Preanalytical Quality Evaluation of Citrate Evacuated Blood Collection Tubes-Ultraviolet Molecular Absorption Spectrometry Confronted with Ion Chromatography.

We previously enabled a direct insight into the quality of citrate anticoagulant tubes before their intended use for specimen collection by introducing an easy-to-perform UV spectrometric method for citrate determination on a purified water model. The results revealed differences between the tubes of three producers, Greiner BIO-ONE (A), LT Burnik (B), and BD (C). It became apparent that tubes C contain an additive, which absorbs light in the ultraviolet range and prevents reliable evaluation of citrate anticoagulant concentration with the suggested method. In this research, we re-evaluate the quality of citrate-evacuated blood collection tubes by complementing UV spectrometry with ion chromatography. (1) Comparable results were obtained for tubes B at 220 nm. (2) Citrate concentrations determined with ion chromatography were lower for tubes A and C. Chromatograms reveal additional peaks for both. (3) Influences of heparin on absorption spectra and chromatograms of citrate were studied. Some similarities with the shape of the anticoagulant spectra of tubes A and C were observed, and the lithium heparin peak in chromatograms is close to them, but a confident judgment was not possible. (4) Contamination of anticoagulant solution with potassium, magnesium, and calcium was confirmed for all the brands, and contamination with lithium for B and C.

Accuracy of Citrate Anticoagulant Amount, Volume, and Concentration in Evacuated Blood Collection Tubes Evaluated with UV Molecular Absorption Spectrometry on a Purified Water Model.

Citrate anticoagulant concentration affects the results of coagulation tests. Until now, the end user had no direct insight into the quality of evacuated blood collection tubes. By introducing an easy-to-perform UV spectrometric method for citrate determination on a purified water model, we enabled the evaluation of (1) the accuracy of the anticoagulant amount added into the tubes by a producer, (2) the accuracy of the volume of anticoagulant solution in the tube at the instant of examination, (3) the anticoagulant concentrations at a draw volume. We examined the Vacuette®, Greiner BIO-ONE, Vacutube, LT Burnik d.o.o., and BD Vacutainer® tubes. The anticoagulant amount added into the tubes during production had a relative bias between 3.2 and 23.0%. The anticoagulant volume deficiency at the instant of examination expressed as a relative bias ranged between -11.6 and -91.1%. The anticoagulant concentration relative bias after the addition of purified water in a volume that equalled a nominal draw volume extended from 9.3 to 25.7%. Draw-volume was mostly compliant during shelf life. Only Vacutube lost water over time. Contamination with potassium, magnesium, or both was observed in all the tubes but did not exceed a 0.21 mmol/L level. This study enables medical laboratories to gain insight into the characteristics of the citrate blood collection tubes as one of the preanalytical variables. In situations that require anticoagulant adjustment for accurate results, this can help make the right decisions. The methodology gives producers additional means of controlling the quality of their production process.

The impact of a hands-on approach to learning visible spectrometry upon students' performance, motivation, and attitudes.

In this paper, the effect of introducing visible spectrometry concepts through hands-on laboratory work upon student learning within four vocational programs are discussed. All together, 118 students, average 18.6 years old, participated in the study. The results showed no correlation between students' motivational components (intrinsic, regulated, and controlled), chemistry self-concept and their achievement on an experiential knowledge test and knowledge gained from this hands-on approach. Statistically significant differences were found for academic achievement among students in a biotechnology technical program (School 1), food processing program (School 2), laboratory biomedicine program (School 3), and a biotechnology general program (School 4). Differences in academic achievement are further reflected in students' perception of particular knowledge gained through their hands-on experiences and in their expressed attitude toward different didactical characteristics. All students, regardless of their study program, highly evaluated the relaxed atmosphere that contributed to their self-confidence in completing their laboratory activities.

Evacuated blood-collection tubes for haematological tests - a quality evaluation prior to their intended use for specimen collection.

BACKGROUND: An inappropriate anticoagulant concentration in a blood sample can cause cell shrinkage and affect the haematocrit and mean corpuscular volume (MCV). In evacuated blood-collection tubes there are two parameters affecting the quality of the product: the anticoagulant amount introduced into the tube during its production and the internal under-pressure at the instant of the blood-specimen collection affecting the draw-volume. No testing procedures that would give an insight into the anticoagulant concentration that can be expected for blood samples after specimen collection have been available up until now. METHODS: The methodology suggested here combines the draw-volume test performed with deionised water using a laboratory made measuring device, and a conductivity measurement. The corrections taking into account the air pressure and ambient temperature provide an insight into the anticoagulant concentration that can be expected for blood samples. Results presented in the form of a nomogram facilitate the routine use of the suggested methodology. RESULTS: Our 338-day study confirmed significant differences and variations in the quality and the anticoagulant concentrations of the K3EDTA and K2EDTA tubes of different producers and identified different examples of non-compliance with the norms during the shelf life of the tubes. CONCLUSIONS: The quality evaluation of the evacuated blood-collection tubes prior to their intended use as suggested here can, in everyday laboratory practice, ensure that the tubes are used only if, and only until, their quality is adequate.

Microdiffusion-based UV-LED spectrometric setup for determining low levels of ethanol in fruit juice.

