Study On Quality Standard Of Chuanhuangbo By HPLC Fingerprint

The dry bark of Rutaceae plant Phellodendron chinense Schneid. is called Huangbo. It's also known as Chuanhuangbo usually. It is one of the most commonly used Traditional Chinese Medicines with its very wide clinical applications in eliminating heat and dampness, purging sthenic fire, reducing asthenia-heat and bone steaming, relieving toxin for pyocutaneous diseases, and etc. The Chinese Pharmacopoeia in 2005 version separated Huangbo from Guanhuangbo that derived from Phellodendron amurense Rupr. for its individual functions as an herbal medicine.The current quality control of the herb Huangbo and its preparation manufacturing is based on the qualitative and quantitative analysis of berberine. Though some known chemical constituents such as palmatine and jatrorrhizine have been recovered and may assist the quality control study, it is still lack of evidence to characterize Huangbo's overall quality properties. In order to use Huangbo as a medicine appropriately and develop its quality control, this paper did a preliminary study on its chemical compositions and fingerprints.Separation and purification were performed by the silica gel Column Chromatography. Two compounds were isolated and identified as Canthin-6-one (I) and Obacunone (II). It's the first time to get Canthin-6-one (I) from the bark of Phellodendron chinense Schneid.The preliminary experiment was performed by ultrasonic extraction and UV diode array detector. Twelve samples of Huangbo purchased in different areas were tested and prepared with 1% acetic acid-methanol. Separation was performed on a Hypersil BDS C₁₈ column for the large pH range tolerance. The mobile phase was composed by 0.3% triethylamine phosphate-water and acetonitrile to do the gradient elution (0～15min, acetonitrile 15%～25%; 15～35min, acetonitrile25%～40%; 35～40min, acetonitrile40%～90%; 40～60min, acetonitrile 90%～100%) at the flow rate of 0.8ml/min. The Injection volume was 5μl. The UV detection wavelength was 305nm, column temperature was 25℃. The eluent was analyzed by HPLC. The HPLC spectra showed nine typical peaks to compose the fingerprint. Palmatine and berberine are identified. The peaks of Canthin-6-one and Obacunone are difficult to see due to lower content in the herbal medicine.The Chuanhuangbo fingerprint presented in this paper may provide a reference for its quality control research. The current study focused on the UV absorption components that were detected by UV diode array detector. Future work needed on other compounds without UV absorption and are not able to be reflected in the fingerprint. In addition, the strong berberine peak for its high concentration may make some other peaks too low to be showed in the HPLC spectra, and then it' s possible that some small concentrated compounds were lost in the fingerprints. So the chemical constituents and quality control of this herbal medicine need to be studied further maybe by another method on the low concentrated compounds and the ones without UV absorption.