IFCC recommended reference method for the determination of the substance concentration of ionized calcium in undiluted serum, plasma or whole blood.

A reference method is described for the determination of the substance concentration of ionized calcium in plasma by which ionized calcium (free or unbound) may be reliably determined on the basis of calibration with aqueous solutions with known concentration of ionized calcium. The composition of the calibration solutions is chosen such that the activity coefficient of the calcium ion is assumed to be identical both in the calibration solutions and in "normal" plasma, i.e. by convention, the ionic strength (Im) is 0.160 mol/kg. The convention is adopted of reporting ionized calcium measurements as concentration expressed as mmol/l. The proposed reference method for ionized calcium measurement in plasma is based on the use of a cell consisting of an external reference electrode with a saturated potassium chloride liquid/liquid junction in combination with a calcium ion-selective membrane electrode of defined construction and performance. Procedures for using the reference cell and a protocol for sample measurement are described. The preparation of the calibration solutions to be used are described in detail in Appendix A, secondary calibration solutions and check standards in Appendix B, and reference cell vessel design in Appendix C.

The slow penetration of enzyme-based biosensors into clinical chemistry analysis.

In 1972 Yellow Springs Instrument Co., Inc. (YSI) introduced the first commercial instrument which could measure glucose in diluted whole blood by use of an enzyme-based biosensor. Until 1986 YSI had a monopoly of this technology for clinical chemistry. Since 1986 another ten companies have commercialized the technology. Enzyme-based biosensors for clinical chemistry are now available for glucose, lactate, and urea. Until 1975 the annual number of papers on enzyme-based biosensors is sparse, but from then there is a significant increase in the number of publications. The commercial interest in glucose is reflected in the number of patents covering this analyte: close to fifty per cent of the patents cover this type of sensor while less than ten per cent of the scientific papers are on glucose sensors. The major problems in developing enzyme-based biosensors for clinical chemistry have been achievement of the necessary linear range and elimination of interferences from the blood. The slow penetration of enzyme-based biosensors into clinical chemistry analysis is due to the fact that it is difficult to develop sensors which meet the high accuracy and reliability requirements, are manufacturable, and have an acceptable shelf life. The number of commercial products utilizing enzyme-based biosensors will probably remain limited in the future due to the limited number of analytes for which a suitable enzyme exists, and for which the use of an enzyme-based biosensor offers the advantages that can justify the costs of development.

Recommendation on sampling, transport, and storage for the determination of the concentration of ionized calcium in whole blood, plasma, and serum. IFC Scientific Division, Working Group on Ion-Selective Electrodes (WGSE).

The substance concentration of ionized calcium (cCa 2+) in blood, plasma, or serum preanalytically may be affected by pH changes of the sample, calcium binding by heparin, and dilution by the anticoagulant solution. pH changes in whole blood can be minimized by anaerobic sampling to avoid loss of CO 2, by measuring as soon as possible, or by storing the sample in iced water to avoid lactic acid formation. cCa 2+ and pH should be determined simultaneously. Plasma or serum: If centrifuged in a closed tube and measured immediately, the pH of the sample will be close to the original value. If there has been a delay between centrifugation and measurement, causing substantial loss of CO 2, equilibration of the sample with a gas mixture corresponding to pCO 2 = 5.3 kPa prior to the measurement is recommended. Conversion of the measured values to cCa 2+ (7.4) is only valid if the pH is in the range 7.2-7.6. Ca 2+ binding by heparin can be minimized by using either of the following: 1) a final concentration of sodium or lithium heparinate of 15 IU/mL blood or less; or 2) calcium-titrated heparin with a final concentration of less than 50 IU/mL blood. Dilution effect can be avoided by use of dry heparin in capillaries or syringes.(ABSTRACT TRUNCATED AT 250 WORDS)

International Federation of Clinical Chemistry (IFCC), scientific division: IFCC recommendation on sampling transport and storage for the determination of the concentration of ionized calcium in whole blood, plasma and serum.

The substance concentration of ionized calcium (cCa2+) in blood, plasma or serum preanalytically may be affected by pH changes of the sample, calcium binding by heparin, and dilution by the anticoagulant solution. pH changes in whole blood can be minimized by anaerobic sampling to avoid loss of CO2, by measuring as soon as possible or by storing the sample in iced water to avoid lactic acid formation. cCa2+ and pH should be determined simultaneously.

