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目的:对二黄汤方药半仿生法(SBE法)≤1 000 Da提取液的指纹图谱各特征峰的来源进行归属研究。方法:采用SBE法分别制备二黄汤全方和各单味药黄芩、黄连、甘草的提取液,绘制相应的HPLC指纹图谱,通过比对,确定其特征峰的来源;通过对照品比对,鉴定部分特征峰的成分。色谱条件:采用Diamonsil C18(250 mm×4.6 mm,5μm)色谱柱,0.02 mol·L~(-1)乙酸铵溶液(冰乙酸调p H 4.0)(A)-乙腈(B)为流动相,梯度洗脱,流速0.5 m L·min~(-1),检测波长265 nm,柱温25℃,进样量20μL。结果:二黄汤的SBE法≤1 000 Da提取液的HPLC指纹图谱有32个特征峰,其中峰9、11、13、16、17、25、26、27、30、31来自黄芩,峰15、20、21来自黄连,峰3、10、12、23、24、32来自甘草,峰2、18、19来自黄芩、黄连和甘草,峰14、28、29来自黄芩和甘草,峰1、4、5、6、7、8、22来自黄连和甘草。以保留时间和紫外吸收曲线为判断依据,进行对照品比对,鉴别出甘草苷、黄芩苷、巴马汀、小檗碱、甘草素、甘草酸、黄芩素、甘草次酸共8个已知成分,其中峰22是含小檗碱和甘草素的混合峰,与归属结果相同。结论:二黄汤SBE法提取液指纹图谱的建立符合要求,归属研究方法合理。与对照品比对,二黄汤SBE法≤1 000 Da提取液中各特征峰的归属结果准确,可为特征峰的成分鉴定和谱效关系研究提供依据。
OBJECTIVE: To study the origin of each characteristic peak of the extraction solution of Erhuang Tang semi-bionic method (SBE method) ≤1 000 Da. Methods: The SBE method was used to prepare the extracts of Radix Scutellariae, Coptis chinensis and Glycyrrhiza uralensis, which were used as a prescription for both Huanghuang Decoction and each single herb. The corresponding HPLC fingerprints were drawn and compared by comparison to determine the source of its characteristic peaks. Part of the composition of the peak. Chromatographic conditions: Diamonsil C18 (250 mm × 4.6 mm, 5 μm) column, 0.02 mol·L -1 ammonium acetate solution (glacial acetic acid p H 4.0) (A) -acetonitrile (B) Gradient elution, flow rate 0.5 m L · min -1, detection wavelength 265 nm, column temperature 25 ℃, injection volume 20μL. Results: There are 32 characteristic peaks in the HPLC fingerprint of SBE method ≤1000 Da for Erhuang Decoction, the peaks 9,11,13,16,17,25,26,27,30,31 come from Scutellaria baicalensis, peak 15, 20,21 From Coptis, Peaks 3,10,12,23,24,32 From Licorice, Peaks 2,18,19 From Scutellaria, Coptis And Glycyrrhiza, Peaks 14,28,29 From Scutellaria And Licorice Peaks 1,4, 5,6,7,8,22 from Coptis and licorice. Based on the retention time and the UV absorption curve, the comparison of the control samples was carried out, and the identification of eight glycosides, baicalin, palmatine, berberine, glycyrrhizin, glycyrrhizin, baicalein and glycyrrhetinic acid Composition, where peak 22 is a mixed peak containing berberine and liquiritigenin, with the same result of assignment. Conclusion: The fingerprint of SBR extract of Erhuangdou decoction meets the requirements and the research method is reasonable. Compared with the reference substance, the eigenvalue results of SBH method ≤1000 Da in Erhuangsu decoction were accurate, which could provide the basis for the identification of the components of the characteristic peak and the relationship between the spectral characteristics and the spectral characteristics.