bluuebunny/crossref_metadata_2025 huggingface dataset:
Coagulation-Time Determination with Automatic Multivariable Analysis, by Use of a Miniature Centrifugal Fast Analyzer
Authors: Bostick W D , Bauer M L , Morton J M , Burtis C A
Published: January 2020

Use of a miniature Centrifugal Fast Analyzer for the parallel photometric monitoring of the coagulation process is shown to have a number of advantages. These include a choice of optical modes, virtually simultaneous initiation and observation of the coagulation process for a number of patient-plasma samples and an on-board control sample, small sample and reagent volume requirements, and automatic determination of a number of diagnostically useful variables (including relative fibrinogen content) from the data recorded in a single run. Also, the system is shown to give results that correlate well with those obtained by conventional techniques for determination of prothrombin time.
Simultaneous high-pressure liquid-chromatographic determination of some anticonvulsants in serum.
Authors: Adams R F , Vandemark F L
Published: January 2020

We describe procedures for simultaneously determining some anticonvulsants (phenobarbital, diphenylhydantoin, primidone, ethosuximide, methsuximide, carbamazepine) in serum by high-pressure liquid chromatography. The drugs, together with an internal standard, phenacetin, are adsorbed from serum onto charcoal and eluted from it with organic solvent. The eluate is analyzed isocratically on a reverse-phase column with a mobile phase consisting of acetonitrile/water (17/83 by volume). The eluted drugs are detected by their absorption at 195 nm, and quantities estimated from their peak areas as compared with those of extracted standards. Results are reproducible to about 6%. Sensitivities, for 0.5-ml serum samples, of 0.1 mg/liter for all the drugs analyzed except ethosuximide (0.5 mg/liter) are attained routinely. Correlation of results with gas chromatography was 0.912 for phenobarbital, 0.982 for diphenylhydantoin, 0.886 for primidone, and 0.966 for ethosuximide. Amobarbital and secobarbital interfere with the analysis; chlordiazepoxide, methaqualone, salicylate, diazepam, and oxazepam do not. Including extraction, analysis time for a single sample is 20 min.
Quantitative fractionation of alkaline phosphatase isoenzymes according to their thermostability.
Authors: PetitClerc C
Published: January 2020

Continuous monitoring of heat denaturation of a mixture of alkaline phosphatase isoenzymes at 60 degrees C and pH 7.5 permits the simultaneous direct identification and quantitation of three isoenzymes: the placental isoenzyme, the L-phenylalanine-sensitive intestinal isoenzyme, and the liver isoenzyme (hepatocytic). The isoenzyme that is principally of bone origin cannot be identified as such without the help of other diagnostic aids and the patient's medical history. All human tissues contain alkaline phosphatase, many organs more than one of the isoenzymes. Liver alkaline phosphatase, which constitutes 40-50% of normal serum alkaline phosphatase activity, was measured in the serum of persons with various liver diseases. Its activity exceeded normal in all types of liver disease; in 80% of cases this increase was accompanied by increased gamma-glutamyl-transferase activity, but the quantitative correlationship (r = 0.54) was not as good as expected if both enzymes come from the same source and are indices of liver dieases. Liver alkaline phosphatase activity increases in the blood early in liver disease, before most liver tests show abnormalities. The other major isoenzyme of normal serum probably represents a mixture of isoenzymes from bone and reticulo-endothelial and vascular tissues, which all contain the same "very heat-labile" alkaline phosphatase. Cord blood and children's sera contain mostly this very heat-labile isoenzyme.
Atypical increase in serum creatine kinase activity in hospital patients.
Authors: Sax S M , Moore J J , Giegel J L , Welsh M
Published: January 2020

We use an ion-exchange column-chromatographic technique for separating creatine kinase isoenzymes in serum, and occasionally observe what appears to be sustained increase in the MB fraction. Most patients whose sera show such behavior have myocardial disease, but not necessarily a recent myocardial infarction. Electrophoretic analysis of a small sampling of such sera revealed that the apparent MB migrates atypically, appearing distinctly between isoezymes MB and MM. In another electrophoretic system, the peak might easily be mistaken for MM. This unusual isoenzyme does not appear to be "macro" creatine kinase. In laboratories that use the ion-exchange technique, the possibility of a falsely positive MB value should be considered in subjects who show persistent increases together with normal or nearly normal values for total creatine kinase activity. A suitable electrophoretic method that clearly demonstrates this unusual isoenzyme should be used in such cases, for confirmation.
Use of cholesterol oxidase for assay of total and free cholesterol in serum by continuous-flow analysis.
Authors: Richmond W
Published: January 2020

Idescribe an assay for total cholesterol in serum, with use of the AutoAnalyzer ii (Technicon), in which cholesterol esters are saponified by alkali, cholesterol is held in aqueous micellar solution with a surfactant (Triton X-100) and oxidized by cholesterol oxidase, and the hydrogen peroxide produced is measured by chelation with Ti4+ and xylenol orange. An assay for free cholesterol in serum, based on similar principles, is also described, and the two can be run simultaneously on a dual-channel AutoAnalyzer II. Standard solutions of cholesterol in isopropanol have poorer carryover characteristics than sera, and therefore do not reach the same continuous-flow steady state as sera of equivalent concentrations. Consequent potential calibration errors are avoided by using micellar solutions of cholesterol containing albumin for standardization. The formation of cholesterol peroxide in solutions of cholesterol in isopropanol is demonstrated, and this constitutes another potential source of error in the calibration of enzymic cholesterol assays. In analyzing patients' sera, results of the total cholesterol assay correlate well with those of a mechanized method in which cholesterol esterase and cholesterol oxidase are used; an automated Abell method, calibrated with solutions of cholesterol in isopropanol, gave slightly higher values. Determinations of the ratio of free cholesterol to total cholesterol by our automated cholesterol exidase assays given values that agree well with published results in which digitonin precipitation is used.
Microcalorimetric determination of glucose in reference samples of serum.
Authors: Goldberg R N
Published: January 2020

