HPLC法测定益心巴迪然吉布亚颗粒中4种化学成分的含量

HPLC determination of four components in Yixin Badiranjibuya granules

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,5μm）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;55~80min,25%A→30%A）,流速1.0 m L·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3.184~31.84μg·m L-1（r=0.999 6）、2.184~21.84μg·m L-1（r=0.999 7）、2.784~27.84μg·m L-1（r=0.999 7）、7.9~39.5μg·m L-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 m L·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·m L- 1（ r =0. 999 6）,2. 184-21. 84 μg·m L- 1（ r =0. 999 7）,2. 784- 27. 84 μg·m L- 1（ r =0. 999 7） and 7. 9-39. 5 μg·m L- 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%,and0. 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. 482mg·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.