目的 建立同时测定经典名方金水六君煎（JLD）中腺苷、鸟苷、5-羟甲基糠醛、阿魏酸、甘草苷、毛蕊花糖苷、芸香柚皮苷、橙皮苷、甘草酸铵、6-姜辣素、藁本内酯11种主要成分的UPLC方法。方法 采用UPLC法，色谱柱为Acquity UPLC HSS T3 C₁₈柱（100 mm×2.1 mm，1.8 μm），0.1%磷酸水溶液-乙腈为流动相，梯度洗脱；体积流量为0.3 mL/min；柱温25℃；进样量1.0 μL；检测波长：腺苷、鸟苷、5-羟甲基糠醛、甘草酸铵为248 nm，阿魏酸为330 nm，甘草苷、毛蕊花糖苷、芸香柚皮苷、橙皮苷、6-姜辣素、藁本内酯为281 nm。采用SIMCA 14.1软件进行偏最小二乘法-判别分析（PLS-DA），以寻找样品间的质量差异成分，同时采用Hotelling's T²和DModX 2种统计量，为不同批次样品的质量设定控制范围。结果 被测定的11种成分色谱峰均有良好的分离度，方法精密度、重复性的RSD均小于3%；在室温条件下24 h内稳定；各成分均有较宽的线性范围和良好的线性关系（r ≥ 0.999 2）；平均加样回收率96.62%~103.15%，RSD均小于2%。将15批不同产地的样品分为3类，通过PLS-DA分析含量测定结果，发现芸香柚皮苷、甘草酸铵、藁本内酯和阿魏酸是影响不同批次JLD物质基准质量贡献较大的4种成分。结论 建立的方法操作简便，结果准确、可靠，可用于JLD及其制剂的质量控制与评价。

Objective To establish a UPLC method for the simultaneous determination of 11 components, including adenosine, guanosine, 5-hydroxymethylfurfural, ferulic acid, glycyrrhizin, verbascoside, rutin naringin, hesperidin, ammonium glycyrrhizinate, 6-gingerol, and ligustilide in Jinshui Liujun Decoction (金水六君煎, JLD). Methods The chromatographic separation was achieved on an Acquity UPLC HSS T3 C₁₈ column (100 mm×2.1 mm, 1.8 μm) with 0.1% phosphate acid solution-acetonitrile as mobile phase at the flow rate of 0.3 mL/min for gradient elution; The column temperature was 25℃. The determination wavelengths were 248 nm for adenosine, guanosine, 5-hydroxymethylfurfural and ammonium glycyrrhizinate, 330 nm for ferulic acid, 281 nm for glycyrrhizin, verbascoside, rutin naringin, hesperidin, 6-gingerol, and ligustilide. The different samples were evaluated with the partial least squares-discriminant analysis (PLS-DA) by SIMCA 14.1 software to find significant components of the quality between the batches. Results The 11 compounds were well separated. The RSD values of precision and reproducibility were all less than 3%. The stability was good in 24 h. The linear relationship between the concentration and peak areas of the 11 compounds was all linear (r ≥ 0.9992). The average recoveries were in range of 96.62%-103.15% and the RSD values were all less than 2%. Fifteen batches of samples from different origins were classified into three categories. Using PLS-DA analysis, the content determination result showed that rutin naringin, ammonium glycyrrhizinate, ligustilide, and ferulic acid were the four components that affected the quality of different batches of JLD. Conclusion The method is simple, reliable, and accurate, and could be used for the quality control and evaluation of JLD.

R286.02

国家“重大新药创制”科技重大专项（2018ZX09721-005-003-006）