Development of a silica monolith modified with Fe3O4 nano‐particles in centrifugal spin column format for the extraction of phosphorylated compounds

In this study, citrate‐stabilised iron oxide nano‐particles (～16 nm) have been immobilised on commercial silica monolithic centrifugal spin columns (MonoSpin) for the extraction of phosphorylated compounds. Two alternative strategies were adopted involving either direct electrostatic attachment to an aminated MonoSpin (single‐layer method) in the first instance, or the use of a layer‐by‐layer method with poly(diallyldimethylammonium) chloride. Field‐emission scanning electron spectroscopy and energy‐dispersive X‐ray spectroscopy was used for confirming notably higher coverage of nano‐particles using the layer‐by‐layer method (2.49 ± 0.53 wt%) compared with the single‐layer method (0.43 ± 0.30 wt%). The modified monolith was used for the selective separation/extraction of adenosine monophosphate, adenosine diphosphate and adenosine triphosphate with elution using a phosphate buffer. A reversed‐phase liquid chromatographic assay was used for confirming that adenosine, as a non‐phosphorylated control was not retained on the modified MonoSpin devices, whereas recovery of 80% for adenosine monophosphate, 86% for adenosine diphosphate and 82% for adenosine triphosphate was achieved.