bluuebunny/crossref_metadata_2025 huggingface dataset:
Microviscosity of amniotic fluid phospholipids, and its importance in determining fetal lung maturity.
Authors: Blumenfeld T A , Cheskin H S , Shinitzky M
Published: January 2020

Fluorescence polarization measurements of microviscosity (apparent viscosity within the hydrophobic center of lipid bilayers) of amniotic fluid correlate well with lecithin/sphingomyelin ratios determined by thin-layer chromatography. In addition to lecithin, phosphatidylglycerol and phosphatidylinositol are important for determining fetal lung maturity, but the lecithin/sphingomyelin ratio gives no information concerning these other phospholipids. The microviscosity of sphingomyelin significantly exceeded that of lecithin over the temperature range 25--37 degrees C; values for phosphatidylglycerol, phosphatidylinositol, and phosphatidylserine were lower. Phosphatidylglycerol, phosphatidylinositol, and phosphatidylserine, added individually, significantly decreased the microviscosity of dispersions with lecithin/sphingomyelin ratios corresponding either to fetal lung immaturity or maturity. Phosphatidylglycerol caused the greatest decrease in both. Mixtures of the three phospholipids in the proportions found in term amniotic fluid decreased the microviscosity of fluids with either mature or immature lecithin/sphingomyelin ratios by 23--27%. When each was present in the proportion found in tracheal aspirate (twice that of term amniotic fluid), the decreases uere 46--50%. This technique quickly and precisely indicates not only fetal lung maturity but also the presence of important phospholipids other than lecithin and sphingomyelin.
Column chromatography and immunoassay compared for measuring the isoenzymes of aspartate aminotransferase in serum.
Authors: Sampson E J , Hannon W H , McKneally S S , McKenzie C , Miller S A , Whitner V S , Burtis C A
Published: January 2020

We compare a column-chromatographic method and a homogeneous immunoassay method for separately measuring the mitochondrial and cytoplasmic isoenzymes of aspartate aminotransferase. Analytical recovery for the two methods averaged 102% (SD, 2%) and 103% (SD, 4%), respectively, for 11 pools prepared by adding the purified isoenzymes to serum and 102% (SD 8.9%) and 89% (SD, 8.1%) for 26 unaltered specimens of human serum. In comparing the results of the immunoassay method (y) to the chromatographic method (x), our measurements agreed closely for the mitochondrial (y = 0.947 X + 7, r = 0.9991, standard error of estimate = 2.9 U/L) and cytoplasmic (y = 0.92x-6, r = 0.9995, standard error of estimate = 2.1 U/L) isoenzymes in pools prepared from the purified isoenzymes. Similar measurements of the 26 human serum specimens yielded the following results for least-squares evaluation; cytoplasmic isoenzyme y = 1.03x-11, r = 0.994, and standard error of estimate = 6.0 U/L; mitochondrial isoenzyme y = 0.75x+0, r = 0.927, and standard error of estimate = 3.9 U/L.
Rapid analysis for iodotyrosines and iodothyronines in thyroglobulin by reversed-phase liquid chromatography.
Authors: Alexander N M , Nishimoto M
Published: January 2020

We describe a 10-min reversed-phase "high-pressure' liquid-chromatographic procedure for measuring tyrosine, monoiodotyrosine, diiodotyrosine, 3,5-diiodothyronine, 3,5,3'-triiodothyronine, 3,3',5'-triiodothyronine, and thyroxine. Resolution and quantitation of a mixture of these amino acids were excellent on LiChrosorb (Altex) RP-8 with isocratic elution (1.5 mL/min) with acetonitrile/water/glacial acetic acid (50/49/1 by vol). As little as 100 ng of each iodoamino acid could be detected and quantitated with a conventional 1-cm, flow-through spectrophotometric (254-nm) detector coupled to a 10-mV strip-chart recorder. Analyses for monoiodotyrosine, diiodotyrosine, 3,5,3'-triiodothyronine, and thyroxine in hog and beef thyroglobulin hydrolysates (sequential digestion with pronase and aminopeptidase) agreed well with results by anion-exchange chromatography and by competitive radioassays. To prevent interference by tryptophan in the analysis for diiodotyrosine, we batch-separated the iodoamino acids by anion-exchange chromatography before the procedure. The procedure we describe seems generally useful for detection and quantitation of thyroid hormones, thyroid hormone metabolites, and iodotyrosines.
Ribonucleic acid in plasma from normal adults and multiple myeloma patients.
Authors: Hamilton T C , Smith A G , Griffin C A , Henderson R J
Published: January 2020

Reports of the presence of RNA in human plasma have been numerous, often suggesting that RNA in plasma is correlated with human disease. We critically examined the methods for determination of RNA in plasma. Lack of method specificity has caused previous workers to overestimate plasma RNA concentrations by more than 50-fold. To isolate RNA from plasma, we used both a phenol-chloroform extraction and a modified Schmidt-Thannhauser procedure. We show that RNA in plasma can be identified and quantified by alkaline hydrolysis of the plasma extract and subsequent separation of the resulting 2'- and 3'-mononucleotides by "high-performance" liquid chromatography. We could not detect RNA in plasma from either apparently healthy, normal adults or multiple myeloma patients, but found 1.1 mg/L in the plasma of a patient with Waldenström's macroglobulinemia. Our method is useful for the specific determination of RNA in plasma and will detect as little as 600 micrograms/L.
Enzymic determination of fructose in seminal plasma by initial rate analysis.
Authors: Anderson R A , Reddy J M , Oswald C , Zaneveld L J
Published: January 2020

