目的 建立超高效液相色谱-四级杆-飞行质谱（UPLC-Q-TOF-MS/MS）方法，结合GNPS分子网络技术，对浙江红山茶花Camellia chekiangoleosa中化学成分进行快速解析，并对其抗炎活性进行初步评价。方法 采用ACQUITY UPLC®HSS T3色谱柱（150 mm×3 mm，1.8 μm），流动相为乙腈-0.1%甲酸水，梯度洗脱，电喷雾离子源（ESI）采用正离子模式扫描。通过对浙江红山茶花70%乙醇提取物进行质谱数据采集，依据分子裂解特征的相似性创建分子网络，总结各类型化合物裂解规律；并采用脂多糖（lipopolysaccharide，LPS）诱导的RAW264.7巨噬细胞炎症模型，考察浙江红山茶花对一氧化氮（nitric oxide，NO）和白细胞介素-6（interleukin-6，IL-6）释放的抑制能力，评价其抗炎效果。结果 从浙江红山茶花70%乙醇提取物中快速表征化合物166个，包括黄酮类、酚类、三萜类、有机酸及其他类型化合物等。抗炎活性结果表明，浙江红山茶花具有较强的抑制LPS诱导的巨噬细胞RAW264.7细胞释放NO和IL-6的作用。结论 UPLC-Q-TOF-MS/MS联合分子网络技术可以系统、高效、全面的鉴定浙江红山茶中的化学成分，包括黄酮、酚类和三萜等，浙江红山茶具有明显抗炎活性，值得进一步的开发利用。

Objective To establish an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) method, combined with GNPS molecular network technology, for the rapid identification of chemical constituents in Camellia chekiangoleosa flower, and then evaluating its anti-inflammatory activity. Methods ACQUITY UPLC®HSS T₃ column (150 mm×3 mm, 1.8 μm) was used. The mobile phases were acetonitrile and 0.1% formic acid water with gradient elution. Electrospray ion source (ESI) scanning in positive ion mode was used. The mass spectrum data of 70% ethanol extract of C. chekiangoleosa flower was collected. The molecular network was established according to the similarity of molecular cracking characteristics, and their cracking laws of various compounds were summarized. Using lipopolysaccharide (LPS) induced inflammation model of RAW264.7 macrophages, the inhibitory ability of C. chekiangoleosa flower on nitric oxide (NO) and interleukin-6 (IL-6) release were investigated to evaluate its anti-inflammatory effect.Results A total of 166 compounds, including flavonoids, phenols, triterpenes, organic acids and other compounds, were rapidly characterized from 70% ethanol extract of C. chekiangoleosa flower. The results of anti-inflammatory activity showed that C. chekiangoleosa flower had a strong effect on inhibiting the release of NO and IL-6 from RAW264.7 macrophages induced by LPS. Conclusion UPLC-Q-TOF-MS/MS combined with molecular network technology can systematically, efficiently and comprehensively identify the chemical constituents of C. chekiangoleosa flower, including flavonoids, phenols and triterpenes, C. chekiangoleosa flower has obvious anti-inflammatory activities and is worth further development and utilization.

R284.1

山茶花抗老修护领域综合利用研究项目（HX20241218）