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M J Peake

Publications and source records attributed to M J Peake.

16 recordsLinked to original sources

[Enzyme spectrophotometry determination of sodium, potassium and chloride ions in serum and urine].

The principles and main features of enzymatic methods for the measurement of sodium, potassium, and chloride are reviewed and their performance compared with current procedures. Each method makes use of a relatively specific enzyme, catalysing a reaction whose rate is sensitive to the ion to be determined. Where the (S)0.5 of the enzyme for the ion is much lower than the assay concentration, the ion concentration may be reduced by a binding agent. Alternatively, a competitive inhibitor may be used to raise the (S)0.5 of the enzyme. In the case of chloride determination with amylase the (S)0.5 of the enzyme is raised by limiting the concentration of free calcium. In the measurement of potassium, interfering ions such as sodium are removed by binding with Kryptofix 221 and improvement in performance is also achieved by use of a bacterial pyruvate kinase less sensitive to sodium. The enzymatic methods are applicable to measurement of sodium, potassium and chloride in blood or urine with good precision, accuracy, and specificity. They can be used on mechanized or manual instruments. There appears to be minimal interferences from compounds found in normal or pathological serum or urine.

Chlorides

Enzymatic determination of potassium in serum.

This is a kinetic assay for measuring K+ in serum, based on the activation of pyruvate kinase (EC 2.7.1.40) by K+. We eliminated interference from Na+ and NH4+ ions, which also activate this enzyme, by including Na+-binding and NH4+-consuming reagents in the reaction mixture. The assay was developed with and evaluated in the Cobas Fara centrifugal analyzer (and has been used in other kinetic analyzers). Within-run and between-run CVs were less than 1.4% and less than 1.6%, respectively. The reaction rate per millimole of K+ per liter (0.05 delta A/min) was more than double that of the reagent blank (0.02 delta A/min). Results correlated well with those by flame photometry, and interference from bilirubin, hemoglobin, lipids, heparin, and other cations was negligible. This method, in conjunction with a previous method we have reported in which beta-galactosidase is used for measuring Na+ in serum, offers a practical alternative to the use of ion-selective electrodes and flame photometry for measuring these clinically important monovalent cations in high-throughput or "stat" biochemical analyzers.

Ammonia

Quantitative method for determining serum alkaline phosphatase isoenzyme activity: estimation of intestinal component.

Intestinal alkaline phosphatase activity was measured using levamisole inhibition, and results were compared with a previously reported method using L-phenylalanine. Sixty two per cent intestinal, 39% placental, and 1.3% of either bone or liver alkaline phosphatase activity remained when alkaline phosphatase activity was inhibited in a 2-amino-2-methyl-1-propanol (AMP) buffer reagent system with 10 mmol/l levamisole (final assay concentration 8.1 mmol/l). The assay imprecision (SD) was 0.6 U/l compared with 3.9 U/l using L-phenylalanine for specimens with total alkaline phosphatase activity less than 250 U/l (reference range 30-120 U/l). In serum pools with raised total alkaline phosphatase activity errors in recovered intestinal activity were small (usually less than 3 U/l) when intestinal alkaline phosphatase was added. Much larger errors and many underestimated results were found using L-phenylalanine. For non-haemolysed specimens it is concluded that an assay based on levamisole inhibition provides a better measure of intestinal alkaline phosphatase activity than L-phenylalanine.

Alkaline Phosphatase

Laboratory evaluation of the Beckman Synchron CX3 clinical chemistry analyzer.

In this evaluation of the Beckman Synchron CX3, the multi-analyte clinical chemistry analyzer exhibited high precision, good linearity, and no carryover for each of the eight analytes measured. Results obtained correlated well with those produced by our routine instrumentation (Beckman Astra, Varian atomic absorption spectrophotometer). The instrument can process up to 75 samples per hour (600 tests per hour if all tests available are requested) and, after calibration, can provide urgent results for the complete panel of tests within 2 1/2 min. The performance characteristics of this instrument make it ideal as a routine or a "stat" analyzer for commonly requested tests in the clinical chemistry laboratory.

