Identification and Standardization of the Anti Food Allergy Chinese Herbal Formula, FAHF-2, by HPLC Chromatographic Fingerprinting

Abstract

RATIONALE: FAHF-2, composed of 9 traditional Chinese herbs, was shown to have potential for treating peanut allergy. However, herbal products containing many chemical constituents makes standardization challenging. Chromatographic fingerprinting has been suggested as a practical and comprehensive approach for evaluating the quality of herbal extracts. This study, aimed at developing a botanical drug for treating food allergy, was undertaken to establish an HPLC fingerprint and chemical markers for standardizing FAHF-2. METHODS: Extracts of FAHF-2 and its 9 individual herbs were analyzed under the same conditions. HPLC analysis was performed on a C18 column. A mixture of acetonitrile and 0.15% phosphoric acid was used as mobile phase in a linear gradient with a flow rate of 1 ml/min. The wavelength range of the photodiode array detector was set from 200 to 600 nm. RESULTS: The HPLC fingerprint of FAHF-2 can be used for standardizing FAHF-2 at three levels: (1) the HPLC fingerprint established 30 peaks, which allow us to monitor batch-to-batch consistency; (2) peak contributions by individual herbs were identified by comparing the FAHF-2 fingerprint with individual herbs. Therefore, if there is alteration of HPLC fingerprint, we will be able to determine which individual herb(s) account for the alteration; (3) the chemical constituents of 10 major peaks were identified using chemical standards. These major chemical constituents can be used as chemical markers, and perhaps ultimately for identifying the active constituents in FAHF-2. CONCLUSIONS: HPLC fingerprinting with chemical markers was established for standardizing the herbal compound, FAHF-2, which is critical in the development of FAHF-2 as a botanical drug.