We describe a spectrophotometric assay for fructose in seminal plasma. The method is based on reduction of fructose by a commercially available preparation of sorbitol dehydrogenase (EC 1.1.1.14), with the concomitant oxidation of NADH. The initial rate of NADH oxidation, which is proportional to the fructose content of seminal plasma, can be measured either with a recording spectrophotometer or by conventional two-point kinetic assay. The method was as accurate, precise, and sensitive as, and more specific and rapid than, currently used colorimetric (resorcinol) methods for fructose determination. Values (mmol/L) for fructose in seminal plasma from several species are: man, 9 +/- 2 (SD); cynamolgus monkey (Macaca fasicicularis)., 108 +/- 19; bull, 30 +/- 1; and rabbit, 13 +/- 4. These values agree with previously published results. We believe the method is appropriate for both research and clinical use.
Radiochemical method for measuring plasma clearance and urinary excretion of pteroylglutamic acid.
Authors: da Costa M , Rothenberg S P , Rosenberg Z
Published: January 2020

A radiochemical procedure is described for specific determination of pteroylglutamate in serum and urine. This method depends on denaturation of methyltetrahydrofolate with peroxide and measurement of the residual folate by a ligand-binding radioassay. The binding determinant for the radioassay is a folate-binding protein, partially purified from chronic myelogenous lekemia cells, that has low affinity for the reduced folates and thus will preferentially measure residual pteroylglutamate rather than any nondenatured residual methyltetrahydrofolate. We used this assay to measure the clearance from plasma and the urinary excretion of pteroylglutamate and a small fraction of serum folate that is stable to this oxidation procedure. The plasma clearance after intravenous injection is characterized by an initial rapid distribution phase followed by a second, slower metabolic phase; after about 2 h all of the administered pteroylglutamate has been cleared from the blood. The peak concentration of total folate in serum 1--2 min after administration of pteroylglutamate exceeded the sum of the endogenous stable and baseline serum folate, indicating that a reduced labile folate was released from the liver and perhaps from other tissues. This reduced folate had a slower metabolic clearance rate and was excreted to some extent in urine. Only 2.3 and 7.9% of the pteroylglutamate administered to two normal subjects was excreted as stable folate.
Intra-individual variation in commonly analyzed serum constituents.
Authors: Morrison B , Shenkin A , McLelland A , Robertson D A , Barrowman M , Graham S , Wuga G , Cunningham K J
Published: January 2020

Concentrations of 27 commonly estimated serum constituents were measured in blood sampled from 20 apparently healthy volunteers at 0830, 1230, and 1630 hours on each of four days, at weekly intervals. Time-dependent statistically significant (p less than or equal to 0.01) variation was observed in 12 of the constituents. The 15 constituents that showed no statistically significant diurnal variation included the immunoglobulins, other specific proteins, enzymes, cholesterol, calcium, copper, and magnesium. On the different days of the study consistent temporal patterns were observed in urea, creatinine, phosphate, zinc, bilirubin, triglyceride, total protein, and albumin. The magnitude of variation was particularly great in potassium, iron, zinc, and bilirubin. In general, day-to-day changes in concentration exceeded within-day changes, except for potassium, phosphate, and zinc, for which greater changes could be observed within-day than from one day to another at the same time.
Adsorption of organic acids from amniotic fluid and urine onto silica gel before analysis by gas chromatography and combined gas chromatography/mass spectrometry.
Authors: Williams V P , Ching D K , Cederbaum S D
Published: January 2020

We describe a method for adsorbing organic acids from amniotic fluid and urine specimens onto a column of silica gel before analysis by gas chromatography and by combined gas chromatography/mass spectrometry. Analytical recoveries of individual organic acids by our adsorption method compare favorably with those obtained by a more laborious manual extraction with ethyl acetate. Results for specimens of amniotic fluid and urine show that our method gives better recoveries of most of the organic acids. Furthermore, because citric acid is also extracted in our method without extracting significant amounts of sulfate or phosphate, the method is a suitable alternative to methods involving the use of DEAE-Sephadex for concentrating these acids.
Washington University Case Conference. Multiple myeloma and hypercalcemia?
Authors: Ladenson J H , McDonald J M , Aguanno J , Goren M
Published: January 2020

We present a case of multiple myeloma with high values for total calcium. The case discussion focuses on the role of the laboratory in diagnosis and monitoring of multiple myeloma and on the causes of the associated hypercalcemia. We present evidence for calcium binding to the paraprotein as a cause for the "hypercalcemia" in this case and we document a difference in the results for total calcium among various methods when this patient's serum was analyzed.
Modification of the DuPont aca as exemplified by a new procedure for neonatal total bilirubin.
Authors: Fraser A D , Lindsay A L
Published: January 2020

Hemoglobin interferes with determination of total bilirubin by the diazo procedure with the DuPont aca. We compared results by that procedure with those by a manual direct spectrophotometric two-wavelength method. Because the manual method was not inhibited by hemolysis as was the DuPont aca diazo procedure, we reprogrammed our DuPont aca to measure the absorbance of diluted serum samples at 452 and 577 nm with use of DuPont absorbance packs as the sample pack and cuvette. Results correlated well with those by the manual spectrophotometric method, and hemolysis no longer interfered. The method is primarily intended for use with serum from neonates.
Competitive enzyme-linked immunoassay for Factor VIII antigen.
Authors: Yorde L D , Hussey C V , Yorde D E , Sasse E A
Published: January 2020