IFCC recommendation on sampling, transport and storage for the determination of the concentration of ionized calcium in whole blood, plasma and serum.

The substance concentration of ionized calcium (c(Ca) (2+)) in blood, plasma or serum preanalytically may be affected by pH changes of the sample, calcium binding by heparin, and dilution by the anticoagulant solution.pH changes in whole blood can be minimized by anaerobic sampling to avoid loss of Co(2), by measuring as soon as possible, or by storing the sample in iced water to avoid lactic acid formation. c(Ca) (2+) and pH should be determined simultaneously.PLASMA OR SERUM: If centrifuged in a closed tube, and measured immediately, the pH of the sample will be close to the original value. If a delay has occurred between centrifugation and the measurement, causing substantial loss of Co(2), equilibration of the sample with a gas mixture corresponding to pCO2= 5.3 kPa prior to the measurement is recommended. Conversion of the measured values to c(Ca) (2+) (7.4) is only valid if the pH is in the range 7.2-7.6.Ca(2+) binding by heparin can be minimized by using either of the following:(1) A final concentration of sodium or lithium heparinate of 15 IU/ml blood or less(2) Calcium titrated heparin with a final concentration of less than 50 IU/ml blood.Dilution effect can be avoided by use of dry heparin in capillaries or syringes. When heparin solutions are used, errors due to dilution or calcium binding can be reduced by using syringes with a heparin solution containing free calcium ions corresponding to the mean concentration of ionized calcium in normal plasma.Conditions for blood collection, storage, and transport to avoid preanalytical errors are described in this paper.

International Federation of Clinical Chemistry (IFCC) scientific division IFCC recommendation. Recommendation on sampling, transport and storage for the determination of the concentration of ionized calcium in whole blood, plasma and serum.

The substance concentration of ionized calcium (cCa2+) in blood, plasma or serum preanalytically may be affected by pH changes of the sample, calcium binding by heparin, and dilution by the anticoagulant solution. pH changes in whole blood can be minimized by anaerobic sampling to avoid loss of CO2, by measuring as soon as possible or by storing the sample in iced water to avoid lactic acid formation. cCa2+ and pH should be determined simultaneously. Plasma or serum: if centrifuged in a closed tube and measured immediately the pH of the sample will be close to the original value. If delay has occurred between centrifugation and the measurement, causing substantial loss of CO2, equilibration of the sample with a gas mixture corresponding to PCO2 = 5.3 kPa prior to the measurement is recommended. Conversion of the measured values to cCa2+ (7.4) is only valid if the pH is in the range 7.2-7.6 Ca2+ binding by heparin can be minimized by using either of the following: a final concentration of sodium or lithium heparinate of 15 IU/ml blood or less, by use of calcium titrated heparin with a final concentration less than 50 IU/ml blood. Dilution effect can be avoided by use of dry heparin in capillaries or syringes. When heparin solutions are used errors due to dilution or calcium binding can be reduced using syringes with a heparin solution containing free calcium ions corresponding to the mean concentration of ionized calcium in normal plasma. Conditions for blood collection, storage, and transport to avoid preanalytical errors are described.

Measurement of free calcium ion in capillary blood and serum.

We describe a new calcium ion-selective electrode for measurement of the substance concentration of free calcium ion [Ca2+] in the plasma phase of whole blood and in serum at 37 degrees C. A sample volume of 50 microliter suffices to obtain simultaneous values of pH and [Ca2+]. We found the within-series analytical standard deviation for serum to be 0.013 mmol/litre (CV, 1.1%) and day-to-day precision to be 0.022 mmol/litre (CV, 1.7%). The reference interval for [Ca2+] (at pH 7.40) in serum was found to be 1.184 +/- 0.054 mmol/litre (2 SD) from measurements on sera from 121 healthy blood donors. Measurements on capillary blood from 29 healthy volunteers gave a mean (+/- 2 SD) value for [Ca2+] (at pH 7.40) of 1.22 +/- 0.072 mmol/litre.