Ireport a detailed series of microcalorimetric measurement of glucose concentrations in five reference samples of serum. The method utilized (a) calibration in the actual medium of analysis and (b) a correction for interferences owing to the nonspecificity of the enzyme hexokinase. The microcalorimetric results are compared with the tentative results of analyses obtained by the spectrophotometric hexokinase/glucose-6-phosphate dehydrogenase method and by isotope-dilution mass spectrometry. In most cases, the microcalorimetric results appear to be in agreement with the results obtained by these latter two methods. A discussion of the basis of the microcalorimetric measurements is presented.
Amino acid composition of human nail, as measured by gas-liquid chromatography.
Authors: Greaves M , Moll J M
Published: January 2020

We describe a quantitative method for determination of histidine and 16 other amino acids in a nail hydrolysate. This is based on the formation of trifluoroacetyl amino acid methyl esters and gas-liquid chromatographic analysis with use of two columns of mixed stationary phases. Histidine is determined as a triple derivative Nalpha-trifluroacetyl-Nim-carbethoxy methyl histidinate. The specificity and reproducibility of the method are illustrated. As little as 1 mg of amino acid per gram of nail can be determined, and 15 hydrolysates can be completely analyzed in 10 working days. Data obtained from multiple analyses of 10 nail hydrolysates by the method described closely parallel those obtained in another laboratory, in which an amino acid analyzer was used.
Direct enzymatic determination of urea in plasma and urine with a centrifugal analyzer.
Authors: Bretaudiere J P , Phung H T , Bailly M
Published: January 2020

A direct enzymatic micromethod (sample volume, 3mul) has been adapted to the centrifugal analyzer (ENI-GEMSAEC) for measurement of urea in plasma and urine. The method is based on urease (urea amidohydrolase, EC3.5.1.5)/glutamate dehydrogenase [l-glutamate:NAD(P)+oxidoreductase (deaminating), EC1.41.3] coupled reactions, and uses a two-point fixed-time (t(1)=20s,t(2)=50s)kinetic scheme for monitoring the rate of comsumption of NADH at 340 nm. Sensitivity and precision of the method are excellent,and results compare well with those from a commonly used continuous-flow method.
Cholesterol oxidase-based determination, by continuous-flow analysis, of total and free cholesterol in serum.
Authors: Lie R F , Schmitz J M , Pierre K J , Gochman N
Published: January 2020

We describe a continuous-flow, automated determination of total cholesterol in serum, which is based on enzymatic hydrolysis of cholesterol esters, oxidation of cholesterol by cholesterol oxidase, and colorimetric measurement of liberated perioxide with 4-aminoantipyrine, phenol, and peroxidase. Free cholesterol is determined with the same AutoAnalyzer II manifold and reagents, except that cholesterol esterase is omitted from the reagent. Cholesterol-in-serum materials that have been assayed by an established method are used for calibration. We found this approach to be necessary because primary cholesterol standards in organic solvents are incompatible with the aqueous reagent. Results of the enzymatic total cholesterol method correlated well with those by an AutoAnalyzer II method which involves an extraction with isopropanol and the Liebermann-Burchard color reaction (total cholesterol, g/liter, yenz= 0991xlb +0.05;r=0.996). Results of the enzymatic free cholesterol procedure agreed satisfactorily with one in which free cholesterol is precipitated as the digitonide and subsequently analyzed colorimetrically with the Liebermann-Burchard reaction (free cholesterol, %, yenz = 0.982xdig -0.7;r= 0.956).
Detection of inhibitors in reduced nicotinamide adenine dinucleotide by kinetic methods.
Authors: Howell B F , Margolis S , Schaffer R
Published: January 2020

Methods are described for detection of lactate dehydrogenase (LDH) inhibitors in preparations of reduced nicotinamide adenine dinucleotide. They are (a) comparison of values by kinetic methods with those measured for highly purified NADH and (b) examination of Lineweaver-Burk plots. Chromatographic inhomogeneities are correlated with deviant values for the kinetic constants of NADH preparations. Lineweaver-Burk plots that curve upward at the high concentrations or have a larger or smaller than normal slope may indicate the presence of inhibitor. As determined in bicarbonate buffer (0.11 mol/liter, pH 7.9) by use of 0.600 mmol/liter pyruvate and NADH freshly separated from impurities by chromatography on diethyl-aminoethyl-cellulose, the Km (apparent) of NADH at 25 degrees C has the value 8.11 +/- 0.71 mumol/liter (SD, n = 28) with LDH-1 (pig heart, 2.48 +/- 0.05 U per milliliter of reaction mixture, or 41.3 +/- 0.8 nmol/liter per second). Under similar conditions, the Km (apparent) of NADH has the value of 8.57 +/- 1.58 mol/liter (SD, n = 21) with LDH-5 (pig muscle, 1.77 +/- 0.03 U/ml of reaction mixture), or 29.4 +/- 0.6 nmol/liter per second). At infinite substrate concentrations with the same pH, buffer, and temperature, the Km (apparent) for NADH was 26.0 +/- 0.63 mumol/liter with LDH-1 and 23.2 +/- 4.6 mumol/liter with LDH-5.
Results for serum cholesterol and triglycerides by gas-liquid chromatography, as compared with a continuous-flow technique.
Authors: Watts R , Carter T , Taylor S
Published: January 2020

