Thermo Scientific TurboFlow automated, online sample preparation technology was investigated, and with careful optimisation, was shown capable of removing the interfering peaks whilst improving the signal-to-noise ratio and peak intensity of the analyte peaks. The advantage for research laboratories is that the analytical sensitivity at low levels of vitamin D metabolites is enhanced due to much lower background and a reduction in potential ion suppression effects. The TurboFlow™ method now in service has been developed and validated to industry recommended guidelines.