We describe a competitive enzyme-linked immunoassay for Factor VIII antigen. Binding of anti-factor VIII to solid-phase Factor VIII antigen is competitively inhibited by the free factor VIII antigen that is to be measured. The amount of anti-Factor VIII bound to solid-phase VIII is measured by applying in sequence a heterologous bridging antibody and a soluble antibody/enzyme immune complex. The soluble complex used was rabbit antiperoxidase/horseradish peroxidase. Peroxidase activity is inversely proportional to the Factor VIII antigen concentration in the original test plasma and is measured spectrophotometrically. The assay can be performed in as little as 4 h with only a microtiter plate, antisera, antigen, and a spectrophotometer. It is sensitive to 0.05 units of Factor VIII antigen per milliliter, and reproducibility, linearity, and normal range are similar to those reported for other techniques.
Gas-chromatographic estimation of urinary oxalate and its comparison with a colorimetric method.
Authors: Farrington C J , Chalmers A H
Published: January 2020

We describe a simple, specific gas-chromatographic method for urinary oxalate. Its specificity was evaluated by precipitating the oxalate as its calcium salt from urine, followed by methylation of the oxalate with boron trifluoride/methanol and subsequent gas-chromatographic separation and quantitation of the dimethyl oxalate. [U-14C]Oxalate and n-decanoic acid are used as internal standards. Analytical recoveries ranged from 93.8 to 97.7% for oxalate-supplemented urine. Twenty replicate analyses of urines containing typical concentrations of oxalate gave CVs of 10.0, 9.1, and 8.3% for low, medium, and high concentrations, respectively. Day-to-day precision (CV) for single analyses repeated on 20 days was 8.6% for the low urinary oxalate concentration. The lower limit of detection of urinary oxalate is 25 mumol/L. Results by our method correlated well (r = 0.95) with those by a colorimetric method (Clin. Chim. Acta 36: 127, 1972) but averaged 68.4% of those obtained colorimetrically (n = 75 samples, p less than 0.001). The expected range for our method is calculated to be 80 to 500 mumol of oxalate per 24-h urine (mean +/- 1 SD: 280 +/- 100).
Novel 125I-labeled nortriptyline derivatives and their use in liquid-phase or magnetizable solid-phase second-antibody radioimmunoassays.
Authors: Kamel R S , Landon J , Smith D S
Published: January 2020

Nortriptyline derivatives prepared by reaction with fluorescein isothiocyanate or conjugation to N-acetyl-L-histidine were radioiodinated and the products purified with Sephadex LH-20 columns to obtain two novel nortriptyline radioligands. Antisera were raised in rabbits by immunization with nortriptyline conjugated to succinylated ovine albumin. By use of the iodinated fluorescein derivative we developed a liquid-phase second-antibody radioimmunoassay that gives results correlating closely (r = 0.98) with those by an established radioimmunoassay of similar specificity in the assay of apparent total amitriptyline and its metabolite nortriptyline in serum or plasma from patients being treated with these drugs. With the iodinated N-acetyl-L-histidine derivative we developed a magnetizable solid-phase second-antibody radioimmunoassay. The cross reactivities of amitriptyline and nortriptyline could be made equal by performing the assay at pH 9.0, which makes it possible to measure true total active drug concentrations in patients receiving amitriptyline.
A statistical method for determining normal ranges from laboratory data including values below the minimum detectable value.
Authors: Tsay J Y , Chen I W , Maxon H R , Heminger L
Published: January 2020

Determination of normal ranges from laboratory data containing undectable values is a frequently encountered problem in the radioimmunoassay of peptide hormones. In the past, such determinations usually have been based on the mid-point method or the one-end Winsorized method. A graphic method involving the use of probability paper has also been reported. We propose that the maximum-likelihood estimation is a more appropriate statistical method for the determination of normal range from this type of data (Type I censored data). With this method, the mean and standard deviation, and hence the tolerance limits, can be estimated. We used the maximum-likelihood estimation method to determine the normal range of serum thyrotropin values obtained from 93 healthy subjects, based on a log normal distribution. Although the serum thyrotropin content was undetectable in 14% of the subjects, a normal range could be calculated. Using tolerance limits for 95% coverage of the population with 90% confidence, we calculated the normal range of thyrotropin to be 0.51-5.75 milliunits/L, with a mean value of 1.71 milliunits/L, and predicted that 91.4% of undetectable serum thyrotropin values will fall within the normal range.
Gas-liquid chromatography of undervatized drugs after chromatographic extraction from blood.
Authors: Werner M , Mohrbacher R J , Riendeau C J
Published: January 2020

We have developed an integrated method that overcomes the two main procedural difficulties of gas-liquid chromatography, namely, solvent-solvent extraction and chemical derivatization. Drugs are extracted from serum by column chromatography on granular diatomaceous earth (kieselguhr). Subsequent gas-liquid chromatography of underivatized samples can be performed on either of two liquid phases. A mixed liquid phase, used for quantitative gas-chromatographic assay on patients with a known therapeutic regimen, has enabled quantitation of 12 drugs in serum. Alternatively, a single liquid phase, used with the mixed liquid phase, permits the gas-chromatographic identification of unknown drugs on the basis of the characteristic pattern of the two relative retention times; by this approach more than 40 drugs have been identified in cases of suspected intoxication, both in serum and in gastric aspirate. Besides providing ease of performance and wide applicability, the proposed procedure offers a degree of precision and accuracy that compares favorably with established methods.
Nephelometry of apolipoprotein B in human serum.
Authors: Heuck C C , Schlierf G
Published: January 2020

