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Vaccimonitor
On-line version ISSN 1025-0298
Abstract
SOTO, Carmen et al. Validation of a colorimetric technique for carbohydrate determination. Vaccimonitor [online]. 2002, vol.11, n.3, pp. 11-14. ISSN 1025-0298.
The appropriate standardization of an analytical assay, followed by the validation of the method and the homogeneity of the chemical and biological reagents used during the different studies are indispensable elements for obtaining optimal results. In this work a methodology was selected for the validation of the analytical method used to determine ribose by the orcinol method using a standard ribose solution (0,025 mg/mL). An analysis of the main validation criteria: linearity, precision, accuracy, sensitivity, specificity, detection limits and robustness were carried out. The usefulness of this procedure was demonstrated and by means of the experimental design and the statistical procedures used, the linearity (r2>0,98), exactness (Fexp. <Ftab. and texp. <ttab.), preciseness (CV<3%), sensitivity (LD = 1 ug/mL and LC = 3.3 ug/mL) and specificity (non significant answer) of the method were verified, for which reasons it is considered reliable. The determination of ribose is a colorimetric assay that quantifies pentoses and is used with the objective of knowing the exact pentose contents in a sample of purified product. It is also used as a final control for Haemophilus influenzae conjugated vaccines according to WHO.
Keywords : Orcinol; ribose; validation; Haemophilus influenzae.