金银花高效液相指纹图谱条件优化与建立

Optimization of conditions and establishment of the fingerprint of Lonicera japonica Thunb. by HPLC

目的优化金银花饮片指纹图谱建立的条件,并建立金银花高效液相指纹图谱,为金银花饮片的质量鉴定提供依据。方法对金银花指纹图谱建立的条件进行优化,包括检测波长、提取溶剂、提取方法、饮片粒度、提取次数、提取时间、色谱条件和色谱柱等,并运用优化的方法建立金银花指纹图谱,选用Kromasil KR100-5C18(4.6 mm×250 mm,5μm)色谱柱,流动相为乙腈-0.4%磷酸水,梯度洗脱,流速为1.0 ml.min-1,检测波长为330 nm,柱温为30°C。结果建立了精密度、稳定性和重复性均良好的HPLC指纹图谱检测方法和金银花指纹图谱,并利用中药色谱指纹图谱相似度评价软件对山东10批金银花饮片的指纹图谱进行相似度评价,10批饮片指纹图谱相似度均大于0.95。结论优化所得方法简便易行,稳定可靠,重复性好,可为金银花指纹图谱的建立提供方法参考,建立的指纹图谱检测标准可以作为金银花饮片质量评价和品种鉴别的重要依据。

Objective To develop an HPLC method for establishment of the fingerprint of Lonicera japonica Thunb.It may provide a basis for identification for quality of the Lonicera japonica. Methods The established conditions were optimized for establishment of the fingerprint of Lonicera japonica,including wavelength,extract solvent,extract method,particle size,extract times,extract time,chromatographic condition and column.And the fingerprint was established on Kromasil KR100-5C18(4.6 mm×250 mm,5 μm) column.Acetonitrile-water was used as gradient mobile phase at the flow rate of 1.0 ml·min-1.The detection wavelength was 330 nm and the column temperature was 30 ℃. Results An HPLC method with good precision,stability and repeatability was developed.The fingerprint of Lonicera japonica was established by the developed method.The similarity of the 10 determined Lonicera japonicas was evaluated on the Similarity evaluation system for chromatographic fingerprint of Traditional Chinese Medicine.And the similarity of the determined batches was larger than 0.95. Conclusion The developed method is simple and accurate,and can provide a method reference for the establishment of the fingerprint of Lonicera japonica.The established fingerprint can be used in quality evaluation and variety discrimination of Lonicera japonica.