山茱萸药材HPLC指纹图谱的优化研究

Optimization of the HPLC fingerprint for Cornus officinalis

目的：优化山茱萸药材的高效液相色谱指纹图谱。方法：采用Phenomenex C18（250 mm×4.6 mm,5μm）色谱柱,以0.1%甲酸溶液（A）-乙腈（B）为流动相,梯度洗脱（0～15 min,2%B→4%B;15～50 min,4%B→16%B;50～57 min,16%B→18%B;57～67 min,18%B→25%B;67～87 min,25%B→90%B;87～95 min,90%B）,流速1.0 mL·min^-1,检测波长260 nm,柱温30℃。采用中药色谱指纹图谱相似度评价系统软件（2004A版）进行指纹图谱分析。结果：分别优化了检测波长、流动相以及HPLC色谱梯度洗脱条件,标定了19个共有指纹特征峰,确定了莫诺苷、马钱苷、獐芽菜苷、山茱萸新苷、没食子酸、5-羟甲基糠醛、原儿茶酸7个化合物,建立了11个不同批次的山茱萸HPLC指纹图谱,相似度高,均处于0.9以上。结论：该方法方法学考察符合指纹图谱技术要求,可作为控制山茱萸质量标准的依据之一。

Objective： To optimize the fingerprint of Cornus officinalis using HPLC technology. Methods： The Phenomenex C18column（ 250 mm × 4. 6 mm,5 μm） and the diode array detector（ DAD） were used for separation and detection,respectively. The separation process was carried out using a gradient elution（ 0- 15 min,2 %B→4 % B; 15- 50 min,4 % B →16 % B; 50- 57 min,16 % B →18 % B; 57- 67 min,18 % B →25 % B; 67- 87 min,25 % B→90 % B; 87- 95 min,90 % B）,which was based on a mobile phase of the mixture aqueous solution with 0. 1 % formic acid（ A）- acetonitrile（ B） at the flow rate of 1. 0 mL·min^-1. The detection wavelength was260 nm,and the column temperature was 30 ℃. The chromatographic fingerprints were analyzed with the similarity evaluation system for chromatographic fingerprint of TCM（ 2004 A edition）. Results： Totally 19 common fingerprint peaks and 7 constituents including morroniside,loganin,sweroside,cornuside,gallic acid,5- hydroxymethyl furfural（ 5- HMF） and protocatechuic acid were identified after optimization of the detection wavelength,mobile phase and HPLC chromatography gradient elution conditions. Furthermore,the similarity of HPLC fingerprints of 11 different batches of Cornus officinalis was high,being over 0. 9. Conclusion： The present methodology meets the technical requirements of fingerprint and could be a basis for the intrinsic quality control standard for Cornus officinalis.