HPLC波长切换法同时测定舒糖络颗粒中4种成分的含量

目的: 建立高效液相色谱波长切换法同时测定舒糖络颗粒中葛根素、大豆苷、毛蕊花糖苷、大豆苷元4个活性成分的含量。方法: 采用Thermo C₁₈色谱柱(250 mm×4.6 mm,5 μm),以甲醇(A)-水(B)为流动相进行梯度洗脱(0~10 min,20%A→30%A;10~20 min,30%A;20~25 min,30%A→40%A;25~30 min,40%A→50%A;30~35 min,50%A→20%A),流速1.0 mL·min^-1,柱温30 ℃,检测波长为250 nm(葛根素、大豆苷、大豆苷元)、330 nm(毛蕊花糖苷)。结果: 葛根素、大豆苷、毛蕊花糖苷、大豆苷元的进样量分别在0.590~7.375 μg(r=0.999 9),0.110~1.375 μg(r=0.999 6),0.081 1~1.015 μg(r=0.999 9),0.033 8~0.422 5 μg(r=0.999 9)范围内与峰面积线性关系良好;检测限(信噪比为3: 1)分别为0.048 5、0.012 6、0.016 2、0.005 6 mg·L^-1,定量限(信噪比为10: 1)分别为0.147 5、0.036 6、0.040 6、0.011 2 mg·L^-1;中间精密度的RSD分别为1.3%、0.80%、1.7%、2.6%;平均加样回收率(n=9)为96.11%、104.0%、99.29%、101.5%。3批样品中葛根素、大豆苷、毛蕊花糖苷、大豆苷元的含量分别为6.970~7.038、1.802~1.871、1.438~1.470、0.399~0.460 mg·g^-1。结论: 本法经方法学验证可用于舒糖络颗粒的多指标性成分的检测。

Objective: To develop an HPLC method for simultaneously determining 4 components(puerarin,daidzin,acteoside,daidzein)in Shutangluo granules. Methods: The Thermo C₁₈ (250 mm×4.6 mm,5 μm)column was adopted with the mobile phase of methanol(A)-water(B)by a gradient elution program(0-10 min,20%A→30%A;10-20 min,30%A;20-25 min,30%A→40%A;25-30 min,40%A→50%A;30-35 min,50%A→20%A)at a flow rate of 1.0 mL·min^-1,the column temperature was set at 30 ℃,and detection wavelengths were 250 nm(puerarin,daidzin,daidzein)and 330 nm(acteoside). Results: The calibration curves showed a good linearity within the ranges of 0.590-7.375 μg(r=0.999 9)for puerarin,0.110-1.375 μg(r=0.999 6)for daidzin,0.081 1-1.015 μg(r=0.999 9)for acteoside and 0.033 8-0.422 5 μg(r=0.999 9)for daidzein.The LODs(S/N=3)of puerarin,daidzin,acteoside,daidzein were 0.048 5 mg·L^-1,0.012 6 mg·L^-1,0.016 2 mg·L^-1,0.005 6 mg·L^-1 and LOQs(S/N=10)were 0.147 5 mg·L^-1,0.036 6 mg·L^-1,0.040 6 mg·L^-1,0.011 2 mg·L^-1 respectively.The RSDs of the intermediate precision were 1.3%,0.80%,1.7%,2.6%.The average recoveries(n=9)were 96.11%,104.0%,99.29% and 101.5%.The content of puerarin,daidzin,acteoside,daidzein in three samples was 6.970-7.038 mg·g^-1,1.802-1.871 mg·g^-1,1.438-1.470 mg·g^-1,0.399-0.460 mg·g^-1. Conclusion: Methodological verification proved this method to be convenient for the determination of multi-indexed ingredients and the quality control of Shutangluo granules.