黄连的3D-HPLC特征指纹图谱研究

Study on 3D-HPLC Characteristics Fingerprints of Coptidis Rhizoma

目的:对不同黄连品种、不同产地味连中生物碱类成分进行比较分析,并建立其3D-高效液相色谱(HPLC)特征指纹图谱。方法:采用HPLC-二极管阵列检测器(DAD)法。色谱柱为XtimateTM C18(250mm×4.6mm,5μm),流动相为30mmol/L碳酸氢铵溶液(含0.1%三乙胺、0.7%氨水)-乙腈(梯度洗脱),检测波长为200～400nm,柱温为30℃,流速为1.0ml/min。利用相似度评价软件和SPSS18.0软件对31批味连、6批雅连、4批云连270nm波长下的指纹图谱进行相似度评价和聚类分析。结果:建立了黄连的3D-HPLC-DAD指纹图谱,共得到11个特征指纹峰,其共有峰面积占总峰面积的90%以上。通过与对照品对照,标定了木兰花碱、盐酸药根碱、盐酸非洲防己碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱7种生物碱类成分。其中,盐酸表小檗碱含量的差异是3种黄连主要的鉴别特征。结论:本方法具有良好的精密度、稳定性和重复性,建立的特征指纹图谱可以作为黄连质量评价的主要依据之一。

OBJECTIVE： To analyze the alkaloids component of various Coptidis Rhizoma from different producing areas, and to establish 3D-HPLC characteristics fingerprints. METHODS： HPLC-DAD was adopted. The determination was performed on Xti- mateTM C18（250 mm×4.6 mm, 5 μm） column with mobile phase consisted of 30 mmol/L ammonium hydrogen carbonate solution （in- eluding 0.1% triethylamine, 0.7% ammonia water）-acetonitrile （gradient elution） at the flow rate of 1.0 ml/min. The detection wavelength was set at 200-400 mn and column temperature was 30 ℃. The similarity evaluation and cluster analysis were carried out to obtained fingerprints of 31 batches of Coptis chinensis, 6 batches of C. deltoidea, 4 batches of C. tecta at 270 nm by similar- ity evaluation software and SPSS18.0 software. RESULTS： 3D-HPLC-DAD fingerprints of Coptidis Rhizoma were set up to obtain 11 characteristic fingerprint peaks; the common peak area took up over 90 % of total peak area. 7 kinds of alkaloids were demarcat- ed by comparing with substance control, such as magnoflorine, jateorrhizine hydrochloride, palmatine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride and berberine hydrochloride. Content difference of epiberberine hy- drochloride was symbolic feature of 3 kinds of Coptidis Rhizoma. CONCLUSION： The method is accurate, stable and reproduc- ible. The established characteristic fingerprints can be taken as one of main criteria of quality evaluation of Coptidis Rhizoma.