Abstract
To solve the problem of carry-over – a persistent chromatographic challenge for bioanalytical assays with highly sensitive detector such as a mass spectrometry (MS) – a new on-line sample pretreatment HPLC/MS system using a column-switching technique was established. This system was designed to reduce carry-over based on a hydrophobic interaction mechanism using a washing function (multi-mobile phase flow system), as well as to remove impurities on-line in a mobile phase for the pretreatment. As a result, a washing function in this system was enabled to reduce carry-over and to remove impurities in a mobile phase by automatic operation. Therefore, concentration levels of donepezil (DH) in dog plasma as low as 10 pg/mL could be determined using this on-line sample pretreatment HPLC/MS system. In addition, method validation results of specificity, linearity, accuracy, precision, lower limit of quantitation (LLOQ), and carry-over demonstrated that this on-line sample pretreatment HPLC/MS system was robust and valid as a practical assay of drugs and metabolites in the biological samples.
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