HPLC法测定益心巴迪然吉布亚颗粒中4种化学成分的含量

目的: 建立同时测定益心巴迪然吉布亚颗粒中木犀草素-7-O-β-D-葡萄糖醛酸苷、迷迭香酸、香叶木素-7-O-β-D-葡萄糖醛酸苷及田蓟苷4种化学成分含量的高效液相色谱方法。方法: 采用Shim-pack ODS(250 mm×4.6 mm,5 μm)色谱柱,以乙腈(A)-0.5%甲酸水溶液(B)为流动相,梯度洗脱(0~30 min,17%A;30~55 min,17%A→25%A;55~80 min,25%A→30%A),流速1.0 mL·min^-1,检测波长330 nm,柱温35 ℃。结果: 木犀草素-7-O-β-D-葡萄糖醛酸苷、迷迭香酸、香叶木素-7-O-β-D-葡萄糖醛酸苷、田蓟苷分别在3.184~31.84 μg·mL^-1(r=0.999 6)、2.184~21.84 μg·mL^-1(r=0.999 7)、2.784~27.84 μg·mL^-1(r=0.999 7)、7.9~39.5 μg·mL^-1(r=0.999 3)范围内呈良好线性关系;益心巴迪然吉布亚颗粒中上述4种有效成分的平均回收率(n=9)分别为99.65%、99.58%、100.1%和99.92%,RSD分别为0.97%、1.4%、1.0%和0.40%。2个厂家6批益心巴迪然吉布亚颗粒测定结果,木犀草素-7-O-β-D-葡萄糖醛酸苷含量为0.390~0.424 mg·g^-1,迷迭香酸含量为0.431~0.482 mg·g^-1,香叶木素-7-O-β-D-葡萄糖醛酸苷含量为0.468~0.532 mg·g^-1,田蓟苷含量为0.614~0.656 mg·g^-1。结论: 本方法经验证,可为益心巴迪然吉布亚颗粒多指标质量评价及控制提供科学依据。

Objective: To establish an HPLC method for simultaneous determination of four components (luteolin-7-O-β-D-glucuronide,rosmarinic acid,diosmetin-7-O-β-D-glucuronide and tilianin)in Yixin Badiranjibuya granules. Methods: The samples were separated on Shim-pack ODS column (250 mm×4.6 mm,5 μm)by gradient elution with acetonitrile (A)-0.5% formic acid aqeous solution (B)(0-30 min,17%A;30-55 min,17%A→25%A;55-80 min,25%A→30%A)as the mobile phase at a flow rate of 1.0 mL·min^-1.The detection wavelength was set at 330 nm and the column temperature was maintained at 35 ℃. Results: The calibration curves of luteolin-7-O-β-D-glucuronide,rosmarinic acid,diosmetin-7-O-β-D-glucuronide,and tilianin were in good linearity over the ranges of 3.184-31.84 μg·mL^-1(r=0.999 6),2.184-21.84 μg·mL^-1(r=0.999 7),2.784-27.84 μg·mL^-1(r=0.999 7)and 7.9-39.5 μg·mL^-1(r=0.999 3)respectively.Their average recoveries were 99.65%,99.58%,100.1%,and 99.92% respectively with RSD values of 0.97%,1.4%,1.0%,a nd 0.40% respectively.The determination results of 6 batches samples from two manufacturers showed that,the content of luteolin-7-O-β-D-glucuronide was 0.390-0.424 mg·g^-1,the content of rosmarinic acid was 0.431-0.482 mg·g^-1,the content of diosmetin-7-O-β-D-glucuronide was 0.468-0.532 mg·g^-1,the content of tilianin was 0.614-0.656 mg·g^-1. Conclusion: The method can be used to provide scientific evidence for the comprehensive quality evaluation and control of Yixin Badiranjibuya Granules through the methodology validation.