HPLC-DAD法同时测定栀子金花丸中10个成分的含量

目的： 建立HPLC-DAD法同时测定栀子金花丸中栀子苷、绿原酸、黄芩苷、芒果苷、小檗碱、黄柏碱、大黄酸、大黄素、芦荟大黄素和大黄酚10个成分的含量。方法： 采用Luna C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.2%醋酸水溶液为流动相，梯度洗脱（0~15 min，8% A→40% A；15~20 min，40% A→60% A；20~30 min，60% A→90% A；30~40 min，90% A→8% A），流速1.0 mL·min^-1，柱温35℃，检测波长258 nm（检测栀子苷、芒果苷、大黄酸、大黄素、芦荟大黄素和大黄酚），280nm（检测黄芩苷和黄柏碱），345 nm（检测绿原酸和小檗碱）。结果： 栀子苷、绿原酸、黄芩苷、芒果苷、小檗碱、黄柏碱、大黄酸、大黄素、芦荟大黄素和大黄酚10个待测成分实现完全分离，并与其他成分达到良好的分离；上述成分线性范围分别为0.66~16.43 μg·mL^-1（r=0.999 7）、0.42~10.45 μg·mL^-1（r=0.999 2）、0.30~7.50 μg·mL^-1（r=0.995 7）、0.64~15.95 μg·mL^-1（r=0.996 5）、0.59~15.06 μg·mL^-1（r=0.999 6）、0.06~1.59 μg·mL^-1（r=0.998 3）、0.27~6.78 μg·mL^-1（r=0.993 8）、0.07~1.82 μg·mL^-1（r=0.989 0）、0.04~1.03 μg·mL^-1（r=0.995 2）和0.10~2.53 μg·mL^-1（r=0.999 8）；平均加样回收率（n=9）分别为100.4%、99.7%、99.0%、99.5%、99.2%、97.8%、97.9%、97.8%、97.8%和99.1%。5批样品中上述10个成分的含量范围分别为3.45~3.72、2.30~2.36、1.68~1.75、3.46~3.58、3.48~3.57、0.34~0.41、1.46~1.55、0.42~0.47、0.22~0.26、0.50~0.56 mg·g^-1。结论： 本法可为栀子金花丸的质量控制和考察提供参考。

Objective: To establish an HPLC method for simultaneous determination of geniposide,chlorogenic acid,baicalin,mangiferin,berberine,phellodendrine,rhein,emodin,aloe emodin and chrysophanol in Zhizi Jinhua pills. Methods: HPLC separation was performed on a Luna C₁₈ column(4.6 mm×250 mm,5 μm),with the mobile phase consisting of acetonitrile-0.2% acetic acid aqueous solution at a flow rate of 1.0 mL·min^-1 in a gradient elution mode(0-15 min,8%A→40%A;15-20 min,40%A→60%A;20-30 min,60%A→90%A;30-40 min,90%A→8%A). The column temperature was at 35℃ and the detection wavelengths were set at 258 nm for geniposide,mangiferin,rhein,emodin,aloe emodin,and chrysophanol,280 nm for baicalin and phellodendrine,and 345 nm for chlorogenic acid and berberine. Results: Geniposide,chlorogenic acid,baicalin,mangiferin,berberine,phellodendrine,rhein,emodin,aloe emodin and chrysophanol were completely separated,and well separated from other constituents. The linear ranges of above-mentioned components were 0.66-16.43 μg·mL^-1(r=0.999 7),0.42-10.45 μg·mL^-1(r=0.999 2), 0.30-7.50 μg·mL^-1(r=0.995 7),0.64-15.95 μg·mL^-1(r=0.996 5),0.59-15.06 μg·mL^-1(r=0.999 6),0.06-1.59 μg·mL^-1(r=0.998 3),0.27-6.78 μg·mL^-1(r=0.993 8),0.07-1.82 μg·mL^-1(r=0.989 0),0.04-1.03 μg·mL^-1(r=0.995 2)and 0.10-2.53 μg·mL^-1(r=0.999 8),respectively. The average recoveries were 100.4%,99.7%,99.0%,99.5%,99.2%,97.8%,97.9%,97.8%,97.8% and 99.1%,respectively. The content ranges of ten ingredients were 3.45-3.72 mg·g^-1,2.30-2.36 mg·g^-1,1.68-1.75 mg·g^-1,3.46-3.58 mg·g^-1,3.48-3.57 mg·g^-1,0.34-0.41 mg·g^-1, 1.46-1.55 mg·g^-1,0.42-0.47 mg·g^-1,0.22-0.26 mg·g^-1 and 0.50-0.56 mg·g^-1,respectively. Conclusion: The established method provides references for the quality control and evaluate of Zhizi Jinhua pills.

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