bluuebunny/crossref_metadata_2025 huggingface dataset:
Simple method for estimating glycosylated hemoglobins, and its application to evaluation of diabetic patients.
Authors: Chou J , Robinson C A , Siegel A L
Published: January 2020

Recent reports have suggested that determination of glycosylated hemoglobin may serve as a clinical aid for long-term blood glucose control in diabetes mellitus. We describe a simple procedure for measuring it by ion-exchange chromatography. Hemolysates were subjected to Bio-Rex 70 chromatographic separation on small columns. Percentages in the normal group ranged from 4.7 to 8.8% of total hemoglobin; the mean +/- standard error was 6.61 +/- 0.31%. Values in the diabetic group ranged from 6.9 to 17.4%; the mean was 10.83 +/- 0.34. Plasma glucose concentrations after fasting, plotted vs. the percent of glycosylated hemoglobin, revealed a linear relationship at normal or moderately high glucose concentrations. However, the values for glycosylated hemolgobin approached a plateau with grossly higher plasma glucose concentrations after fasting. Our results support the view that, due to its long half-life, the estimation of glycoylated hemoglobin reflects the integrated glucose concentrations to which the erythrocytes have been previously exposed.
Thyroid hormone contamination in crystalline diiodotyrosine.
Authors: Alexander N M , Nishimoto M
Published: January 2020

Crystalline preparations of diiodotyrosine contain trace quantities of thyroxine that can be detected by several competitive radioassays and by chromatography on Sephadex. Spontaneous coupling of diiodotyrosine to thyroxine during iodination of tyrosine and coprecipitation of hormone with diiodotyrosine crystals are likely explanations for the trace contamination. We discuss the significance of these findings in relation to studies on the biosynthesis of thyroxine.
Determination of a trivariate reference region for free thyroxine index, free triiodothyronine index, and thyrotropin from results obtained in a health survey of middle-aged women.
Authors: Kågedal B , Sandström A , Tibbling G
Published: January 2020

We measured free thyroxine index, free triiodothyronine index, and thyrotropin in serum in a community survey of the female population 39--60 years old. A statistical method using Mahalanobis' distance was applied to the data, to identify and eliminate highly abnormal values, "outliers." There was a small but statistically significant increase in each of the three hormones with age. After correction for age dependency, significant but rather small correlations were found between the hormones. The material was used to calculate univariate reference intervals and a trivariate 0.95-tolerance region.
Modified erythrocyte uroporphyrinogen I synthase assay, and its clinical interpretation.
Authors: Piepkorn M W , Hamernyik P , Labbé R F
Published: January 2020

Assay of erythrocyte uroporphyrinogen I synthase is an accepted diagnostic test for acute intermittent porphyria, particularly in those individuals who are asymptomatic or in whom the disease is not biochemically manifested by excretion of excess porphyrin precursor. The assay described is based upon a coupled-enzyme procedure in which added delta-aminolevulinic acid and its dehydratase present in erythrocytes are used to generate porphobilinogen as substrate for uroporphyrinogen synthase. Zinc and dithiothreitol are added with preincubation to give maximum activity and reproducibility. These agents also prevent inhibition by lead. Healthy young women had a mean activity of 40 nmol of porphyrin formed per milliliter of erythrocytes per hour, men and activity of 38 nmol/ml/h. Preparation of control specimens is described. This assay gave within-day CVs ranging from 1.9 to 2.8%. Precautions in interpretation of results are discussed.
Characterization of the principal human prostatic acid phosphatase isoenzyme, purified by affinity chromatography and isoelectric focusing. Part II.
Authors: Vihko P
Published: January 2020

The principal enzyme of human prostatic acid phosphatase [orthophosphoric monoester phosphohydrolase (acid optimum), EC 3.1.3.2], which had been highly purified by affinity chromatography, isoelectric focusing, and gel filtrations, was shown to be homogeneous at pH 5.0 by sedimentation equilibrium analysis. The amino acid composition was determined and the sedimentation coefficient of the native molecule measured. The relative molecular mass was 89,000 at pH 5.0, as measured by analytical ultracentrifugation. The Km-value of the enzyme for p-nitrophenyl phosphate as substrate is 1.8-10(-4) mol/liter. I also examined substrate speificity, different inhibitors, and the effects of pH, temperature, and serum on the enzyme activity.
Gas--liquid chromatographic microdetermination of underivatized ethosuximide (alpha-ethyl-alpha-methyl succinimide) in plasma or serum.
Authors: Fellenberg A J , Pollard A C
Published: January 2020

A gas-liquid chromatographic procedure for the microdetermination of ethosuximide is described. Ethosuximide is extracted from acidified plasma or serum into chloroform containing an internal standard alpha,alpha-dimethyl-beta-methyl succinimide. Part of the initial chloroform extract is gently evaporated, the residue redissolved in n-heptane at 60 degrees C, and an aliquot analyzed by gas-liquid chromatography. The procedure is rapid, reliable, sensitive, and specific. It requires a 25--50 microliter sample for a single estimation, has a detection threshold of less than 10 micromol/liter, and is suitable for routine clinical use.
Distribution of thioridazine and its metabolites in human tissues and fluids obtained postmortem.
Authors: Dinovo E C , Bost R O , Sunshine I , Gottschalk L A
Published: January 2020

