基于植物化学分析和抗炎筛选的质量标志物：UHPLC-Q-Orbitrap HRMS 用于降香的质量控制的综合策略。
Quality markers based on phytochemical analysis and anti-inflammatory screening: An integrated strategy for the quality control of Dalbergia odorifera by UHPLC-Q-Orbitrap HRMS.
机构信息
School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong, Yantai University, Yantai 264005, China.
School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong, Yantai University, Yantai 264005, China.
出版信息
Phytomedicine. 2021 Apr;84:153511. doi: 10.1016/j.phymed.2021.153511. Epub 2021 Feb 12.
BACKGROUND
Quality control, key for the clinical application of traditional Chinese medicines (TCMs), should be connected to the authentication and efficacy of TCMs. The heartwood of Dalbergia odorifera has been widely used to treat inflammation-related diseases. However, in the Chinese pharmacopeia, only the total volatile oil, which does not sufficiently reflect the clinical efficacy, is used as a quality control indicator.
PURPOSE
Establishing a "phytochemical-specificity-effectiveness-Q-marker" analytical strategy to improve the quality control of D. odorifera.
METHODS
Combined with biosynthetic pathway analysis, phytochemical compositions identified by UHPLC-Q-Orbitrap HRMS were used to build substantial phytochemical groups and further discover specific Q-markers. Then, lipopolysaccharide-stimulated RAW 264.7 cells were used to screen effective anti-inflammatory ingredients. Finally, a UHPLC-HRMS method was developed and validated to quantify the selected Q-markers in D. odorifera samples.
RESULTS
Along the constructed biosynthetic pathways, 93 phytochemical components were identified in D. odorifera, including 7 chalcones, 13 flavanones, 21 isoflavones, 21 isoflavanones, 3 flavonols, 19 neoflavones, etc. Among them, 31 compounds representing these 6 categories were further evaluated for their anti-inflammatory activities. It revealed that the extract of D. odorifera and nine flavonoids in the noncytotoxic range could alleviated lipopolysaccharide-stimulated inflammation in RAW 264.7 cells by decreasing the production of proinflammatory mediators such as nitric oxide and interleukin-6. Notably, neoflavones, as species-specific components, exhibited superior anti-inflammatory activities among the representative compounds. Finally, 12 Q-markers (butin, liquiritigenin, eriodictyol, melanettin, naringenin, butein, genistein, 4'-hydroxy-4-methoxydalbergione, isoliquiritigenin, 2,4-dihydroxy-5-methoxybenzophenone, medicarpin, and pinocembrin), which reflect specificity and effectiveness, were successfully quantified in 10 batches of samples from different origins. The origins and consistency of D. odorifera could be efficiently discriminated by hierarchical cluster analysis (HCA).
CONCLUSION
The analysis strategy that combines phytochemical analysis with anti-inflammatory screening clarified the therapeutic material basis and discovered Q-markers, which possibly offers a more comprehensive quality assessment of D. odorifera.
背景
质量控制是中药（TCM）临床应用的关键，应与 TCM 的鉴定和疗效相关联。降香的心材被广泛用于治疗炎症相关疾病。然而，在中国药典中，仅使用总挥发油作为质量控制指标，而总挥发油并不能充分反映临床疗效。
目的
建立“化学成分-特异性-功效-质量标志物（Q-标志物）”分析策略，以提高降香的质量控制水平。
方法
结合生物合成途径分析，采用 UHPLC-Q-Orbitrap HRMS 鉴定的化学成分用于构建实质性化学成分群，并进一步发现特异性 Q-标志物。然后，利用脂多糖刺激 RAW 264.7 细胞筛选具有抗炎作用的有效成分。最后，建立并验证 UHPLC-HRMS 方法，以定量测定降香样品中选定的 Q-标志物。
结果
沿着构建的生物合成途径，在降香中鉴定出 93 种化学成分，包括 7 种查尔酮、13 种二氢黄酮、21 种异黄酮、21 种异黄烷酮、3 种黄酮醇、19 种新黄烷酮等。其中，进一步评估了代表这 6 类的 31 种化合物的抗炎活性。结果表明，降香提取物和非细胞毒性范围内的 9 种黄酮类化合物可通过降低一氧化氮和白细胞介素-6 等促炎介质的产生来减轻脂多糖刺激的 RAW 264.7 细胞炎症。值得注意的是，新黄烷酮作为种特异性成分，在代表性化合物中表现出更好的抗炎活性。最后，成功定量了来自不同产地的 10 批样品中的 12 个 Q-标志物（布替、甘草素、圣草酚、黑色素、柚皮苷、布替、染料木素、4'-羟基-4-甲氧基降二氢黄牛木酮、异甘草素、2,4-二羟基-5-甲氧基二苯甲酮、甘草次酸和芹菜素），这些 Q-标志物反映了特异性和功效。层次聚类分析（HCA）可有效区分降香的产地和一致性。
结论
结合化学成分分析和抗炎筛选的分析策略阐明了治疗物质基础，并发现了 Q-标志物，这可能为降香提供更全面的质量评估。
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