We assessed the usefulness of gas-liquid chromatography forthe routine and reference measurement of cholesterol and triglycerides (triacylglycerols). Compared with results obtained by an AutoAnalyzer II method, correlation was good with both serum specimens (triglycerides, r = 0.88; cholesterol, r = 0.90) and lipoprotein fractions isolated by ultracentrifugation (triglycerides, r = 0.98; cholesterol, r = 0.98). However, none of the comparisons was completely free of analytical bias, and in general the AutoAnalyzer method underestimated the cholesterol value and overestimated triglycerides. The obvious advantage of the AutoAnalyzer is its greater analytical speed, but the chromatographic method appeared to be more accurate and precise and to suffer less from interfering substances, and in addition gave information about the cholesterol ester and triglyceride composition. We expect gas-liquid chromatography to be of major importance in the calibration of other analytical methods.
Sarcolemmal membrane changes related to enzyme release in the imipramine/serotonin experimental animal model.
Authors: Silverman L M , Gruemer H D
Published: January 2020

We report specific findings in the imipramine/serotonin animal model that are consistent with sarcolemmal membrane alterations. Among these findings are cytoplasmic enzyme release, diminished uptake of alpha-aminoisobutyrate (an amino acid analog), decreased oxygen consumption in isolated rat diaphragm, and ribosuria. Furthermore, we describe for the first time the release of the MB isoenzyme of creatine kinase from a source other than cardiac tissue; that is, isolated diaphragms from imipramine/serotonin-treated animals release increased amounts of MB isoenzyme as compared to diaphragms from control animals. We believe the similarities between this animal model and the human disease (Duchenne muscular dystrophy) support a genetically determined generalized membrane abnormality in the pathogenesis of this form of muscular dystrophy.
Interference by acetaminophen in the glucose oxidase-peroxidase method for blood glucose determination.
Authors: Kaufmann-Raab I , Jonen H G , Jähnchen E , Kahl G F , Groth U
Published: January 2020

Acetaminophen, p-aminophenol, and oxyphenbutazone interfere with the glucose oxidase/peroxidase method for glucose. Structurally related compounds that lack a free phenolic hydroxyl group (acetanilide, aniline, and phenylbutazone) do not interfere. During the analytical procedure acetaminophen is consumed. One mole of acetaminophen leads to an apparent loss of four moles of glucose. The hexokinase/glucose-6-phosphate dehydrogenase method (Boehringer Hexokinase method) is not affected by these substances.
Simple modification of a liquid scintillation counter to decrease cost of and time for radioimmunoassay.
Authors: Dixon R , Cohen E
Published: January 2020

A simple, inexpensive modification in the elevator mechanism of a liquid scintillation counter allows for radioimmunoassay tubes (12 X 75 mm) containing the antibody-bound radioligand to be counted directly, thereby obviating the need for counting vials and considerably decreasing the amount of fluor used. Furthermore, because assay and counting are all done in the same tube, the modification shortens assay time and increases precision by avoiding what is often the most time consuming and inaccurate step, the transfer of antibody-bound radioligand to a counting vial. We find that use of this modification results in about a 75% decrease in the overall cost for materials and a substantial decrease in assay time.
Use of gas chromatography and mass spectrometry to analyze underivatized volatile human or animal constituents of clinical interest.
Authors: Politzer I R , Dowty B J , Laseter J L
Published: January 2020

The study of volatilizable components in human and animal samples by gas chromatography and mass spectrometry is reviewed for the years 1970--1975. Only naturally volatile, underivatized compounds are considered, from sources such as body fluids, tissues, and breath. Where possible, the clinical samples are discussed in terms of normal profiling, aberrations associated with disease states, and specific compound analyses. A brief descriptive technical section is included, stressing types of sample preparation currently being used.
Laboratory confirmation of the diagnosis of cystic fibrosis.
Authors: Tocci P M , McKey R M
Published: January 2020

The recent commercial introduction of a method for detecting albumin in meconium makes screening for cystic fibrosis feasible for many hospitals. If the tests is adopted, confirmatory tests should be available. Quantitative analyses of sweat for sodium by flame photometry and for chloride by silver titration and ion-sleective electrodes are now used as confirmatory tests. We compare results of these confirmatory methods applied to presons with cystic fibrosis, respiratory disorders, or digestive disorders, and to control subjects.
Gas-chromatographic identification of drugs in gastric aspirate samples: a rapid screening procedure for emergency toxicology.
Authors: Abramson F P
Published: January 2020

Gastric aspirate has not been used frequently in the identification of ingested drugs. Because there is only qualitative significance in such a sample, a new method is described that facilitates identification. A 1-ml gastric aspirate sample is extracted with 100 mul of chloroform and analyzed directly by gas chromatography. Seventeen drugs commonly involved in overdose cases are included in this 25-min procedure. The gastric drug screen has been applied to more than 500 patients, and results are described.
Radioimmunuassay compared to thin-layer and gas--liquid chromatography for detecting methadone in human urine.
Authors: Roerig D L , Wang R I , Mueller M M , Lewand D L , Adams S M
Published: January 2020

