基于序贯代谢和UPLC-HRMS的陈皮入血成分分析  

Analysis on Components Absorbed into Blood of Citri Reticulatae Pericarpium in Rats Based on Sequential Metabolism and UPLC-HRMS

作　　者：陈洪娇 刘伟 李雪岩 祁东盈 于爽 刘慧宁 刘海波[2] 刘洋[1] CHEN Hongjiao;LIU Wei;LI Xueyan;QI Dongying;YU Shuang;LIU Huining;LIU Haibo;LIU Yang(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]中国医学科学院&北京协和医学院药用植物研究所,北京100193

出　　处：《中国实验方剂学杂志》2023年第9期179-187,共9页Chinese Journal of Experimental Traditional Medical Formulae

摘　　要：目的:采用序贯代谢方法和超高效液相色谱-高分辨质谱法(UPLC-HRMS)研究陈皮化学成分在大鼠体内不同部位的代谢情况。方法:以SD雄性大鼠为实验对象,在体肠灌流给予陈皮醇提物后分别制备肠代谢、肝代谢血液样品,灌胃给药后收集综合代谢血液样品。运用UPLC-HRMS分析样品,以乙腈(A)-0.1%甲酸水溶液(B)为流动相进行梯度洗脱(0~10 min,10%~30%A;10~30 min,30%~95%A;30~31 min,95%~10%A;31~35 min,10%A),流速0.35 mL·min^(-1),采用加热电喷雾离子源,正、负离子模式扫描,扫描范围m/z 100~1 500。在此检测条件下,比较陈皮醇提物、空白血浆及不同含药血浆的图谱差异,根据保留时间、精确相对分子质量、二级碎片离子及对照品信息,分析鉴别各样品的化学成分。结果:从陈皮醇提物中共鉴定出44个化学成分,包含黄酮氧苷、黄酮碳苷和多甲氧基黄酮类化合物等。序贯代谢研究结果表明,在肠代谢样品中检测到陈皮中的22个化学成分,肝代谢样品中检测到18个化学成分,3种代谢样品中均可以检测到9个相同的化学成分(芸香柚皮苷、橙皮苷、橙皮内酯、5,7,8,3’,4’,5’-hexamethoxy-flavone、异甜橙黄酮、甜橙黄酮、川陈皮素、3,5,6,7,8,3’,4’-七甲氧基黄酮、橘皮素),共有22个化合物以原型形式入血。结论:鉴定的21个结构明确且以原型入血的成分可能是陈皮潜在的活性成分,不同代谢部位的成分差异可为阐明陈皮代谢成分的体内生物转化过程提供实验依据。Objective:To study the metabolism of chemical components from Citri Reticulatae Pericarpium(CRP)in different parts of rats by sequential metabolism and ultra performance liquid chromatography-high resolution mass spectrometry(UPLC-HRMS).Method:SD male rats were employed as experimental subjects,and blood samples of intestinal metabolism and hepatic metabolism were prepared after administration of CRP ethanol extract by in situ intestinal perfusion,and comprehensive metabolic samples were collected after intragastric administration.UPLC-HRMS was used to analyze the samples with acetonitrile(A)-0.1%formic acid aqueous solution(B)as the mobile phase for gradient elution(0-10 min,10%-30%A;10-30 min,30%-95%A;30-31 min,95%-10%A;31-35 min,10%A)at a flow rate of 0.35 mL·min^(-1),using a heated electrospray ionization with positive and negative ion mode scanning in the range of m/z 100-1500.Under these conditions,the differences in the profiles of CRP ethanol extract,blank plasma and drug-containing plasma under different treatment groups were compared,and the chemical components of each sample were analyzed and identified based on the retention time,accurate relative molecular mass,primary and secondary ion fragments,and the information of reference substances.Result:A total of 44 chemical components were identified in the CRP ethanol extract,including flavone-O-glycosides,flavone-C-glycosides and polymethoxyflavonoids,etc.The results of sequential metabolism showed that 22 chemical components in CRP were detected in the intestinal metabolic sample,18 chemical components were detected in the hepatic metabolic sample,and 9 identical chemical components(narirutin,hesperidin,meranzin,5,7,8,3ʹ,4ʹ,5ʹ-hexamethoxy-flavone,isosinensetin,sinensetin,3,5,6,7,8,3ʹ,4ʹ-heptamethoxyflavone,nobiletin and tangeretin)could be detected in all three metabolic samples,with a total of 22 compounds entering the blood in prototype form.Conclusion:The identified 21 components with well-defined structures entering the blood as prototypes ma

关 键 词：陈皮 序贯代谢方法 超高效液相色谱-高分辨质谱法(UPLC-HRMS) 入血成分 黄酮氧苷 黄酮碳苷 多甲氧基黄酮 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] R96[理学—分析化学] O657[理学—化学]