Thioridazine and metabolites were measured in brain, liver, and kidney specimens, obtained postmortem from two subjects whose deaths were related to acute intoxication with thioridazine, by gas-liquid chromatography. Although the absolute concentration measured for thioridazine and metabolites differed in the two cases, the metabolic pattern for each tissue, expressed in terms of the percentage of total drug in each tissue, was quite similar. The brain, liver, and kidney metabolic patterns, however, are in sharp contrast to the plasma metabolite patterns observed for subjects on a therapeutic regimen of thioridazine. As this example demonstrates, postmortem specimens are a valuable (but seldom used) source of human pharmacological data.
Plasma estriol vs. estrogen assays in 24-h urines as an index to fetal status.
Authors: Kirkish L S , Barclay M L , Parra J B , Compton A A , McCann D S
Published: January 2020

Conventionally, urinary estrogens are measured to monitor fetal well-being in the third trimester of pregnancy. There are many theoretical and practical advantages to the use of recently introduced plasma estriol assays; consequently, we undertook to compare the results of a spectrophotometric assay for total urinary estrogen with values for plasma total and free (unconjugated) estriol obtained by radioimmunoassay with commercially available kits. The data correlate well, r = 0.70 for plasma total estriol with urinary estrogen (expressed as milligrams of estrogens per gram of creatinine [E/C]), r = 0.79 for plasma free estriol with E/C, and r = 0.71 between the two plasma values. Minute-to-minute fluctuations during 90 min and day-to-day fluctuations followed during five days suggest that fluctuations of up to +/- 45% are within the normal range (means +/- SD of the day-to-day fluctuations were 16.9 +/- 7.7% and 15.6+/- 8.2%). Turn-around time is a minimum of 29 h for the urinary estrogen assay, 4.5 h for the plasma total estriol, and 3 h for the plasma free estriol.
Alternative mechanized method for analysis for inorganic phosphate without reduction.
Authors: Roberts N B
Published: January 2020

The mechanized measurement of inorganic phosphate in plasma can be achieved without reduction by only minor alterations to an existing continuous-flow reduction procedure. The method is based on the enhancement of the extinction of the phosphomolybdate complex by the detergent Triton X-100. Results obtained agree well with those obtained by the standard reduction (Fiske-SubbaRow) procedure. The correlation coefficient between the two methods was 0.996 and statistical analysis of the data confirmed very good agreement between the two sets of values.
Modified Jendrassik--Grof method for bilirubins adapted to the Abbott Bichromatic Analyzer.
Authors: Mori L
Published: January 2020

I describe a semi-micro method for use with a discrete analyzer, the Abbott ABA-100, requiring 75 microliter of sample for determination of both the total and direct bilirubins. This method utilizes a serum blank. It is simple and reasonably rapid. The azobilirubin formed by both the total and the direct bilirubins produces a reddish-violet color, which is measured at 550 nm. Values obtained are comparable to those by the automated Jendrassik and Grof procedure adapted by Gambino and Schreiber (total r = 0.999, direct r = 0.990). The method is linear to 300 mg/liter. Day-to-day precision (CV) for the total bilirubin was 8.8% for a 8 mg/liter sample (n = 45) and 4.7% for a 57 mg/liter sample (n = 46). The absorbance of hemoglobin and its derivatives (540 nm) caused a false decrease in the measured bilirubin content, the direct being more affected than the total. This problem was alleviated by the use of caffeine and acid blanks.
Mass-fragmentographic determination of catecholamine metabolites in amniotic fluid and its possible clinical usefulness.
Authors: Muskiet F A , Jeuring H J , Nagel G T , de Bruyn H W , Wolthers B G
Published: January 2020

We describe the determination of 3-methoxy-4-hydroxy-phenylacetic acid, 3-methoxy-4-hydroxyphenylamandelic acid, and 3-methoxy-4-hydroxyphenylethylene glycol in amniotic fluid by means of mass fragmentography, with use of deuterated internal standards. The results expressed in terms of absolute concentration and creatinine concentration, are given as a function of gestational age. In the 15th to 17th week, concentrations in amniotic fluid are a reflection of those in the mother's serum, whereas in the 32nd to 40th week, these concentrations, expressed in terms of creatinine, are similar to those found in the urine of newborns. We discuss the possible usefulness of the determination of catecholamine metabolites in amniotic fluid in the diagnosis of congenital neuroblastoma, maternal pheochromocytoma, and underdevelopment.
New enzymatic method for serum uric acid at 500 nm.
Authors: Trivedi R C , Rebar L , Berta E , Stong L
Published: January 2020

We describe a manual method, well suited to mechanization, for quantitating serum uric acid at 500 nm. In the assay mixture (0.10 ml of sample and 3.00 ml of reagent) the hydrogen peroxide produced from uric acid by uricase is coupled with p-hydroxybenzoate and 4-aminoantipyrine in the presence of peroxidase to form a colored complex, which is measured. A separate sample blank is obviated by taking an initial absorbance measurement 20 s after the sample is added. The reaction is complete within 5 min; its sensitivity is 0.001 deltaA/mg per liter. Absorbances are linearly related to uric acid concentrations up to 120 mg/liter. Many substances that may be present in normal serum do not interfere, but bilirubin in moderately above-normal concentrations will interfere. The procedure can be modified to largely correct for this, when necessary. The proposed method (y) correlated well (r = 0.979) with the uric acid 293 nm reference method (x) and the relation is described by the equation y = 0.998x + 2.42.
Bisalbuminemia. A new molecular variant, albumin Vancouver.
Authors: Frohlich J , Kozier J , Campbell D J , Curnow J V , Tárnoky A L
Published: January 2020

