The increased performance of white cell (WBC) filters makes it difficult to count precisely the number of residual WBCs. Concentrations as low as 0.01 WBC per microL cannot be determined with electronic cell counters, conventional hemocytometers, or the flow cytometric techniques currently being used. This article describes a simple, manual method using a Nageotte hemocytometer with a large-volume chamber (50 microL) to count the number of WBCs contained in red cell (RBC) suspensions (preparations A, B, and C) and in platelet suspensions (preparation D) diluted 1 in 10 pure, or concentrated two fold. To validate the method, several reference ranges, prepared by successively adding mononuclear cells to a suspension of pure RBCs or platelets, were used. Among the different series, validation ranges varied from 0.2 to 12 to 0.01 to 0.5 WBCs per microL and correlation coefficients ranged from 0.929 to 0.996. To determine the limit of accurate detection, accuracy tests (n = 160) were carried out by two experienced operators on samples with WBC concentrations of about 5, 10, and 120 times the concentration at the theoretical limit of detection (1 WBC/chamber). No significant difference was observed in the various types of preparations (A, B, C, D) in the tests performed by the two operators. However, intra-assay coefficients of variation were 18, 9.5, and 2.2 percent, respectively, at WBC concentrations of 5, 10, and 120 times that at the theoretical limit of detection. These observations show that a limit of accurate detection (10%) seems to be reached when 10 cells are observed in a Nageotte hemocytometer.(ABSTRACT TRUNCATED AT 250 WORDS)

After testing several chromatographic systems described in the literature, the optimal operating conditions were retained to develop a simple and rapid method grouping TLC and HPLC/UV for separation, identification and dosage of kojic acid (CAS RN 501-30-4) or arbutine (CAS RN 497-76-7) in skin-whitening cosmetic products. The screening is carried out by TLC on cellulose plate and by TLC on silicagel plate with UV indicator and polar mobile solvent. Regarding identification and quantitative determination by HPLC/UV, the best results were obtained by direct phase chromatography, using a polar stationary phase greffed with diol groups and a polar buffered mobile phase at PH 2,5. Intralaboratory prevalidation tests were carried out on the HPLC method: detection limit, linearity and intralaboratory repeatability of standard curves and repeatability of samples quantitative determination.

To evaluate accurately the current performance of filtration, the French Produits Sanguins Labiles study group, composed of 21 transfusion teams, conducted a large-scale 6-month study involving over 1400 filtrations and 3000 controls. Some 745 standard red cell concentrates (RBC concentrates) and 690 concentrates previously white cell (WBC)-reduced by removal of buffy coat (BC-poor RBC concentrates) were filtered using six commercially available filters: at least 170 results were collected per filter, spread among a minimum of three teams. Prefiltration controls show that the removal (manual and automated) of the buffy coat results in an initial WBC reduction of approximately 63 percent, along with a hemoglobin loss of 4 g (7%). After filtration, residual WBCs were counted in the Nageotte manual counting chamber. The reliability of this counting method, which is simple and adapted to low WBC concentrations, was characterized in this study by a 25-percent coefficient of variation (CV) for a concentration of 2.5 WBCs per microL (i.e, 0.6 x 10(6) WBCs/filtered unit). The analysis of the results shows that, for five of six filters (1 filter was excluded), the postfiltration median value of residual WBCs was 1.1 x 10(6) in filtered RBC concentrates (n = 590), whereas it was 0.34 x 10(6) in filtered BC-poor RBC concentrates (n = 581). The difference is significant (p less than 10(-8), Wilcoxon test). Hemoglobin loss due to filtration varies according to the filter, from 5.7 +/- 2.2 to 17.3 +/- 2.5 g.(ABSTRACT TRUNCATED AT 250 WORDS)

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