A new kind of ambient ionization method named superhydrophobic preconcentration paper spray ionization mass spectrometry (SHPPSI MS) is introduced, where superhydrophobicity and paper spray mass spectrometry (PS MS) are coupled. The SHPPSI MS requires only microliter amounts of analyte solutions, allows easy sampling procedure, and provides high sensitivity for a diverse array of analytes. It can be used to detect food adulteration at extremely low concentrations. The experimental methodology involves modifying one of the surfaces of a triangularly cut filter paper to make it acquire low surface energy by drop casting a green and ecofriendly superhydrophobic coating material over it followed by drying. A micrometer scale defect was made at close proximity to one of the tips of the paper using a pin. Preconcentration of the sample was accomplished by allowing a 10 μL droplet of an aqueous solution of the analyte to stand at the defect followed by drying naturally. The dried paper was used as the substrate for paper spray mass spectrometry by eluting the analyte with a suitable solvent. This novel technique was used to detect melamine in adulterated milk, whose detection at the ppt level in milk normally needs sophisticated instruments, a larger amount of sample, and a complex sampling procedure, including further purification and separation. The SHPPSI MS detects melamine directly from milk at the sub-ppb level by simply putting a microdroplet of adulterated milk at the substrate and eluting the sample with methanol. This paper-based technique can be a promising tool for direct sensing of analytes such as drugs in body fluids, pesticides in water and soil, etc.