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Biomedical subjects

M Lever

Publications and source records attributed to M Lever.

12 recordsLinked to original sources

Same-day batch measurement of glycine betaine, carnitine, and other betaines in biological material.

Glycine betaine, carnitine, carnitine esters, butyrobetaine, and proline betaine (stachydrine) concentrations in biological materials can be reliably measured in 100-microliters samples, with a detection limit below 1 mumol/liter. The procedure is suitable for batches of more than 30 specimens and it is possible to obtain a single result within 2 h. The betaines are extracted into an acetonitrile:methanol mixture, dried with anhydrous disodium hydrogen phosphate containing argentous oxide. The 4-bromophenacyl ester derivatives are formed using 4-bromophenacyl triflate as reagent, in the presence of solid magnesium oxide as base. The derivatives are separated by high-performance chromatography on a silica column, in a mixed partition and ion-exchange mode.

Animals

Factors in the choice of detergent in automated oestriol estimation.

Detergent must be added to the strongly acidic Kober reagent to obtain reproducible and well separated peaks in continuous flow analytical methods for oestriol in pregnancy urine. Brij 35 and other polyether detergents are not satisfactory because they or their impurities interfere in the reaction causing high blank response in colorimetric reactions and diminished fluorescence responses. In a single phase fluorometric method these detergents also caused turbidity or phase separation in the solution which passed through the fluorometer. Some anionic sulphate and sulphonate detergents were also unsatisfactory being immiscible with Kober reagent. Selected cationic and betaine detergents are shown to be possible alternatives, although one which is satisfactory in the kober reagent may be unsuitable for a dilution reagent. The betaine Empigen BB was found to be suitable for inclusion in both of these reagents.

Autoanalysis

Routine serum lipid analysis in New Zealand.

For two years pairs of serum specimens with a wide range of cholesterol and triglyceride concentrations were regularly dispatched each month to all New Zealand medical laboratories known to measure blood lipids. Four weeks after specimen dispatch each laboratory received a report which displayed all results along with the overall means, standard deviations, and results from "reference laboratories". Six-monthly summaries were prepared for each laboratory in which the previous 12 results were compared with the corresponding "target values" by regression analysis. This allowed classification of inaccuracy into one or more of three categories. Random error (imprecision) explained most of the discrepancies, but systematic errors also contributed strongly to the observed interlaboratory variation. No single class of laboratories performed significantly differently from any others. Approximately 60 percent of the 16000 cholesterol analyses done each month in New Zealand, and 40 percent of the 10000 triglyceride analyses, are performed with precision thought to be adequate for clinical usage.

Cholesterol

Automated continuous flow determination of urine albumin by competition with dye-detergent binding.

A continuous flow automated method for urine albumin was developed based on the ability of albumin to displace bromophenol blue from a bromophenol blue detergent complex. The method is almost specific for albumin, giving a slight response with an alpha-globulin fraction from serum. Results agreed closely with rocket electrophoresis albumin estimates on urine samples, but significantly less closely with "total urine protein" estimates by an acid protein precipitation, followed by biuret colour reaction, procedure. This method is being used to replace the indefinite "total protein" test for most routine purposes.

Albuminuria

Regression analysis in interlaboratory surveys: a case study with cholesterol and triglycerides.

1. A new interlaboratory survey design, that uses regression analysis to compare results from each laboratory with target values, was tested using cholesterol and triglyceride analyses. The fifty New Zealand laboratories involved showed considerable interlaboratory variation (CV = 8% to 27% for cholesterol, 13% to 113% for triglycerides), 30% and 40% of which was associated with systematic differences between laboratories. 2. End-of-period summaries using regression analysis confirmed the presence of systematic errors. These were either simple types caused apparently by incorrect standardisation (regression slope, B not equal to 1.0) or inappropriate blank correction (intercept, A not equal to zero) or complex types presumably due to nonlinearity or nonspecificity. Graphical display of results from each laboratory aided fault diagnosis and allowed the detection of between-run standardisation differences. 3. Method comparison studies were made: the only highly significant result being lower precision achieved by enzymatic cholesterol methods compared with other colorimetric methods.

Blood Chemical Analysis

Simulation of laboratory errors and their effects on interlaboratory quality-control programs.

The random errors in an analytical method are additive and can be classified into analytical response-dependent and -independent terms. Non-random errors, caused by systematic faults in the analytical procedure, are not always distriguishable from the random errors, but some cases of non-linear assay response and unsuitable standardisation can be studied usefully in models without random error. Interlaboratory quality control programs cannot distinguish systematic and random error until the pattern of results on a number of specimens, or pairs of specimens, can be studied. In this case linear regression analysis is a powerful method for distinguishing different forms of error especially when response-dependent random errors do not predominate. The range of concentrations used for regression whould be as wide as that in which quantitative distinctions are used in clincal diagnosis and treatment. Preliminary reports, of the results on which the regression analysis is based, are most suitably presented on Youden diagrams with paired specimens.

Analysis of Variance

A design for interlaboratory quality-control programs.

A design for interlaboratory quality-control programs is described. Speimens are despatched in pairs to participating laboratories. The results returned by laboratories are compared with each other and with reference results, but the frequent brief reports which are prepared are not regarded as the main laboratory assessments, although these reports can alert laboratories to gross imprecision and inaccuracy. When sufficient results have been returned, a linear regression analysis is carried out between results from each laboratory and the reference results. The statistics obtained from the regression data provide a concise source of information about the form of inaccuracy (imprecision and systematic error) present.

Chemistry, Clinical

Contributions of other sterols to the estimation of cholesterol.

The responses of 5alpha-cholestan-3beta-ol, 5alpha-cholest-7-ene-3beta-ol and cholesta-5,7-dien-3beta-ol, normally found in human serum, were examined by: (1) the Liebermann-Burchard reaction, (2) the Zak (ferric chloride) reaction, (3) an enzymatic cholesterol method monitored by estimating the amount of hydrogen peroxide produced, (4) an enzymatic cholesterol method monitored by observing the change in absorbance at 240 nm, and (5) gas chromatography. The results show that none of these methods is specific for cholesterol; contributions from the sterols examined range from zero to more than 150% relative to cholesterol. For the first four methods contributions depend on the conditions under which each test is performed.

Cholestadienols