We studied the development of light scattering in the reaction between anti-apolipoprotein B and apolipoprotein B in intact very-low-density lipoproteins (I) and low-density lipoproteins (II) as well as in lipoproteins treated with lipases, and found considerable differences in the kinetics of the immunoreaction for the two lipoprotein classes. Pre-incubation with triglyceride lipase and cholesterol esterase caused a decrease of final light scattering in I but only minimal changes in the reaction with II. Non-ionic detergent not only decreased the original light scattering in hyperlipemic serum samples, but also accelerated the immunoreaction. Under standardized conditions, results of quantitative nephelometry correlated highly significantly with quantitative determination of apolipoprotein B by radial immunodiffusion, both for normolipemic and hyperlipoproteinemic serum samples. The nonspecific light scattering caused by neutral lipids in intact lipoproteins could be minimized when samples were pre-incubated with lipolytic enzymes.
Rapid fluorometry of estrogens in nonpregnancy urine, with use of chloroform extraction and purification by anion-exchange chromatography.
Authors: Goutte-Coussieu C , Habrioux G , Eichenberger D , Jayle M F
Published: January 2020

We describe a new method of extraction and purification of estrogens in low concentration in urine, involving a rapid enzymic hydrolysis of a 10-mL sample, automatic extraction (in a tube) with CHCl3/ethyl acetate, purification by chromatography on a disposable "mini-column" of AG 1-X2, and fluorometry by continuous flow according to Itrich's procedure [Acta Endocrinol. (Copenhagen) 35, 34 (1960)]. Conditions of hydrolysis, extraction, and purification were studied. Within-day precision (CV) was 7.5%, the mean between-day precision 8.5%. The sensitivity was 6 microgram of total estrogens per liter of urine. The specificity was assessed particularly by comparison with the results obtained by Schöller et al. [Acta Endocrinol. (Copenhagen) 57, suppl. 107 (1966)] and by gas-chromatography-mass-spectrometry. The normal limits as calculated by this technique were identical to values reported by others. The technique is rapid; results are obtained in 3 h. It is quite suitable for routine determinations: 100 assays can be done by a team of three technicians in one working day.
Continuous-flow fluoroimmunoassay of serum gentamicin, with automatic sample blank correction.
Authors: Shaw E J , Watson R A , Smith D S
Published: January 2020

We describe the continuous-flow automation of a quenching fluoroimmunoassay for determination of the antibiotic gentamicin in serum. In the flow system, sample is mixed with fluorescein-labeled gentamicin, followed by antiserum, and after a short incubation the fluorescence is measured. The intrinsic blank signal of each sample is corrected for during its single passage through the system by pumping antiserum discontinuously, so as to divide the final stream into sections corresponding to conventional separate assay and blank mixtures. Analyses time for each sample is 6 min, and 20 samples can be run per hour. Sensitivity, precision, and accuracy are adequate for clinical purposes, and assays of patients' samples correlate satisfactorily with the results of manual quenching and polarization fluoroimmunoassays, and plate diffusion bioassay.
An enzymic and centrifugal method for estimating high-density lipoprotein cholesterol.
Authors: Allen J K , Hensley W J , Nicholls A V , Whitfield J B
Published: January 2020

Enzymic measurement of high-density lipoprotein cholesterol with a centrifugal analyzer is described. We used polyethylene glycol (Mr 6000), final concentration 100 g/L, to precipitate low-density and very-low-density lipoproteins, thereby eliminating the difficulties of the commonly used heparin/Mn2+ precipitation method and facilitating the use of ethylenediaminetetraacetate-stabilized plasma. As measured by rocket immunoelectrophoresis, this final concentration of polyethylene glycol completely precipitates beta-lipoproteins, leaving the alpha-lipoproteins in solution. Between-run reproducibility (CV) was 3.6%, within-run reproducibility (CV) 0.8%. Reagent costs currently are $US 0.13 per test and large numbers of samples can be handled conveniently. Normal ranges were compiled for 539 men and 444 women. The high-density lipoprotein cholesterol for men was 1.20 +/- 0.31 (SD) mmol/L and for women 1.52 +/- 0.38 (SD) mmol/L.
Measurement of alpha-, beta-, and gamma tocopherol in serum by liquid chromatography.
Authors: De Leenheer A P , De Bevere V O , Claeys A E
Published: January 2020

A liquid-chromatographic assay for alpha, beta, and gamma-isomeric tocopherols in human serum is reported. The tocopherols and the internal standard (tocol) are absorbed into a silica gel column and are eluted in less than 10 min with n-hexane/isopropanol (99.4/0.6, by vol) at a flow rate of 1 mL/min. The complete analysis requires no longer than 30 min. Within-day precision (CV) was 1.4% (mean = 13.18 mg/L, n = 24), 7.4% (mean = 0.214 mg/L, n = 14), and 1.3% (mean = 1.01 mg/L, n = 14) for alpha-, beta-, and gamma-tocopherol, respectively. Day-to-day precision (CV) was 4.4% (mean = 9.85 mg/L, n = 10) for alpha-tocopherol, 9.1% (mean = 0.222 mg/L, n = 20) for beta-tocopherol, and 3.8% (mean = 1.00 mg/L, n = 20) for gamma-tocopherol. Extraction recoveries for alpha-, beta-, and gamma-tocopherol averaged 92.4 +/- 2.9% (n = 5), 91.4 4/- 7.6% (n = 5), an 92.0 +/- 4.1% (n = 4), respectively. The smallest injected amount detectable is estimated to be 0.03 micrograms for alpha-, and 0.04 micrograms for beta- and gamma-tocopherol.
Effects of wavelength error and spectral band width on measurement of alkaline phosphatase activity.
Authors: Schifreen R S , Burnett R W
Published: January 2020

We re-examined the effects of wavelength error and spectral band width on the measurement of alkaline phosphatase activity. The methods we studied is relatively insensitive to these two factors, a conclusion we base on both experimental results and theoretical analysis. These findings are in conflict with a recently published report [Lott et al., Clin. Chem. 24, 938 (1978)], and we suggest a possible explanation for this.
Incorporation of a SMAC analyzer into the data-processing procedures of a computer-assisted laboratory.
Authors: Undrill P E , Stroud R E , Paterson N
Published: January 2020

