Validación y estimación de la incertidumbre del método enzimático GOD-PAP, para la determinación de glucosa en suero humano
No Thumbnail Available
Date
2023
Journal Title
Journal ISSN
Volume Title
Publisher
Universidad Nacional de Trujillo
Abstract
El presente trabajo de investigación tiene como objetivo validar y estimar la incertidumbre de la medición del método enzimático glucosa oxidasa - peroxidasa (GOD-PAP) para cuantificar glucosa en sangre. Se elaboró curva de calibración en el rango de 20 a 400 mgdL-1 con la cual se cuantifico glucosa en muestra de suero, y en estándares de glucosa (20, 200 y 380 mgdL-1) durante cinco días de validación. Para la validez del método se calculó la linealidad, rango de trabajo, límite de detección, límite de cuantificación, exactitud, precisión e incertidumbre de la medición. En el estudio de la linealidad se encontró la ecuación de regresión y = 0,0031x + 0,0007 con un coeficiente de determinación de 0,9999. Los límites de detección (LOD) y cuantificación (LOQ) fueron 2,2 mgdL-1 y 7,4 mgdL-1 respectivamente. La exactitud se evaluó mediante el porcentaje de recuperación, siendo satisfactoria, con porcentaje de recuperación de 98,4-104% (20 mgdL-1), 99-103,1% (200 mgdL-1) y 98,1-102,2% (380 mgdL-1). De igual manera la precisión evaluada mediante la repetibilidad (%CVr) y reproducibilidad (%CVR) fueron menores a los criterios de los coeficientes de variación de Horwitz. Finalmente, en la estimación de la incertidumbre en los tres niveles de concentración, se determinaron valores de 20,2 ± 2,5 mgdL-1; 202,3 ± 6,2 mgdL-1 y 380,7 ± 11,0 mgdL-1 utilizando un factor de cobertura de k=2 con un nivel de confianza del 95%. En Conclusión, se obtuvo la evidencia de que el método enzimático glucosa GOD-PAP es lineal, exacto y preciso garantizado la determinación precisa y confiable de la glucosa en sangre.
The objective of this research work is to validate and estimate the uncertainty of the measurement of the glucose oxidase - peroxidase (GOD-PAP) enzymatic method to quantify blood glucose. Calibration curve was prepared in the range of 20 to 400 mgdL-1 with which glucose was quantified in serum sample, and in glucose standards (20, 200 and 380 mgdL-1) during five days of validation. For the validity of the method, the linearity, working range, detection limit, quantification limit, accuracy, precision and uncertainty of the measurement were calculated. In the study of linearity, the regression equation y = 0.0031x + 0.0007 was found with a coefficient of determination of 0.9999. The limits of detection (LOD) and quantification (LOQ) were 2.2 mgdL-1 and 7.4 mgdL-1 respectively. The accuracy was evaluated by the recovery percentage, being satisfactory, with a recovery percentage of 98.4-104% (20 mgdL-1), 99-103.1% (200 mgdL-1) and 98.1-102. 2% (380 mgdL-1). Likewise, the precision evaluated through repeatability (%CVr) and reproducibility (%CVR) were lower than the Horwitz coefficients of variation criteria. Finally, in the estimation of the uncertainty at the three concentration levels, values of 20.2 ± 2.5 mgdL-1 were determined; 202.3 ± 6.2 mgdL-1 and 380.7 ± 11.0 mgdL-1 using a coverage factor of k=2 with a confidence level of 95%. In conclusion, evidence was obtained that the GOD-PAP glucose enzymatic method is linear, accurate and precise, ensuring accurate and reliable determination of blood glucose.
The objective of this research work is to validate and estimate the uncertainty of the measurement of the glucose oxidase - peroxidase (GOD-PAP) enzymatic method to quantify blood glucose. Calibration curve was prepared in the range of 20 to 400 mgdL-1 with which glucose was quantified in serum sample, and in glucose standards (20, 200 and 380 mgdL-1) during five days of validation. For the validity of the method, the linearity, working range, detection limit, quantification limit, accuracy, precision and uncertainty of the measurement were calculated. In the study of linearity, the regression equation y = 0.0031x + 0.0007 was found with a coefficient of determination of 0.9999. The limits of detection (LOD) and quantification (LOQ) were 2.2 mgdL-1 and 7.4 mgdL-1 respectively. The accuracy was evaluated by the recovery percentage, being satisfactory, with a recovery percentage of 98.4-104% (20 mgdL-1), 99-103.1% (200 mgdL-1) and 98.1-102. 2% (380 mgdL-1). Likewise, the precision evaluated through repeatability (%CVr) and reproducibility (%CVR) were lower than the Horwitz coefficients of variation criteria. Finally, in the estimation of the uncertainty at the three concentration levels, values of 20.2 ± 2.5 mgdL-1 were determined; 202.3 ± 6.2 mgdL-1 and 380.7 ± 11.0 mgdL-1 using a coverage factor of k=2 with a confidence level of 95%. In conclusion, evidence was obtained that the GOD-PAP glucose enzymatic method is linear, accurate and precise, ensuring accurate and reliable determination of blood glucose.
Description
Keywords
Validación, método enzimático GOD-PAP, incertidumbre de medición