HPLC法测定姜状三七5种皂苷含量

目的:建立测定姜状三七中人参皂苷Rg₁、人参皂苷Rb₁、人参皂苷R₀、竹节参皂苷Ⅳ和竹节参皂苷Ⅳ_a含量的HPLC方法。方法:采用Agilent ZORBAX SB-C₁₈(150 mm×4.6 mm, 5μm)色谱柱,以0.1%磷酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0~5 min, 15%B→20%B; 5~35 min, 20%B→40%B; 35~45 min, 40%B→90%B),流速1.5 mL·min^-1,检测波长203 nm,柱温20℃。结果:人参皂苷Rg₁、人参皂苷Rb₁、人参皂苷R₀、竹节参皂苷Ⅳ和竹节参皂苷Ⅳ_a进样量分别在0.398~7.960、0.049~1.477、1.000~12.500、0.206~5.150和0.200~4.000μg范围内线性关系良好(r>0.9999),平均回收率在99.91%~103.41%之间,RSD≤1.16%。5种皂苷总量:根茎 >块根 >须根,根皮>木质部,六年 >五年 >四年 >三年。结论:建立的方法简便快速,分离效果好,可用于姜状三七的质量检测;测定不同部位、不同组织和不同年限皂苷含量,便于了解姜状三七的化学特性,为姜状三七的质量评价以及合理开发利用提供依据。

Objective: To establish an HPLC method for determining ginsenoside Rg₁, ginsenoside Rb₁, ginsenoside R₀, chikusetsusaponin Ⅳ and chikusetsusaponin Ⅳ_a in P.zinginberensis.Methods: The analysis was carried out on an analytical column Agilent ZORBAX SB-C₁₈(150 mm×4.6 mm, 5μm) with gradient elution by 0.1% phosphoric acid solution(A) -acetonitrile(B) solution(0-5 min, 15%B→20%B;5-35 min, 20%B→40%B;35-45 min, 40%B→90%B), at the detection wavelength of 203 nm and a flow rate of 1.5 mL·min^-1.The column temperature was 20℃.Results: Linearity of ginsenoside Rg₁, ginsenoside Rb₁, ginsenoside R₀, chikusetsusaponinⅣ and chikusetsusaponinⅣ_a standards was established within the concentration range of 0.398-7.960μg, 0.049-1.477μg, 1.000-12.500μg, 0.206-5.150μg and 0.200-4.000μg with r>0.9999, respectively. The average recoveries(n=5) of the ginsenoside Rg₁, ginsenoside Rb₁, ginsenoside R₀, chikusetsusaponinⅣ and chikusetsusaponinⅣ_a were between 99.91%-103.41% with RSD≤1.16%.The total contents of 5 saponins in different parts, different tissues and different years were ranged as follows:rhizome >tuberous >fiberous root; root bark>xylem; 6 years >5 years >4 years >3 years.Conclusion: The developed method is simple, well separated and precise, which can be used to control the quality of P.zingiberensis.Determination of 5 saponins gave an approach to understand characteristics of P.zingiberensis and provided a chemical basis of reasonable exploitation and utilization of P.zingiberensis.