Radioimmunoassay was compared to thin-layer and gas--liquid chromatographic methods for detection of methadone in the urine of patients undergoing methadone maintenance therapy as treatment of heroin abuse. With urine samples known to contain methadone, 84% were positive by thin-layer chromatography as compared to 99% positives by the other two methods. This difference is attributed to the difference in sensitivity of the three methods. All three methods gave consistently positive results with urine samples from patients receiving 25 mg of methadone per day or more. With smaller daily doses the percentage of positive results obtained with thin-layer chromatography decreased. Analysis of urine samples not containing methadone showed no incidence of cross reaction of other drugs with the methadone radioimmunoassay. The methadone radioimmunoassay appears to be both sensitive and reliable; however, certain other factors limit its use as a primary screening method.
Relationship between erythrocyte 2,3-diphosphoglycerate and age in a normal population.
Authors: Kalofoutis A , Paterakis S , Koutselinis A , Spanos V
Published: January 2020

The concentration of 2,3-diphosphoglycerate in erythrocytes was determined in 369 male and 376 female normal subjects, 1 to 86 years of age, derived from the Greek population. It was inversely correlated (P less than 0.01) with age. Persons over age 60 showed a decrease (P less than 0.05) in 2,3-diphosphoglycerate concentrations as compared those 1-20 years of age.
Interferences with the starch-iodine assay for serum amylase activity, and effects of hyperlipemia.
Authors: Gitlitz P H , Frings C S
Published: January 2020

Recent reports imply or claim that amylase activity is inhibited in hyperlipemic sera, because diluted samples showed greater activities when assayed by a starch-iodine method. We find that dilutions of both clear and lipemic samples give higher-than-expected activities when assayed by a starch-iodine method, an effect attributable to the variable effects of protein, turbidity, and triglycerides in the hyperlipemic samples. Thus the starch-iodine method is unsuitable for assessing the effects of hyperlipemic samples on amylase activity. To do so, we used an alternative method, in which soluble dyed amylopectin is used as the substrate. This method exhibits apparent zero-order kinetics, and we detected no interfering factors. Plots of sample volume (x) vs. activity (y) for clear and hyperlipemic (triglycerides up to 80 g/liter) sera gave straight lines with y-intercepts of zero. Evidently hypertriglyceridemia does not inhibit amylase activity.
Screening for erros in galactose metabolism with the erythrocyte.
Authors: Rabinow B E , Wong P W , Maschgan E R , Natelson S
Published: January 2020

We propose determination of the ratio of the rate of galactose metabolism to glucose metabolism by erythrocytes as a screening test for abnormalities in glucose and galactose metabolism. Packed erythrocytes (20 mul) are incubated for 1 h at 37 degrees C in 0.42 ml of a solution comprising phosphate buffer (pH 7.4), 0.4 mg of glucose, 60 mg of methylene blue, and 50 nCi of either [1-14C]glucose or [1-14C]galactose. Metabolism is then stopped by injecting dilute H2SO4 through a rubber septum sealing the flasks. On incubating the acidified solution for 1 h, the evolved CO2 is trapped in a well containing ethanolamine, which is suspended from the septum. The radioactivity of the well and its contents is measured in a scintillator, and from these data CO2 is calculated. (The scintillation medium is preheated with ethanolamine to eliminate chemiluminescence.) For normal adults mean values for CO2 are 0.468 mumol/liter of erythrocytes per minute for galactose and 37.8 mumol/liter of erythrocytes per minute for glucose. Homozygous galactosemics exhibit no galactose metabolism but the rate for flucose metabolism is normal. Results for parents of homozymotes are described. We review various causes for galactosemia and point out that transfer of galactose through the cell wall and into the erythrocyte is markedly reduced in certain cats and, although unreported, may possibly be a cause for galactosemia in humans.
Evaluation of a new blood-collecting device ("microtainer") that is suited for pediatric use.
Authors: Hicks J M , Rowland G L , Buffone G J
Published: January 2020

We evaluated the prototype of a "Microtainer" (800-mul) tube (Becton-Dickinson Co., Rutherford, N.J. 07070), In which a semi-solid silicone material is used to separate serum from erythrocytes on centrifugation. We compared results for 18 serum analytes with those obtained for specimens collected by the "dripping" method of capillary blood collection. The serum was examined microscopically for platelets and leukocytes from both normal and leukemic specimens. The cellular elements were all contained in the erythrocyte layer. Hemolysis was negligible. Means for paired samples did not differ significantly, except for Na+, for which the difference was not clinically significant. Values for potassium and lactate dehydrogenase were unchanged 24 h after collection, even though the serum was left in the collecting device.
Improved batch and column separation in the assay of hemoglobin A2.
Authors: Morin L G
Published: January 2020

Hemoglobin A2 is a batch fractionated on a column containing diethylaminoethyl-cellulose (DE52, Whatman) with a discontinuous buffer system at pH 8.9 and 7.3. A short micro-column method is also presented, which allows separation in less than 30 min with no interference from hemoglobin S. Both methods clearly distinguish normal from thalassemia specimens, and are simpler and more rapid than previously published methods.
Automated enzymatic determination of L-lactate in serum, with use of a miniature centrifugal analyzer.
Authors: Hadjiioannou T P , Hadjiioannou S I , Brunk S D , Malmstadt H V
Published: January 2020

