世界中医药

目的：探讨采用高效液相色谱法（HPLC）分析三黄汤传统煎法和现代煎药机法的指纹图谱差异。方法：选取10个不同批次的三黄汤配方，配制传统煎法三黄汤供试品溶液和现代煎药机三黄汤供试品溶液，色谱条件采用4.6 mm×250 mm的DIAMONSIL C18保护柱（5 μm），以30 ℃为柱温，进样量为15 μL，以甲醇为流动相A，1%磷酸水为流动相B，检测波长为277 nm，进行HPLC指纹图谱对比研究。结果：三黄汤专属性良好。线性关系考察显示黄芩苷回归方程Y=75.64X+21.31，r=0.999 8(n=5)，黄芩苷对照品范围在0.5~12.0 μg/mL，线性考察结果显示线性关系良好。精密度结果最终取大于总峰面积5%的色谱峰相对峰面积RSD平均值1.776%，三黄汤供试品溶液精密度良好。稳定性结果显示色谱峰相对峰面积RSD平均值1.843%，三黄汤供试品溶液24 h内稳定性良好。重复性试验结果显示色谱峰相对峰面积RSD平均值1.911%，三黄汤的重复性良好。加样回收率检测结果显示平均回收率99.81%，RSD 1.79%，高相液相检测方法所测加样回收率结果可靠。样品测定结果显示在同一色谱洗脱梯度下，传统煎法的三黄汤指纹图谱120 min即完成检测，而现代煎药机法制备的三黄汤指纹图谱在131.87 min才完成检测；另外从图谱中可见现代煎药机制备的三黄汤中色谱峰更多，可能与现代煎药机的高压煎煮设备能析出更多共有峰有关。从各自的指纹图谱相似度中可得，10批传统煎法三黄汤的相似度和10批现代煎药机法三黄汤的相似度均高于0.9，其中传统煎法平均相似度为0.971 4，现代煎药机法平均相似度为0.981 6，略高于传统煎法。结论：传统煎法和现代煎药机法制备的三黄汤的HPLC指纹图谱存在一定差异。

English Summary:

To compare the fingerprints of traditional decocting method and modern decocting machine methods of Sanhuang Decoction by high performance liquid chromatography(HPLC).Methods:Ten different batches of Sanhuang Decoction were randomly selected by random digital table method to prepare Sanhuang decoction solution of traditional decocting method and Sanhuang Decoction solution of modern decocting machine.The chromatographic conditions were as follows：DIAMONSIL C18 protective column(5 μm）of 4.6 mm×250 mm，column temperature 30 ℃，injection volume 15 μL，methyl alcohol as mobile phase A and 1% phosphoric acid water as mobile phase B，detection wavelength of 277 nm，for HPLC fingerprint comparison.Results:1）The specificity of Sanhuang Decoction was tested by HPLC.The results showed that the specificity of Sanhuang Decoction was good. 2） Linear relationship test showed that the regression equation of baicalin y=75.64X+21.31，r=0.999 8(n＝5).The results showed that the range of baicalin reference substance was 0.5~12.0 ug/mL，and the linear relationship was good. 3）The RSD average value of relative retention time of chromatographic peaks larger than 5% of the total peak area was 1.776%.The results showed that the solution of Sanhuang Decoction had good precision. 4）The results of stability showed that the RSD value of relative retention time of chromatographic peaks was 1.843%，suggesting that the solution of Sanhuang Decoction had good stability within 24 hours. 5）The results of repeatability test showed that the RSD value of relative retention time of chromatographic peaks was 1.911%，suggesting that the repeatability of Sanhuang Decoction was good. 6）The average recovery was 99.81% and RSD was 1.79%.The results showed that the HPLC was reliable. 7）Sample determination results showed that under the same chromatographic elution gradient，the fingerprint of traditional decocting method of Sanhuang Decotion was completed within 120 minutes，and the fingerprint of modern decocting machine prepared Sanhuang Decotion was completed within 131.87 minutes; In addition，from the chromatogram，it can be seen that there were more chromatographic peaks in the modern decocting machine prepared Sanhuang Decoction，which may be compared with that more common peaks can be separated from the high pressure decocting equipment of the modern decocting machine.From the similarity of fingerprints，the similarity of 10 batches of Sanhuang Decoction prepared by traditional decocting method and 10 batches of Sanhuang Decoction prepared by modern decocting machine method were higher than 0.9，of which the average similarity of traditional decocting method was 0.9714，and that of modern decocting machine method was 0.9816，slightly higher than that of traditional decocting method.Conclusion:The HPLC fingerprint of Sanhuang Decoction prepared by traditional decocting method and modern decocting machine method is different.

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