Of 18 members of a Fiji Indian family investigated, eight of the 12 males and two of the six females had an electrophoretically slow-type bisalbuminemia (alloalbuminemia). The albumin was characterized by the hiterto unique ratio of the two bands (Al A 35%: variant 65%), and by dye-binding studies and electrophoretic mobility in different media. The data suggest that this is a new variant, which we propose to call albumin Vancouver (Al Va).
High-performance liquid-chromatographic assay of aminoglycoside antibiotics in serum.
Authors: Anhalt J P , Brown S D
Published: January 2020

We describe procedures for the high-performance liquid-chromatographic assay of gentamicin, amikacin, and tobramycin in serum. The aminoglycoside antibiotic is extracted from serum by using a CM-Sephadex column and is measured by reversed-phase, ion-pair chromatography. Continuous-flow, post-column derivatization with o-phthalaldehyde is used to form fluorescent products for detection. An internal standard is used in each assay. Synthesis of 1-N-acetylgentamicin C1, the internal standard for the gentamicin assay, is described. For each procedure, concentration and instrument response are linearly related in the range of therapeutically important concentrations. The methods are precise and results correlate well with results obtained by bioassay. The between-day CV (n = 10) for each procedure was less than 4%. No substances have been detected in clinical specimens that interfere with these chromatographic procedures. The possible use of these methods for analysis for other aminoglycoside antibiotics is discussed.
High-performance liquid-chromatographic simultaneous determination of commonly used tricyclic antidepressants.
Authors: Proelss H F , Lohmann H J , Miles D G
Published: January 2020

We report a method for simultaneous measurement of five commonly used tricyclic antidepressant drugs (doxepin, desipramine, nortriptyline, imipramine, and amitriptyline) in serum by paired-ion high-performance liquid chromatography, with use of a reversed-phase column and ultraviolet detection at 254 nm. The drugs are extracted from 2 ml of serum at pH 14 into hexane/isoamyl alcohol (99/1 by vol) and re-extracted into 200 microliter of 0.1 mol/liter HCI. An aliquot of the aqueous acid phase is chromatographed with use of a methanol/acetonitrile/water (41/15/44) solvent system, containing 5 mmol of pentanesulfonic acid per liter of phosphate buffer (0.1 mol/liter, pH 6.5), at a flow rate of 1,5 ml/min. Analytical recoveries of the drugs from serum increase with increasing concentration, from 62% at 25 microgram/liter to 93% at 300 microgram/liter. Linear response is observed for drug concentrations up to 1500 microgram/liter and the detection limit is 2-3 microgram/liter. Within-run precision ranges from 1.4 to 2.9% and day-to-day precision from 1.7 to 7%, depending on the specific drug. The entire procedure can be completed within 45 min and is well adapted to the routine clinical laboratory. Of 48 common basic and several neutral drugs tested for possible interferences, only three benzodiazepines, three phenothiazines, and three antihistamines interfere with the assay of doxepin, desipramine, and nortriptyline, respectively.
Effect of in vitro hemolysis on chemical values for serum.
Authors: Frank J J , Bermes E W , Bickel M J , Watkins B F
Published: January 2020

Hemolysis in serum specimens is commonplace. This study examines the effect of hemolysis on results of selected chemical assays. Hemolysis was simulated by adding hemolysate to serum to give hemoglobin concentrations of 90 to 2800 mg/liter and a rating by technologists of 0 to 4 + hemolyzed. The effect of hemolysis on values for some serum constituents, particularly acid phosphatase and creatine kinase, was shown to be method dependent. Not unexpectedly, hemolysis most affects results for lactate dehydrogenase.
Continuous-flow enzymic method evaluated for measurement of serum triglycerides with use of an improved lipase reagent.
Authors: Whitlow K , Gochman N
Published: January 2020

We compared a modified enzymic continuous-flow method for serum triglycerides, in which a microbial lipase is used without protease, to an extraction-fluorometric method. Results (for total--blank) correlated well with those by the comparison procedure: enzymic = 1.002 fluorometric--14 mg/liter; r = 0.9968. Serum blanks were measured for each sample in both methods, and the concentration of free glycerol measured by the enzymic method was shown to be as much as 15 to 71% of the total triglyceride result in 7% of the samples analyzed. The dual-channel continuous-flow system provides automatic free-glycerol blank subtraction for the enzymic method.
Measurement of plasma nortriptyline concentrations: radioimmunoassay and gas-chromatography compared.
Authors: Robinson J D , Braithwaite R A , Dawling S
Published: January 2020

We compared measurement of plasma nortriptyline by a recently developed radioimmunoassay technique with values obtained by traditional gas chromatography. The coefficient of correlation (r) was 0.98 for 84 samples from 21 controlled patients; for a separate series of 45 samples from 34 patients who were receiving in addition other medication, r was 0.96. Use of radioimmunoassay in the routine determination of plasma nortriptyline concentration is discussed.
Continuous-flow analysis for serum albumin, by use of the immediate reaction with bromcresol green.
Authors: Plesher C J
Published: January 2020

