HPLC-MS/MS法测定芪葵颗粒中25个成分的含量  

Simultaneous determination of 25 components in Qikui granules by HPLC-MS/MS

作　　者：陆明霞 欧阳冰琛 廖健城 桂婉玉 向杰 邹建东 李长印 LU Ming-xia;OUYANG Bing-chen;LIAO Jian-cheng;GUI Wan-yu;XIANG Jie;ZOU Jian-dong;LI Chang-yin(Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Hospital of Traditional Chinese Medicine,Clinical Pharmacology,Nanjing 210029,China)

机构地区：[1]南京中医药大学附属医院/江苏省中医院临床药理科,南京210029

出　　处：《药物分析杂志》2022年第10期1708-1719,共12页Chinese Journal of Pharmaceutical Analysis

基　　金：江苏省自然科学基金面上项目(BK20201502);“333高层次人才培养工程”科研资助项目(BRA2020384);江苏省卫健委医学科研项目面上项目(H2019092);江苏省中医院高峰人才项目(y2018rc22,y2021rc31);江苏高校优势学科建设工程资助项目(苏政办发[2018]87号);南京中医药大学中医学优势学科三期项目开放课题(ZYX03KF030)。

摘　　要：目的:建立高效液相色谱-串联质谱(HPLC-MS/MS)法测定芪葵颗粒中25个成分(7,2’-二羟基-3’,4’-二甲氧基异黄烷、L-苯丙氨酸、毛蕊异黄酮-7-O-葡萄糖苷、芦丁、2,3,5,4’-四羟基二苯乙烯-2-O-葡萄糖苷、美迪紫檀苷、异黄芪皂苷Ⅱ、杨梅素、无水柠檬酸、紫檀烷、黄芪皂苷Ⅲ、黄芪甲苷、黄芪皂苷Ⅰ、环黄芪苷Ⅱ、异黄芪皂苷Ⅰ、L-色氨酸、黄芪皂苷Ⅱ、异槲皮苷、刺芒柄花苷、金丝桃苷、没食子酸、槲皮素、毛蕊异黄酮、芒柄花素、大黄素)的含量。方法:采用Agilent Poroshell 120 SB-C_(18)色谱柱(100 mm×3.0 mm,2.7μm),以含0.1%甲酸和1 mmol·L^(-1)甲酸铵的水溶液(A)-甲醇(B)为流动相,梯度洗脱,流速0.4 mL·min^(-1),柱温45℃,进样体积2μL;采用电喷雾离子源(ESI),多反应监测(MRM)模式,负离子模式扫描。结果:25个成分在测定的质量浓度范围内线性关系良好,相关系数(r)均>0.9950,精密度、重复性、稳定性均良好,平均加样回收率在89.5%~105.7%,RSD≤4.0%;3批芪葵颗粒中上述25个成分的含量范围分别为0.005~0.006、0.102~0.104、0.107~0.155、0.073~0.074、0.016~0.019、1.058~1.120、0.032~0.047、0.040~0.052、5.600~6.617、0.026~0.030、0.009~0.011、0.009~0.013、0.077~0.085、0.018~0.026、0.131~0.154、0.061~0.068、0.032~0.038、0.108~0.127、0.043~0.064、0.167~0.185、0.084~0.110、0.021~0.026、0.186~0.203、0.091~0.094、0.020~0.033 mg·g^(-1)。结论:所建立的方法可准确测定芪葵颗粒中多个主要成分,适用于芪葵颗粒制剂的质量评估与控制。Objective:To establish a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for the simultaneous determination of 25 components(7,2’-dihydroxy-3’,4’-dimethoxyisoflavan,L-phenylalanine,calycosin-7-O-glucoside,rutin,isoastragalosideⅡ,2,3,5,4’-tetrahydroxyl diphenylethylene-2-O-glucoside,methylnissolin-3-O-glucoside,myricetin,citric acid,3-hydroxy-9,10-dimethoxypterocarpan,astragalosideⅢ,astragalosideⅣ,astragalosideⅠ,cyclocephalosideⅡ,isoastragalosideⅠ,L-tryptophan,astragalosideⅡ,isoquercitrin,formononetin-7-O-glucoside,hyperoside,gallic acid,quercetin,calycosin,formononetin and emodin)in Qikui granules.Methods:Chromatographic conditions:the chromatographic column was Agilent Poroshell 120 SB-C_(18)(100 mm×3.0 mm,2.7μm).The mobile phase was composed of 1 mmol·L^(-1)ammonium formate containing 0.1%formic acid(A)and methanol(B)in gradient elution at a flow rate of 0.4 mL·min^(-1)and the column temperature was set at 45℃.The injection volume was 2μL.Mass spectrum conditions:the analytes were determined by electrospray ionization source(ESI)combined with multiple reaction monitoring(MRM)in negative ion mode.Results:The linear relationships of 25 components were good in corresponding ranges with correlation coefficients(r)all above 0.9950.The precision,repeatability and stability were good.The average recoveries varied between 89.5%and 105.7%with RSDs≤4.0%.And the average contents of the above-mentioned 25 components in 3 batches of Qikui granules were 0.005-0.006,0.104-0.104,0.107-0.155,0.073-0.074,0.016-0.019,1.058-1.120,0.032-0.047,0.040-0.052,5.600-6.617,0.026-0.030,0.009-0.011,0.009-0.013,0.077-0.085,0.018-0.026,0.131-0.154,0.061-0.068,0.032-0.038,0.108-0.127,0.043-0.064,0.167-0.185,0.084-0.110,0.021-0.026,0.203-0.186,0.091-0.094,and 0.020-0.033 mg·g^(-1),respectively.Conclusion:The established method can accurately determine several main components in Qikui granules and is suitable for quality evaluation and control of Qikui granules.

关 键 词：高效液相色谱-串联质谱法 芪葵颗粒 多反应监测模式 7 2’-二羟基-3’ 4’-二甲氧基异黄烷 L-苯丙氨酸 毛蕊异黄酮-7-O-葡萄糖苷 芦丁 异黄芪皂苷 2 3 5 4’-四羟基二苯乙烯-2-O-葡萄糖苷 美迪紫檀苷 杨梅素 无水柠檬酸 黄芪紫檀烷 黄芪皂苷 黄芪甲苷 环黄芪苷 L-色氨酸 异槲皮苷 刺芒柄花苷 金丝桃苷 没食子酸 槲皮素 毛蕊异黄酮 芒柄花素 大黄素 

分 类 号：R917[医药卫生—药物分析学]