[Lipemia and bilirubin influences for twenty-four biochemical parameters measurement]. / Interférences de la lipémie et de l'ictère sur le dosage de 24 paramètres biochimiques.

The study of the influence of the lipemia and icterus was performed experimentally for twenty-four biochemistry parameters on the Roche Cobas 6000 CE analyzer. Overloads in Intralipid(®) or ditaurate of bilirubin were performed on several plasma pools. The limit of 10% was chosen to define interference on the measurement. The parameters studied were classified into different categories depending on their measurement is affected or not. Knowledge of these data allows the biologist to adapt its reporting procedures in the case of lactescent and/or icteric samples.

[Comparison of calculated LDL cholesterol (LDL-C) versus measured LDL cholesterol (LDL-M) and potential impact in terms of therapeutic management]. / Évaluation d'une méthode de dosage direct du LDL-cholestérol et de son impact potentiel en termes de prise en charge thérapeutique.

LDL-cholesterol value is one of the criteria used by the Haute autorité de santé (HAS) in the management of patients in primary and secondary prevention with the aim to reduce cardiovascular mortality. In this respect, the recommendations have been established based on target to achieve LDL-cholesterol. Currently in France, the determination of LDL-cholesterol is mainly carried out by the Friedewald formula whose limits are well known. However, reliable methods for the determination of LDL-cholesterol exist. We compared the results of calculated and measured LDL-cholesterol obtained from 444 patients presenting normal triglyceridemia values in terms of ranking relative to the thresholds of the HAS. The correlation between the two methods is quite good, but a significant difference (p <0.0001) was observed between the calculated and measured values of LDL-cholesterol. On the other hand in 17% of cases the classification of subjects will be different, with a majority so overestimation of calculated LDL-cholesterol with respect to measured LDL-cholesterol. This overestimation is not proportional, in fact most values measured LDL-cholesterol, the higher the calculate-measured difference is important. The rating difference is particularly important when subjects have between 1 and 3 factors of cardiovascular risk where the target LDL-cholesterol to achieve is between 1.3 and 1.9 g/L. The management of patients with lipid lowering may potentially be dependent on the method used for the determination of LDL-cholesterol.

[Hemolysis influence on twenty-two biochemical parameters measurement]. / Interférence de l'hémolyse sur le dosage de vingt-deux paramètres biochimiques.

The study of the influence of hemolysis was determined experimentally for twenty two biochemical parameters on the analyzer Cobas 6000 ce (Roche Diagnostics). The addition method of hemolysate was used to create an increasing concentration of hemoglobin ranging from 0 to 2000 µmol/L. The limit of 10% variation was chosen to define the influence of hemolysis on the measurement. The parameters studied were classified into several categories: the parameters for which hemolysis does not influence the measurement: albumin, uric acid, calcium, C-reactive protein, myoglobin, NT -pro BNP, S100 protein, and urea; parameters impacted positively leading to an overestimation of the result: aspartate aminotransferase, total cholesterol, creatine kinase, creatinine, lactate dehydrogenase, magnesium, magnesium, total protein, triglycerides; and negatively impacted settings so causing an underestimation of the result: alanine amino- transferase, gamma glutamyl transferase, lipase, alkaline phosphatase, troponin T hypersensitive. Certain parameters influence of hemolysis varies depending on the magnitude of the measured parameter this interference being observed for normal values but disappearing for pathological values: creatinine, cholesterol, alkaline phosphatase, triglycerides, or the inverse interference is greater than for conventional pathological values: lipase, alanine amino-transferase. Knowledge of this variability interference allows the biologist to adapt its methods of reporting in the case of haemolysed samples.

[Influence of anticoagulant on the plasma level of fifteen biochemical parameters]. / Influence de la nature de l'anticoagulant sur le dosage plasmatique de quinze paramètres biochimiques.

The study of the influence of the anticoagulant used in blood collection tubes to obtain plasma was performed for fifteen biochemical parameters measured with automated Cobas 6000 (Roche Diagnostics). For each parameter tested the entire measurement domain was studied. The comparison of results obtained on plasma blood sample obtained by lithium heparin and EDTA include: correlation, the limits of acceptability in the standards of monitoring and interpretation standards regression defined by the SFBC and analysis of Bland-Altman. The parameters studied were classified into three categories. The parameters for which the assay is not influenced by the nature of the anticoagulant used: apolipoprotéin A1, apolipoprotein B, alanine amino-transferase, creatine kinase, creatinine, total cholesterol, HDL-cholesterol, lipase, NT-Pro BNP, troponine T and urea. The parameters for which the results are underestimated EDTA plasma, including those for which the impact is moderate and for which the interpretive standards are not changed: triglycerides, and those for which performance standards are changed on one or more levels: aspartate aminotransferase and lactate dehydrogenase; and finally the not practicable EDTA plasma parameters: alkaline phosphatase.

