Tian Xiang, Ding Rui-Xue, Ba Gen-Na, Bai Yu-Xia, Laxi Na-Mu-Ji-la, Rao Xiao-Yong, Luo Xiao-Jian
Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China.
Mongolian Medicine College, Inner Mongolia University for Nationalities, Tongliao 028000, China.
Zhongguo Zhong Yao Za Zhi. 2018 Oct;43(19):3962-3969. doi: 10.19540/j.cnki.cjcmm.20180726.009.
To establish the high performance liquid chromatography (HPLC) fingerprint for Digeda-4 decoction (DGD-4D), determine the contents of aesculetin, geniposide, picroside Ⅰ, picroside Ⅱ and ellagicacid in DGD-4D, and provide the scientific foundation for quality control of DGD-4D. The analysis was performed on Diamonsil(2) C₁₈ (4.6 mm×250 mm,5 μm) column, with methanol-0.1% phosphoric acid aqueous solution as mobile phase for gradient elution. The flow rate was 1.0 mL·min⁻¹; injection size was 10 μL; temperature was maintained at 30 °C, and the detection wavelength was set at 254 nm. The common mode of DGD-4D HPLC fingerprint was established, and the hidden information was analyzed by Chemometrics. Chromatographic peaks for DGD-4D were identified by HPLC and quantitative analysis was conducted for characteristic peaks. There were 17 common peaks in the fingerprints and the similarity of the fingerprints was over 0.9 in all 15 batches. The samples were broadly divided into four kinds by principal component analysis and clustering analysis. Four marker compounds were verified by partial least squares discriminant analysis, and No. 9, 12 and 14 peaks were identified as geniposide, picroside Ⅱ, and picroside Ⅰ respectively. The average recoveries were in the range of 95.91%-97.31%. The HPLC fingerprint method for content determination is reliable, accurate, rapid, simple, and reproducible, and can be used as one of the effective methods to control the quality of DGD-4D.
建立地黄4号方（DGD - 4D）的高效液相色谱（HPLC）指纹图谱，测定DGD - 4D中秦皮乙素、栀子苷、龙胆苦苷Ⅰ、龙胆苦苷Ⅱ和鞣花酸的含量，为DGD - 4D的质量控制提供科学依据。采用Diamonsil(2) C₁₈（4.6 mm×250 mm，5 μm）色谱柱，以甲醇 - 0.1%磷酸水溶液为流动相进行梯度洗脱。流速为1.0 mL·min⁻¹；进样量为10 μL；柱温保持在30℃，检测波长设定为254 nm。建立了DGD - 4D HPLC指纹图谱的共有模式，并采用化学计量学方法对其中隐藏的信息进行分析。通过HPLC对DGD - 4D的色谱峰进行指认，并对特征峰进行定量分析。指纹图谱中共有17个共有峰，15批样品指纹图谱相似度均大于0.9。通过主成分分析和聚类分析将样品大致分为四类。采用偏最小二乘法判别分析对4种指标性成分进行验证，确定9号、12号和14号峰分别为栀子苷、龙胆苦苷Ⅱ和龙胆苦苷Ⅰ。平均加样回收率在95.91% - 97.31%之间。该HPLC指纹图谱含量测定方法可靠、准确、快速、简便、重复性好，可作为控制DGD - 4D质量的有效方法之一。
