基于“质-量”双标的丹参质量分析方法研究

Study on quality analysis method of Salviae Miltiorrhizae Radix et Rhizomabased on“quality-quantity”double standard

目的建立以对照药材为基准物质的定性和不依赖多种对照品定量的丹参药材“质-量”双标控制方法。方法采用HPLC法,以对照药材为基准物质建立丹参特征图谱,通过四极杆-飞行时间质谱(quadrupole-timeofflightmass spectrometry,Q-TOF-MS)技术鉴定共有峰的化学成分,以对照药材和供试药材特征峰的相似度,明确药材真伪,即“质”;对内标成分丹酚酸B进行准确定量,以内标成分计,计算不同批次供试品中各特征峰的相对含量,取“平均数-标准差”作为特征峰化学成分相对于内标物质化学成分的相对含量下限,依据特征峰相对含量下限明确丹参药材优劣,即“量”。结果建立的特征图谱及含量测定方法符合方法学考察要求;确定了6个共有色谱峰,分别为迷迭香酸、紫草酸、丹酚酸B、隐丹参酮、丹参酮Ⅰ、丹参酮Ⅱ_(A),各供试药材与对照药材的相似度均大于0.90;确定了丹参药材特征峰化学成分相对含量下限。结论该方法不依赖多种对照品,能清晰、快速地判断药材的真伪优劣,为丹参的质量控制提供参考。

Objective To establish a“quality-quantity”double standard control method for Danshen(Salviae Miltiorrhizae Radix etRhizoma),which was qualitative based on the reference traditional Chinese medicinal materials(TCMM)as standard substances and quantified without relying on multiple reference substances.Methods HPLC technology was used to establish the characteristic chromatogram of Salviae Miltiorrhizae Radix etRhizomabased on the reference TCMM as standard substances.The chemical constituents of common peaks were identified by quadrupole-time of flight mass spectrometry(Q-TOF-MS)technology,and the authenticity of Salviae Miltiorrhizae Radix etRhizomawas determined by the similarity of characteristic peaks of the reference TCMM and the tested TCMM.The above statement is“quality”.The relative content of each characteristic peak in different batches of the tested TCMM was calculated by accurate quantition of internal standard component salvianolic acid B.The“average value-standard deviation”was taken as the lower limit of the relative content of characteristic peak chemical components relative to chemical composition of internal standard substance,and the merits of Salviae Miltiorrhizae Radix etRhizomawas determined according to the lower limit of the relative content of characteristic peak.The above statement is“quantity”.Results The characteristic chromatogram and content determination method met the requirements of methodology investigation.A total of six common chromatographic peaks were determined,which were rosmarinic acid,alkannic acid,salvianolic acid B,cryptotanshinone,tanshinone Ⅰ,tanshinone Ⅱ_(A).The similarity between the tested TCMM and reference TCMM was greater than 0.90.The lower limit of relative content of chemical components of Salviae Miltiorrhizae Radix et Rhizoma characteristic peak was defined.Conclusion The method can judge the authenticity of medicinal materials clearly and quickly without many reference substances,and provide reference for quality control of Salviae Miltiorrhizae Radix et Rhizoma.