This paper describes the incorporation of a SMAC (Technicon) analyzer into data-processing techniques that have been developed on existing computer hardware during several years. The SMAC system is interfaced directly to a small computer, and suitable peripherals produce a manageable form of result tabulation for subsequent reporting, as well as provide quality-control information to the SMAC operators in real time. The design is such as to facilitate the performance analysis of the SMAC system during its initiation period and during normal service operation.
Radioimmunoassay of "pregnancy-specific"-beta 1-glycoprotein (SP1).
Authors: Kaminska J , Calvert I , Rosen S W
Published: January 2020

A radioimmunoassay for SP1 ("pregnancy-specific" beta 1-glycoprotein) is described in which a sensitivity of 0.1 microgram/L was achieved by use of an antiserum with Ka approximately equal to 1.7 x 10(11) L/mol and delayed addition of a 125I-SP1 tracer (specific acty, 25 Ci/g) that was stable for use in the assay for at least six to seven weeks. We also report an additional anti-SP1 serum, from an early bleeding, that displayed "apparent positive cooperativity" in the IgG but not the IgM fraction.
Comparison of current methods for high-density lipoprotein cholesterol quantitation.
Authors: Warnick G R , Cheung M C , Albers J J
Published: January 2020

We compared six precipitation methods for high-density-lipoprotein cholesterol quantitation with an ultracentrifugation method, the accuracy of which was improved by correcting for 4% manipulative loss in the d greater than 1.063 fractions. For purposes of comparison, the apoprotein B-associated cholesterol (average 56 mg/L) measured by immunoassay in the d greater than 1.063 fractions was subtracted. In 65 plasma samples from men, women, and children, a heparin-Mn2+ procedure with Mn2+ at 46 mmol/L produced results slightly higher (+16 mg/L), while results with Mn2+ at 92 mmol/L averaged slightly lower (-8 mg/L) than the comparison ultracentrifuge method. Results that were about 5% low were obtained by the dextran sulfate 500-Mg2+ (-25 mg/L) and phosphotungstate-Mg2+ (-31 mg/L) methods. A heparin-Ca2+ method produced results 10% high (+58 mg/L). Results by a polyethylene glycol-6000 precipitation method were 12% low (-64 mg/L). Precision was better with the two heparin-Mn2+ and the dextran sulfate 500-Mg2+ procedures, with CVs of 4%, intermediate with phosphotungstate-Mg2+ and polyethylene glycol-6000 (CV 6-7%), and poorest with heparin-Ca2+ (CV 10%). Precipitation by phosphotungate-Mg2+ appeared more sensitive to reagent concentration and temperature variations than either the heparin-Mn2+ or dextran sulfate 500-Mg2+ methods. We conclude that these precipitation methods are not equivalent, but give rise to significant systematic differences in high-density-lipoprotein cholesterol quantitation.
Enzyme immunoassay for serum lidocaine in antiarrhythmic therapy.
Authors: Lehane D P , Wissert P J , Menyharth P , Levy A L , Kukucka M A
Published: January 2020

We have adapted the commercially available EMIT kit [Clin. Chem. 23, 1161 (1977)] to the Gilford System 3500 Analyzer. Sample volume is 10 microliter. We compare reagent blank-corrected absorbance changes at 340 nm between 15 and 55 s for samples and a series of calibrators, and calculate results with use of a logarithmic transformation. Within-run precision (CV) for a serum pool with 4.0 mg of added lidocaine per liter was 2.7% (n = 45); day-to-day precision was 3.3% (n = 15). Analytical recoveries of 2 to 6 mg of lidocaine per liter were 90-102% (average, 97.3%). Results correlated significantly with those by a gas-chromatographic technique. No clinically significant interferences by concomitantly administered medications were observed. The procedure is rapid (42 samples per hour) and is well suited to the fast response required in monitoring lidocaine therapy. Usefulness of the assay data in the management of arrhythmias in the coronary care unit is discussed.
Preparation of cytosol for steroid receptor determinations with the Beckman "Airfuge".
Authors: Heidemann P H , Wittliff J L
Published: January 2020

Preparation of cytosols from several estrogen target organs was compared, with use of the Beckman Airfuge and a preparative ultracentrifuge. The specific estrogen-binding capacities of cytosols prepared with these instruments were indistinguishable. Similarly, the sedimentation profiles of estrogen-receptor species as determined by sucrose gradient centrifugation were identical. We conclude that the Airfuge provides a rapid, accurate, and inexpensive means of preparing cytosols for measurements of steroid receptors in the clinical laboratory.
Elemental composition of platelets. Part II. Water content of normal human platelets and measurements of their concentrations of Cu, Fe, K, and Zn by neutron activation analysis.
Authors: Kiem J , Borberg H , Iyengar G V , Kasperek K , Siegers M , Feinendegen L E , Gross R
Published: January 2020

We determined the elements Cu, Fe, K, and Zn in normal human platelets by neutron activation analysis. The platelets were obtained from seven donors and treated as described in Part I. The elemental composition is expressed on a wet-weight basis for plasma-free platelets. The following results were obtained (+/- values are 1 SD); "Pure" platelets: trapped plasma = 378 +/- 35 mg/g, water content = 715 +/- 15 mg/g, mean weight of the single platelet (by two different methods) = 9.9 +/- 1.1 pg and 11.2 +/- 1.7 pg, K = 4.39 +/- 1.06 mg/g, Zn = 49.23 +/- 10.97 microgram/g, Fe = 12.28 +/- 2.94 microgram/g, and Cu = 1.39 +/- 0.25 microgram/g. "Impure" platelets: trapped plasma = 349 +/- 31 mg/g, water content = 736 +/- 12 mg/g, K = 3.26 +/- 0.78 mg/g, Zn = 35.71 +/- 7.99 microgram/g, Fe = 17.11 +/- 5.10 microgram/g, and Cu = 1.39 +/- 0.21 microgram/g. To our knowledge, no data on Fe and Cu in platelets have hitherto been reported.
Improved liquid-chromatographic determination of propranolol in plasma, with fluorescence detection.
Authors: Jatlow P , Bush W , Hochster H
Published: January 2020

