Goji berry, the fruits of Lycium barbarum L., in Chinese name of gouqizi, are very popular edible materials and traditional Chinese medicine (TCM) in China and overseas. In previous work, over 100 chemical constituents were characterized from Goji berry by systematical chromatographic separation and capillary electrophoresis, etc. However, it is not common on studying the glycosidic components of spermine alkaloids which may relate to the increasing sweet taste of varieties of Goji berry. This paper provided the discovery of thirty alkaloids including twenty-five glycosidic components from the aqueous extract of latest planted variety of Goji berry. Of which nineteen structures bear novel pentose glycosyl groups. The molecular networking based on the high performance liquid chromatography quadrupole flight ion mass spectrometry (HPLC-Q-TOF MS) data analysis were firstly used in the chemical studies of Goji berry, which should lay a foundation for the in-depth study of Goji berry drug development.