We describe an automated enzymatic reaction-rate method for spectrophotometric determination of lactate in serum with a miniature centrifugal analyzer. The L(+) -lactate is selectively oxidized in the presence of lactate dehydrogenase (EC 1.1.1.27) and NAD+ to form NADH, which is measured from its absorption. Reaction rates are determined automatically, and unknown concentrations are calculated from a computer-generated calibration curve with aqueous lithium lactate standards. Lactate concentrations in the range 0.32-1.6 mug/4 mul (80-400 mg/liter) of sample were determined with relative errors and coefficient of variation of 4.8%. Analytical recovery of lactate added to pooled serum was 89-112% (average, 101%). Comparison with a kit ("Rapid Lactate") method gave a correlation coefficient squared of 0.979 over a concentration range of 39-779 mg/liter.
Colorimetry of carbohydrates with m-aminophenol, and its proposed use in measurement of serum amylase activity.
Authors: Goodwin J F
Published: January 2020

In the reaction of m-aminophenol with carbohydrates in a thiourea/acetic acid reagent, addition of borate suppresses chromogen formation for hexoses and pentoses, and enhances it for disaccharides. All reaction products absorb maximally at 520 nm. The relatively greater intensity of the m-aminophenol/maltose reaction product was used in a saccharogenic method for amylase, with low blank readings. This method is compared with two dye/starch procedures.
Creatine kinase isoenzyme patterns in human tissue obtained at surgery.
Authors: Tsung S H
Published: January 2020

Chromatography on DEAE-Sephadex A-50 was applied to study the quantitative distribution of creatine kinase isoenzymes in extracts of human tissue obtained during surgery. The results are compared with those determined by an immunological method [Clin. Chim. Acta 58, 223 (1975)]. Conflicting results for some organs as reported by the two methods are probably attributable to postmortem autolysis.
An integrated approach to lipid profiling: enzymatic determination of cholesterol and triglycerides with a centrifugal analyzer.
Authors: Wentz P W , Cross R E , Savory J
Published: January 2020

We describe enzymatic methods for determination of cholesterol and triglycerides (triacylglycerols), adapted for use with a centrifugal analyzer. Triglycerides are determined by a two-point kinetic method, which utilizes a single glycerol standard for calibration and provides for a within-run reagent blank correction. The endpoint determination of cholesterol combines the unique blanking capabilities of the centrifugal analyzer to correct for sample and reagent blank interferences. Results correlate well with those obtained by procedures standardized by the Lipid Research Clinics Program for use with a continuous-flow system.
Evaluation of a dipstick test for glucose in urine.
Authors: Dyerberg J , Pedersen L , Aagaard O
Published: January 2020

As an example of qualitative tests, a dipstick analysis for glucose in urine has been tested for the influence of modifying factors on the test result. Two different types of dipsticks were examined, "Clinistix" and "S-Gluko-test." Used according to manufacturer's instructions, the latter is more sensitive and selective. By multivariance analysis the following variables were examined: urine samples, inter- and intra-analyst, exposure to light, and dipstick batch. The first three contributed significantly to the total variation in results, inter-specimen variation being the most important. With knowledge of the frequency of testing urines with a given glucose concentration and the probability of the result at that concentration, an expression of the probability of the glucose content of a urine sample can be obtained. Even with the tests of the type examined having a sensitivity and specificity exceeding 95%, 14 of 100 patients suspected of having diabetes mellitus on the basis of a dipstick examination will be found to have a urinary glucose concentration of less than 2 mmol/liter. These figures were found when the prevalence of urines with a glucose concentration exceeding 2 mmol/liter was 17.5%.
Effect of halothane anesthesia on muscle, liver, thyroid, and adrenal-function tests in man.
Authors: Johnstone R E , Kennell E M , Brummund W , Shaw L M , Ebersole R C
Published: January 2020

Seven healthy men volunteers received 6.6 +/- 1.3 (SD) percent-hours of halothane oxygen anesthesia without surgery. Serum bilirubin, alanine aminotransferase, and aspartate aminotransferase significantly increased after anesthesia, which may indicate subclinical liver-cell damage. Creatine kinase of skeletal muscle origin increased above 90 U/liter in six subjects, indicating subclinical muscle-cell damage. Cortisol, triiodothyronine uptake, thyroxine, and free thyroxine index increased significantly immediately after anesthesia. Serum bromide concentrations had increased by fivefold on the second day after anesthesia, and on the ninth day was still elevated fourfold. Oral temperatures increased 0.7 degrees C 6 h post-anesthesia, possibly because of increased thyroxine activity. Lactate dehydrogenase, hydroxybutyrate dehydrogenase and gamma-glutamyltransferase activities did not change significantly. No drugs administered during the course of this study chemically interfered with any of the test methods used.
Cholesterol in amniotic fluid, determined by gas chromatography.
Authors: Jones Owen V M , Ho F K , Mazzuchin A , Doran T A , Liedgren S , Porter C J
Published: January 2020

Cholesterol was extracted from amniotic fluid, saponified, converted to its trimethylsilyl derivative, and gas chromatographed, with cholesteryl acetate as the internal standard. The method is sufficiently accurate and precise for use with the range of concentrations of cholesterol found in amniotic fluid (5 to 48 mg/litre). Total cholesterol was measured in amniotic fluids collected at different stages of gestation. No significant trend or change was observed nor was cholesterol in the amniotic fluid and the mother's serum correlated at any stage of gestation. Thus we conclude that cholesterol is not a useful indicator of fetal age or maturity. Cholesterol concentrations in amniotic fluid from complicated pregnancies were within the range found for normal pregnancies.
Enzymatic method for determination of CO2 in serum.
Authors: Forrester R L , Wataji L J , Silverman D A , Pierre K J
Published: January 2020

