目的 建立决明子Cassiae Semen的指纹图谱和多指标成分含量测定方法，结合化学计量学进行不同产地决明子的质量评价。方法 采用Waters HSS T3 C₁₈（150 mm×3.0 mm，1.8 μm）色谱柱，以乙腈-甲醇（3∶1）为流动相A，0.1%磷酸水溶液为流动相B，梯度洗脱，体积流量0.4 mL/min，柱温40 ℃，检测波长285 nm，进样量2 μL。采用中药指纹图谱相似度评价软件进行相似度评价，运用化学计量学对不同产地决明子进行分析，同时测定决明子中的11个指标成分含量。结果 建立的指纹图谱共标识出26个共有峰，经对照品指认出13个成分；经聚类分析和主成分分析（principal component analysis，PCA）可将不同决明子聚为4类，并通过正交偏最小二乘判别法（orthogonal partial least squares-discriminant analysis，OPLS-DA）以变量权重值（variable importance projection，VIP）＞1为指标筛选出峰3（大黄酚-1-O-β-D-四吡喃葡萄糖苷）、峰6（决明子苷）、峰7（红链霉素-6-O-β-龙胆二糖苷）、峰8（橙黄决明素-6-O-葡萄糖苷）、峰9（决明子苷C）、峰10、峰17（橙黄决明素）、峰21和峰23（红链霉素）作为区分不同产地决明子药材的差异标志物。18批决明子中异红镰霉素龙胆二糖苷、决明子苷B2、决明子苷、红链霉素-6-O-β-龙胆二糖苷、橙黄决明素-6-O-葡萄糖苷、决明子苷C、决明子苷B、橙黄决明素、黄决明素、红链霉素和大黄酚的质量分数分别为0.425～1.315、4.129～8.790、0.453～1.849、2.617～4.739、0.792～2.372、1.424～3.533、0.496～1.439、0.630～1.359、0.483～2.298、2.130～9.516、2.006～3.841 mg/g；经方法学考察，各成分呈现良好的线性关系。结论 建立的决明子指纹图谱结合化学模式识别可有效区分不同产地的决明子，多成分含量测定方法稳定、可靠，为决明子药材的质量评价与控制提供参考。

Objective To establish a fingerprint and multi-indicator component content determination method for Juemingzi (Cassiae Semen), and combine it with chemometrics to evaluate the quality of Cassiae Semen from different origins. Methods Waters HSS T3 C₁₈ (150 mm × 3.0 mm, 1.8 μm) was used as the chromatographic column, acetonitrile-methanol (3∶1) was used as mobile phase A, 0.1% phosphoric acid solution was used as mobile phase B, and gradient elution was performed. The flow rate was 0.40 mL/min, the column temperature was 40 ℃, the detection wavelength was 285 nm, and the injection volume was 2 μL. Traditional Chinese medicine fingerprint similarity evaluation software was used for similarity evaluation, chemometrics was applied to analyze Cassiae Semen from different origins, and the content of 11 indicator components in Cassiae Semen was determined. Results The established fingerprint identified a total of 26 common peaks, and 13 components were identified through reference standards. Through cluster analysis and principal component analysis (PCA), different Cassiae Semen samples can be clustered into 4 categories, and through the application of orthogonal partial least squares-discriminant analysis (OPLS-DA) method with VIP > 1 as the criterion, peaks 3 (emodin-1-O-β-D-glucoside), 6 (cassiaside), 7 (rubrofusarin-6-O-β-gentiobioside), 8 (aurantio-obtusin-6-O-D-glucoside), 9 (cassiaside C), 10, 17 (aurantio-obtusin), 21, and 23 (rubrofusarin) were selected as differential markers for distinguishing Cassiae Semen from different origins. The content ranges of isorhamnetin-3-O-β-D-glucopyranoside, cassiaflavone B2, cassiaflavone, isorhamnetin-6-O-β-D-glucopyranoside, chrysin-6-O-glucoside, cassiaflavone C, cassiaflavone B, chrysin, chrysin-3-O-β-D-glucopyranoside, isorhamnetin, and chrysophanol in 18 batches of Cassiae Semen were 0.425-1.315, 4.129-8.790, 0.453-1.849, 2.617-4.739, 0.792-2.372, 1.424-3.533, 0.496-1.439, 0.630-1.359, 0.483-2.298, 2.130-9.516, and 2.006-3.841 mg/g, respectively. According to the methodology, each component exhibited a good linear relationship. Conclusion The established fingerprint of Cassiae Semen, combined with the chemical pattern recognition, can effectively distinguish Cassiae Semen from different origins. The multi-component content determination method is stable and reliable, providing a reference for the quality evaluation and control of Cassiae Semen.

R282.6

2022年佛山市南海区重点领域科技攻关专项（南科[2023]20号-18）