We describe an analysis for propranolol in plasma, with use of reversed-phase "high-pressure" liquid chromatography and fluorescence detection. Pronethalol is used as the internal standard. The procedure, which involves extraction into an organic solvent, evaporation, and clean-up by micro-scale back extraction from hexane into an aqueous phase, is specific and sensitive. The detection limit is less than 6 microgram/L (2.3 X 10(-8) mol/L). Within-day and between-day coefficients of variation are 1.8 and 4.4%, respectively. Commonly used drugs, including procainamide, N-acetylprocainamide, and quinidine, do not interfere.
A simple method for the intravenous fat-tolerance test.
Authors: Redheendran R , Edwards J A , Schnatz J D
Published: January 2020

A simple method for measuring intravenous fat tolerance with an artificial fat emulsion (intralipid) was studied and compared with the currently used method of Carlson and Rossner [J. Lab. Invest. 29, 271-280 (1972)]. The proposed method uses as little as 3 mL of whole blood to follow, by nephelometry, the elimination of intravenously administered fat emulsions. There is a high correlation between the K2 values obtained by the current method for plasma and those obtained by the proposed method (r = 0.903). The latter is simpler, quicker, and more acceptable to patients.
Interference of iodine-125 ligands in radioimmunoassay: evidence implicating thyroxine-binding globulin.
Authors: Painter K , Vader C R
Published: January 2020

Although there is abundant published evidence that radioiodinated antigens interfere in digoxin radioimmunoassays, other radioimmunoassays are similarly affected. We investigated the relationship of radioiodinated antigen structure to its function in the immunoassay. Carrier-free 125I-labeled iodotyrosine and iodohistamine derivatives were incubated with human serum, and the bound and free fractions were separated. We demonstrated that diiodotyrosyl analogs bind avidly to serum proteins. Because protein binding could be reduced with competitors that inhibit thyroxine-binding globulin, such as 1,8-anilinonaphthalene sulfonate and thyroxine, thyroxine-binding globulin was clearly implicated. Diiodotyrosyl compounds also bound to solutions of purified thyroxine-binding globulin, and this binding was inhibited by the same two competitors. We postulate that thyroxine-binding globulin is the major source of the heretofore unexplained interference of radioiodinated haptens. We present recommendations for eliminating or minimizing such interference.
Glycohemoglobin and glucose tolerance tests compared as indicators of borderline diabetes.
Authors: Dix D , Cohen P , Kingsley S , Senkbeil J , Sexton K
Published: January 2020

We concurrently measured glycohemoglobin and performed 3-h oral (100 g) glucose tolerance tests on 69 ambulatory patients suspected of having abnormal carbohydrate metabolism. The patients were divided into two groups: (a) The 37 patients for whom the results were normal had plasma glucose concentrations of 0.70--1.15 milligram during fasting and 0.70--1.23 g/L 2 h after glucose ingestion. (b) Borderline diabetics exceeded one or both of these limits. The range of glycohemoglobin in the normal group was 3.0--4.7% of total hemoglobin. Of the 21 borderline diabetics, 11 had increased glycohemoglobin (4.8--8.0%). The difference in tolerance test results between borderline diabetics with and without increased glycohemoglobin was insufficient to predict the status of glycohemoglobin. We suggest a tentative definition for latent diabetes: increased glycohemoglobin in the presence of normal or borderline-abnormal glucose concentration in plasma collected during fasting.
Clinical biochemistry of epilepsy. I. Nature of the disease and a review of the chemical findings in epilepsy.
Authors: Natelson S , Miletich D J , Seals C F , Visintine D J , Albrecht R F
Published: January 2020

In idiopathic or generalized epilepsy, serum glucose and cholesterol concentrations tend to be low, especially just before the seizure. Glucose tolerance curves are abnormal and variable. The electrolyte balance is disturbed, and epileptics tend to go readily into alkalosis. Serum [Na+] is usually unaffected, but [K+] is normal to low between attacks and increases during and after the seizure. Serum [Cl-] is usually high just before the seizure. Epileptics are generally mildly hypocalcemic, especially in the period before the seizure. Serum urea and nonprotein nitrogen values are low between paroxysms but increase after the seizure. Serum protein concentration is usually normal. Stress, which releases epinephrine and corticotropin, results in high serum citrate concentration, which probably contributes to decreased serum [Ca2+] just before a seizure. In the healthy individual, any increase in serum citrate is accompanied by increasing [Ca2+]. In the rabbit, convulsions can be induced with corticotropin, a result of increased serum citrate concentration coupled with a decrease in [Ca2+]. The net result is severe hypo-ionic-calcemia. A similar phenomenon has been reported in a few humans. Administration of insulin causes serum citrate concentrations to decrease. Apparently, the dynamic system that controls glucose and lipid metabolism, and thus electrolyte balance, through the hormones epinephrine, corticotropin, insulin, glucagon, calcitonin, and parathormone, is abnormal in the epileptic.
Clinical biochemistry of epilepsy. II. Observations on two types of epileptiform convulsions induced in rabbits with corticotropin.
Authors: Natelson S , Miletich D J , Seals C F , Visintine D J , Albrecht R F
Published: January 2020