Albumin reacts with bromcresol green reagent almost immediately, other serum proteins more slowly. This phenomenon can be exploited to improve the specificity of this routine method for determining serum albumin. I describe a 30-s reaction as adapted to continuous-flow analyzers, with no interference from other serum proteins. The method shows linearity to 72 g/liter. Results of this method correlate very well with those by the Laurell "rocket" and manual (30-s) bromcresol green methods. This procedure gives a considerable improvement in specificity for continuous-flow analyzers as compared with current bromcresol green methods.
Enzymatic determination of plasma ammonia: evaluation of Sigma and BMC Kits.
Authors: Pesh-Imam M , Kumar S , Willis C E
Published: January 2020

We compared two kit methods for the enzymatic determination of plasma ammonia. The procedures involved measuring the decrease in absorbance at 340 nm resulting from the consumption of NADH in the Sigma kit, NADPH in the BMC kit. Both kits gave acceptable results for aqueous samples, but for plasma specimens values with the Sigma kit significantly exceeded those obtained with the BMC kit. In our hands, values with the BMC kit were more accurate and precise. On the basis of these results, we conclude that the BMC kit is the better choice.
Automated development of a kinetic method for the continuous-flow determination of creatinine.
Authors: Olansky A S , Deming S N
Published: January 2020

A "stopped-flow" method for the kinetic Jaffé determination of creatinine was developed, with the use of a computer-controlled continuous-flow system. Simplex optimization was used to find conditions of hydroxide and picrate giving maximum sensitivity for creatinine. We used a modified central composite experimental design to evaluate creatinine sensitivity and albumin, glucose, and acetone interferences as functions of hydroxide and picrate concentrations. More importantly, this work illustrates that the automated development of clinical chemical methods offers an efficient means of obtaining optimized, well-understood analytical procedures for subsequent routine use in the clinical chemistry laboratory.
Evaluation of a new multichannel analyzer, "Astra-8".
Authors: Finley P R , Williams R J , Lichti D A , Thies A C
Published: January 2020

We evaluated a new multiple-channel chemistry analyzer, the Beckman "Astra-8," which is controlled by a microprocessor with 52K bytes of memory. The instrument we tested performed the following tests: Na, K, CO2, chloride, urea nitrogen, glucose, and creatinine. We compared the Astra-8 to a continuous-flow (SMA 6/60) and a discrete (Du Pont aca, urea nitrogen only) analyzer. The Astra-8 demonstrated excellent precision, linearity, accuracy, analytical recovery, lack of interference, ease of operation, and satisfactory comparison to values obtained by the comparison methods.
Interlaboratory comparison of results of erythrocyte protoporphyrin analysis.
Authors: Jackson K W
Published: January 2020

Each of 65 laboratories analyzed 10 whole-blood samples for erythrocyte protoporphyrin by one or more of several analytical procedures. These procedures were of two types: (a) extraction of protoporphyrin from the erythrocytes into ethyl acetate/acetic acid, re-extraction into hydrochloric acid, and fluorometric measurement; or (b) direct reading in a portable fluorometer (hematofluorometer), with no pretreatment of the blood sample. Interlaboratory correlation was generally poor, especially between laboratories using extraction procedures. Hematofluorometric results intercorrelated better, but they had a low bias as compared to the extraction approach. Nationwide standardization of the test is required to assure satisfactory interlaboratory performance and to identify laboratories whose results are sufficiently accurate to be used for interpretations according to guidelines set forth by the Center for Disease Control for erythrocyte protoporphyrin testing.
Interference by tubing material in continuous-flow systems.
Authors: de Jong E B , Goldschmidt H M , Loog J A , van Alphen C C
Published: January 2020

An anomalous behavior in the measurement of calcium with the smac multi-channel high-speed analyzer was eliminated by taking the diluted serum directly from the dilution loop of the instrument before it enters the stainless-steel riser.
High-density lipoproteins estimated by an enzymatic cholesterol procedure, with a centrifugal analyzer.
Authors: Ash K O , Hentschel W M
Published: January 2020

We describe an assay for high-density lipoprotein cholesterol, adapted to a centrifugal analyzer, the GEMSAEC System 3, which includes use of an increased Mn2+concentration (91 mmol/liter) [J. Lipid Res. 19, 65 (1978)] and ethylenediaminetetraacetate [Clin. Chem. 22, 98 (1976)]. Modifications to the GEMSAEC system include reducing the mixing burst and preconditioning the sample tip. Accuracy of this procedure, as assessed by analysis of a control pool from the Center for Disease Control, was 99.2%. Day-to-day precision for two control pools was 320 +/- 13 and 506 +/- 17 mg/liter. Serum sample volume was decreased to 0.5 ml. In blanks with heparin/Mn2+ present, the pseudocholesterol concentrations resulting from a reaction of the enzymatic cholesterol reagent and the heparin/Mn2+ precipitating reagent depend on the source of the enzymatic reagent and appear to be enhanced slightly by the use of ethylenediaminetetraacetate. Pseudocholesterol concentrations reach a maximum at heparin/Mn2+ concentrations well below those needed to completely precipitate the low-density and very-low-density lipoprotein fractions. Population reference values were obtained from analyses done on 224 local physicians (mean: male, 500 mg/liter; female, 620 mg/liter) and 156 ambulatory patients (mean: male, 463 mg/liter; female, 553 mg/liter).
Comparison of radioassay and microbiological assay for serum folate, with clinical assessment of discrepant results.
Authors: Baril L , Carmel R
Published: January 2020