In vivo removal of high- and low-molecular-weight compounds in hemodiafiltration with on-line regeneration of ultrafiltrate.

BACKGROUND: Current methods of renal replacement therapy, combining convection and diffusion, are largely unsatisfactory in removing uremic toxins. Adsorption is a third mechanism that has been applied in extracorporeal therapy. This study evaluates the impact of hemodiafiltration with on-line regeneration of ultrafiltrate, a new two-step integrated sorbent system, on in vivo removal of a wide spectrum of solutes with different molecular weights. METHODS: Pre- and post-dialysis concentrations of small, medium-size, and large molecules were determined in ten patients undergoing regular hemodiafiltration treatments with on-line regeneration of the ultrafiltrate. We also analyzed, at different times of the same dialysis session, the inlet and outlet ultrafiltrate; the latter had been regenerated by the sorbent cartridge and was used as reinfusion liquid. The mean dialysis time was 260 +/- 21.2 min with a blood flow of 361 +/- 33.3 ml/min and a reinjection volume of 3.6 +/- 0.2 l/h. RESULTS: Urea, creatinine and phosphate reduction ratio were respectively 69.8 +/- 8.2, 61.9 +/- 5.5, and 40.2 +/- 17.3%. Removal of medium-size markers such as calcitonin, osteocalcin, beta2-microglobulin, cystatin C, myoglobin and prolactin varied between 24 and 60%. The percentage of reduction for retinol binding protein and alpha1-microglobulin was negligible and we were unable to demonstrate any removal of alpha1-acid glycoprotein, pre-albumin, and albumin in the regenerated ultrafiltrate. CONCLUSION: The hemodiafiltration with on-line regeneration of ultrafiltrate is a new hemodialysis system, which allows uremic toxin removal over a wide molecular-weight spectrum.

Automated multicapillary electrophoresis for analysis of human serum proteins.

BACKGROUND: We evaluated a new, automated multicapillary zone electrophoresis (CE) instrument (Capillarys), 4.51 software version; Sebia) for human serum protein analysis. METHODS: With the Capillarys beta1-beta2+ reagent set, proteins were separated at 7 kV for 4 min in 15.5 cm x 25 micro m fused-silica capillaries (n = 8) at 35.5 degrees C in a pH 10 buffer with online detection at 200 nm. Serum samples with different electrophoretic patterns (n = 265) or potential interference (n = 69) were analyzed and compared with agarose gel electrophoresis (AGE; Hydrasys)-Hyrys, Hydragel protein(e) 15/30 reagent set; Sebia). RESULTS: CVs were <3.5% for albumin, <11% for alpha(1)-globulin, <4.1% for alpha(2)-globulin, <7.4% for beta-globulin, and <5.8% for gamma-globulin (3 control levels); measured throughput was 60 samples/h. In patients without paraprotein (n = 116), the median differences between CE and AGE were -5.4 g/L for albumin, 4.0 g/L for alpha(1)-globulin, 0.7 g/L for alpha(2)-globulin, 0.6 g/L for beta-globulin (P <0.001 for all fractions), and -0.1 g/L for gamma-globulin (not significant). More samples had at least one gamma-migrating peak detected by CE (n = 135 vs 130; paraprotein detection limit, approximately 0.5-0.7 g/L), but fewer were quantified (n = 84 vs 91) because of gamma- to beta-migration shifts. There was a 1.2 g/L median difference between CE and AGE for gamma-migrating paraprotein quantification (n = 69; P <0.001). Several ultraviolet-absorbing substances (lipid emulsion, hemoglobin) or molecules (contrast agent, gelatin-based plasma substitute) induced CE artifacts. CONCLUSIONS: The Capillarys instrument is a reliable CE system for serum protein analysis, combining advantages of full automation (ease of use, bar-code identification, computer-assisted correction of alpha(1)-globulins) with high analytical performances and throughput.