酸枣仁合欢方中5个皂苷成分含量的高效液相-二极管阵列法同时测定

HPLC-DAD simultaneous determination of contents of 5 saponin constituents in SHF

目的建立酸枣仁合欢方(SHF)皂苷部位5个指标成分的含量测定方法,实现一次进样即完成5个指标成分的含量测定的目的。方法利用高效液相-二极管阵列法(HPLC-DAD)以HPLC-DAD法结合线性梯度洗脱的方式进行含量测定。色谱柱:Agilent TC-C_(18)柱(4.6 mm×250 mm,5 m);流动相:乙腈(A)-0.1%磷酸水;流速:1 ml·min^(-1);运行时间:60 min;柱温:30℃,进样量:20μl;检测波长:204 nm。线性梯度洗脱程序为:0~14.5 min,14%~20%A;14.5~32min,20%A;32~39.9 min,20%~29%A;39.9~42 min,29%~90%A;42~57 min,90~100%A;57~60 min,100%A。结果SHF抗抑郁有效部位中6个指标成分峰均可得到良好分离,且在测定浓度范围内具有良好的线性关系(R^2≥0.9991),回收率均大于99%。结论实验所用测定方法简便快速,灵敏度与重现性俱佳,可满足含量测定要求,可实现一次进样同时测定SHF皂苷部位5个指标成分含量的目的,为制定完备的SHF抗抑郁部位的质量控制标准打下基础,为研制质量可控的创新中药提供重要依据。

Objective This research is to establish an HPLC method for the simuhaneous determination of Albiflorin, （ - ） - Sy- ringaresnol -4 - O - β - D - apiofuranosyl - （ 1→2 ） - β - D - glueopyranoside, Betulinic acid, Betulin, Eleutheroside A in Su- anzaorenhehuan Formula （SHF） Saponin. Methods HPLC -DAD analysis was performed on a Agilent TC -C18 column （4.6 mm × 250 mm, 5 μm）. The mobile phase： acetonitrile （A） -0. 1% phosphoric acid aqueous solution. The flow rate： 1 mLmin - 1. Elution time ： 60 min. The column temperature ： 30 ℃. The injection volume ： 20 μL. The detection wavelength ： 204 nm. The elution mode is gradient elution： 0 - 14.5 min, 14% -20% A; 14.5 - 32 min, 20% A; 32 -39.9 min, 20% - 29%A; 39.9 -42 min, 29% -90%A; 42-57 min, 90- 100%A; 57 -60 min, 100%A. Results Under this condition, each peak was well separated and the 6 constituents had good linear relation within their determination of the concentration （R2 〉 0. 9991 ）. The recovery rate is more than 99%. Conclusion The method was investigated in the methodological aspect. Then it is found that the method is rapid, with good sensitivity and reproducibility, which can meet the requirements of content determina- tion. This study providesa scientific basis for the quality control standards of SHF Saponin and the development of effective, quali- ty controllable and innovative traditional Chinese medicine （TCM）.