基于UPLC-Q-Exactive-MS技术鉴定接骨七厘片的化学成分及血中移行成分  

Identification of the compounds of Jiegu Qili tablets and constituents absorbed into blood based on UPLC-Q-Exactive-MS technology

作　　者：方晓洋 周融融 沈冰冰[1,2] 何维维 肖洁 徐瑾 曾宏亮 张水寒 FANG Xiaoyang;ZHOU Rongrong;SHEN Bingbing;HE Weiwei;XIAO Jie;XU Jin;ZENG Hongliang;ZHANG Shuihan(Academy of Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Institute of Chinese Medicine Resources,Hunan Academy of Chinese Medicine,Changsha410013,China;Center Laboratory,Affiliated Hospital of Hunan Academy of Chinese Medicine,Changsha 410006,China;Center of Medical Laboratory Animals,Hunan Academy of Chinese Medicine,Changsha 410013,China;Hunan Jinsha Pharmaceutical Co.,Ltd.,Changsha 410015,China)

机构地区：[1]湖南中医药大学中医药研究院,长沙410208 [2]湖南省中医药研究院中药资源研究所,长沙410013 [3]湖南省中医药研究院附属医院中心实验室,长沙410006 [4]湖南省中医药研究院医学实验动物中心,长沙410013 [5]湖南金沙药业有限责任公司,长沙410015

出　　处：《药学前沿》2024年第12期605-616,共12页China Pharmacist

基　　金：湖南省自然科学基金面上项目(2021JJ30416)。

摘　　要：目的采用超高效液相色谱串联四极杆静电场轨道阱质谱(UPLC-Q-Exactive-MS)技术对接骨七厘片的化学成分及大鼠血中移行成分进行定性分析,为该制剂的体内有效物质研究提供参考。方法体外色谱条件:Hypersil GOLD C18色谱柱(50 mm×2.1 mm,1.9µm),流动相为0.1%甲酸水溶液和甲醇(梯度洗脱),柱温为30℃,进样量为5µL;体外质谱条件:样品经电喷雾电离,采用正负离子双模式检测,扫描范围为m/z100~1500。体内色谱条件:ACQUITYHPLCHSST3色谱柱(100mm×2.1mm,1.8µm),流动相为95%水+5%乙腈和47.5%乙腈+47.5%异丙醇+5%水(梯度洗脱),柱温为40℃,进样量为8µL。体内质谱条件:样品经电喷雾电离,采用正负离子双模式检测,扫描范围为m/z 70~1050。连续7 d灌胃给予SD大鼠接骨七厘片,通过腹主动脉取血获得含药血清及空白血清样品(雌、雄性接骨七厘片含药血清组和雌、雄性空白血清组)。采用非靶向LC-MS技术并利用Progenesis QI软件进行对比分析,鉴定大鼠血中移行成分。结果体外共鉴定出75种化合物,体内共鉴定出98种化合物,结合体内外鉴定的化学成分,共鉴定出7种入血成分,分别为甲基异茜草素、花生四烯酸、柠檬酸、焦谷氨酸、3-吲哚丙酸、16-羟基棕榈酸、油酸酰胺。结论该方法高效、准确灵敏,为今后在接骨七厘片药效物质基础和质量控制研究提供科学参考。Objective To conduct the qualitative analysis of the chemical composition and mobile components in rat blood of Jiegu Qili tablets using ultra performance liquid chromatography-quadrupole-exactive-mass spectrometry(UPLC-Q-Exactive-MS)technology,and to provide reference for the in vivo study of effective substances in this preparation.Methods In vitro chromatographic condition was as follows:Hypersil GOLD C18 chromatography column(50 mm×2.1 mm,1.9μm)was used,the mobile phase was 0.1%formic acid aqueous solution and methanol(gradient elution),the column temperature was 30℃,and the injection volume was 5μL.In vitro mass spectrometry condition was as follows:the sample was ionized by electric spray and detected by positive and negative ion dual mode,with scanning range of m/z 100~1500.In vivo chromatographic condition was as follows:ACQUITY HPLC HSS T3 chromatography column(100 mm×2.1 mm,1.8μm)was used,the mobile phase was 5%water+5%acetonitrile and 47.5%acetonitrile+47.5%isopropanol+5%water(gradient elution),the column temperature was 40℃,and the injection volume was 8μL.In vivo mass spectrometry condition was as follows:the sample was ionized by electric spray and detected by positive and negative ion dual mode,with scanning range of m/z 70~1050.SD rats were given Jiegu Qili tablets by intragastric administration for 7 days,and drug-containing serum and blank serum samples(female and male Jiegu Qili tablets serum group,and female and male blank serum group)were obtained through abdominal aorta.The non-targeted LCMS technique and Progenesis QI software were used for comparative analysis to identify the transitional components in the blood of rats.Results 75 compounds were identified in vitro,and 98 compounds were identified in vivo.Based on the chemical composition identified in vitro and in vivo,a total of 7 blood components were identified,including methyl isoquercetin,arachidonic acid,citric acid,pyroglutamic acid,3-indole propionic acid,16 hydroxypalmitic acid,and oleic acid amide.Conclusion This metho

关 键 词：接骨七厘片 成分分析 含药血清 超高效液相色谱串联四极杆静电场轨道阱质谱 化学成分 代谢产物 入血成分 

分 类 号：R286.0[医药卫生—中药学]