用双标定量指纹法定桂附地黄丸平行五波长高效液相色谱对照指纹图谱标准

Determination of the reference fingerprint criteria of Guifu Dihuang Wan parallel five wavelength fingerprints by dual referential fingerprint quantified method

目的用双标定量指纹法研究桂附地黄丸(Guifu Dihuang Wan,GFDHW)对照指纹图谱(RFP)来实现指纹图谱标准的复原。方法采用HPLC等吸收双波长法测定了桂附地黄丸中没食子酸、5-羟甲基糠醛、芍药苷、马钱苷和丹皮酚含量。在210、230、246、265和280nm 5个特征检测波长处建立GFDHW平行五波长指纹谱;以没食子酸和丹皮酚为双参照物峰,采用双标定量指纹法恢复GFDHW-RFP。结果以GFDHW中5组分含量测定结果和平行五波长指纹谱鉴定结果分别聚类分析比较不同厂家产品质量差异,结果以后者更为准确可靠。建立了GFDHW双标定量指纹法技术标准。结论基于多指标定量分析和平行五波长高效液相指纹谱的双标定量指纹法能够确定RFP的标准制剂和实现实时随行定量检测GFDHW指纹图谱,这是GFDHW现代化的有效新方法。

Objective To establish a dual referential fingerprint quantified method（DRFQ） to determine the referential fingerprints（RFP）,and to realize the recovery of a fingerprint standard.Methods Five components of Guifu Dihuang Wan（GFDHW） were determined by RP-HPLC with dual wave length-equal absorbance,which was callic acid（GA）,5-hydroxymethyl-2-furaldehyde（5-HMF）,paeoniflorin（PNF）,loganin（LGN）,and paeonol（POL）.The parallel 5 wavelengths HPLC fingerprints of GFDHW were established under 210,230,246,265 and 280 nm to recover GFDHW-RFP with DRFQ by choosing GA and POL peak as the dual referential peaks.Results With cluster analyzing the 5 components determination and five-wavelength fingerprints,the differences in product quality among different manufacturers were compared,and the latter was more accurate and reliable.The DRFQ technology standard of GFDHW was established.Conclusion Based on multi-target quantitative analyses and parallel multi-wavelength high performance liquid chromatography fingerprints,the standard preparation and its RFP can be found and determined and the real-time accompanying quantitative detection of GFDHW fingerprint is thoroughly realized by DRFQ.This can be a new effective way for GFDHW modernization.