Biological fingerprinting analysis of the traditional Chinese prescription Longdan Xiegan Decoction by on/off-line comprehensive two-dimensional biochromatography

A comprehensive two-dimensional biochromatography method using a silica-bonded human serum albumin (HSA) column and a RP-HPLC column was developed for the biological fingerprinting analysis of bioactive components in a traditional Chinese medicine (TCM) prescription, Longdan Xiegan Decoction (LXD). The biochromatography with HSA-immobilized stationary phase was applied to study the interaction of multiple components in LXD with HSA in the first dimension, and fractions of HSA column were further separated by a silica monolithic ODS column (on-line)/an ODS column (off-line) coupled with a diode array detector and an atmospheric pressure chemical ionization mass spectrometer (APCI-MS). More than 100 compounds in LXD that interacted with the immobilized HSA were separated and analyzed. Among them 19 compounds were identified based on their retention values, UV spectra, molecular weights and mass spectra. The results show that the developed comprehensive two-dimensional biochromatography system reported here is capable of being used for biological fingerprinting analysis of natural products in complex matrices such as extracts of TCMs and their prescriptions.