一测多评法测定龙血竭中5种有效成分

Determination of five active components in Draconis Resina by QAMS method

目的建立龙血竭中5种有效成分的一测多评法,探讨一测多评法在龙血竭质量控制中的应用。方法采用高效液相色谱法,使用Fortis Xi C18(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈(A)-1.0%冰醋酸水溶液(B),梯度洗脱,0~10 min,25%~30%A;10~60 min,30%~50%A,体积流量1.0 mL/min;检测波长278 nm;柱温30℃;进样量10μL。以紫檀茋为内参,分别建立7,4′-二羟基黄酮、白藜芦醇、龙血素A和龙血素B相对于紫檀茋的相对校正因子,分别采用外标法和一测多评法测定5种成分的量,并比较二者结果的相对误差。结果 7,4′-二羟基黄酮质量浓度在10.23~102.27μg/mL、白藜芦醇质量浓度在11.01~110.14μg/mL、龙血素A质量浓度在9.47~94.72μg/mL、龙血素B质量浓度在11.59~115.90μg/mL、紫檀茋质量浓度在24.35~243.52μg/mL线性关系良好;4种化学成分相对于紫檀茋的相对校正因子分别为0.626、1.064、1.154、0.837;且在不同实验条件下耐用性良好(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龙血竭中5种有效成分的测定方法,一测多评法为龙血竭质量控制提供了新的模式与方法。

Objective To establish a quantitative analysis of multi-components by single marker（QAMS） for determination of five active components in Draconis Resina and discuss application of QAMS in quality control of ethnic medicines.Methods Using the method of HPLC,the Fortis Xi C18 column（250 mm × 4.6 mm,5 μm） was used.The mobile phase was composed of acetonitrile（A）-1.0% acetic acid（B） with gradient elution（0—10 min,A：25%→30%; 10—60 min,A：30%→50%） at the flow rate of 1.0 mL/min.The detection wavelength was 278 nm,the column temperature was 30 and the sample size was 10℃ μL.Pterostilbene was selected as an internal standard to establish the relative correction factors（RCFs） of 7,4′-dihydroxyflavone,resveratrol,loureirin A,and loureirin B with reference to pterostilbene so as to achieve simultaneous determination of multi-indexed components.The contents of five active components were determined by both external standard method（ESM） and QAMS.Meanwhile,relative error（RE） between QAMS and ESM was analyzed to evaluate QAMS method.Results There were good linearities in the range of 10.23—102.27 μg/mL for 7,4′-dihydroxyflavone,11.01—110.14 μg/mL for resveratrol,9.47—94.72 μg/mL for loureirin A,11.59—115.90 μg/mL for loureirin B and 24.35—243.52 μg/mL for pterostilbene,RCFs of 7,4′-dihydroxyflavone,resveratrol,loureirin A and loureirin B with reference to pterostilbene were 0.626,1.064,1.154,and 0.837 respectively,and repeatability was good in different experimental conditions（RSD 3.0%）.There were no significant difference between the quantitative results of the two methods.Conclusion QAMS method is feasible,credible,and can be used to determine multiple components in Draconis Resina.QAMS can be adopted as a novel strategy for quality control of ethnic medicines.