RESUMO
In the process of drug development, preclinical safety studies are to be performed that require the analysis of the compound at very low concentrations with high demands on the performance of the analytical methods. In the current study, a UPLC-MS/MS method was developed and validated to quantify hydroxyzine hydrochloride in an extracellular solution used in a hERG assay in concentrations ranging from 0.01 to 10µM (4.5ng/ml-4.5µg/ml). Chromatographic separation was achieved isocratically on an Acquity BEH C18 analytical column. The assay was validated at concentrations of 0.11-1.1ng/ml in end solution for hydroxyzine hydrochloride. Linearity was demonstrated over the range of concentrations of 0.06-0.17ng/ml and over the range of concentrations of 0.6-1.7ng/ml in end solution with the coefficient of correlation r>0.99. Accuracy of the achieved concentration, intra-run, and inter-run precision of the method were well within the acceptance criteria (being mean recovery of 80-120% and relative standard deviation ≤10.0%). The limit of quantification in extracellular solution was 0.09ng/ml. Hydroxyzine hydrochloride in extracellular solution proved to be stable when stored in the fridge at 4-8°C for at least 37 days, at room temperature for at least 16 days and at +35°C for at least 16 days. The analytical method was successfully applied in hERG assay.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Líquido Extracelular/química , Hidroxizina/análise , Espectrometria de Massas em Tandem/métodos , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Canais de Potássio Éter-A-Go-Go/genética , Humanos , Limite de Detecção , Modelos Lineares , Padrões de Referência , Reprodutibilidade dos Testes , SoluçõesRESUMO
BACKGROUND: The high-throughput analysis of strontium and calcium in plasma, tissue and bone (femur) using inductively coupled plasma-MS is described. Method validation, the results and experience obtained during sample analysis are highlighted. RESULTS: The different matrices were destructed with concentrated nitric acid by heating at 60°C overnight. Using this approach it was possible to analyze large numbers of samples in parallel. CONCLUSION: Inductively coupled plasma-MS proved to be a highly sensitive, robust and efficient technique for the analysis of several different biological samples. A total of 6767 samples were analyzed, and the performance of the method was illustrated by the fact that only 0.2% of the samples had to be reanalyzed due to anomalous results and ISR for all matrices fulfilled the acceptance criteria.
