^(1)H-NMR定量测定黄芪注射液中亲水性成分和疏水性成分  

Quantitative determination of hydrophilic and hydrophobic components in Astragalus Injection based on^(1)H-NMR

作　　者：杨嘉誉 徐绍静 张琪 应旭辉 李文竹 潘坚扬[1] 瞿海斌[1] YANG Jiayu;XU Shaojing;ZHANG Qi;YING Xuhui;LI Wenzhu;PAN Jianyang;QU Haibin(Institute of Pharmaceutical Informatics,College of Pharmaceutical Sciences,Zhejiang University,Hangzhou 310058,China;Chiatai Qingchunbao Pharmaceutical Co.,Ltd.,Hangzhou 310023,China)

机构地区：[1]浙江大学药学院,药物信息学研究所,浙江杭州310058 [2]正大青春宝药业有限公司,浙江杭州310023

出　　处：《中草药》2024年第10期3321-3330,共10页Chinese Traditional and Herbal Drugs

摘　　要：目的建立黄芪注射液中亲水性成分和疏水性成分的^(1)H-NMR定量分析方法。方法以10%氘代水(10%D2O)和氘代甲醇(CD3OD)为氘代溶剂制备黄芪注射液亲水性成分和疏水性成分的^(1)H-NMR样品。使用Bruker Avance Ⅲ 600型核磁共振波谱仪采集^(1)H-NMR图谱,具体实验参数如下:Noesygppr1D脉冲序列;温度为298 K,数据点32 K,采集时间(AQ)为2.980 s,脉冲宽度(SW)为7211.54 Hz,弛豫延迟时间(D1)分别为15.0 s和20.0 s,频率偏移(O1)分别为2816.65 Hz和2930.31 Hz,接收器增益(RG)值为57,扫描次数(NS)32次。结果在^(1)H-NMR图谱中归属出33种成分,分别是异亮氨酸、亮氨酸、缬氨酸、丙氨酸、乳酸、焦谷氨酸、天冬酰胺、γ-氨基丁酸、脯氨酸、甲酸、乙酸、富马酸、苹果酸、琥珀酸、丙二酸、蔗糖、果糖、半乳糖、葡萄糖、胆碱、甜菜碱、葫芦巴碱、尿苷、腺苷、胞苷、鸟苷、腺嘌呤、黄芪皂苷Ⅲ、黄芪甲苷、芒柄花苷、毛蕊异黄酮苷、9,10-二甲氧基紫檀烷-3-O-葡萄糖苷以及异黄烷苷。对其中27种化学成分(异亮氨酸、缬氨酸、丙氨酸、乳酸、焦谷氨酸、天冬酰胺、γ-氨基丁酸、脯氨酸、甲酸、乙酸、富马酸、苹果酸、琥珀酸、丙二酸、蔗糖、果糖、葡萄糖、胆碱、甜菜碱、葫芦巴碱、尿苷、腺苷、鸟苷、腺嘌呤、黄芪皂苷Ⅲ、黄芪甲苷、毛蕊异黄酮苷)进行定量,可定量的成分占注射液固含量90%以上。该方法的精密度、重复性、样品稳定性和耐用性良好,各个成分线性关系良好,r 2大于0.9990,加样回收回收率为98.19%~101.40%。结论建立了一种简单、快速、可靠的^(1)H-NMR定量方法用于测定黄芪注射液中的亲水性成分和疏水性成分,为黄芪注射液的质量评价提供了新的分析手段。Objective To establish a ^(1)H-NMR quantitative method for the hydrophilic and hydrophobic components in Astragalus Injection(黄芪注射液).Methods ^(1)H-NMR samples of hydrophilic and hydrophobic components in Astragalus Injection were prepared with 10%deuterated water(10%D2O)and deuterated methanol(CD3OD),respectively.Then Bruker Avance Ⅲ 600 NMR spectrometer was used to collect ^(1)H-NMR spectra.The parameters were set up as follows:Noesygppr1D pulse sequence,temperature was 298 K,data point was 32 K,acquisition time(AQ)was 2.980 s,pulse width(SW)was 7211.54 Hz,relaxation delay time(D1)was 15.0 s and 20.0 s,respectively,frequency offset(O1)were 2816.65 Hz and 2930.31 Hz,respectively,receiver gain(RG)was 57,and the number of scans(NS)was 32.Results A total of 33 components were assigned,namely isoleucine,leucine,valine,alanine,lactate,pyroglutamate,asparagine,γ-aminobutyric acid,proline,formate,acetate,fumaric acid,malic acid,succinic acid,malonic acid,sucrose,fructose,lactose,glucose,choline,betaine,trigonelline,uridine,adenosine,cytidine,guanosine,adenine,astragaloside III,astragaloside IV,ononin,calycosin-7-glucoside,9,10-dimethoxypterocarpan-3-O-glucoside and isomucronulatol 7-O-glucoside.Quantitative analysis was conducted for 27 chemical constituents,including isoleucine,valine,alanine,lactate,pyroglutamate,asparagine,γ-aminobutyric acid,proline,formate,acetate,fumaric acid,malic acid,succinic acid,malonic acid,sucrose,fructose,glucose,choline,betaine,trigonelline,uridine,adenosine,guanosine,adenine,astragaloside III,astragaloside IV and calycosin-7-glucoside.The components that can be quantified by this method account for more than 90%of the solid content in the injection.The methodological validation results showed that the method’s precision,repeatability,sample stability,and robustness were good.The linear of each component was good,the correlation coefficient was more than 0.9990,and the recovery rate ranged from 98.19%to 101.40%.Conclusion A simple,rapid and reliable ^(1)H-NMR quantitative

关 键 词：质子核磁共振 黄芪注射液 含量测定 中药注射剂 亲水性成分 疏水性成分 ^(1)H-NMR定量分析方法 异亮氨酸 缬氨酸 丙氨酸 乳酸 焦谷氨酸 天冬酰胺 γ-氨基丁酸 脯氨酸 甲酸 乙酸 富马酸 苹果酸 琥珀酸 丙二酸 蔗糖 果糖 葡萄糖 胆碱 甜菜碱 葫芦巴碱 尿苷 腺苷 鸟苷 腺嘌呤 黄芪皂苷Ⅲ 黄芪甲苷 毛蕊异黄酮苷 

分 类 号：R283.6[医药卫生—中药学]