Simultaneous determination of three curative flavonoids and neurotoxic anisatin in star anise fruits by ultra-high performance liquid chromatography-tandem mass spectrometry

Chinese star anise is an anise-scented star-shaped fruit of the evergreen Illicium verum tree. It is generally used as a spice and folk medicine for treating digestive illness. Chinese star anise may be adulterated with Japanese star anise (fruit of Illicium anisatum tree with similar morphology to the former), containing high content of toxic anisatin leading to severe physical disorders. In this work, a UHPLC-MS/MS method has been developed to determine three quality markers (quercetin, kaempferol, and luteolin) and toxicity marker (anisatin) of star anise using quercetin-d3, an exogenous heavy isotopic compound, as internal standard. Chromatographic separation of quality and toxicity marker compounds was achieved using a T3 column within two minutes. The method was successfully developed, validated, and used to quantify the multiple marker compounds in both botanical reference materials and star anise samples. This method can be used for quality control and assessment of Chinese star anise products.