目的建立清肝颗粒提取液、醇沉液、浓缩液的UPLC指纹图谱及8种指标成分含量测定方法，研究其在提取、醇沉、浓缩关键工序中的量质传递规律。方法采用Waters CORTECS T3 C18色谱柱，以0.2%磷酸水溶液-乙腈为流动相梯度洗脱，分别建立10批清肝颗粒提取液、醇沉液、浓缩液的UPLC指纹图谱，进行共有峰归属与相似度评价；同步测定(R,S)-告依春、新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、甘草酸铵8种指标成分的含量，计算各成分在“提取液→醇沉液→浓缩液”工序间的转移率；结合主成分分析（PCA）与正交偏最小二乘-判别分析（OPLS-DA）筛选影响中间体质量差异的成分。结果 10批样品共标定21个共有峰，各批次指纹图谱相似度均＞0.925；经与对照品比对指认出8个色谱峰，依次为(R,S)-告依春（峰1）、新绿原酸（峰3）、绿原酸（峰5）、隐绿原酸（峰6）、异绿原酸B（峰12）、异绿原酸A（峰13）、异绿原酸C（峰15）、甘草酸铵（峰20）；经主成分分析（PCA），提取出3个主成分，累积方差贡献率为90.470%；以变量投影重要性（VIP）值＞1为阈值，共筛选出峰14、峰4、新绿原酸、甘草酸铵、峰16、绿原酸6个潜在质量差异成分。量质传递研究显示，8种指标成分工序间转移率均在平均值± 30%范围内，均值水平上，提取液→醇沉液步骤中新绿原酸、绿原酸、隐绿原酸转移率低于异绿原酸A/B/C；而醇沉液→浓缩液步骤相反，异绿原酸A/B/C转移率更低。结论基于UPLC指纹图谱与多指标定量，初步构建清肝颗粒生产中间体指标成分的质量控制体系，为清肝颗粒关键生产工序的质量控制与评价体系构建提供科学依据。

Objective To establish UPLC fingerprint profiles and quantitative methods for eight marker compounds in the extract, alcohol-precipitated solution, and concentrated solution of Qinggan Granules, and to investigate the quantity-quality transfer patterns during key processing steps including extraction, alcohol precipitation, and concentration. Methods A Waters CORTECS T3 C18 column was used with a gradient elution system consisting of 0.2% phosphoric acid aqueous solution and acetonitrile. UPLC fingerprint profiles were established for ten batches of extract, alcohol-precipitated solution, and concentrated solution of Qinggan Granules, followed by identification of common peaks and similarity evaluation. The contents of eight marker compounds—(R,S)-goitrin, neo-chlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and ammonium glycyrrhizinate—were simultaneously determined, and transfer rates of each compound across the “extract→alcoholprecipitated solution→concentrated solution” process were calculated. Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were applied to identify components contributing to differences in intermediate product quality. Results A total of 21 common peaks were identified across the ten batches, with all fingerprint similarities exceeding 0.925. Eight peaks were confirmed by comparison with reference standards: R,S)-goitrin (peak 1), neo-chlorogenic acid (peak 3), chlorogenic acid (peak 5), cryptochlorogenic acid (peak 6), isochlorogenic acid B (peak 12), isochlorogenic acid A (peak 13), isochlorogenic acid C (peak 15), and ammonium glycyrrhizinate (peak 20). PCA revealed three principal components accounting for 90.470% of the cumulative variance. Using variable importance in projection (VIP) values >1 as threshold, six potential quality-differentiating components were selected: peak 14, peak 4, neo-chlorogenic acid, ammonium glycyrrhizinate, peak 16, and chlorogenic acid. The mass transfer study showed that the inter-process transfer rates of the eight index components were all within ± 30% of the mean. At the mean level, in the extract→alcohol precipitation liquid step, the transfer rates of neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid were lower than those of isochlorogenic acids A/B/C; whereas in the alcohol precipitation liquid→concentrated liquid step, the opposite was observed. Conclusion Based on UPLC fingerprinting and multi-index quantification, a quality control system for the key component indicators of the production intermediate of Qinggan Granules was established, providing a scientific basis for the construction of the quality control and evaluation system for the key production processes of Qinggan Granules.

R284.1

山东省重点研发计划（竞争性创新平台）项目（2024CXPT074）