Simultaneous HPLC analysis of neurotransmitters can be challenging due to the similarity of their structures.
Simultaneous HPLC analysis of neurotransmitters can be challenging due to the similarity of their structures. Furthermore, since neurotransmitters are small polar molecules, they can be difficult to retain on a standard C18 column. Imtakt's multi-mode Scherzo SS-C18 column overcomes these challenges through the innovative nature of its stationary phase. The Scherzo SS-C18 stationary phase contains a large amount of strong ionic ligands, which enables the column to retain polar ionic compounds better than a standard C18 column (see Figure 1).
Figure 1: Three novel multi-mode stationary phases, SS-C18 far left.
All data was generated with a semi-micro HPLC system equipped with UV or ELS detection.
Separation of 13 key neurotransmitters was performed using gradient elutions on Scherzo SS-C18 (150 × 3 mm) and Unison UK-C18 (150 × 3 mm) HPLC columns (see Figure 2). The Scherzo SS-C18 multi-mode column separated the neurotransmitters in well resolved peaks within a run time of 25 min. The more traditional C18 column, Unison UK-C18 column, could not retain these compounds sufficiently, as the peaks were clustered together and eluted close to the void volume. The Scherzo SS-C18 multi-mode column outperformed the more traditional C18 column.
Figure 2: Scherzo SS-C18 simultaneously separates 13 key neurotransmitters
More targeted separation of monoamine/amino acid neurotransmitters was performed using isocratic elution on a Scherzo SS-C18 column of dimensions 250 × 3 mm (see Figure 3). This data shows baseline separation of neurotransmitters that have similar structures, which would be difficult to accomplish on a traditional C18 column. All peaks were well resolved, and the use of ammonium phosphate buffer resulted in sharper peaks than did the use of phosphoric acid buffer.
Figure 3: Scherzo SS-C18 retains monoamine/amino acid neurotransmitters.
The new multi-mode Scherzo SS-C18 column showed better separation of neurotransmitters than a standard C18 column. Thirteen key neurotransmitters can be separated simultaneously with this column without any coelution. Moreover, neurotransmitters of similar structure, such as L-adrenaline and L-noradrenaline, are well resolved on this column. The unique ionic/hydrophobic stationary phase on the Scherzo SS-C18 provides a better alternative to scientists seeking to retain and resolve small polar neurotransmitters.
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