Folate assays by use of radiolabeled folate provide obvious practical advantages over the standard microbiological assay, but remain incompletely tested. We therefore compared results for 415 sera with a kit involving 3H-labeled folate and the Lactobacillus casei microbiological method. We examined the patients' data when there were discrepancies between the two methods. Although the correlation overall was satisfactory, results were discrepant in 25% of cases. In 74% of the latter, the radioassay result appeared to be the correct one, primarily because L. casei results were suppressed by antibiotics being taken by the patient. The radioassay occasionally gave falsely high values for patients with liver disease and falsely low ones for patients who had received isotopes for scanning purposes. Several assay kits that make use of 125I- or 75Se-labeled folate were also tested. Although these results correlated with the results of 3H-labeled folate assay, various problems appeared, including the possible need for serum-supernate control tubes in one kit. Answers to these and other questions and careful clinical correlation of results are needed for any folate radioassays before their adoption for routine clinical use.
Determination of pyruvate oxidation rate and citric acid cycle activity in intact human leukocytes and fibroblasts.
Authors: Willems H L , de Kort T F , Trijbels F J , Monnens L A , Veerkamp J H
Published: January 2020

We measured pyruvate oxidation in intact leukocytes and fibroblasts by measuring 14C02 production. The optimal pyruvate concentration appeared to be higher than that usually applied. Activities remained constant during the incubation and were proportional to the amount of tissue protein added. Man values (+/-SD) were 2.8 +/- 0.9 nmol/h per 10(6) cells and 37 +/- 14 nmol/h per mg of protein for leukocytes and fibroblasts, respectively, for [1-14C]pyruvate oxidation; and 2.1 +/- 0.8 nmol/h per 10(6) cells and 18 +/- 7 nmol/h per mg of protein, respectively for [2-14C]pyruvate oxidation. We compared oxidation rates of pyruvate and 2-oxoglutarate by intact cells with those of isolated mitochondria. The ratio of 14CO2 production vs. activity of mitochondrial marker enzyme demonstrated that the rate of pyruvate oxidation can adequately be assayed in intact cells, but that the permeability of the cell membrane is rate-limiting in the oxidation of 2-oxoglutarate. No significant oxidation of other intermediates of the citric acid cycle was found, presumably owing to a low rate of transport of these substances across the cell membrane.
Variance components of serum constituents in healthy individuals.
Authors: Van Steirteghem A C , Robertson E A , Young D S
Published: January 2020

Analytical, intra-individual, and inter-individual components of variance were estimated in 10 volunteers for 34 assays encompassing 22 different constituents. Replicate blood specimens were obtained four times weekly. Nine constituents were measured with more than one instrument [Technicon AutoAnalyzer II and SMAC System, Perkin-Elmer Kinetic Enzyme Analyzer KA-150, and (or) the Abbott Bichromatic Aanalyzer ABA-100]. For those constituents that were stored for randomized batch analysis, results were examined for evidence of changes during storage and within-run analytical drift. Alkaline phosphatase, cholesterol, dopamine-beta-hydroxylase, and SMAC-measured lactate dehydrogenase had a ratio of intra-individual to interindividual variance of less than 0.10, thus exhibiting a high individuality. This ratio was greater than one for sodium, carbon dioxide, and creatine kinase. Analytical variance was more than 30% of total variance for sodium, carbon dioxide, calcium, total protein, and chloride. Analytical variance was less than 20% of intra-individual variance for bilirubin, creatine kinase, KA-150-measured lactate dehydrogenase, triglycerides, and urea nitrogen. Limits for the expected difference between measurements of the same constituent for the same individual on different days are also presented.
Hemopyrrole and kryptopyrrole are absent from the urine of schizophrenics and normal persons.
Authors: Gendler P L , Duhan H A , Rapoport H
Published: January 2020

We describe a method for detecting hemopyrrole and kryptopyrrole in urine, with a detection limit of 100 mug/liter (1 part in 10(7)). Urine is thoroughly extracted with methylene chloride and the extract is concentrated and examined by gas chromatography and gas chromatography-mass spectrometry. No hemopyrrole or kryptopyrrole could be detected in 52 samples, from 17 controls, 29 schizophrenics, and six persons with acute intermittent porphyria.
Automated assay of gamma-aminobutyric acid in human cerebrospinal fluid.
Authors: Böhlen P , Schechter P J , van Damme W , Coquillat G , Dosch J C , Koch-Weser J
Published: January 2020

We describe an automated amino acid analyzer with fluorescence detection (o-phthalaldehyde) which permits sensitive and rapid determinations of gamma aminobutyric acid in human cerebrospinal fluid. Concentrations as low as 50 nmol/liter can be accurately determined in 100 mul samples at the rate of one sample per hour. Concentrations in untreated cerebrospinal fluid increase rapidly after sampling by lumbar puncture. The concentration in immediately deproteinized samples from 38 patients with intervertebral disc disorders was 220 +/- 81 nmol/liter (mean +/- SD).
Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate at 25, 30, and 37 degrees C.
Authors: Buhl S N , Jackson K Y , Graffunder B
Published: January 2020

Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate were determined for isoenzymes 1 and 5 at 25, 30, and 37 degrees C. Three of the nine different buffers examined--imidazole, triethanolamine, and N-tris(hydroxymethyl)-methyl-2-aminoethane sulfonic acid--are satisfactory. Beta-NADH, pyruvate, and hydrogen ion concentrations were chosen to measure both isoenzymes with maximal-equal-sustainable efficiency at the lowest substrate concentrations. Approximately 95% of each isoenzyme is measured, for activities up to threefold the upper normal limit, if the measurements are made immediately after the reaction is initiated. The Arrhenius relationship for each isoenzyme is unique. Interconversion of results from one temperature to another is practical only with reservations. Results at 37 degrees C are not as reliable as those at 25 degrees C.
Determination of quinidine by high-performance liquid chromatography.
Authors: Powers J L , Sadee W
Published: January 2020

