摘要
目的建立基于UPLC-Q-Orbitrap HRMS的复方血栓通胶囊中多种活性成分定量分析方法,并采用主成分分析法对其质量进行综合评价。方法采用Acquity UPLC~?BEH C_(18)色谱柱(50 mm×2.1 mm,1.7μm),以乙腈(A)-0.1%甲酸水(B)为流动相进行梯度洗脱,以实现化合物的前期分离;通过Q-Orbitrap MS先进的正、负离子同时监测、一级全扫描及自动触发二级质谱扫描的模式捕捉目标成分的精确相对分子质量及碎片离子信息,以实现对待测物的准确定性和定量;最后将定量结果与主成分分析法相结合对不同批次药物进行科学的质量评价分析。结果在优化的色谱、质谱条件下,甜菜碱、琥珀酸、丹参素钠、丹参素、原儿茶酸、原儿茶醛、咖啡酸、芦丁、人参皂苷Rg_1、迷迭香酸、丹酚酸A、丹酚酸B、汉黄芩素、毛蕊异黄酮、芒柄花黄素、黄芪皂苷Ⅱ、黄芪皂苷Ⅰ、人参皂苷Rg_3、二氢丹参酮Ⅰ、丹参酮Ⅰ、隐丹参酮、丹参酮Ⅱ_A和齐墩果酸分别在0.009 8～0.314 5、0.067 8～2.170 7、0.044 2～1.413 3、0.059 6～1.907 2、0.003 3～0.104 4、0.002 8～0.089 9、0.001 2～0.038 3、0.006 3～0.203 2、0.960 5～30.735 5、0.022 2～0.709 0、0.083 7～2.679 5、0.593 8～19.002 6、0.000 2～0.005 3、0.012 3～0.394 4、0.004 5～0.143 5、0.009 2～0.293 4、0.066 0～2.113 3、0.033 0～1.055 0、0.004 5～0.145 5、0.015 9～0.508 1、0.024 1～0.772 0、0.009 3～0.297 8、0.002 5～0.078 8μg/mL线性关系良好(r≥0.999 0);精密度、重复性及稳定性良好(RSD≤5%);加样回收率在98%～101%,RSD均小于3%;定量分析结果表明大多数批次药物质量较为稳定,其中人参皂苷Rg_1、丹酚酸B、琥珀酸、丹酚酸A、丹参素和丹参素钠对药物质量具有较大影响,可对其进行重点监控以保证药物批次质量。结论建立的定量方法灵敏度高且准确性好,方法学考察结果符合测定要求,可用于复方血栓通胶囊中多种活性成分的快速测定,并为其质量评价提供新的科学依据和参考。
Objective To establish a quantitative analysis method of multiple active components in Compound Xueshuantong Capsules（CXC） based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry（UPLCQ-Orbitrap HRMS）, and make a quality assessment using principal component analysis. Methods The column was Acquity UPLC~？BEH C_（18）（50 mm × 2.1 mm, 1.7 μm） and the mobile phase was consisted of acetonitrile（A）-water（B）（containing 0.1% formic acid） with gradient elution; The information of accurate mass and fragment ions was obtained by the novel ＂monitored simultaneously for positive and negative ions, Full MS scan and automatic trigger secondary mass spectrometry＂ mode of Q-Orbitrap MS technology to realize the accurate qualitation and quantitation; Using the quantitative results combined with the principal component analysis to achieve the scientific assessment of the drug in different batches. Results Under the optimized conditions, betaine, succinic acid, salvianic acid A sodium, danshensu, protocatechuate, protocatechuic aldehyde, caffeic acid, rutin, ginsenoside Rg_1, rosmarinicacid, salvianolic acid A, salvianolic acid B, wogonin, calycosin, formononetin, astragaloside Ⅱ, astragaloside I, ginsenoside Rg_3, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone Ⅱ_A, and oleanic acid all showed good liner relationship（r ≥ 0.999 0） in the range of 0.009 8—0.314 5, 0.067 8—2.170 7, 0.044 2—1.413 3, 0.059 6—1.907 2, 0.003 3—0.104 4, 0.002 8—0.089 9, 0.001 2—0.038 3, 0.006 3—0.203 2, 0.960 5—30.735 5, 0.022 2—0.709 0, 0.083 7—2.679 5, 0.593 8—19.002 6, 0.000 2—0.005 3, 0.012 3—0.394 4, 0.004 5—0.143 5, 0.009 2—0.293 4, 0.066 0—2.113 3, 0.033 0—1.055 0, 0.004 5—0.145 5, 0.015 9—0.508 1, 0.024 1—0.772 0, 0.009 3—0.297 8, 0.002 5—0.078 8 μg/mL, respectively; The results of the accuracy, repeatability, and stability all reached the standards（RSD ≤ 5%）; The recoveries ranged from 98%—101% and RSDs were all below 3%; the analysis results showed that the quality of the most batches was stable, the ginsenoside Rg_1, salvianolic acid B, succinic acid, salvianolic acid A, danshensu, and salvianic acid A sodium had a great influence on the quality of the medicine, which could be specially monitored to ensure the quality of different batches of the medicine. Conclusion The methods established in this paper have a high sensitivity and accuracy; The results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in CXC. The research also provides a new scientific basis and reference for the quality assessment at the same time.
作者
李宁
LI Ning(Department of Pharmacy,Affiliated Cancer Hospital of Zhengzhou University,Henan Cancer Hospital,Zhengzhou 450008,China)
出处
《中草药》
CAS
CSCD
北大核心
2018年第19期4552-4560,共9页
Chinese Traditional and Herbal Drugs
作者简介
李宁，男，副主任药师，研究方向为中药质量控制。Tel：（0371）65587016 E-mail：lining7621@126．com
