Reversed-Phase High Pressure Liquid Chromatography Separation of Deuterated Compounds of Biological Importance from Their Protio Analogs

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Abstract
A rapid and a practical HPLC method with UV-VIS detection was developed for the separation and analysis of deuterated carotenoids from their protio analogs. Four different chromatography systems were developed. The results showed that with reversed-phase C18 columns it was possible to baseline resolve fully deuterated carotenoids from the nondeuterated analogs. In all instances the deuterated compound eluted ahead of its protio analog indicating that van der Waals forces are operational during the separation process. Specificity, sensitivity, and reproducibility make these methods particularly suitable in plant chemistry for semi-preparative purification processes and methodologies.