Quinidine in serum was measured at 330 nm, after protein precipitation, with an alkyl phenyl reversed-phase chromatographic column by peak-height determination (external standards.) Sensitivity was 0.3 mg/liter, with linear response to at least 10 mg/liter serum. Interassay precision, measured on 20 consecutive days, gave a CV of 8.2% at 2 mg/liter of serum and 5.1% at 5 mg/liter. Quinine and primaquine are not separable from quinidine under the assay conditions, and dihydroquinidine and chloridazepoxide are only partly resolved. No assay interference was encountered with a series of control serum samples obtained from patients with various diseases, who were being treated with various drugs other than quinidine, except in one serum sample with a high bilirubin concentration (300 mg/liter). If such an assay interference is present, an alkaline extraction of quinidine before analysis has to be used. Results by our assay and those by a single-extraction fluorescence method correlated poorly (r=0.87), and the fluorescence assay gave 50% +/- 7% (SEM) higher values than our method, due to chromatographic separation of a major fluorescent non-phenolic metabolite of quinidine, which can also be measured by our assay.
An enzymatic triglyceride method that is suitable for long-term population studies.
Authors: Koehler D F , Steele B W , Azar M M , Kuba K , Dempsey M E
Published: January 2020

An enzymatic triglyceride method has been shown to be a suitable alternative to the Lipid Research Clinics' extraction/fluorometry method in long-term population studies. Correlation of results obtained with this method by this laboratory (y-axis) and by the Minneapolis Lipid Research Clinic Laboratory (x-axis) during a nine-week standardization period produced a curve with an intercept of -72 mg/liter, a slope of 1.019, and a correlation coefficient of r=0.996 (n=47). During this standardization period certain methodological problems were observed and corrected. An increase in background in certain clinical specimens, caused by spontaneous degradation of NADH, was observed, accurately measured, and taken into account when appropriate.
Solid-phase radioimmunoassay for morphine, with use of an affinity-purified morphine antibody.
Authors: Steiner M , Spratt J L
Published: January 2020

Morphine antibody purified by affinity chromatography was used to develop a solid-phase radioimmunoassay for morphine in polystyrene tubes. The tubes are coated with an appropriate concentration of the purified antibody, rinsed three times with buffered saline, and stored at -15 degrees C. Using tritiated dihydromorphine, we determined competitive morphine binding by difference when the radioactivity in the assay supernates was measured after incubation (1 h, 37 degrees C). Five standard curves, with use of serum equivalents of morphine ranging from 0 to 6 mug/liter, were linear and had a mean correlation coefficient of 0.98. Uncer conditions of the assay, levorphanol was comparable to morphine in its inhibitory effect on binding of labeled dihydromorphine, whereas dextrorphan was essentially inactive. Morphine-3-glucuronide, a major metabolite, is 55-fold less inhibitory in terms of its capacity to displace the radiolabel. We believe that the sensitivity of the technique, coupled with the simplicity of nonseparatory sampling, renders the system suitable for rapid determination of morphine and related compounds in biological fluids.
Automated measurement of serum thyroxine with the "ARIA II," as compared with competitive protein binding and radioimmunoassay.
Authors: Reese M G , Johnson L R
Published: January 2020

Two conventional serum thyroxine assays, run in separate laboratories, one by competitive protein binding and one by radioimmunoassay, were used to evaluate the automated ARIA II (Becton Dickinson Immunodiagnostics) serum thyroxine assay. Competitive protein binding as compared to ARIA II with 111 clinical serum samples gave a slope of 1.04 and a correlation coefficient of 0.94. The radioimmunoassay comparison to ARIA II with 53 clinical serum samples gave a slope of 1.05 and a correlation coefficient of 0.92. The ARIA II inter-assay coefficient of variation for 10 replicates of low, medium, and high thyroxine serum samples was 6.2, 6.0, and 2.9%, respectively, with an inter-assay coefficient of variation among 15 different assays of 15.5, 10.1, and 7.9%. The automated ARIA II, with a 2.2-min cycle per sample, gives results that compare well with those by manual methodology.
High-speed automated atomic absorption spectrophotometry.
Authors: McMurray C H , Blanchflower W J
Published: January 2020

We describe an integrated automated system for the atomic absorption spectrophotometer. The system makes use of a light-dependent resistor to detect the presence of a marker ion which is present along with the sample ion in the flame. An electronic circuit has been developed which enables the marker ion to trigger the read signal of the spectrophotometer. The system has been tested by determining magnesium and calcium in serum. Alternative modes of operation and different potential applications are discussed.
Improved ultraviolet spectrophotometry of serum theophylline.
Authors: Carter P , Wallace J E , Blum K
Published: January 2020

We present an improved method for ultraviolet spectrophotometry of theophylline in serum. We studied various extraction techniques aimed at eliminating interferences from co-extractable serum constituents. In the resulting modified procedure, 1 ml of serum is required and a salt-solvent pair of ammonium sulfate and chloroform/hexane is used for extraction. The solvent forms the top phase after extraction, the lower phase after back-extraction, thereby permitting easy removal of the appropriate phase from culture tubes. The use of ammonium sulfate coupled with the added specificity of the extraction solvent results in an extract with low background absorption and a well-defined spectrum for the extracted theophylline.
Spectrophotometry of theophylline-7-acetic acid and theophylline.
Authors: Owen J A , Nakatsu K
Published: January 2020

