Chinese herbal formula is a complex mixture of chemical components. Chromatographic fingerprint has been suggested to be a practical and comprehensive approach for identifying authenticity and evaluating the quality, consistency and the stability of raw herbal materials and herbal extracts. We recently generated a simplified anti-food allergy herbal formula FAHF-5. This study was undertaken to establish the chromatographic fingerprint HPLC analysis of FAHF-5.

The RP-HPLC- DAD system was applied for identification and quality assessment of FAHF-5, containing extracts of 3 traditional Chinese medicines, Ganjiang, Huangbai and Lingzhi. HPLC analysis was performed on a Zorbax SB-C18 column and the column temperature was 30°C. The mixture of acetonitrile (A) and (B) 0.2M phosphoric acid was used as mobile phase in linear gradient mode (5%-30% of A for 050min, 30%75% of A for 5080min), with a flow-rate of 1ml/min.

The chromatographic fingerprint of FAHF-5 was setup by RP-HPLC with DAD detection(scan from 200 to 400nm). When absorbance at 280nm, more peaks and better separation could be obtained. From the chromatographic fingerprint of FAHF-5, 31 diagnostic peaks were observed. The ratios of major peak area between peaks 9, 10+11 and 21 were 1:2.1:4.6. On the basis of the retentive time and the on-line UV spectra of chemical markers, peak 21 in the HPLC fingerprint profile of FAHF-5 was confirmed as berberine and palmatine. Peak 30 was identified as 6-gingerol.

Chromatographic fingerprint with marker compounds can be used as a mean of identification and standardization of FAHF-5 herb product.