一测多评法测定藏药金腰草中4种黄酮类化学成分的含量

目的: 建立藏药金腰草中4 种黄酮类化学成分含量测定的一测多评法,探讨一测多评法在民族药质量控制中的应用。方法: 采用高效液相色谱法,使用Fortis Xi C₁₈(250 mm×4.6 mm,5 μm)色谱柱,流动相为乙腈(A)-1.0% 醋酸水溶液(B),梯度洗脱(0~15 min,10%A→28%A;15~30 min,28%A;30~40 min,28%A→55%A;40~55 min,55%A;55~60 min,55%A→90%A),流速1.0 mL·min^-1,检测波长270 nm,柱温30 ℃,进样量10 μL。以Penduletin 为内参物,分别建立Chrysosplenoside D、Pendulin、ChrysosplenosideB 相对于Penduletin 的相对校正因子,分别采用外标法和一测多评法测定4 种化合物的含量,并比较二者结果的相对误差。结果: Chrysosplenoside D 质量浓度在28.05~140.24 μg·mL^-1,Pendulin 质量浓度在16.53~82.64 μg·mL^-1,Chrysosplenoside B 质量浓度在48.08~240.40 μg·mL^-1,Penduletin 质量浓度在32.53~162.64 μg·mL^-1 范围内线性关系良好;3 种化学成分相对于Penduletin 的相对校正因子分别为0.588、0.629、0.489;且在不同实验条件下重现性良好(RSD < 3.0%);一测多评法的计算结果与外标法测得结果无显著差异。结论: 建立的一测多评法可作为金腰草中4 种黄酮类化学成分的含量测定方法,一测多评法为民族药质量控制提供了一个新的模式与方法。

Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) for determination of four flavones in Chrysosplenium nudicaule Bunge,and discuss application of QAMS in quality control of ethnic medicines. Methods: With the method of HPLC,a Fortis Xi C₁₈ (250 mm×4.6 mm,5 μm) column was adopted. The mobile phase was composed of acetonitrile(A)-1.0% acetic acid solution(B) with gradient elution(0-15 min,10%A→28%A;15-30 min,28%A;30-40 min,28%A→55%A;40-55 min, 55%A;55-60 min,55%A→90%A) at the flow rate of 1.0 mL·min^-1. The detection wavelength was 270 nm,Chrysosplethe column temperature was 30 ℃,and the sample size was 10 μL. Penduletin was selected as an internal standard to establish the relative correction factors (RCFs) of Chrysosplenoside D,Pendulin and Chrysosplenoside B with reference to Penduletin respectively. Both external standard method (ESM) and QAMS were employed for the determination of the contents of four flavones. Meanwhile,relative error (RE) between QAMS and ESM was analyzed to evaluate QAMS method. Results: There were good linearities in the range of 28.05-140.24 μg·mL^-1 for Chrysosplenoside D,16.53-82.64 μg·mL^-1 for Pendulin,48.08-240.40 μg·mL^-1 for Chrysosplenoside B and 32.53-162.64 μg·mL^-1 for Penduletin. RCFs of Chrysosplenoside D,Pendulin and Chrysosplenoside B with reference to Penduletin were 0.588,0.629 and 0.489 respectively,and repeatability was good in different experimental conditions (RSD < 3.0%);There were no significant differences between the quantitative results of the two methods. Conclusion: QAMS method is feasible,and credible,which can be used to determine multiple components in Chrysosplenium nudicaule Bunge. QAMS can be applied as a new strategy for quality control of ethnic medicines.