Quality Of Arnebia Euchroma Royle Johnst

Objective:Study on the quality of radix Arnebeia euchroma Royle Johnst was conducted by TLC, UV and HPLC. A fingerprint established through HPLC. The effective method was chosen to evaluate the quality of different sources of Arnebeia euchroma Royle Johnst.Methods:The extraction effects of petroleum ether, ethyl acetate, chloroform, dichloromethane, normal hexane and ethyl alcohol were investigated for naphthoquinone in Arnebia euchroma Royle Johnst. L9(34) orthogonal experiments were taken for alcohol extraction process optimization. Ten batches of Arnebia euchroma Royle Johnst from different markets in different districts of Xinjiang were collected, identified by thin layer chromatography. UV spectrophotometry was used to measure the hydroxyl naphthoquinone pigment contents and reversed-phase HPLC for the contents of L-shikonin, Acetylshikonin, Deoxyshikonin and β,β’-dimethyl acryloyl shikonin. The conditions of HPLC were as follows:chromatographic column Scienhome Kromasil C18(4.6mm×200mm,5μm), flowing phase acetonitrile-0.3%phosphoric acid, gradient elution, flow rate1.0mL/min, column temperature35℃, wavelength274nm and elution time50min.The national pharmacopoeia committee of Chinese medicine chromatographic fingerprint similarity evaluation software system was used to analyze data and establish HPLC fingerprint common model of Arnebia euchroma Royle Johnst. Results:1. Optimum extraction process was as follows:coarse powder of Arnebia euchroma Royle Johnst quenched8h in12times the volume of95%ethyl alcohol;2. Identified by thin layer chromatography, four reference substances of L-shikonin, Acetylshikonin, Deoxyshikonin and β,β’-dimethyl acryloyl shikonin showed distinct spots under UV light (366nm), those ten batches of Arnebia euchroma Royle Johnst either.3. The hydroxyl naphthoquinone pigment contents of those ten batches were included in the range of0.6-17.3mg measured by UV spectrophotometry;4. The linearity was good for L-shikonin (r=0.9998,0.175-2.8μg/mL),Acerylshikonin(r=0.9999,25-400μg/mL), Deoxyshikonin (r=0.9999,0.375-6mg/mL) and β,β’-dimethyl acryloyl shikonin(r=0.9999,25-400μg/mL) measured by HPLC. The content range of L-shikonin, Acetylshikonin, Deoxyshikonin and β,β’-dimethyl acryloyl shikonin was0.01～0.96mg/g,0.39-24.1mg/g,0.77～31.1mg/g and0.25-13.2mg/g, respectively;5. The HPLC fingerprint common model of Arnebia euchroma Royle Johnst was established,14common peaks were calibrated and4chromatographic peaks were attributed. There was great similarity difference between those ten batches and the contrast calculated by fingerprint similarity software.Conclusion: l.The fingerprint method is simple and feasible;2.The ten batches of Arnebia euchroma Royle Johnst from different resources are identified as Arnebia euchroma Royle Johnst by thin layer chromatography;3.The inherent quality of Arnebia euchroma Royle Johnst can be shown more thoroughly by measuring the content of naphthoquinone in Arnebia euchroma Royle Johnst;4.Scientific evidence can be provided for QC research of Arnebia euchroma Royle Johnst by using the fingerprint method.