地黄中5种成分含量测定及HPLC指纹图谱的建立

Content determination of five compounds in Semen Rehmanniae and establishment of its HPLC fingerprint

建立地黄中地黄苷D、益母草苷、松果菊苷、肉苁蓉苷A、毛蕊花糖苷5种成分含量测定方法,并建立地黄高效液相色谱(HPLC)指纹图谱,为地黄质量控制提供科学依据。采用资生堂CAPCELL PAK C_(18)色谱柱,以乙腈-0.1%甲酸溶液为流动相,梯度洗脱,柱温为30℃,变波长测定,流速为1.0 mL/min;通过“中药色谱指纹图谱相似度评价系统(2012年版)”软件分析地黄的HPLC指纹图谱,对其质量一致性进行评价。结果显示,按照所建立的含量测定方法,5种成分在各自浓度范围内具有良好的线性关系(R2>0.999 0),精密度试验、重复性试验、稳定性试验RSD为0.12%~1.58%,平均加样回收率为96.5%~98.6%;建立的地黄HPLC指纹图谱共识别23个共有峰,地黄样品与对照指纹图谱的相似度为0.904~0.950。结果提示,建立的含量测定方法及HPLC指纹图谱适用于地黄质量的评价及生地黄与熟地黄的识别,准确度高,重现性好,可为地黄药材的质量控制提供技术支撑。

The purpose was to establish a content determination method for detecting 5 compounds rehmannioside D,leonuride,echinacoside,castanoside A and acteoside in Semen Rehmanniae,and simultaneous establish its HPLC fingerprint in order to provide scientific support for its quality control.Determination was carried out on CAPCELL PAK C18column(250.0 mm×4.6 mm,5μm),using acetonitrile and 0.1%formic acid solution as mobile phase for gradient elution at a flow rate of 1.0 mL/min under changing wavelength,and column temperature was 30℃.Furthermore,the qualities of Semen Rehmanniae samples were evaluated through analyzing their HPLC fingerprints by using“Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine(2012)”.The results showed that five components presented good linearity in their concentration range(R2>0.9990),and the RSD values of precision,stability and repeatability were between 0.12%to 1.58%,and the average recoveries were within 96.5%-98.6%.The established HPLC fingerprint identified 23 common peaks,and the similarity between samples and the generated reference chromatogram ranged from 0.904 to 0.950.The established content determination method and HPLC fingerprint are accuracy and reproducible,which are suitable for quality evaluation of Semen Rehmanniae and identification cooked Semen Rehmanniae from raw samples,also can provide an important support for the quality control of Semen Rehmanniae.