Analysis of albendazole and metabolites by RAM-cLC-MS/MS using online direct pre-concentration of biofluids

Lima, Adriel M.; Lanças, Fernando M.; Santos-Neto, Álvaro J.

Palavras-chave:  Column switching, Restricted-access media (RAM), Capillary liquid chromatography, Antihelminthics, Sample preparation automation.

Resumo:  Analysis of drugs and other related molecules in biological fluids is essential in the pharmaceutical field. Currently, the demand for faster and more complex analysis drives analytical chemistry to develop innovative solutions. Multidimensional liquid chromatography columns for coupling with direct injection of biological fluids has gained attention in recent years. At the same time, the coupling between liquid chromatography and mass spectrometry gave remarkable scientific development in biochemical and biomedical area. This study evaluated the reduction in scale of column switching systems using RAM column for the analysis of anthelmintics in biological fluids. A column switching system was developed with capillary columns with 200 μm of inner diameter. This system was coupled to tandem mass spectrometry providing highly sensitive and simultaneous analysis, and low sample consumption. The determination of albendazole and some of its biotransformation products was validated in biofluids, consuming, per analysis, less than 8 min and only 1 μL of diluted sample. This capillary system contrasts with the conventional system commonly used, which consume between several hundred and a thousand times more sample to achieve the same detectability. The quantification limits obtained for the anthelmintics were between 2 to 5 ng ml-1. The values for intra-day and inter-day precision were less than 20% for LQs and less than 14.5% for the other levels. The accuracy for LQs was within ± 20% and for the other concentration levels within ± 13.1%. The linearity in plasma was obtained between LQ and 500 ng mL-1 for the studied analytes with correlation coefficients (r) greater than 0.996 for all compounds. Importantly, the method used only sample of 333 nL to reach the quantification level of a few nanograms per milliliter, while methods in conventional scale consumed between 100 μL and 1 mL of sample to achieve the same values.

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