Autoanalysis

Enzymatic determination of sodium in serum.

This is a kinetic assay for measuring serum Na+ concentration based on determination of Na+-dependent beta-galactosidase (EC 3.2.1.23) activity. The method, sufficiently sensitive to measure sub-millimolar concentrations of Na+, was modified by including a Na+-binding agent (cryptand) to provide a linear assay for serum Na+ concentrations between 110 and 160 mmol/L. The assay was developed with and evaluated in the Cobas Fara centrifugal analyzer (and has been used in other kinetic analyzers). Within-run and between-run CVs were less than 1%. The reaction rate for normal serum samples (0.20 delta A/min) is about 10-fold that of the reagent blank. Results correlated well with flame photometry. Interference from bilirubin, hemoglobin, lipemia, heparin, and other cations was negligible. The method offers a practical alternative to the use of ion-selective electrodes and flame photometry for measuring serum Na+ in high-throughput or "stat" biochemical analyzers.

Bridged Bicyclo Compounds

Quantitative method for determining serum alkaline phosphatase isoenzyme activity I. Guanidine hydrochloride: new reagent for selectively inhibiting major serum isoenzymes of alkaline phosphatase.

The potential use of the protein denaturant guanidine hydrochloride to inhibit selectively the enzyme activity of serum alkaline phosphatase isoenzymes from liver, bone, intestine, and placenta was investigated. Inhibition of each isoenzyme was shown to be dependent on time and concentration of inhibitor. In the presence of 0.3 mol/l (28.7 g/l) guanidine hydrochloride for 170 seconds 14%, 47%, and 90% of the total alkaline phosphatase activity remained in samples of bone, liver, and intestinal origins, respectively. In contrast, the activity of the placental isoenzyme increased by 24%. The degree of inhibition was shown to be independent of total alkaline phosphatase activity. Investigations were performed at 37 degrees C using the Cobas Bio centrifugal analyser. We conclude that this reagent has several practical advantages over urea as a selective inhibitor of alkaline phosphatase isoenzymes, including a faster and more reproducible inhibition at a much lower reagent concentration.

Alkaline Phosphatase

Quantitative method for determining serum alkaline phosphatase isoenzyme activity II. Development and clinical application of method for measuring four serum alkaline phosphatase isoenzymes.

A method for quantitating the liver, bone, intestinal and placental alkaline phosphatase activity of serum, using an algorithm for converting selective inactivation by guanidine hydrochloride, L-phenylalanine, and heat into equivalent isoenzyme activity is described. The method can individually quantify mixtures of isoenzymes to within a margin of 3%; it has acceptable reproducibility and has been used to develop both age and sex related reference ranges. Analysis time is about 30 minutes. The clinical reliability of this method has been shown in a study of 101 patients, in 79% of whom isoenzyme results were compatible with the final clinical diagnosis; in 10% a clinical diagnosis resulted from isoenzyme analysis, and in a further 11% the source of the increased alkaline phosphatase activity was identified and supported by electrophoresis, with a definite clinical diagnosis yet to be made.

Adult

Immunoturbidimetric measurement of transferrin.

This paper describes the development of an automated immunoturbidimetric assay for transferrin on a centrifugal analyser. Regression analysis of transferrin values measured immunoturbidimetrically demonstrates good agreement with data obtained by radial immunodiffusion (y = 0.997 + 0.024 g/l, r = 0.980, n = 50). The assay has a detection limit of 1.0 g/l and working range of approximately 1.0 to 6.0 g/l of transferrin. Day-to-day coefficient of variation is less than 3.5%. Immunoturbidimetric transferrin (g/l) and total iron binding capacity values (mumol/l) were compared using an established total iron binding capacity method (y = 0.050 X - 0.030 g/l, r = 0.967, n = 50). Minimal interference was found for lipaemic, haemolysed or icteric samples. Transferrin reference values with a mean of 3.05 g/l and 95% limits from 2.45 to 3.65 g/l were derived using serum from 300 apparently healthy subjects (150 males, 150 females). We conclude that the proposed transferrin method is more reliable and easier to perform than presently available total iron binding capacity methods.

