建立清肝润肺止咳露高效液相色谱（HPLC）特征图谱，测定其中吗啡、可待因、罂粟碱、那可丁 的含量，并结合化学模式识别方法，综合评价该制剂的质量。方法 采用HPLC 法，以甲醇- 5 μmoL·L-1 庚烷 硫酸钠溶液（用磷酸调节pH 至2.6）为流动相进行梯度洗脱，流速：1.0 mL·min-1，柱温：20 ℃，检测波长： 220 nm，进样量：10 μL。利用指纹图谱相似度评价软件，结合聚类分析（CA）、主成分分析（PCA）和偏最小二 乘法判别分析（PLS-DA）对10 批清肝润肺止咳露的特征图谱进行评价。结果 10 批清肝润肺止咳露特征图谱 共确定16 个共有峰，指认了5 个共有峰，分别为吗啡、可待因、蒂巴因、罂粟碱、那可丁。其中4 个成分在 线性范围内（吗啡0.70~8.70 μg、可待因0.68~8.50 μg、罂粟碱0.60~7.55 μg、那可丁0.38~4.80 μg）均有良好的 线性关系（r≥0.999 7）；精密度、稳定性、重复性试验的RSD 均小于3.0%；平均加样回收率在97.93%~99.43% 之间（RSD＜3.1%）；含量测定结果：吗啡43.58~76.47 μg·mL-1，可待因16.64~78.92 μg·mL-1，罂粟碱29.93~ 41.22 μg·mL-1，那可丁14.44~33.22 μg·mL-1。通过CA、PCA 和PLS-DA 分析，可将10 批清肝润肺止咳露分 为2 类（S3、S4、S8、S10 聚为一类，其余聚为一类），并筛选出峰2、峰4、峰13、峰11（吗啡）、峰1、峰 10、峰8、峰12、峰16（那可丁）9 个差异性成分。结论 该研究建立的HPLC 特征图谱及含量测定方法简单、 准确，结合化学模式识别法，可用于清肝润肺止咳露的质量控制。

To establish the HPLC fingerprint of Qinggan Runfei Zhike Distillate and determine the contents of morphine，codeine，papaverine and narcodine，and carry out chemical pattern recognition in order to evaluate the quality of the preparation comprehensively. Methods HPLC was carried out by using methanol- 5 μmoL·L-1 sodium heptanesulfonate solution （adjust pH to 2.6 with phosphoric acid） as the mobile phase for gradient elution. The flow rate was 1.0 mL·min-1，the column temperature was 20 ℃，the detection wavelength was at 220 nm，and the injection amount was 10 μL. Similarity evaluation combined with hierarchical clustering analysis （CA）， principal components analysis （PCA） and discriminant analysis of partial least squares （PLS- DA） were used to evaluate the HPLC chromatographic fingerprint of 10 batches of Qinggan Runfei Zhike Distillate. Results There were 16 common peaks in the fingerprint of 10 batches of Qinggan Runfei Zhike Distillate， and 5 common peaks were identified， which were morphine， codeine， thebaine， papaverine and nacodine. The linear range of morphine，codeine， papaverine and nacodine in the content determination results are 0.70-8.70， 0.68-8.50， 0.60-7.55 μg， 0.38-4.80 μg （r≥0.999 7）. The RSDs of precision，stability and repeatability tests are less than 3.0%. The average sample recovery rate ranges are 97.93%-99.43%（RSD＜3.1%）. The contents in Qinggan Runfei Zhike Distillate are morphine 43.58-76.47 μg · mL-1， codeine 16.64-78.92 μg · mL-1， papaverine 29.93-41.22 μg · mL-1， nacodine 14.44-33.22 μg·mL-1. The results of CA， PCA and PLS-DA showed that ten batches of Qinggan Runfei Zhike Distillate could be clustered into two categories，S3，S4，S8，S10 were gathered into one category，and the rest were clustered into another category. Nine differential components were screened out， including peak 2， peak 4， peak 13， peak 11 （morphine），peak 1，peak 10，peak 8，peak 12，and peak 16 （nacodine）. Conclusion The HPLC fingerprint and content determination method established in this study are simple and accurate. The above method combined with chemical pattern recognition can be used for the quality control of Qinggan Runfei Zhike Distillate.

R284.1

广州市科技计划项目（2023A03J0297）。