We describe an enzymatic method, requiring only 10 mul of serum, for determining CO2 as bicarbonate or dissolved gas. Phosphoenolpyruvate carboxylase catalyzes the reaction of HCO3- with phosphoenolypyruvate to give oxalacetate. The resulting NADH, in the presence of malate dehydrogenase, is oxidized to NAD+, and the decrease in absorbance at 340 nm is directly proportional to the amount of CO2 present in the sample. Reaction is complete in 3 to 6 min under assay conditions, and is linearly related to CO2 concentrations between 8 and 65 mmol/liter. Analytical recovery is 95-110% (average, 101%). Two laboratories compared values obtained by continuous-flow analysis. The resulting correlation coefficients were 0.966 and 0.987, values for the t-test were t(paired) equals 0.473 and t(paired) equals 0.334, and average day-to-day precisions (three concentrations) were 3.9% and 4.2%.
Parallel electrophoretic fractionation of alkaline phosphatase and serum protein on cellulose acetate strips: clinical evaluation.
Authors: Burlina A , Galzigna L
Published: January 2020

We describe here a simple, time-saving method for separating alkaline phosphatases on cellulose acetate, with a parallel detection of serum protein pattern. The staining procedure is based on a coupling reaction between the reaction product of alpha-naphthol with Fast Red Violet LB. The enzymatic reaction is done by bringing the cellulose acetate strip containing the phosphatase band into contact with the surface of a solid agar/substrate mixture. The whole procedure requires 80 min. The method has some features in common with other recently proposed procedures but is more reproducible and rapid because of the choice of the reagents and an improved yield of the coupling step.
P-Hydroxybenzoic acid hydrazide procedure for serum glucose adapted to the Technicon "SMA 12/60," and compared with other glucose methods.
Authors: Ferraro J J , Caccavo F A , Saifer A
Published: January 2020

We adapted the p-hydroxybenzoic acid hydrazide procedure for serum glucose for use with the Technicon SMA 12/60 AutoAnalyzer. Like the o-toluidine method, this method is based on a general carbohydrate reaction except that it occurs in a mildly alkaline medium and the intense yellow color formed is measured at 400 nm. Advantages of this reagent over o-toluidine include lower cost, less toxicity, and higher purity. Aside from those carbohydrates that are present in serum in insignificant quantities, there are no interferences from various physiological compounds or drugs (hypoglycemic agents) found either in normal persons or diabetics. Within-run and day-to-day values had coefficients of variation of 1.39% and 3.44%, respectively; recoveries ranged from 100 to 102% (mean, 101%). Comparative data showed excellent agreement with the hexokinase (r equals 0.998; y equals 0.950x + 5.91) and glucose oxidase (r equals 0.996; y equals 0.986x + 5.34) enzymatic ("true") glucose methods, and with the o-toluidine procedure (r equals 0.998; y equals 0.979x + 3.14).
Lipoamide dehydrogenase in serum: a preliminary report.
Authors: Pelley J W , Little G H , Linn T C , Hall F F
Published: January 2020

Lipoamide dehydrogenase was identified in serum and the optimal conditions for its assay at 30 degrees C were defined. The pH optimum in tris(hydroxymethyl)aminomethane buffer is 7.8, and activity is inhibited if buffer concentration exceeds 100 mmol/liter. Saturating concentrations of the substrates NAD+ and lipoamide are 3 mmol/liter and 5 mmol/liter, respectively. Activity is decreased eightfold when lipoic acid is substituted for lipoamide. Activity is linearly related to enzyme concentration up to limiting absorbance change of 0.300 at 340 nm, and both within-day and day-to-day precision are satisfactory. Data suggest a normal range (2 SD) of 3-19 kU/liter. The highest value measured in serum was 473 kU/liter. A correlation with direct bilirubin concentrations (r equals 0.435, P less than 0.01) was found.
Modified colorimetric method for procainamide in plasma.
Authors: Sitar D S , Graham D N , Rangno R E , Dusfresne L R , Ogilvie R I
Published: January 2020

We describe a modified colorimetric method for determining procainamide in plasma by use of a diazotization and coupling reaction with N-(1-naphthyl)-ethylenediamine dihydrochloride (Marshall's reagent). The sensitivity of the assay is increased twofold over the originally reported colorimetric method, and the lower limit of detection for the assay of a 1.0-ml plasma sample has been decreased to 0.5 mg/liter.
Adaptation of blank-corrected methods for measurement of total and conjugated bilirubin and uric acid to the centrifugal analyzer.
Authors: Cross R E , Heintges M G , Savory J , Wentz P W
Published: January 2020

The blank-corrected determination in serum of total and conjugated bilirubin and of uric acid has been adapted to the centrifugal analyzer. Bilirubin is measured by a modified Jendrassik and Grof procedure; uric acid is determined kinetically by using a reaction based on reduction of ferric ion, including a uricase-treated blank. Sensitivity and precision of both methods are excellent, and results compare well with those obtained by commonly used procedures.
New techniques for ion-selective measurements of ionized calcium in serum after pH adjustment of aerobically handled sera.
Authors: Schwartz H D
Published: January 2020