A spectrophotometric assay procedure is described for individually measuring theophylline-7-acetic acid and theophylline in 2.0 ml of serum. Absorbance is a linear function of concentration over a range of 0.5-40 mg/liter for theophylline-7-acetic acid and 0.5-40 mg/liter for theophylline. No endogenous or exogenous compounds were found that interfere with theophylline-7-acetic acid determinations. As with other spectrophotometric theophylline assays, theobromine and, to a slight extent, phenobarbital interfere with theophylline determinations.
An enzymatic assay for determining free and total cholesterol in tissue.
Authors: De Hoff J L , Davidson L M , Kritchevsky D
Published: January 2020

We describe a method for determining free and total cholesterol in extracts of rat-liver lipids by use of a cholesterol esterase/cholesterol oxidase enzymatic kit (Boehringer Mannheim Corp). The lipids are solubilized (as micelles) directly into the aqueous reagent mixture. Results of the enzymatic method correlate well with those obtained by an acid/ferric chloride assay of digitonin precipitates of the extracts. The enzymatic assay of free and total cholesterol in tissue offers advantages of simplicity over previously used methods.
Quantitative toxicology: interlaboratory and intermethod evaluation in New York State.
Authors: Buhl S N , Kowalski P , Vanderlinde R E
Published: January 2020

The New York State Department of Health has conducted a proficiency evaluation program in quantitative toxicology since 1974. Serum samples containing a barbiturate and phenytoin, together with either glutethimide, procainamide, or theophylline, are sent to participating laboratories quarterly. Within the first two years of the program the percentage of laboratories able to quantitate 75% of the test samples to within 25% of the gravimetric values increased from 25 (1974-1975) to 40% (1975-1976). This improvement was partly due to licensure requirements, improved technology for sample preparation and analysis, and the availability of better quality-control practice. An obstacle to obtaining uniform accuracy is the lack of adequate calibration or testing materials. To overcome these obstacles, pure drugs are weighed into a bovine serum matrix, and the weights are confirmed by reference laboratories and used as the target values in the testing program. Comparison of the methods used by participants in this program for barbiturate and phenytoin yielded equations different from those found in other method evaluations.
Amperometric nonenzymatic determination of serum glucose by means of a nickel-catalyst electrode.
Authors: Schick K G , Magearu V G , Huber C O
Published: January 2020

We describe a new, nonenzymatic method for determining serum glucose. This method is based upon the direct electrochemical oxidation of glucose by means of stable nickel-catalyst that is electrodeposited onto a lead dioxide electrode surface. Linear calibration plots are obtained for glucose in the 0.1 to 4500 mg/liter range. Values for 23 sera with glucose concentrations ranging from 300 to 4300 mg/liter showed a linear correlation coefficient of 0.996 when compared with values reported by a hospital laboratory using the hexokinase method. Typically a 25-microliter sample is used, but samples as small as 5 microliter have been successfully analyzed. The sample is added to 25.00 ml of pH 13 electrolyte containing 1 mmol of NiSO4 per liter; the amount of increase in anodic current comproses the analytical signal. For 20 repetitive analyses of a serum specimen, the CV was less than 4%. Ascorbic acid is rapidly decomposed in the electrolyte used and thus does not interfere. Uric acid is inactive at the concentrations present in serum samples. The sensitivity and simplicity of the new method suggest it as an alternative to currently available procedures.
Some factors affecting the utility of bovine blood as a control material in lead-screening programs.
Authors: Penton Z , Bissell G
Published: January 2020

Bovine blood is used frequently as control material in interlaboratory comparisons for blood-lead analysis, but an erythrocyte protoporphyrin control material is needed. We present data on the rate of changes in blood lead and erythrocyte protoporphyrin in steers fed lead acetate. We also show, by comparison of fluorescence spectra with those of known compounds, that most of the porphyrin in lead-burdened steers probably exists as free protoporphyrin IX along with some zinc-complexed protoporphyrin IX. Bovine blood is suitable as an erythrocyte protoporphyrin control material for laboratories using acid extraction procedures, but not for the proficiency testing of laboratories using hematofluorometers.
Method-specific variations in the calibration of a new immunoglobulin standard suitable for use in nephelometric techniques.
Authors: Whicher J T , Hunt J , Perry D E , Hobbs J R , Fifield R , Keyser J , Kohn J , Riches P , Smith A M , Thompson R A , Ward A M , White P
Published: January 2020

The putative International Federation of Clinical Chemistry immunoglobulin standard, IFCC 74/1, was calibrated against the World Health Organization (WHO) immunoglobulin standard 67/99 by three different methods: automated immunoprecipitation, Laurell rocket immunoelectrophoresis, and radial immunodiffusion. The same antisera were used in all the assays, which were performed in five expert laboratories. With the aid of linearizing transformations for the dilution curves of both materials and use of a carefully weighted statistical evaluation, values in International Units for IgG, IgA, and IgM were ascribed to IFCC 74/1. The values achieved by the three different methods were not statistically different, except in the case of IgA. IgM assays by automated immunoprecipitation were excluded owing to a high degree of imprecision.
Serum chromium and angiographically determined coronary artery disease.
Authors: Newman H A , Leighton R F , Lanese R R , Freedland N A
Published: January 2020

