RESUMEN
As part of the formulation of a cell-based pharmaceutical product, cells were harvested from mice and incubated in a cocktail containing cell culture media and high levels of trifluoroacetic acid (TFA). The cells were washed with a phosphate-buffered saline solution to remove residual cell culture media and other reagents before the cells were infused back into the mice from which they originated. Because of the potentially toxic nature of the TFA, the cells were washed multiple times and the final wash was monitored for residual TFA in order to demonstrate the efficient removal of the reagent before the cell product could be reintroduced into the test animal. This report describes the method that was developed incorporating anion-exchange chromatography with suppressed conductivity detection for the analysis of residual TFA (down to 50 ng/ml) in the presence of high concentrations of phosphate and chloride interferences. The ultimate sensitivity of the method was improved by selectively removing halide anions using a silver cartridge before sample analysis. The method proved to be rugged and reproducible enough to be validated and used to monitor residual TFA levels in cell washes in support of an acute toxicological study. Results demonstrating the method's sensitivity, selectivity, precision and linearity were reported.
Asunto(s)
Cromatografía por Intercambio Iónico/métodos , Fosfatos/química , Cloruro de Sodio/química , Ácido Trifluoroacético/análisis , Tampones (Química) , Estándares de Referencia , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Analytical methods were developed for the separation of the enantiomers of four cyclopentenone precursors of prostaglandins. The resolution obtained is correlated with the chemical environment around the chiral center of the cyclopentenones. The analytical methods were scaled up to preparative loadings and the chromatographic parameters were varied to determine their effect on the preparative separations. The correlation between analytical resolution and preparative resolution was also investigated. In addition to the precursors, the preparative resolution of the enantiomers of a synthetic prostaglandin analogue was investigated.