A novel setup is described in which we combined the separation of a volatile substance from a sample with a complex matrix on the basis of a microdiffusion process with a kinetic on-line spectrometric monitoring of the reaction in the receptive medium at 365 nm. The fruit juice was selected as a model for testing the performance of the setup in real-life applications. The ethanol content in fruit juice can be considered as an indicator of the fruit-juice quality and should not exceed the regulatory limiting values. After optimising the microdiffusion process, blackcurrant, orange and two varieties of apple juice were analysed. The sample analysis lasted 15 min at 35°C. The ethanol concentrations were found to be between 0.9 and 4.0 mmol/L, and were comparable to the results obtained using the SIST:ISO 2448:1998 standard method, which is time consuming, labour intensive and requires high sample volumes. The setup can easily be adapted for determining other volatile substances in low concentrations in complicated samples of different types by introducing different chemistry and replacing the light source if the light of a different wavelength is required. The measuring characteristics of the setup were critically assessed, the main sources of uncertainty recognised and the possibilities for further improvements of the setup and the procedure considered.

Detailed uncertainty budget for major and minor ions in stock combined calibration standards--influence of impurities in chemicals.

A general methodology for a systematic evaluation of the uncertainty was derived for each particular ion in stock combined calibration standards in which concentrations of different ions extend over up to five orders of magnitude resulting in detailed uncertainty budgets with the aim of recognising the major contributions to combined uncertainties. This work confirmed that it is justifiable that the mass fraction of impurities in other chemicals is taken into account when calculating the mass concentration of an ion in combined calibration standard solution similarly to what is already common practice in accounting for the purity of chemical. It was proven that impurities in chemicals which are sources of major ions have significant effect on uncertainty budget of minor ions; already if the major ion exceeds 25 times the minor ion's concentration. For several ions it was confirmed that mass fraction of the impurities was the major source of uncertainty.

Ionic composition of seawaters and derived saline solutions determined by ion chromatography and its relation to other water quality parameters.

Ion chromatography (IC) presents new possibilities for assessing information about environmental samples, namely waters of various compositions, ranging from high-purity water to highly saline ones. Constant proportion between major ions present in seawater, has been assumed in the past, from which the first practical equation relating chlorinity and salinity has been developed, being later substituted by a practical salinity scale, derived from conductivity measurements relative to a standard seawater, according to internationally accepted recommended procedures. Seawaters are characterized by salinity values around 35 while derived saline solutions may present considerable changes in ionic composition, conductivity, hence on salinity. Natural and anthropogenic phenomena may introduce new issues requiring clarification for which qualitative and quantitative information from additional sources is useful, e.g. ionic composition from IC. The different ranges of concentration of major and minor species present in seawater and derived saline solutions are a challenge for the optimization of a practical methodology for composition assessment in two single IC runs, one for anions and another one for cations, which has been attained in this work. Composition of saline solutions determined by IC was critically assessed in terms of anion-cation balance and further related to conductivity and salinity measurements aiming to evaluate the quality/completeness of ion chromatographic analyses performed at preselected conditions and to search for other meaningful relations for efficient recognition/distinction between saline solutions of different types.

A new type of a spectrometric microtitration set up.

A spectrometric microtitrator was developed from a spectrometer with a microreaction chamber and a tri-colour light-emitting diode (LED) as the light source. A novel, vertical, optical geometry of the spectrometric microtitration chamber was introduced and tested. This novel geometry also required a new method for mixing the titrated solution. A laboratory-made 50mul syringe pump was used for the addition of the titration reagent. The 10-channel module for light effects, which makes possible a low-cost hardware approach to changing the titration protocols, was used for coordinating the operation of the microtitration set up. The system, with 10 channels and a regulated speed of operation, is flexible enough to allow an operator to generate different titration protocols. The performance test showed that the speed of titration-reagent addition can be regulated in the range from 0.87 to 21.8mulmin(-1). The smallest achievable volume addition is equal to 35nl. The mixing rate can be continuously regulated by an electrical pulse that initiates the mixing cycle. The quickest rate is every 1.6s, and the slowest rate is every 4.8s. The spectrometric microtitration set up was successfully tested for several different real-life spectrometric titrations, including an iodometric titration, a determination of CO(2) in deionised water, and EDTA titrations of copper(II) ions with no indicator. The volume of the examined solution can be as small as 220mul. The titration-reagent consumption is usually between 10 and 35mul. Coefficients of variation of the end point volume determination (n = 5) at different experimental conditions and different average volumes of consumed reagents (7.06, 12.17 and 22.88mul) were 2.4, 1.3 and 1.2%, respectively. The novel geometry of the spectrometric microtitration chamber proved to be useful for real-life applications.

Spectrometer with microreaction chamber and tri-colour light emitting diode as a light source.

This paper describes a tri-colour light emitting diode (LED)-photoresistor (PR)-based in situ spectrometer with geometry that differs from the previously presented and allows for small volumes of test solution (350mul) and effective homogenisation of reagents. The emission maximums of the tri-colour LED (TC-LED) are at 470, 565 and 660nm. The basic measuring characteristics of the prototype were evaluated. The prototype was tested for determination of calcium in natural waters with the o-cresolphtalein complexon and was proved to be useful for real life applications. The in situ spectrometer with microreaction chamber enables the rapid optimisation of experimental conditions, as was demonstrated on the example of oxidation of alcohols with potassium dichromate. The drop-based experimental approach to the ruggedness test is fast, economic and gives reliable results with the minimum waste production. The prototype, connected to the computer, functions as a kinetic microreactor suitable also for following rapid changes. In spite of the manual addition of the initialising reagent, the data are lost only during the first 3s.