Human aortas sampled from populations where there is little advanced atheromatous plaque formation contain higher concentrations of chromium than do aortas from populations in which atheromatosis is prevalent. In the present study serum cholesterol, triacylglycerols, and chromium (Cr3+) concentrations were measured in 32 subjects in whom coronary artery disease was assessed by cineangiography. The distribution of subjects with diseased and normal arteries overlapped below 5.50 microgram of chromium per liter. Only subjects free of coronary artery disease had chromium concentrations greater than or equal to 5.50 microgram/liter. The role of chromium was assessed in the context of the selected risk factors: cholesterol, triacylglycerols, and systolic and diastolic blood pressure. The group with coronary artery disease had significantly lower serum chromium concentrations than did the group with normally patent arteries.
Computer-controlled instrument system for sequential chemical testing III. Application to liver assessment.
Authors: Cembrowski G S , Larson F C , Huntington R W , Selliken J H , Toren E C
Published: January 2020

We used the previously described [Clin. Chem. 19, 1114 (1973)] and evaluated [Clin. Chem. 19, 1122 (1973)] computer-controlled instrument system for sequential chemical testing to select and perform tests of hepatic status, to aid the clinician in the diagnosis of liver disease. Results for total bilirubin, aspartate aminotransferase, and alkaline phosphatase obtained from the continuous-flow analysis (SMA 12/60) admission screen were used by the instrument system to determine selectively the values for gamma-glutamyltransferase, alanine aminotransferase, creatine kinase, and total and direct bilirubin. Kit methods for the latter four tests were evaluated on the system; results were similar to manual procedures. A software, enzymatic ratemeter was found to be better than the previously described hardware ratemeter. The follow-up tests of serum prescribed by the system are compared to clinician-prescribed follow-up tests and discharge diagnoses. In 10 of 19 cases, the system and clinician ordered similar follow-up tests; in three cases follow-up differed, and in six cases, the system ordered follow-up tests and the clinician ordered none.
Characteristics of statistical parameters used to interpret least-squares results.
Authors: Davis R B , Thompson J E , Pardue H L
Published: January 2020

This paper discusses properties of several statistical parameters that are useful in judging the quality of least-squares fits of experimental data and in interpreting least-squares results. The presentation includes simplified equations that emphasize similarities and dissimilarities among the standard error of estimate, the standard deviations of slopes and intercepts, the correlation coefficient, and the degree of correlation between the least-squares slope and intercept. The equations are used to illustrate dependencies of these parameters upon experimentally controlled variables such as the number of data points and the range and average value of the independent variable. Results are interpreted in terms of which parameters are most useful for different kinds of applications. The paper also includes a discussion of joint confidence intervals that should be used when slopes and intercepts are highly correlated and presents equations that can be used to judge the degree of correlation between these coefficients and to compute the elliptical joint confidence intervals. The parabolic confidence intervals for calibration cures are also discussed briefly.
Analysis for potassium in human erythrocytes by use of a standard-additions method and an ion-selective electrode.
Authors: Mangubat E A , Hinds T R , Vincenzi F F
Published: January 2020

A method based on standard additions to the sample was used to measure potassium (K+) in human erythocytes with an ion-selective electrode. A computer program was developed for rapid analysis of data obtained with the electrode. The analysis takes into account factors such as temperature that may affect electrode behavior. Results obtained by the method agree with those obtained by flame-emission spectroscopy. The coefficient of correlation between the two methods is 0.96. Our method is simple and rapid. The computer program is applicable to other analyses with ion-selective electrodes.
A technique for determining the probability of abnormality.
Authors: Grannis G F , Lott J A
Published: January 2020

Histograms of the results of quantitative clinical laboratory analyses are usually broadly skewed because some of the patients have normal values with respect to the analyte being measured while values for others are below or above normal. We describe a technique for calculating the maximum possible number of individuals in a patient population who will give normal values and, by difference, the minimum possible number of those who will give below-normal and above-normal values. The technique permits estimation of the probability that any given result represents a member of a particular subpopulation, as well as selection of decision limits for patient classification, and estimates the degree of change required for a subject with a normal value to start giving below- or above-normal values.
Simultaneous liquid-chromatographic determination of 12 common sedatives and hypnotics in serum.
Authors: Kabra P M , Koo H Y , Marton L J
Published: January 2020

We present a method for simultaneously determining 12 hypnotics and sedatives (primidone, methyprylon, phenobarbital, butabarbital, butalbital, ethchlorvynol, pentobarbital, amobarbital, phenytoin, glutethimide, secobarbital and methaqualone) in 200 microliter of serum. Serum proteins are precipitated with an acetonitrile solution containing 5-(4-methylphenyl)-5-phenylhydantoin, the internal standard. The drugs are eluted from a reversed-phase column with a mobile phase consisting of an acetonitrile/phosphate buffer, at a flow rate of 3.0 ml/min. The eluted drugs are detected by their absorption at 195 nm; their quantities are estimated from their peak heights. Each analysis requires no longer than 30 min at the optimum column temperature of 50 degrees C. The lower limit of detection for all of these drugs is less than 10 ng/sample for drug standard. A sensitivity of 1.0 mg/liter of serum is attained routinely for each of the drugs. Analytical recoveries for the 12 drugs varied from 94 to 112%, with good day-to-day precision (CV between 3.8 and 10.4%). Of more than 35 drugs tested for possible interference, only ethotoin interferes with the analysis of phenobarbital.