| Rhizoma Chuanxiong. is used in clinic as an important Traditional Chinese Medicine(TCM).14 samples of Rhizoma Chuanxiong. were purehased from TCM shops in different places of China. The present study provided the methods to isolate constituents and determine the content of the compounds in Rhizoma Chuanxiong. by HPLC. HPLC fingerprint chromatogram was established and the systematic quality evaluation of Rhizoma Chuanxiong. was also developed.Rhizoma Chuanxiong. was extracted with 80% ethanol. by solvent extraction、various chromatographic techniques of isolation(Si gel CC, Sephadex LH-20, Semi-preparative Chromatography) and recrystallization,8 compounds were isolated and purified from CHCl3 and EtOAc extract. By means of spectroscopic analysis and physiochemical properties,8 of them were identified as:ferulic acid (Ⅰ)、6,7-di-hydroxyligustilide (Ⅱ)、4-hydroxy-3-butylphthalide (Ⅲ)、vanillic acid (Ⅳ)、senkyunolide C (Ⅴ)、caffeic acid (Ⅵ)、β-sitosterol (Ⅶ) and daucosterol (Ⅷ).Ferulic acid、6,7-di-hydroxyligustilide and 4-hydroxy-3-butylphthalide in Rhizoma Chuanxiong. were determined by RP-HPLC. The result showed the calibration curves of ferulic acid、6,7-di-hydroxyligustilide and 4-hydroxy-3-butylphthalide were linear in the range of 7.140～71.40μg·mL-1 (r= 0.9999)、39.90～399.0μg·mL-1 (r= 0.9997) and 0.2880～2.880μg·mL-1(r= 0.9998) respectively. The average recovery of ferulic acid、6,7-di-hydroxyligustilide and 4-hydroxy-3-butylphthalide were 98.0%(RSD= 3.4%)、91.0%(RSD= 1.7%) and 98.5%(RSD= 1.5%) respectively. Above methods were simple, rapid and with good reproducibility, It provide a quantitative basis for the quality assessment for Rhizoma Chuanxiong.Ferulic acid and Caffeic acid in Rhizoma Chuanxiong. were determined by RP-HPLC. The result showed the calibration curves of Ferulic acid and Caffeic acid were linear in the range of 13.62-245.2μg·mL-1 (r=0.9995)、29.40～529.2μg·mL-1 (r= 0.9996) respectively. The average recovery of Ferulic acid and Caffeic acid were 90.2%,97.3%, and the RSD were 1.9%,2.0%, respectively. Above methods were simple, rapid and with good reproducibility, It provide a quantitative basis for the quality assessment for Rhizoma Chuanxiong.The RP-HPLC fingerprint analysis for the quality control of Rhizoma Chuanxiong. was established. HPLC with SHINWA-ODS column (250 mm×4.6 mm,5μm). The mobile phase was methanol (A) and 0.5% acetic water solution (B) with gradient elution mode at the flow rate of 1.0 mL·min-1. The gradient condition was 0-5 min, A:10%, B:90%; 5-20 min, A:10%'30%, B:90 %'70%; 20-40 min,A:30%'60%, B:70%'40%; 40-60min, A:60%'70%, B:40%'30%; 60-75 min,A:70%'100%,B:30%'0%. The detection wavelength was 280 nm. Column temperature:25℃.14 batches of Rhizoma Chuanxiong. were determined under the condition described above. With the Within-groups linkage method and the Cosine used as the measurement index, the Hierarchical cluster was applied to the fingerprints of Rhizoma Chuanxiong. and the 14 batches of samples were divided into two grades. Among them the grade I was commendatory. The similarity data were also obtained. These results of the commendatory should be all over 0.90. The results of similarity snalysis were the same as that of cluster analysis. The HPLC chromatographic fingerprints similarity analysis method can be used to control the quality of Rhizoma Chuanxiong. effectively. When unknown samples are analyzed by this method, identification, classification and assessment can be carried out by compared the similarity of standard samples with the unknown ones. A new methodology for the quality control of Rhizoma Chuanxiong. samples has been established in this study. The method should be helpful explore as a new quality control method for traditional Chinese medicine. |
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