We propose than an alarm mechanism is operative in animals, designed to regulate neuromuscular irritability by regulating [Ca2+]. Epinephrine or corticotropin (ACTH), injected intramuscularly into animals, causes a hypercitricemia, resulting in decreased [Ca2+]. This increases muscular excitability to facilitate escape. To avoid over reaction, [Cl-] is shifted into the plasma without a concomitant shift of Na+, thus generating an acidosis and an increase in ionization of Ca. Plasma pH, pCO2, total CO2, and [K+] decrease, and [Mg2+] increases. The acidosis, decrease in K+, and increase in [Mg2+] serve to counteract the effect of the decrease in [Ca2+], to protect against tetany. In the rabbit the hypercitricemia observed upon ACTH administration is accompained by a severe hypocalcemia and drop in blood pressure, resluting in tetanic convulsions. This seems to indicate calcitonin release, independent of the hypercitricemia. Thyroidectomized rabbits show only mild hypocalcemia when given ACTH, but develop a severe acidosis and typical grand mal epileptiform seizures. Administration of ACTH and then calcitonin to the goat, an animal resistant to the effects of ACTH alone, simulates the effect observed in the rabbit with respect to changes in blood components and blood pressure. Changes in the blood in the goat and rabbit resemble those in humans before an epileptic seizure. alpha-Melanotropin, containing a portion of the ACTH sequence, reacts in a manner similar to ACTH but more rapidly.
Determination of serum 5'-nucleotidase with a centrifugal analyzer.
Authors: Harvey M S , van der Stoel A G , Backer E T
Published: January 2020

We describe a multi-point method for the determination of the serum enzyme activity 5'-nucleotidase by means of an NADH sensor reaction. Studies show that the assay may be carried out in one step, without preincubation, by using a GEMSAEC Fast Analyzer.
Glucosuria: accuracy and precision of laboratory diagnosis by dip stick analysis.
Authors: James G P , Bee D E
Published: January 2020

Two urine dip-stick assays for glucose were studied and compared to a quantitative hexokinase procedure. We determined sensitivity, specificity, predictive value of "normal" and "abnormal" dip stick results, assay efficiency, technologist precision, and method association. Precision was poor. Attempts to estimate urine glucose concentration with the dip sticks frequently produced gross inaccuracies. We believe the dip stick glucose assay should be considered qualitative only; as a qualitative test, the dip sticks are fairly efficient at distinguishing glucose concentrations of less than 0.3 g/L from concentrations greater than 0.3 g/L.
Gas-chromatographic analysis for chlorpromazine and some of its metabolites in human serum, with use of a nitrogen detector.
Authors: Bailey D N , Guba J J
Published: January 2020

We describe a gas-chromatographic method for measuring chlorpromazine and its metabolites chlorpromazine sulfoxide, mono-N-desmethylchlorpromazine, and di-N-desmethylchlorpromazine at therapeutic concentrations in human serum, with use of a nitrogen detector. The compounds are extracted from serum at pH 10.5 into hexane/isoamyl alcohol, re-extracted into dilute HCl, and then extracted into hexane after alkalinization of the HCl. The N-desmethylated metabolites are measured as their respective N-trifuloracetyl derivatives; the parent drug and its sulfoxide are measured as the unchanged bases. Promazine is the internal standard. As little as 5 micrograms of chlorpromazine, 20 micrograms of chlorpromazine sulfoxide, 20 micrograms of mono-N-desmethylchlorpromazine, and 10 micrograms of di-N-desmethylchlorpromazine per liter can be measured in 2-mL samples of serum. The within-run coefficients of variation for assays of these drugs at 100 micrograms/L are 2.7%, 5.6%, 5.1%, and 5.3%, respectively. The procedure was applied to patients receiving therapeutic doses of chlorpromazine and to patients who had ingested an overdose of chlorpromazine.
Non-chromatographic radioimmunoassay procedure for urinary aldosterone.
Authors: Walsh P R , Wang M C , Turner E A
Published: January 2020

We describe a sensitive, specific, and simple procedure for measuring aldosterone in human urine, which requires no chromatographic purification before quantification by radioimmunoassay but does include hydrolysis and extraction steps. Rabbit anti-aldosterone serum is sued, generated against aldosterone-18,21-dihemisuccinate coupled to human serum albumin. The antibody cross reacted little with other structurally related steroids that are in human urine. Our procedure was validated by comparing values for urinary aldosterone in human urine, with and without preliminary purification by chromatography on either paper (y = 0.92x + 2.9; r = 0.99; p less than 0.01) or (Sephadex LH-20) column (y = 0.98x + 0.6; r = 0.99; p less than 0.01). Values by our procedure also correlated well (y = 1.03x - 0.8; r = 0.99; p less than 0.01) with those obtained with use of a validated commercial "kit" for urinary aldosterine. All reagents for the proposed method are available commercially.
Urinary 3-methoxy-4-hydroxymandelic acid as measured by liquid chromatography, with on-line post-column reaction.
Authors: Flood J G , Granger M , McComb R B
Published: January 2020