I report further experience in measuring ionized calcium (Ca2+) with the AMT Electron System and its serum standards and solid-state, dip, calcium-selective electrodes. With this system, serum pH can be adjusted with CO2 gas and Ca2+ and pH simultaneously measured; when 5.2% CO2 (40 mm pco2) is used for sample equilibration, the standard bicarbonate concentration is also provided. I measured serum Ca2+ as a function of pH between pH 7.0 and 9.0 and found the relationship to be reproducible, with no evidence of irreversible complexing of Ca2+. When the pH of aerobically exposed, mailed sera was restored to the original values, their values for Ca2+ were the same as for the fresh sera. Measurement of Ca2+ in routinely (aerobically) handled sera after pH restoration with CO2 gas was therefore validated, both samples from within an institution and mailed specimens. Standardization to pH 7.40 is recommended for routine measurements, is generally more accurate than use of heparin or quasianaerobic techniques, and is a practical approach. In patients with possible uncompensated acid-base disturbance (which may be indicated by an abnormal standard bicarbonate concentration if not suspected clinically), patient pH should be measured independently as part of the usual strict, anaerobic blood-gas-analysis procedures. Abnormal patient pH must be considered in the interpretation of Ca2+ results determined at pH 7.40 which are borderline or slightly abnormal; most accurately, Ca2+ may be measured in the separated sera at the previously determined patient pH value. Studies of aqueous solutions with the currently used Ca2+ electrodes showed a selectivity coefficient (the constant which relates the activity of an interfering ion to the activity of calcium that would contribute the same emf) KNa=0.0031 +/- 0.0003 (SE) and KMg=0.046 +/- 0.004 (SE). At physiological concentrations of Ca2+, physiologically encountered variation in Na+ is of no significance in resulting Ca2+, but extreme variation in Mg2+ may cause an error of approximately 1%.
Automated separation and measurement of lactate dehydrogenase isoenzymes.
Authors: Busby M G , Kudirka P J , Carey R N , Toren E C
Published: January 2020

We describe a totally automated, computer-controlled system for separating and measuring the activity of lactate dehydrogenase (EC 1.1.1.27) isoenzymes. These isoenzymes in tissue extracts were separated on disposable DEAE-cellulose mini-columns. Resolution was complete except that LD-1 was not resolved from LD-2.
On the design of clinical chemistry quality-control sera.
Authors: Grannis G F , Miller W G
Published: January 2020

Quality-control specimens having quantitave interspecimen relationships were prepared by (a) making admixtures of two serum pools and (b) making various dilutions of the admixtures. The specimens were analyzed for many constituents to illustrate how the data obtained could be manipulated and used. The inter-related specimens permit the reduction of quality-control data to a few target values and should be particularly useful for the evaluation of multi-channel analyzers, and for use in inter-laboratory proficiency surveys. Aspects of the design of quality control and proficiency test specimens are also discussed.
Effect of tobramycin on urinary gamma-glutamyltransferase activity: Studies in a case of renal carcinoma.
Authors: Beck P R , Chaudhuri A K
Published: January 2020

Gamma-Glutamyltransferase activity was studied in a man presenting with recurrent septicemia owing to pyonephrosis and renal carcinoma. Increased activity in the urine was ascribable to administration of the aminoglycoside antibiotic, tobramycin. That the renal carcinoma did not contribute to the increased values was confirmed by homogenization and enzyme histochemistry of the tumor. Although the activity of this enzyme in serum was greater than normal, this persisted postoperatively, and thus was not related to the renal carcinoma.
The anion gap: its use in quality control.
Authors: Witte D L , Rodgers J L , Barrett D A
Published: January 2020

Because the anion gap is useful in classifying acid-base disorders, quantitative information on this variable should be readily available. Three populations were studied and means and central 90% ranges were determined for the anion gap, calculated as sodium-(chloride + bicarbonate). In 1047 hospitalized patients the mean was 12 mEq/liter, 86 faster volunteers had a mean of 13 mEq/liter with a 90% range of 9--17 mEq/liter, and 203 plasmaphoresis donors had a mean of 11 mEq/liter with a 90% range of 7--16 mEq/liter. Most of this variability can be accounted for by analytical rather than physiological variation. This quantitation of the anion gap is useful for interchecking electrolyte values as an additional and inexpensive means of laboratory quality control. In 73 of 75 abnormal anion gaps, for which the data were verified to be correct, the clinical information provided a reasonable explanaation for the abnormality.
Decreased radioassay values for folate after serum extraction when pteroylglutamic acid standards are used.
Authors: Mitchell G A , Pochron S P , Smutny P V , Guity R
Published: January 2020

Radoiassay values for folate in protein-denautured (extracted) sera are compared with values obtained by individual serum blank correction. Lower folate values are reported for sera extracted at pH 1.3, 7.6, 10.5, and 12.4 when pterolyglutamic acid (PGA) calibration standards are used. The results are unchanged when ascorbate added to the sera is increased from 1 to 20 g/liter and when [3H]PGA is substituted for 125I-labeled PGA. However, a higher mean value for extracted serum is reported when N-5-methyltetrahydrofolic acid calibration standards are substituted for PGA. Because both patients' samples and assay calibrators were subjected to the same extraction methods, our results suggest that serum folate behaves differently from PGA during the extractions.
Efficacy of activated charcoal hemoperfusion in removing lethal doses of barbiturates and salicylate from the blood of rats and dogs.
Authors: Hill J B , Palaia F L , McAdams J L , Palmer P J , Maret S M
Published: January 2020

