摘要
目的以制备反相高效液相-蒸发光散射检测(RP-HPLC-ELSD)法为基础,大量分离制备达玛烷-3β,12β,20,25-四醇(25-OH-PPD)差向异构体,比较间隔连续进样法和离线制备法大量分离制备25-OH-PPD差向异构体的制备效率,探寻出最佳的分离制备色谱法。方法分别对间隔连续进样法以及离线制备法在不同比例的流动相和不同进样量下对25-OH-PPD差向异构体分离制备情况进行了考察,通过比较制备效率、转移率等筛选出最佳的制备方法。结果 2种方法最佳分离制备色谱条件和制备结果:间隔连续进样法流动相为甲醇-水(83∶17),体积流量20 m L/min,进样质量浓度20 mg/m L,进样量1 m L,20(S)-25-OH-PPD的制备效率为18.01 mg/h,20(R)-25-OH-PPD的制备效率为35.36 mg/h;离线制备法流动相为甲醇-水(81∶19),体积流量20 m L/min,进样质量浓度200 mg/m L,进样量2.5 m L;20(S)-25-OH-PPD的制备效率为50.55mg/h,20(R)-25-OH-PPD的制备效率为51.93 mg/h。结论离线制备法分离制备效率优于小质量浓度间隔连续进样法,离线制备法操作简单,重现性好,制备量大,为25-OH-PPD差向异构体混合物的大量分离制备奠定了良好的基础。
Objective A method of reversed phase high performance liquid chromatogram-light scattering detector（RP-HPLC-ELSD） was developed to separate and prepare a large number of 25-OH-PPD epimeric mixture, in order to explore the best separation conditions between method of continuous sampling interval and off-line preparation method. Methods The separation consequence of the method of continuous sampling interval and off-line preparation method were investigated respectively, under different proportion of mobile phase and concentration of sample volume, the optimum preparation method was screened by comparing the efficiency and transfer rate. Results The suitable operation conditions of the two methods and the result of the preparation were obtained： method of continuous sampling interval as follow： the mobile phase is methanol and water（83∶17）, and the volume flow and injection volume are 20 mg/m L and 1 m L, respectively, while the flow rate is 20 m L/min; Under above conditions the preparation efficiency of 20（S）-25-OH-PPD and 20（R）-25-OH-PPD is 18.01 and 35.36 mg/h, respectively; Off-line preparation method, the mobile phase is methanol and water（81∶19）, and the volume flow and injection volume is 200 mg/m L and 2.5 m L, while the flow rate is 20 m L/min, The preparation efficiency of 20（S）-25-OH-PPD and 20（R）-25-OH-PPD is 50.55 and 51.93 mg/h, respectively. Conclusion Preparation efficiency of off-line preparation method is higher than that of the method of continuous sampling interval; This method is convenient and reliable and has large amount of 20（S）-25-OH-PPD and 20（R）-25-OH-PPD, which can establish a good foundation for the separation and preparation of 25-OH-PPD isomer.
出处
《中草药》
CAS
CSCD
北大核心
2016年第7期1126-1131,共6页
Chinese Traditional and Herbal Drugs
基金
"十一五"国家重大新药创制项目(2009ZX09102-114)
国家自然科学基金资助项目(81273389)
中药标准提取物的工业化色谱制备平台建设(2009ZX09301-012-105B)
