Fingerprint of Marker Substances in Gami-Honghwa-Tang(KH-19) by HPLC-DAD

High Performance Liquid Chromatography-Diode Array Detector(HPLC-DAD)에 의한 가미홍화탕 (KH-19)의 지문 분석

Objectives : This study was aimed to evaluate marker substances in Gami-Honghwa-Tang (KH-19) by high performance liquid chromatography-photodiode array detector (HPLC-DAD). Gami-Honghwa-Tang is composed of nine crude herbs, Rehmanniae Radix Preparata, Angelicae Gigantis Radix, Cnidii Rhizoma, Paeoniae Radix, Corni Fructus, Moutan Cortex Radicis, Lycii Fructus, Carthami Flos and Glycyrrhizae Radix. Methods : The separation was performed on an Aquasil C18 (4.6×250mm) column by gradient elution with 0.05% TFA in H2O - 0.05% TFA in acetonitrile (0 min 100:0, 20 min 90:10, 40 min 70:30, 60 min 50:50, 80 min 0:100, 90 min 100:0) as the mobile phase at a flow-rate of 1.0 ml/min with detection at 190-800nm. Also we examined the contents for bacteria, pesticide residue and harmful heavy metals. Results : HPLC-DAD was employed to determine the quantities and the qualities of several marker substances such as 5­hydroxymethyl-2-furaldehyde (5-HMF), paeonol, loganin, paeoniflorin, glycyrrhizin, and decursin in the KH-19. There were no bacterial contents, pesticide residues, or harmful heavy metals. Conclusions : We suggest these results could be a useful evidence for quality control of KH-19. This method permits fingerprints of selected individual marker substances from herbal prescriptions without derivatization, multiple purification steps, or lengthy separation times.