Simple Estimation of Uncertainty in the Quantification of Cefazolin by HPLC and Bioassay

Abstract

This work proposes a procedure to estimate the measurement uncertainty for the quantification of cefazolin by high-performance liquid chromatography (HPLC) and microbiological assay (bioassay). HPLC method was performed according to United States Pharmacopeia (USP) and bioassay according to Brazilian Pharmacopeia (FB). The quantification of cefazolin by high-performance liquid chromatography was assess through the calibration curve equation (Y=0.0199 X-0.0323). The HPLC uncertainty was estimated based on the error of the slope and the intercept (between 4.4% and 5.3%). The bioassay uncertainty was estimated based on the standard deviation of inhibition zones of samples and standards solutions (between 6.2 and 6.7%). The uncertainties for bioassay are expected to be higher those obtained by HPLC. Both HPLC method and bioassay estimated uncertainties are reasonable for the scope of each method.

An alternative capillary electrophoresis method for the quantification of caspofungin in lyophilisate powder and its measurement uncertainty

“…The main sources of uncertainty should be considered in the final uncertainty estimation, including sampling (Wunderli, 2003), matrix effects and interferences, environmental conditions (Leito et al, 2002;Lourenço et al, 2012), uncertainties of mass and volumetric equipment, uncertainties of spectrophoto-metric (Sooväli et al, 2006;Hsu and Chen, 2010) and chromatographic equipment (Anglov et al, 2003;Lourenço, 2012), uncertainties of biological and microbiological responses (Niemi and Miemelã, 2001;Lourenço, 2012), purity of reagents and chemical reference substances (Weitzel, 2012), method validation (Brüggemann and Wennrich, 2002) and random variability (Chui et al, 2002).…”

“…Several works regarding the measurement uncertainty from high performance liquid chromatograph methods (Okamoto et al, 2013;Ghisleni et al, 2014b), UV spectrophotometric methods (Saviano and Lourenço, 2013) and microbiological assays (Lourenço, 2012;Ghisleni et al, 2014a) have been reported in literature. However, as found in literature, there is no work describing the identification and quantification of main sources of measurement uncertainty from capillary electrophoresis methods.…”

Métodos espectrofotométrico, RP-UPLC e microbiológico para determinação de linezolida em formas farmacêuticas

“…Na técnica cromatográfica, quanto menor o tamanho da partícula interna que reveste a coluna cromatográfica, maior o número de pratos teóricos, ou seja, é mais eficiente a separação das substâncias que eluem próximas. (CIOLA, 1998, SKOOG et al, 2002 (ANGLOV et al, 2003, LEITO et al, 2002, LOURENÇO et al, 2005, LOURENÇO 2012, LOURENÇO et al, 2004, LOURENÇO et al, 2007, NIEMI et al, 2001, SOOVÄLI et al, 2006, LOURENÇO 2013, OKAMOTO et al, 2013, HSU 2010.…”

“…As variáveis comuns, que podem influenciar o espectro de absorção de uma substância, incluem a natureza do solvente, o pH da solução, a temperatura, as concentrações de eletrólitos e a presença de substâncias interferentes (SKOOG et al, 2002 (CHAMBERS 2006, BUTLER et al, 2011. Os métodos reportados na literatura para a determinação de linezolida incluem espectrofotometria em UV, HPLC usando detecção em UV e fluorescência, eletroforese capilar, TLC seguida de análise densiométrica e ensaios microbiológicos (LOPES et al, 2010, BORNER et al, 2001, MOHAPATRA et al, 2011, LOPES et al, 2009 (ANGLOV et al, 2003, LEITO et al, 2002, LOURENÇO et al, 2005, LOURENÇO 2012, LOURENÇO et al, 2004, LOURENÇO et al, 2007, NIEMI et al, 2001, SOOVÄLI et al, 2006, LOURENÇO 2013, OKAMOTO et al, 2013, HSU 2010.…”

Desenvolvimento de métodos microbiológicos rápidos (MMRs) para avaliação da potência de agentes antimicrobianos e suas incertezas de medição

“…EURACHEM/CITAC, 2012). A incerteza de medição de um resultado pode surgir de muitas fontes, incluindo fatores como amostragem, efeitos da matriz e interferentes, condições ambientais, incerteza proveniente de balanças e equipamentos volumétricos, incerteza proveniente de equipamentos espectrofotométricos e cromatográficos, incertezas provenientes da resposta biológica e microbiológica, pureza de reagentes e substâncias químicas de referência, validação de métodos e erros aleatórios (NIEMI e NIEMELÃ, 2001;BRÜGGEMANN e WENNRICH, 2002;CHUI et al, 2002;LEITO et al, 2002;ANGLOV et al, 2003;WUNDERLI, 2003;LOURENÇO et al, 2004, LOURENÇO et al, 2005, SOOVÄLI et al, 2006LOURENÇO et al, 2007, HSU e CHEN, 2010LOURENÇO, 2012;LOURENÇO et al, 2012, WEITZEL, 2012LOURENÇO, 2013;SAVIANO e LOURENÇO, 2013;GHISLENI et al, 2014; OSTRONOFF e LOURENÇO, 2015).…”