Abstract:A high performance liquid chromatography-diode array detection-mass spectrometry method with atmospheric pressure chemical ionization mode (HPLC-DAD-MS-APCI) was developed to determine carotenoids in Scutellaria barbata D. Don (Banzhilian). Results showed that a total of 19 carotenoids including internal standard were separated within 37 min by employing a YMC C30 column and a gradient mobile phase composed of methanol–acetonitrile (86:14, v/v) and methylene chloride. The various carotenoids in S. barbata were identified and quantified using internal standard β-apo-8′-carotenal, in which all-trans-lutein and its cis isomers constituted the largest portion, followed by all-trans-β-carotene and its cis isomers. A high recovery of 93.0–100.9% and a high reproducibility were obtained with this method. This would allow determination of carotenoids in herbal samples, and provide a basis for possible production of functional foods with S. barbata D. Don as raw material.