目的 建立不同海拔大黄药材的HPLC指纹图谱，明确海拔对大黄泻下作用的影响，筛选与海拔相关的泻下活性差异成分，为大黄药材质量评价提供参考。方法 采集甘肃礼县不同海拔人工种植的大黄样品，采用HPLC技术构建其指纹图谱；结合主成分分析（PCA）与正交偏最小二乘判别分析（OPLS-DA），筛选不同海拔大黄的差异性成分；以小鼠5 h内排便粒数为药效评价指标，比较不同海拔大黄的泻下活性；通过灰色关联度分析（GRA）、偏最小二乘回归分析（PLS-RA）及变量投影重要性（VIP）分析，将HPLC指纹图谱数据与泻下药效数据进行关联，筛选调控大黄泻下作用的关键质量指标。结果 成功构建大黄HPLC指纹图谱，共确定29个共有峰，其中12个峰经指认明确对应成分，6号峰儿茶素、7号峰表儿茶素、9号峰大黄酸-8-O-β-D-葡萄糖苷、10号峰番泻苷B，18号峰大黄素-8-β-D-吡喃葡萄糖苷、20号峰大黄酚-8-O-β-D-吡喃葡萄糖苷、21号峰大黄素-6-O-葡萄糖苷、25号峰芦荟大黄素、26号峰大黄酸、27号峰大黄素、28号峰大黄酚、29号峰大黄素甲醚。化学计量学分析显示，19批大黄样品经PCA可清晰分为高海拔与低海拔2类，OPLS-DA结果与PCA分类一致，进一步验证了海拔对大黄化学成分的影响；泻下作用实验表明，高海拔组大黄的促排便作用显著优于低海拔组（P＜0.05）；谱效关系综合分析结果显示，峰14、峰17、峰25（芦荟大黄素）与大黄泻下作用的关联性最强，是调控大黄泻下作用的关键成分。结论 建立的大黄HPLC指纹图谱稳定可靠，且通过谱-效关系明确了与泻下作用相关的核心成分及海拔对药效的影响，为大黄药材的质量控制、品质评价及合理应用提供了重要参考。

Objective To establish HPLC fingerprints of Rheum palmatum from different altitudes, clarify the influence of altitude on the laxative effect of R. palmatum and screen the laxative activity-related components associated with altitude, providing a reference for the quality evaluation of R. palmatum. Methods R. palmatum samples from artificial cultivation at different altitudes in Li County, Gansu Province were collected. HPLC was used to construct their fingerprint profiles. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were combined to screen the differential chemical components of R. palmatum at different altitudes. The number of fecal pellets excreted by mice within five hours was used as the pharmacodynamic evaluation index to compare the laxative activities of R. palmatum at different altitudes. Grey relational analysis (GRA), partial least squares regression analysis (PLS-RA), and variable importance in the projection (VIP) analysis were used to correlate the HPLC fingerprint profile data with the laxative activity data and screen the key quality indicators regulating the laxative effect of R. palmatum. Results The HPLC fingerprint profile of R. palmatum was successfully established, with a total of 29 common peaks identified. Among them, 12 peaks were identified as corresponding components: peak 6 catechin, peak 7 epicatechin, peak 9 rhein-8-O-β-Dglucoside, peak 10 sennoside B, peak 18 emodin-8-β-D-glucopyranoside, peak 20 chrysophanol-8-O-β-D-glucopyranoside, peak 21 emodin-6-O-glucoside, peak 25 aloe-emodin, peak 26 rhein, peak 27 emodin, peak 28 chrysophanol, and peak 29 physcion. Chemometric analysis showed that 19 batches of R. palmatum samples could be clearly divided into high-altitude and low-altitude groups by PCA, and the OPLS-DA results were consistent with the PCA classification, further verifying the influence of altitude on the chemical components of R. palmatum. The laxative effect experiments indicated that the laxative effect of R. palmatum in the highaltitude group was significantly better than that in the low-altitude group (P < 0.05). The comprehensive analysis of the spectrum-effect relationship showed that peaks 14, 17, and 25 (aloe-emodin) had the strongest correlation with the laxative effect of R. palmatum and were the key potential components regulating the laxative effect. Conclusion The HPLC fingerprint profile of R. palmatum established in this study was stable and reliable. Through the spectrum-effect relationship, the core components related to the laxative effect and the influence of altitude on the efficacy were clarified, providing an important reference for the quality control, quality evaluation, and rational application of R. palmatum.

R285.5

国家药品监督管理局药品监管科学体系建设重点项目（RS2024Z006）；中检院关键技术研究基金项目（GJJS20220702）；中检院中药所学科建设项目（1020050090118）；国家药典委员会国家药品标准制修订研究课题（BZ20230277）；新疆维吾尔自治区重点研发计划项目（2024B02024）