目的: 建立25(R,S)-鲁斯可皂苷元及其光学异构体的1H核磁共振定量法(1H qNMR)含量测定方法。方法: 采用外标法,以鲁斯可皂苷元对照品为对照,分别以δ 5.42、3.45和3.79处的信号为25(R,S)-鲁斯可皂苷元、25R鲁斯可皂苷元和25S鲁斯可皂苷元的定量峰,以δ 6.63处的信号为内标富马酸的定量峰进行校准,对25(R,S)-鲁斯可皂苷元及其光学异构体进行定量研究。结果: 25(R,S)-鲁斯可皂苷元的含量分别为97.88%(RSD=0.93%,n=5),与外标法(97.88%)、质量平衡法(97.98%),紫外分光光度法(98.40%)基本一致;25R-鲁斯可皂苷元含量为49.63%(RSD=1.5%,n=5)和25S-鲁斯可皂苷元含量为46.47%(RSD=1.5%,n=5),测定结果之和与25(R,S)-鲁斯可皂苷元总量基本一致。结论: 1H qNMR方法简便易行,可用于25(R,S)-鲁斯可皂苷元及其光学异构体的含量测定,为光学异构体的含量测定提供了新的技术方法;同时开展了外标法核磁定量测定方法的研究,为今后核磁定量方法的广泛使用进行了新的尝试。
Objective: To establish an 1H quantitative nuclear magnetic resonance (1H qNMR) method for determining the contents of 25(R,S)-ruscogenin, 25R-ruscogenin and 25S-ruscogenin. Methods: The external standard methods were applied with ruscogenin reference as external standard. The quantitative peaks of δ 5.42, δ 3.45 and δ 3.79 for 25(R,S)-ruscogenin, 25R-ruscogenin and 25S-ruscogenin, respectively and δ 6.63 for fumaric acid as the internal substance,were used to be calculated. Results: The content of 25(R,S)-ruscogenin was 97.88%(RSD=0.93%, n=5), which was nearly consistent with the external method (98.60%), the mass balance method(97.98%) and UV spectrophotometer method(98.40%). The content of 25R-ruscogenin was 49.63% (RSD=1.5%, n=5) and the content of 25S-ruscogenin was 46.47%(RSD=1.5%, n=5), the total of which were almost identical with the amount of 25(R,S)-ruscogenin. Conclusion: 1H qNMR is a simple and convenient method for the determination of 25R and 25S-ruscogenin. It provides a new method for the determination of optical isomers. At the same time, the exploration of the external standard method for nuclear magnetic quantitative determination has been carried out, which provides a new method for the future use of nuclear magnetic quantitative methods.
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