目的 建立牵牛子中6个酚酸类成分（新绿原酸、绿原酸、咖啡酸、隐绿原酸、异绿原酸A和异绿原酸C）的HPLC含量测定方法，研究炒制前后牵牛子中这些成分的含量变化规律。方法 采用HPLC法同时测定新绿原酸、绿原酸、咖啡酸、隐绿原酸、异绿原酸A和异绿原酸C的含量，运用化学计量学方法对含量测定结果进行分析，并通过模拟炮制的方法对此类成分在不同温度下的转化进行初步研究。色谱条件为Kromasil C₁₈（250mm×4.6mm，5μm）色谱柱，流动相为甲醇-0.1%甲酸水溶液，梯度洗脱（0～20min，10%甲醇；20～25min，10%～15%甲醇；25～45min，15%甲醇；45～50min，15%～30%甲醇；50～75min，30%甲醇；75～80min，30%～40%甲醇；80～110min，40%甲醇），检测波长330nm，体积流量 1 mL/min，进样量10μL，柱温30℃。结果 与生品比较，炒制后牵牛子中绿原酸的含量无明显变化，新绿原酸、隐绿原酸和异绿原酸 C 在炒制后含量显著上升；咖啡酸和异绿原酸A在炒制后含量显著下降。层次聚类分析（hierarchical clusteranalysis，HCA）将30批样品分为生品与炮制品2类，主成分分析（principal component analysis，PCA）和正交偏最小二乘-判别分析（partial least squares-discriminant analysis，PLS-DA）均标记了4个差异性成分。各单体化合物在模拟炮制过程中发生了相互转化，转化程度与加热温度相关。结论 牵牛子与炒牵牛子的化学成分含量发生了一定程度改变，其中新绿原酸、隐绿原酸、异绿原酸A和异绿原酸C可作为牵牛子炒制前后质量评价的关键性指标，可为牵牛子与炒牵牛子质量标准的建立和深入研究提供科学依据。

Objective To establish the HPLC method for the determination of six phenolic acids (neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, isochlorogenic acid A, and isochlorogenic acid C) in Qianniuzi (Pharbitidis Semen) and to study the content changes of the compounds in Pharbitidis Semen before and after stir-frying. Methods The contents of neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, isochlorogenic acid A, and isochlorogenic acid C were determined simultaneously by HPLC, and analyzed by chemometrics. The conversions among them at different temperatures were preliminary studied by simulated processing method. Kromasil C₁₈ (250 mm×4.6 mm, 5 μm) was employed, and the mobile phase was methanol-0.1% formic acid aqueous solution for gradient elution (0—20 min, 10% methanol; 20—25 min, 10%—15% methanol; 25—45 min, 15% methanol; 45—50 min, 15%—30% methanol; 50—75 min, 30% methanol; 75—80 min, 30%—40% methanol; 80—110 min, 40% methanol). The detection wavelength was set at 330 nm, flow rate was 1 mL/min, the injection volume was 10 μL, and the column temperature was 30 ℃. Results Compared with raw Pharbitidis Semen, the content of chlorogenic acid had no significant change after processing, while the contents of neochlorogenic, cryptochlorogenic acid and isochlorogenic acid C were increased significantly. And the contents of caffeic acid and isochlorogenic acid A were decreased significantly. The 30 batches of Pharbitidis Semen were divided into two categories, raw and processed products, by hierarchical cluster analysis (HCA). Four components that caused the differences were marked by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). Six monomers transformed into each other in the process of simulated processing, and the degree of transformation was related to temperature. Conclusion The contents of chemical components in Pharbitidis Semen and fried Pharbitidis Semen have changed to a certain extent. Among them, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A and isochlorogenic acid C can be used as the key indicators for quality evaluation of Pharbitidis Semen before and after processing. It can provide a scientific basis for the establishment and in-depth study of quality standards for Pharbitidis Semen and fried Pharbitidis Semen.

R283.1

国家重点研发计划项目(2018YFC1707106);山东省重点研发计划(2021SFGC1202)