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The tea-like beverage <i>Stevia rebaudiana</i> Bertoni (<i>Stevia</i>) is popular in China because it reduces blood glucose and has a sweet taste. In this work, a comprehensive quality assessment of <i>Stevia</i> led to the discovery of five phenylethanoid glycosides, namely steviophethanoside (<b>1</b>), cuchiloside (<b>2</b>), salidroside (<b>3</b>), icariside D (<b>4</b>), and tyrosol (<b>5</b>). Of them, compound <b>1</b> is a novel compound. Mass spectrometry and NMR spectroscopy were employed to confirm the absolute configuration. A hydrolytic step with 4 N TFA at 95 &#176;C for 4 h was used to confirm the monosaccharides. In addition, Discovery Studio 4.0 was used to predict the ADME and toxicity activity of compound <b>1</b>. The results suggested that compound <b>1</b> was biocompatible and had poor toxicity, which was verified by rat INS-1 islet &#946; cells through an MTT assay. Meanwhile, a significant stimulatory effect on INS-1 cells was observed, which indicated a hypoglycemic effect of compound <b>1</b>. This is the first report that describes a natural, novel, and hypoglycemic phenylethanoid glycoside in <i>Stevia</i>.