传统煎法和现代煎药机制备补阳还五汤的HPLC指纹图谱对比研究

Comparative Study on HPLC Fingerprint of Buyang Huanwu Decoction Prepared by Traditional Decocting Method and Modern Decoction Mechanism

目的:采用高效液相(HPLC)对比研究补阳还五汤传统煎法和现代煎药机法的指纹图谱差异。方法:首先采用随机数字表法随机选取不同产地的药材配出10个不同批次的补阳还五汤配方,配制传统煎法补阳还五汤供试品溶液和现代煎药机补阳还五汤供试品溶液后,以优化的色谱条件进行HPLC指纹图谱对比研究。结果:1)采用HPLC色谱法对补阳还五汤进行专属性试验,结果显示补阳还五汤专属性良好。2)线性关系考察显示阿魏酸回归方程y=259.33X+48.708,r=0.999 9(n=5),结果表明阿魏酸对照品范围0.997~9.97μg/μL,线性考察结果显示线性关系良好。3)精密度试验结果最终取大于总峰面积5%的色谱峰相对峰面积的RSD平均值2.014%,色谱峰相对保留时间RSD平均值0.29%,研究结果显示补阳还五汤供试品溶液精密度良好。4)稳定性试验结果最终取大于总峰面积5%的色谱峰相对峰面积的RSD平均值1.839%,色谱峰相对保留时间RSD平均值0.02%,研究结果显示补阳还五汤供试品溶液24 h内稳定性良好。5)重复性试验结果最终取大于总峰面积5%的色谱峰相对峰面积的RSD平均值1.901%,色谱峰相对保留时间RSD平均值0.72%,结果显示补阳还五汤的重复性良好。6)回收率试验结果显示平均回收率99.76%,RSD 2.14%,表明这一色谱条件下的HPLC方法检测加样回收率结果可靠。7)样品测定结果显示传统煎法的补阳还五汤指纹图谱131.88 min即完成检测,而现代煎药机法制备的补阳还五汤指纹图谱在140 min才完成检测;另外从图谱中可见现代煎药机制备的补阳还五汤中色谱峰更多。从各自的指纹图谱相似度中可得,其中传统煎法平均相似度为0.956,现代煎药机法平均相似度为0.974,略高于传统煎法。结论:传统煎法和现代煎药机法制备的补阳还五汤的HPLC指纹图谱存在一定差异。

Objective:To compare the fingerprint difference of Buyang Huanwu Decoction between traditional decoction and modern decoction machine by high performance liquid chromatography(HPLC). Methods:Firstly,random number table method was used to randomly select the prescription of 10 different batches of Buyang Huanwu Decoction of different batches,prepared the traditional decoction of Buyang Huanwu Decoction,and the modern decocting machine Buyang Huanwu Decoction.After the optimized chromatographic conditions,the HPLC fingerprint was compared. Results:1)HPLC chromatography was used to make a specific test of Buyang Huanwu Decoction,the results showed that the Buyang Huanwu decoction had good specificity.2)The linear relationship showed the regression equation of ferulic acid y=259.33X+48.708,r=0.999 9( n =5).The results showed that the range of ferulic acid control was in 0.997~9.97 μg/μL,and the linear investigation showed that the linear relationship was good.3)The results of the precision test were 5% of the total peak area of the peak area of 5%,the average value of the relative peak area of the peak area was 2.014%,and the relative retention time of the chromatographic peak was 0.29% of the average value of RSD.The results showed that the precision of the solution of Buyang Huanwu decoction was good.4)Finally,the results of the stability test were 5% of the total peak area and the RSD average 1.839%.The chromatographic peak relative retention time RSD average value was 0.02%,and the research results showed that Buyang Huanwu decoction solution 24 h stability.5)The results of repeatability test were 5% of the total peak area,and the average value of the peak area was 1.901%,and the relative retention time of the chromatographic peak was 0.72%.The results of this study showed that the repeatability of Buyang Huanwu decoction was good.6)The average recovery was 99.76% and RSD was 2.14%.The results showed that the HPLC method under this chromatographic condition was reliable.7)The test results showed that the traditional decoction of Buyang Huanwu decoction fingerprint 131.88 min was completed,while the Buyang Huanwu decoction fingerprint made by the modern decocting machine was detected in 140 min,and the chromatogram peak of Buyang Huanwu decoction prepared by the modern decocting mechanism could be seen from the atlas.The average similarity of the traditional decocting method was 0.956 and the average similarity of the modern decocting machine method was 0.974,which is slightly higher than the traditional decocting method. Conclusion:The HPLC fingerprint of Buyang Huanwu decoction prepared by traditional decocting method and modern decocting machine is different.