Pérez CM, Suárez PY, González RH, Achong GL, Borrell CY

Language: Spanish
References: 7
Page: 232-244
PDF size: 258.87 Kb.

ABSTRACT

Introduction: complexity and diversity in the structure of Streptococcus pneumoniae capsular makes the analytical methodology for characterization and quality control a troublesome aspect. The colorimetric m/hydroxibiphenil method devised by Blumenklrantz and Asboe-Hansen in 1973 allows reducing the disadvantages of the suggested method in the European Pharmacopea 6th edition, 2011.
Objective: to validate the m-hydroxybiphenil method for quantitation of uronic acid in purified Streptococcus pneumonia polysaccharides.
Methods: the length of validity of a sodium tetraborate solution, responsible for generating the analytical response, was estimated, and additionally, a methodology for quantitation of uronic acid by the m-hydroxybiphenil method was selected in which a standard galacturonic acid solution at 1mg/ml was used. The validation parameters were evaluated as follows: linearity, accuracy, precision, range and specificity, according to the present requirements. This method was compared with the carbazol method described in the European Pharmacopea 6th edition, 2011 for quantitation of uronic acid.
Results: it was demonstrated that the useful lifetime of the sodium tetraborate solution was 24 hours. The m-hydroxybiphenil method was specific, linear, accurate and precise in the range of 2 to 20 µg/mL because the set acceptance criteria were satisfactorily complied with for each of them. The comparison of the suggested method with the standardized method yielded that no statistically significant differences exist between them.
Conclusions: the colorimetric m-hydroxybiphenil method proved to be valid for the quality control of purified Streptococcus pneumonia polysaccharide samples and the results are comparable to those of the recommended method in the European Pharmacopea, 2011.

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