Gingival samples treated with toothpastes containing tocopherols (vitamin E) were investigated employing HPLC chromatography. The aim was to verify that vitamin E is actually enriched in the tissue, which could have beneficial effects on oral health. After determination of the tocopherols available in the toothpastes, control samples from healthy test persons and subjects suffering from gingivitis were analyzed. Subsequently, gingival tissues from diseased test persons who treated their teeth with the toothpastes containing tocopherols using various kinds of concentrations or applications were investigated. The first step of the analysis was a fast and careful extraction employing matrix solid-phase dispersion (MSPD). Afterward, the separation of the different tocopherol homologues existing was performed by HPLC chromatography on highly selective C30 RP phases. The identification of the tocopherol homologues was performed using the on-line coupling of HPLC with NMR spectroscopy and mass spectrometry.