Quality and antioxidant activity detection of Crataegus leaves using on-line high-performance liquid chromatography with diode array detector coupled to chemiluminescence detection

An on-line high-performance liquid chromatography–diode array detector combined with chemiluminescence detection (HPLC–DAD–CL) method was developed to evaluate the antioxidant activity and quality of Crataegus leaves derived from different species and habitats. The chromatographic fingerprint and active profiles of Crataegus leaves were simultaneously achieved by HPLC–DAD–CL. Main antioxidants in Crataegus leaves were identified by HPLC–DAD–CL and HPLC/ESI/MS. Data analysis based on the total antioxidant activities of 10 batches of samples indicated that there were obvious differences among samples from different species and habitats. Good precision and reproducibility were obtained for DAD and CL detection with relative standard deviations less than 1.0% for retention time and 4.0% for intra and inter-day variations. The results suggested that the on-line method might provide potential information for the identification and quantitation of active markers, which were directly associated with the quality of herbal medicines.