UPLC-MS/MS法同时测定复方血栓通胶囊中9种成分的含量  

Simultaneous Determination of 9 Components in Compound Xueshuantong Capsules by UPLC-MS/MS

作　　者：孙志[1] 胡玉荣[2] 左莉华[1] 周霖[1] 姜晓芳[1] 刘新[1] 吕晓景 包晓悦 康建[1] 张晓坚[1] SUN Zhi HU Yurong ZUO Lihua ZHOU Lin JIANG Xiaofang LIU Xin LYU Xiaojing BAO Xiaoyue KANG Jian ZHANG Xiaojian(Dept. of Pharmacy, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China College of Pharmacy, Zhengzhou University, Zhengzhou 450001, China)

机构地区：[1]郑州大学第一附属医院药学部,郑州450052 [2]郑州大学药学院,郑州450001

出　　处：《中国药房》2017年第21期2959-2963,共5页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81272563);郑州大学第一附属医院院内青年创新基金项目

摘　　要：目的:建立同时测定复方血栓通胶囊中丹参素、咖啡酸、迷迭香酸、丹酚酸B、丹酚酸A、丹参酮Ⅰ、隐丹参酮、丹参酮ⅡA和熊果酸含量的方法。方法:采用超高效液相色谱-串联质谱法。色谱柱为ACQUITY UPLC~?BEH C_(18),流动相为乙腈-0.1%甲酸(梯度洗脱),流速为0.2 mL/min,柱温为40℃,进样器温度为10℃,分析时间为7 min,进样量为5μL;离子化模式为电喷雾电离,离子源温度为150℃,毛细管电压为3.5 kV,锥孔气流量为50 L/h,脱溶剂气温为350℃,脱溶剂气流量为650 L/h,雾化气压力为7×105Pa,工作模式为正、负离子结合的多反应监测模式。结果:丹参素、咖啡酸、迷迭香酸、丹酚酸B、丹酚酸A、丹参酮Ⅰ、隐丹参酮、丹参酮ⅡA和熊果酸检测质量浓度线性范围分别为10.0~100.0μg/mL(r=0.999 8)、0.1~1.0μg/mL(r=0.999 8)、4.0~40.0μg/mL(r=0.999 9)、10.0~100.0μg/mL(r=0.999 9)、15.0~150.0μg/mL(r=0.999 7)、8.0~80.0μg/mL(r=0.999 8)、10.0~100.0μg/mL(r=0.999 7)、50.0~500.0μg/mL(r=0.999 7)、6.0~60.0μg/mL(r=0.999 8);定量限分别为40.0、9.6、38.0、88.0、130.0、39.0、4.4、3.2、10.0 ng/mL,检测限分别为12.0、3.0、11.0、26.0、39.0、12.0、1.3、1.0、3.0 ng/mL;精密度、稳定性、重复性试验的RSD<3%;加样回收率分别为97.34%~103.20%(RSD=2.19%,n=6)、97.22%~102.39%(RSD=2.03%,n=6)、98.51%~101.70%(RSD=1.32%,n=6)、97.86%~102.49%(RSD=2.09%,n=6)、96.75%~103.12%(RSD=2.36%,n=6)、98.43%~101.65%(RSD=1.25%,n=6)、97.59%~101.50%(RSD=1.50%,n=6)、96.45%~102.88%(RSD=2.58%,n=6)、97.02%~103.11%(RSD=2.38%,n=6)。结论:该方法操作简单、结果准确,可用于复方血栓通胶囊中9种成分含量的同时测定。OBJECTIVE： To establish a method for simultaneous determination of tanshinol, caffeic acid, rosmarinic, salvianolic acid B, salvianolic acid A, tanshinoneⅠ, cryptotanshinone, tanshinone ⅡA and ursolic acid in Compound xueshuantong capsules. METHODS： UPLC-MS/MS method was adopted. The determination was performed on ACQUITY UPLC？ BEH C18 column with mobile phase consisted of acetonitrile-0.1% formic acid （gradient elution） at the flow rate of 0.2 mL/min. The column temperature was 40 ℃, and the temperature of injector was 10 ℃. Analysis time was 7 min, and sample size was 5 μL. The electrospray ionization source （ESI） was used; ion source temperature was 150 ℃; capillary voltage was 3.5 kV; cone flow was 50 L/h; desolvation temperature was 350 ℃; desolvation gas flow was 650 L/h; nebuliser pressure was 7×105 Pa; ion monitoring and multiple reaction monitoring （MRM） was performed. RESULTS： The linear ranges of tanshinol, caffeic acid, rosmarinic, salvianolic acid B, salvianolic acid A, tanshinoneⅠ, cryptotanshinone, tanshinone ⅡA and ursolic acid were 10.0-100.0 μg/mL （r=0.999 8）, 0.1-1.0 μg/mL （r=0.999 8）, 4.0-40.0 μg/mL （r=0.999 9）, 10.0-100.0 μg/mL （r=0.999 9）, 15.0-150.0 μg/mL （r=0.999 7）, 8.0-80.0 μg/mL（r=0.999 8）, 10.0-100.0 μg/mL （r=0.999 7）, 50.0-500.0 μg/mL （r=0.999 7） and 6.0-60.0 μg/mL （r=0.999 8）, respectively. The limits of quantitation were 40.0, 9.6, 38.0, 88.0, 130.0, 39.0, 4.4, 3.2 and 10.0 ng/mL, separately. The limits of detection were 12.0, 3.0, 11.0, 26.0, 39.0, 12.0, 1.3, 1.0 and 3.0 ng/mL, respectively. RSDs of precision, stability and reproducibility tests were all lower than 3%. The recoveries were 97.34%-103.20% （RSD=2.19%,n=6）, 97.22%-102.39% （RSD=2.03%,n=6）, 98.51%-101.70% （RSD=1.32%,n=6）, 97.86%-102.49% （RSD=2.09%,n=6）, 96.75%-103.12% （RSD=2.36%,n=6）,98.43%-101.65%（RSD=1.25%,n=6）, 97.59%-101.50%（RSD=1.50%,n=6）, 96.45%-102.88% （RSD=2.58%,n=6）, 97.02%-103.11% （RSD=2.38%,n=6�

关 键 词：超高效液相色谱-串联质谱法 复方血栓通胶囊 含量测定 丹参素 咖啡酸 迷迭香酸 丹酚酸B 丹酚酸A 丹参酮Ⅰ 隐丹参酮 丹参酮ⅡA 熊果酸 

分 类 号：R917[医药卫生—药物分析学]