Tandem mass spectrometric analysis of novel caffeine scaffold-based bifunctional compounds for Parkinson's disease

Novel bifunctional compounds composed of a caffeine scaffold attached to nicotine (C₈ -6-N), 1-aminoindan (C₈ -6-I), or caffeine (C₈ -6-C₈ ) were designed as therapeutics or diagnostics for Parkinson's disease (PD). In order to probe their pharmacological and toxicological profile, an appropriate analytical method is required. The goal of this study is to establish a tandem mass spectrometric fingerprint for the development of quantitative and qualitative methods that will aid future assessment of the in vitro and in vivo absorption, distribution, metabolism, excretion (ADME) and pharmacokinetic properties of these lead bifunctional compounds for PD.

Accurate mass measurement was performed using a hybrid quadrupole orthogonal time-of-flight mass spectrometer while multistage MS/MS and MS³ analyses were conducted using a triple quadrupole linear ion trap mass spectrometer. Both instruments are equipped with an electrospray ionization (ESI) source and were operated in the positive ion mode. The source and compound parameters were optimized for all three tested bifunctional compounds.

The MS/MS analysis indicates that the fragmentation of C₈ -6-N and C₈ -6-I is driven by the dissociation of the nicotine and 1-aminoindan moieties, respectively, but not caffeine. A significant observation in the MS/MS fragmentation of C₈ -6-C₈ suggests that a previously reported loss of acetaldehyde during caffeine dissociation is instead a loss of CO₂ .

The collision-induced tandem mass spectrometry (CID-MS/MS) analysis of these novel bifunctional compounds revealed compound-specific diagnostic product ions and neutral losses for all three tested bifunctional compounds. The established MS/MS fingerprint will be applied to the future development of qualitative and quantitative methods.