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Dimethyl sulfoxide (DMSO) has wide biomedical applications such as cryoprotectant and hydrophobic drug carrier. Here, we report for the first time that DMSO can generate a distinctive chemical exchange saturation transfer (CEST) signal at around −2 ppm. Structural analogs of DMSO, including aprotic and protic solvents, also demonstrated CEST signals from −1.4 to −3.8 ppm. When CEST detectable barbituric acid (BA) was dissolved in DMSO solution and was co‐loaded to liposome, two obvious peaks at 5 and −2 ppm were observed, indicating that DMSO and related solvent system can be monitored in a label‐free manner via CEST, which can be further applied to imaging drug nanocarriers. With reference to previous studies, there could be molecular interactions or magnetization transfer pathways, such as the relayed nuclear Overhauser enhancement (rNOE), that lead to this detectable CEST contrast at negative offset frequencies of the Z‐spectrum. Our findings suggest that small molecules of organic solvents could be involved in magnetization transfer processes with water and readily detected by CEST magnetic resonance imaging (MRI), providing a new avenue for detecting solvent–water and solvent–drug interactions. [ABSTRACT FROM AUTHOR]