摘要
目的:采用多种技术手段首次建立能够用于含异荭草苷的中药材及其成药质量控制的异荭草苷对照品。方法:采用紫外光谱、红外光谱、核磁共振波谱(包括1H-NMR和13C-NMR)和串联质谱法进行结构确证,用薄层色谱法和高效液相色谱法进行纯度检测,用质量平衡法(异荭草苷量+有机杂质量+水分的量+残留溶剂量+无机杂质量=100%)进行定量分析,并用定量核磁技术对赋值结果进行验证。其中,为了保障赋值准确性,首先用2种专属性不同的薄层色谱系统对供试品的杂质分布、总量情况进行整体、直观的考察,然后采用2种不同选择性的高效液相色谱系统对其色谱纯度进行准确测定;在上述考察中,用异荭草苷与其异构体荭草苷的分离情况验证所用色谱系统的分离能力,以保障所得色谱纯度结果的准确性。结果:首批异荭草苷对照品的定性鉴别结果与文献报道均一致;采用的2个液相色谱系统和2个薄层色谱系统均能将异荭草苷和荭草苷较好地相互分离,供试品的纯度大于98%;质量平衡法的赋值结果为94.0%,定量核磁共振法的验证结果为92.4%。结论:首批异荭草苷对照品的定性鉴别结果符合要求,定量赋值结果准确、可靠,能够用于含异荭草苷的中药材及其成药的质量控制。
Objective：To establish primary chemical reference substance（CRS） of isoorientin for quality control of isoorientin-containing herbal medicines by various techniques.Methods：The structure of the candidate material was characterized by ultraviolet-visible（UV）,infrared（IR）,liquid chromatography-mass spectrum（MS） and nuclear magnetic resonance（NMR,including1H-NMR and13C-NMR）,and its chromatographic purity was determined by high-performance liquid chromatographic（HPLC） and thin-layer chromatographic（TLC） systems.Mass balance method（the total amount of isoorientin,organic impurities,water,residual solvents plus inorganic impurities was 100 percent） was applied for its quantification while quantitative NMR technique was used to confirm accuracy of the assigned value.In these tests,to guarantee the accuracy of value assignment,two specific TLC systems were adopted to make an overall and visual investigation on the impurity distribution and the total quantity of the candidate material,and then two different HPLC systems were used to determine the accuracy of the chromatographic purity.In the above investigation,the resolution ability of the chromatographic system was verified based on the separation status of isoorientin and its isomer orientin to ensure the accuracy of the chromatographic purity result.Results：The results of qualitative identification of the first batch of isoorientin reference substance were consistent with those reported in literatures and the purity of the candidate material was greater than 98 percent.Isoorientin could get baseline separation from orientin by both of the two HPLC systems adopted or by the two TLC conditions applied.The assigned value by mass balance method was 94.0 %,in accordance with that obtained by quantitative NMR（92.4%）.Conclusion：The structure and purity of isoorientin candidate material met the requirements of CRS for assay.The accuracy of the assigned value obtained by mass balance method was accurate and reliable.Thus,the primary
出处
《药物分析杂志》
CAS
CSCD
北大核心
2015年第6期1072-1077,共6页
Chinese Journal of Pharmaceutical Analysis
基金
国家十二五"重大新药创制"专项(2014ZX09304307-002)
关键词
异荭草苷
对照品标定
定性鉴别
标准物质分析
质量平衡法
定量核磁共振技术
isoorientin reference substance calibration structural characterization standard material analysis mass balance method quantitative NMR technique
