenzoxy-L-lysine thiobenzyl ester, a substrate for both enzymes. We used a centrifugal analyzer system for rapid data acquisition and interactive analysis. Optimized conditions for assay of a particular enzyme are not constant for different substrates in the same buffering agent. For example, in 1,4-piperazine diethanesulfonic acid buffer at 37 degrees C, thrombin-catalyzed hydrolysis of Chromozym TH is optimal at 125 mmol/L buffer, 100 mmol/L NaCl, and pH 8.2, whereas substitution of S-2238, also a tripeptide p-nitroanilide, yields optimal hydrolysis at 85 mmol/L buffer, 300 mmol/L NaCl, and pH 7.2. We conclude that optimized assay conditions are best obtained by an extensive survey of available buffers and a detailed investigation of the effects of variation in pH and in the concentrations of the buffer and auxiliary reagents through use of both one-factor-at-a-time and multivariate response surface experimentation.
Performance of the Hitachi 705 evaluated.
Authors: Douville P , Forest J C 
Published: January 2020

Abstract
We evaluated a new discrete analyzer, the Hitachi 705. The instrument can process sequentially as many as 19 different enzymes or other analytes, with a throughput of 180 tests per hour. We evaluated the spectrophotometer, the sample and reagent probes, and the performance for 11 assays: creatine kinase, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyltransférase, amylase, glucose, urea nitrogen, creatinine, albumin, calcium, and phosphorus. The total imprecisions (within-day plus day-to-day component of imprecision) (CV) were: for the photometer (six concentrations of potassium dichromate), 0.14 to 0.4%; for the serum probe (55 to 20 microL), 0.4 to 1.0%; for the reagents probes (50 to 350 microL), 0.3 to 1.2%; for the enzymatic procedures, 2.0 to 8.0%; and for the other tests (glucose, urea nitrogen, creatinine), 1.0 to 4.0%. Good linearity was obtained in the proposed range for the 11 tests, except for calcium. Comparison of the total analytical error with the medically allowable error at a decision value showed that the test results were acceptable, except for calcium and phosphorus. The instrument showed good reliability and practicability.
Studies of the Regan alkaline phosphatase isoenzyme in plasma from a lung-carcinoma patient.
Authors: Cleeve H J , Tua D C 
Published: January 2020

Abstract
We compared an alkaline phosphatase (EC 3.1.3.1) isoenzyme from a plasma sample of a patient with carcinoma of the lung, who was suspected of having the Regan-type isoenzyme, with placental serum alkaline phosphatase and with tissue preparations from placenta, liver, bone, and intestine. The placental and Regan phosphatases responded similarly to heat treatment and modification by various inhibitors, except that the Regan isoenzyme was more resistant to EDTA inactivation than was the placental serum alkaline phosphatase, but more sensitive than placental tissue alkaline phosphatase. Isoelectric focusing and Cellogel electrophoresis showed the Regan isoenzyme to have a mobility similar to that of the placental type. We conclude that the isoenzyme was of the Regan type and support the contention that the placental and Regan isoenzymes are products of the same structural gene, with small differences due to post-genetic modification.