目的 建立附子理中丸（Fuzi Lizhong Pills，FLP）处方的大蜜丸（Dami Pills，FLP-D）、水蜜丸（Shuimi Pills，FLP-S）及浓缩丸（Nongsuo Pills，FLP-N）的HPLC指纹图谱，同时测定7种药效成分含量，寻找FLP处方与剂型关联的化学特征关键质量属性。方法 采用分层抽样法结合研究实际，随机抽取市售FLP-D 30批，FLP-S 12批，FLP-N 12批，建立以上54批FLP的HPLC指纹图谱，并同时测定甘草苷、苯甲酰新乌头原碱、党参炔苷、6-姜酚、白术内酯III、白术内酯I、10-姜酚7种有效成分含量；利用主成分分析（principal component analysis，PCA）和偏最小二乘法-判别分析（partial least squares-discriminant analysis，PLS-DA）方法对FLP-D、FLP-S和FLP-N各自的HPLC指纹图谱特征峰及7种有效成分进行分析。结果 FLP-D、FLP-S和FLP-N的HPLC指纹图谱分别标定出20、25、20个特征峰，通过PCA和PLS-DA，筛选出14个差异性标志物，其中8、9、12、17、18、25号峰来自甘草，29、32号峰来自干姜，22号峰来自白术。含量测定结果表明，甘草苷、苯甲酰新乌头原碱、6-姜酚、白术内酯III、白术内酯I在FLP-D、FLP-S和FLP-N中的含量差异显著，其中6-姜酚为变量重要性投影（variable importance in projection，VIP）值＞1.0的差异性标志物。结论 建立的HPLC指纹图谱与多成分同时测定方法能有效评价FLP-D、FLP-S和FLP-N的化学成分差异；识别出的14个差异性标志物可作为区分FLP-D、FLP-S和FLP-N的潜在化学特征关键质量属性；甘草苷、苯甲酰新乌头原碱、白术内酯III、白术内酯I可作为区分FLP-D与FLP-N或区分FLP-S与FLP-N的化学特征关键质量属性，6-姜酚可作为同时区分FLP-D、FLP-S和FLP-N 3种不同剂型的化学特征关键质量属性。为进一步完善FLP系列制剂的质量标准提供了参考，同时为解决同方异剂中药的质控指标同质化问题提供了新思路。

Objective To find the chemical critical quality attributes (CQAs) associated with the dosage form characteristics of Fuzi Lizhong Pills (FLP, 附子理中丸) by establishing HPLC fingerprints of Dami Pills (FLP-D), Shuimi Pills (FLP-S) and Nongsuo Pills (FLP-N) and determining contents of seven effective components. Methods According to the stratified sampling method and research requirement, 30 batches of FLP-D, 12 batches of FLP-S and 12 batches of FLP-N were randomly selected from the market. The HPLC HPLC fingerprints of 54 batches of FLP were establishe. The contents of seven effective components, including liquiritin, benzoylmesaconine, lobetyolin, 6-gingerol, atractylenolide III, atractylenolide I, and 10-gingerol were simultaneously determined. Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were used to analyze the HPLC fingerprint characteristic peaks of FLP-D, FLP-S and FLP-N and seven active components. Results There were 20 characteristic peaks in the fingerprints of FLP-D, 25 characteristic peaks in FLP-S, and 20 characteristic peaks in FLP-N. A total of 14 differential markers were determined by PCA and PLS-DA. Among them, peaks 8, 9, 12, 17, 18 and 25 originated from Gancao (Glycyrrhizae Radix et Rhizoma), peaks 29 and 32 originated from Ganjiang (Zingiberis Rhizoma), and peak 22 originated from Baizhu (Atractylodis Macrocephalae Rhizoma). The contents of liquiritin, benzoylmesaconine, 6-gingerol, atractylenolide III and atractylenolide I in FLP-D, FLP-S and FLP-N were significantly different. The compound 6-gingerol was a differential marker with variable importance in projection (VIP) value > 1.0. Conclusions The established HPLC fingerprint and multi-component simultaneous determination method can effectively evaluate the chemical composition differences of FLP-D, FLP-S and FLP-N. The identified14 differential markers could be the potential CQAs of FLP-D, FLP-S and FLP-N. Liquiritin, benzoylmesaconine, atractylenolide III and atractylenolide I could be used as CQAs to distinguish FLP-D and FLP-N, or FLP-S and FLP-N. 6-gingerol could be used as CQAs for differentiating FLP-D, FLP-S and FLP-N simultaneously. Our research provides a reference for further enhancing the quality standards of FLP, and presents a novel idea to address the homogeneity issue in quality control for traditional Chinese medicine with the same prescription but different dosage forms.

R283.6

四川省科技计划重点研发项目（2020YFS0567）；四川省科技厅四川省科技计划项目（2021YJ0251）；四川省中医药管理局科学技术研究专项（2023MS600）；成都中医药大学杏林学者青基人才专项（QJRC2022027）