Adult

Rapid cholesterol measurement: patient classification in heart risk evaluation clinics.

Plasma cholesterol levels of patients attending a heart risk evaluation clinic were measured both in the clinic at the time of attendance, and later in a reference laboratory. The workflow of evaluation clinics may be improved, and patient satisfaction enhanced, by in situ measurement, since more than 60% of patients can be given a complete risk score at the time of attendance.

Cholesterol

Assessment of the Beckman oxygen rate analyzer for the enzymatic assay of cholesterol.

We describe an evaluation of the Beckman oxygen rate analyzer for the determination of cholesterol. The precision and accuracy obtained did not fulfill the short-term criteria of the Center for Disease Control but the precision met the criteria of the College of American Pathologist for analysis of cholesterol in screening programs. Good overall correlation between the oxygen rate method and a continuous-flow method with prior extraction was found for patient samples. The results obtained by the oxygen rate method had net positive bias up to the limit of linearity of 8.6 mmol cholesterol/liter. The instrument is not recommended for laboratory or epidemiological studies but may prove useful in screening programs.

Autoanalysis

Evaluation of a kit for the colorimetric determination of inorganic phosphate.

An evaluation of a colorimetric kit method for the determination of inorganic phosphate (Pierce Phosphorus Auto/Stat Kit) is described. The within-batch and between-batch precisions were shown to fulfil current criteria, and recovery experiments, linearity studies, analyses of quality control materials, and studies of possible interfering substances evidenced good accuracy. Comparison of the results obtained on samples from patients with those obtained by the vanadate/molybdate continuous-flow method showed that the test method had a comparative positive bias. The use of a calibration reference serum as standard is recommended. The kit method is technically simple, requiring no protein precipitation, and analyses can be performed rapidly.

Colorimetry

Manual and centrifugal analyzer determination of cholesterol and triglycerides with single-vial enzymatic reagents.

We have assessed Calbiochem single-vial enzymatic reagent systems for the analysis for cholesterol and triglycerides, using manual and centrifugal analyzer techniques. Analysis for cholesterol by both techniques had acceptable performance by the short-term criteria of the Center for Disease Control (CDC) during this study carried out under optimal conditions of variance. Analysis for triglycerides by the manual technique did not meet CDC criteria, and we do not recommend use of this method. Analysis for triglycerides by the centrifugal analyzer technique had acceptable short-term precision but had unacceptable accuracy at high concentrations of triglycerides; this method may be considered to have acceptable performance for routine laboratory use.

Centrifugation

Effectiveness of an outpatient urine screening program.

We evaluated the effectiveness of a routine outpatient urinalysis screening program on a sample population of 2600 patients. The 189 abnormal urine results found in 182 patients were followed up by study of any new clinical and laboratory investigations or therapeutic modifications initiated on the basis of any abnormal test result. The urinalysis screening program appeared to have significant bearing on diagnosis or treatment in only 13 patients. Abnormalities found in 150 of the 182 patients were either not noted or no further positive action was taken. Thus we concluded that under the conditions of this study the urine screening program added to hospital costs without significant benefit to the patient.

Bilirubin

Urinalysis by use of multi-test reagent strips: two dipsticks compared.

We compared Ames' "N-Multistix" with Boehringer's "Combur-8" ("Chemstrip-8") multi-test urine reagent strips by analysis of contrived urine specimens, testing accuracy, precision, specificity, and limits of detection of both products. Relative cost and ease of use were also examined. Each brand of urinary dipstick had specific advantages but it is unlikely that patient care would be adversely affected by preferential use of either product.

Acetoacetates