Rats were injected intraperitoneally with lethal doses of sodium pentobarbital (115 mg/kg) or a lethal mixture of sodium salicylate (500 mg/kg) and sodium acetazolamide (25 mg/kg). Within about 20 min, part of each group was connected to an extracorporeal circuit containing uncoated activated charcoal and part to an empty control circuit. After a 90-min hemoperfusion, the treated groups showed a significantly decreased mortality (58% to 14% for pentobarbital; 100% to 0% for salicylate). Dogs were injected intravenously with lethal doses of sodium phenobarbital (175 mg/kg). One group was treated by hemoperfusion through an empty device in a control extracorporeal circuit, a second group was treated with loose-bed activated charcoal devices, and a third group with fixed-bed activated charcoal devices. For both the fixed and loose-bed devices, a 5-h hemoperfusion markedly decreased mortality (100% to less than or equal to 15%). The lethal combination of salicylate and closed-circuit methoxyflurane anesthesia was also successfully treated in dogs. This study clearly demonstrates the lifesaving potential of uncoated activated charcoal hemoperfusion.
Rapid radioisotopic procedure for determination of nortriptyline in plasma.
Authors: Maguire K P , Burrows G D , Coghlan J P , Scoggins B A
Published: January 2020

With the widespread use of tricyclic antidepressant drugs, the relationship between the concentration of the drug in the plasma and the therapeutic response is of considerable interest. We describe a double-isotope derivative dilution procedure for measuring plasma nortriptyline. In the method, [14C]nortriptyline is used for estimating procedural losses and [3H]acetic anhydride for derivative formation. The assay is rapid and adequately specific, sensitive, precies, and reproducible for routine clinical use. We used it to investigate the variation in steady-state drug concentrations in plasma of persons who were on a 150 mg/day dose of nortriptyline. Intra-individual variation from day to day was 10-14%. This variation was not significantly affected by the dosage schedule, the time of sampling after an oral dose, or the storage of the plasma samples. For 19 patients on 150 mg of nortriptyline per day, the mean concentration in plasma was 181 +/- 22 (SE) mug/liter, a value that compares well with our previous findings and those of other groups.
Serum quinidine concentrations: comparison of fluorescence, gas-chromatographic, and gas-chromatographic/mass-spectrometric methods.
Authors: Huffman D H , Hignite C E
Published: January 2020

Serum quinidine concentrations were determined in patients on chronic therapeutic doses. Although results were higher by a protein precipitate-fluorescence method as compared to a specific extraction fluorescence method, there was substantial correlation between results by the two methods (r = 0.945, P less than 0.001). We established the specificity of the extraction method by a methylation gas-chromatographic method in which the base peak in the mass spectra of the methylated products of both quinidine and cinchonidine, the internal standard, was monitored. We conclude that the protein precipitate method should be discarded.
Rapid radioimmunoassay for methotrexate in biological fluids.
Authors: Hendel J , Sarek L J , Hvidberg E F
Published: January 2020

We report a radioimmunoassay for the cytostatic folic acid antagonist methotrexate. Antiserum was raised in rabbits immunized with a conjugate of methotrexate and methylated bovine serum albumin. The calibration curve of the assay is linear from 1 to 100 ng methotrexate per 100-mul sample, 0.1 ng methotrexate still inhibiting the binding of the tracer by 20%. The coefficient of variation ranged from 4.04% to 11.58%. The reaction of the antiserum proved to be specific for methotrexate, as only insignificant interference could be demonstrated with folic acid, folinic acid (leucovorin), or tetrahydrofolic acid. With the method, results are available about 20 h after blood sampling at a daily capacity of at least 48 samples (i.e., about 15 determinations), depending on the equipment. Concentrations of methotrexate in plasma and erythrocytes were determined in five patients who were on continuous treatment with the drug. In one patient the concentration/time curves were followed in plasma and erythrocytes by frequent determinations for 24 h after a parenteral dose. The results indicate that the procedure is well suited for clinical investigations.
Pharmacokinetic interpretation of data gathered during therapeutic drug monitoring.
Authors: Dvorchik B H , Vesell E S
Published: January 2020

We review some pharmacokinetic principles that can facilitate interpretation of data obtained during therapeutic drug monitoring: the one- and two-compartment models, volume of drug distribution, drug clearance, organ clearance, bioavailability, first-pass effect, chronic or repetitive dosing, and use of urine and saliva to measure drug clearance and drug binding to plasma proteins, respectively. We also describe use of saliva to estimate rapidly, conveniently, and noninvasively the concentration of the free, pharmacologically active form of the drug as well as the fraction of drug bound to plasma protein.
Measurement of clomipramine, N-desmethyl-clomipramine, imipramine, and dehydroimipramine in biological fluids by selective ion monitoring, and pharmacokinetics of clomipramine.
Authors: Dubois J P , Küng W , Theobald W , Wirz B
Published: January 2020

To quantitatively determine tricyclic antidepressant agents, we used a combined gas chromatograph/mass spectrometer system, and deuterium-labeled internal standards. Recovery exceeds 95% and the coefficient of variation is less than 4% for human whole-blood samples supplemented with 5 to 15 ng of clomipramine hydrochloride or 20 to 60 ng of dehydroimipramine hydrogen fumarate per milliliter. For both amines, the detection limit is 0.3 mug/liter; Six healthy volunteers who received a single oral dose of 50 mg of clomipramine hydrochloride showed peak drug concentrations in the blood 3 to 5 h after administration, ranging between 14.4 and 30.1 mug/liter. Plasma/whole blood concentration ratios varied from 0.70 to 1.20, and the cumulative renal elimination from 0 to 72 h is less than 0.2% of the dose. This method is suitable for in vivo bioavailability studies of unchanged clomipramine, dehydroimipramine, and imipramine after a single oral dose of as little as 25 mg.