Mass spectrometry (MS) and Raman spectroscopy are complementary analytical techniques used to provide information related to chemical structures and functional groups of target analytes. Each instrument provides specific chemical information. If these two analytical tools are coupled online, comprehensive structural information can be simultaneously collected from the analytes of interest without losing any important chemical information. Nevertheless, exploring a suitable interface for coupling of these analytical tools, which are governed with different operation principles, remains challenging. In this study, we used a small piece of tissue paper as an interface for hyphenating a Raman spectroscope and a mass spectrometer online. The paper played multiroles as sample loading substrate and an emitter to generate electrospray. Furthermore, it can facilitate surface-enhanced Raman spectroscopic analysis to improve analyte signals in Raman spectra. A sample droplet was placed on the tissue paper located close to the laser of the Raman spectroscope and the inlet of mass spectrometer. Raman spectra were first collected by the Raman spectroscope through laser irradiation followed by generation of electrospray on the edge of the paper for MS analysis. Positional isomers were used as model samples to demonstrate the effectiveness of the hyphenated analytical tool in distinguishing isomers. The feasibility of using this Raman-MS hyphenated technique for monitoring chemical reactions online in real time was also investigated.