HPLC法同时测定桃儿七中8个成分的含量

目的：建立HPLC同时测定桃儿七药材中苦鬼臼毒素葡萄糖二苷、山柰酚葡萄糖苷、4'-去甲鬼臼毒素、鬼臼毒素葡萄糖苷、槲皮素、鬼臼毒素、鬼臼毒酮、山柰酚含量的方法。方法：采用HyPURITY C₁₈色谱柱（250 mm×4.6 mm，5 μm），以甲醇-0.4%磷酸水溶液为流动相，进行梯度洗脱，流速0.8 mL·min^-1，柱温25℃，检测波长290 nm。结果：苦鬼臼毒素葡萄糖二苷、山柰酚葡萄糖苷、4'-去甲鬼臼毒素、鬼臼毒素葡萄糖苷、槲皮素、鬼臼毒素、鬼臼毒酮、山柰酚的质量浓度分别在3.6~36、3.84~38.4、12.96~129.6、39.69~396.9、18.8~188、113.92~1132.9、5.6~56、14.4~144 μg·mL^-1范围内与峰面积呈良好的线性关系；平均加样回收率（n=9）均高于98%，RSD小于3.0%；22批桃儿七样品中上述8个成分的含量范围分别为0.309~2.047、0.229~1.730、1.536~7.467、3.171~22.631、2.071~9.724、11.727~66.420、0.688~3.177、1.730~7.331 mg·g^-1。结论：本方法适用于桃儿七中有效成分的含量测定。

Objective: To establish an HPLC method for simultaneous determination of picropodophillotoxin-4-O-β-D-glucopyranosyl-(1 → 6)-β-D-glucopyranoside, kaempferol-3-O-β-D-glucoside,4'-demethylpodophyllotoxin,podophyllotoxin-4-O-β-D-glucoside,quercetin,podophyllotoxin,podophyllotoxone and kaempferol in Sinopodophylli Radix et Rhzoma.Methods: The HPLC separation was performed on an HyPURITY C₁₈ (250 mm×4. 6 mm,5 μm)column with the mobile phase of methanol-0. 4% phosphoric acid solution wtih gradient elution at a flow rate of 0. 8 mL·min^-1. The column temperature was 25℃,and the detection wavelength was 290 nm.Results: The linear ranges of picropodophillotoxin-4-O-β-D-glucopyranosyl-(1 → 6)-β-D-glucopyranoside,kaempferol-3-O-β-D-glucoside,4'-demethylpodophyllotoxin,podophyllotoxin-4-O-β-D-glucoside,quercetin,podophyllotoxin,podophyllotoxone and kaempferol were 3. 6-36 μg·mL^-1,3. 84-38. 4 μg·mL^-1,12. 96-129. 6 μg·mL^-1,39. 69-396. 9 μg·mL^-1,18. 8-188 μg·mL^-1,113. 92-132. 9 μg·mL^-1, 5. 6-56 μg·mL^-1 and 14. 4-144 μg·mL^-1,within the scope of good linear with peak area present relationship, respectively;the average recoveries(n=9)for the sample preparation of the markers were more than 98. 0% and RSDs were less than 3. 0%;the content ranges of-above 8 compounds in 22 samples were 0. 309-2. 047 mg·g^-1,0. 229-1. 730 mg·g^-1,1. 536-7. 467 mg·g^-1,3. 171-22. 631 mg·g^-1,2. 071-9. 724 mg·g^-1,11. 727-66. 420 mg·g^-1, 0. 688-3. 177 mg·g^-1 and 1. 730-7. 331 mg·g^-1,respectively.Conclusion: The established method was suitable for determination of active components in Sinopodophylli Radix et Rhzoma.