We describe a method for measurement of 3-methoxy-4-hydroxymandelic acid (vanillylmandelic acid, VMA) in urine. After the pH of the urine is adjusted to 2.7 and the sample is filtered, exactly 15 microL is injected onto a C-18 reversed-phase column. VMA is eluted from the column with 10 mmol/L phosphate buffer, pH 2.7, containing 30 mL of acetonitrile per liter. The eluate stream is combined with alkaline periodate and then passed through a 60 degrees C water bath. The VMA is completely oxidized to vanillin, which is detected and quantitiated by its absorbance at 360 nm. No deterioration of the column was noted after 167 such injections of urine samples. Long-term control data indicate a CV of 12 and 10% at VMA concentrations of 1.5 and 5.8 mg/L, respectively. Although results correlate well (r = 0.976) with those by the method of Pisano et al. [Clin. Chim. Acta 7, 285 (1962)], they average 10% lower. Of 20 compounds tested, only methyl dopa interfered with the procedure as described.
Liquid-chromatographic measurement of taurine in cerebrospinal fluid of normal individuals and patients with meningitis.
Authors: Shihabi Z K , White J P
Published: January 2020

Taurine was measured in cerebrospinal fluid by reacting it with fluorescamine to form a fluorescent derivative, followed by separation on a reversed-phase column and fluorometric detection and evaluation. The assay is rapid (17 min) and sensitive to as little as 1 mumol/L. The mean value for 27 cerebrospinal fluid samples collected from patients free from meningitis and aneurysm was 5.7 +/- 1.8 mumol/L. Twenty-two patients with bacterial meningitis showed a 0- to 20-fold increase in cerebrospinal fluid taurine, with a return to normal values after antibiotic treatment.
Hepatic function assessed (in rats) during chemotherapy with some anti-cancer drugs.
Authors: Capel I D , Jenner M , Dorrell H M , Williams D C
Published: January 2020

Using rats, we studied how best to assess hepatic damage after administering therapeutic doses of each of five anti-cancer drugs or of the hepatotoxin, carbon tetrachloride. As indexes, we compared measurement of the concentration of administered antipyrine in plasma with measurement in serum of alpha-fetoprotein or of the activities of five enzymes that reportedly best reflect hepatic damage. The biological half-life of antipyrine in the plasma was increased more than threefold on pretreating the rats with any of the five cytotoxic drugs or with carbon tetrachloride. In contrast, the concentrations of alpha-fetoprotein, alkaline phosphatase, gamma-glutamyltransferase, or glutamate dehydrogenase were not consistently increased. Of the enzymes tested in serum, aspartate aminotransferase and ornithine carbamoyltransferase best indicated hepatic impairment resulting from the treatment with anti-cancer drugs. Our results imply that determination of the pharmacokinetics of marker drugs such as antipyrine better indicates hepatic dysfunction induced by cytotoxic agents than does measurement of the enzymes liberated into serum as a result of damage to liver mitochondria.
Inter- and intra-individual variations in the saliva/blood alcohol ratio during ethanol metabolism in man.
Authors: Jones A W
Published: January 2020

The inter- and intra-individual components of variation in the saliva/blood alcohol ratio have been calculated from experiments with 48 male subjects after they drank 0.72 g of ethanol per kilogram of body weight as neat whisky after a short fast. Saliva and blood ethanol profiles were monitored at 30--60 min intervals for up to 7 h after intake. The analytical component of variation inherent in an estimate of the saliva/blood alcohol ratio, expressed as coefficient of variation, was 1.75%. I calculated saliva/blood ethanol ratios for each subject at each sampling time by taking the antilogarithm of the difference (log saliva alcohol--log blood alcohol). The mean ratio between 60 and 360 min after drinking was 1.077 (n = 336) with 95% confidence limits of 1.065 and 1.088. Moreover, the individual ratios showed no systematic variation throughout the absorption, distribution, and elimination phases of ethanol metabolism. Using a two-way analysis of variance and allowing for analytical sources of variation, I determined that the inter- and intra-subject variance components were 53 and 47% of the total biological variation. The saliva/blood alcohol ratio during ethanol metabolism, determined once in a single individual, had a biologically derived coefficient of variation of 10%.
Gas-chromatographic analysis of patterns of fatty acids of cholesteryl esters and phosphatidylcholine.
Authors: Rockerbie R A , Dobson R D , Frohlich J
Published: January 2020

We have developed a quick, reliable method for compositional analysis of the C14 to C20 fatty acids of cholesteryl esters, based on the use of a double extraction-solvent system and whisker-walled capillary columns with a new polar phase. Fatty acids of phosphatidylcholine, after separation by thin-layer chromatography, were determined on the same column. Our normal values are in good agreement with those published previously. We found marked changes in the fatty acid composition of both cholesteryl esters and phosphatidylcholine in cord blood serum, in various disease states, and in post-mortem specimens. The previously reported correlation between oleic (18:1) and linoleic (18:2) acids in serum cholesteryl esters was confirmed for normal adults, but no such correlation exists in cord blood specimens. We also found a significant correlation between the relative amounts of fatty acids in cholesteryl esters and phosphatidylcholine for all unsaturated fatty acids and for palmitate in all groups of subjects.
Solid-phase enzyme immunoassay for serum ferritin.
Authors: Anaokar S , Garry P J , Standefer J C
Published: January 2020

We describe a solid-phase enzyme immunoassay for ferritin in human serum, with use of a horseradish peroxidase-labeled antibody and a highly sensitive chromogen, 2,2'-azino-di(3-ethyl-benzthiazoline-6-sulfonate). The assay requires only 10 microL of serum per assay, relatively less time and labor than other assys for ferritin, and as little as 10 pg of ferritin can be measured. We believe this assay offers a reliable alternative to radioimmunoassay for serum ferritin.
Heparin interferes with albumin determination by dye-binding methods.
Authors: Bonvicini P , Ceriotti G , Plebani M , Volpe G
Published: January 2020

Heparin, when added in vitro at rather high concentrations (400--1000 kilounits/L) as an anticoagulant, interferes with albumin determination by bromcresol green by forming an insoluble precipitate with this dye. The interference is much less with bromcresol purple. It can be abolished by addition of hexadimethrine bromide.