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目的建立高效液相色谱法同时测定黑参中稀有人参皂苷20(S)-Rg_3、20(R)-Rg_3、Rk1、Rg_54种有效成分的含量。方法采用COSMOSIL C_(18)-PAQ(4.6 mm×250 mm,5μm)色谱柱,流动相为水-乙腈梯度洗脱,检测波长203 nm,柱温30℃,流速1.0 m L·min~(-1),进样体积10μL。结果稀有皂苷20(S)-Rg_3、20(R)-Rg_3、Rk1、Rg_5得到良好的分离效果,分别在0.051~0.256 mg·m L~(-1)(r=0.999 9)、0.009~0.280 mg·mL~(-1)(r=0.999 7)、0.051~0.303 mg·m L~(-1)(r=0.999 9)和0.093~0.279 mg·m L~(-1)(r=0.999 9)内与色谱峰面积呈良好的线性关系。平均加样回收率分别为103.9%、99.2%、97.0%和100.6%,RSD分别为0.71%、0.73%、1.97%和0.57%。结论本实验所建立的黑参中皂苷含量测定方法准确、简单、可靠、重复性好,可用于黑参药材中4种主要化学成分含量的测定,其结果可为其质量控制、合理用药及进一步研究开发提供参考。
OBJECTIVE To establish a HPLC method for the simultaneous determination of the active constituents of ginsenoside 20 (S) -Rg_3,20 (R) -Rg_3, Rk1 and Rg_54 in ginseng. Methods The column was COSMOSIL C1818PAQ (4.6 mm × 250 mm, 5 μm). The mobile phase was water-acetonitrile gradient. The detection wavelength was 203 nm, the column temperature was 30 ℃, the flow rate was 1.0 m L · min ~ (- 1), injection volume 10μL. Results The results showed that the best separation efficiencies were obtained at 0.051 ~ 0.256 mg · mL -1 (r = 0.999 9), 0.009 ~ 0.280 (R), Rg 3 (r = 0.999 7), 0.051-0.303 mg · m L -1 (r = 0.999 9) and 0.093-0.29 mg · m L -1 (r = 0.999 9) and the chromatographic peak area showed a good linear relationship. The average recoveries were 103.9%, 99.2%, 97.0% and 100.6%, respectively, with RSDs of 0.71%, 0.73%, 1.97% and 0.57%, respectively. Conclusion The method for determination of saponin in black ginseng established in this experiment is accurate, simple, reliable and reproducible. It can be used for the determination of four main chemical components in Radix Ginseng. The results can be used for its quality control, rational drug use and further Research and development for reference.