基于UHPLC-ESI-QqQ-MS/MS技术的中药复方新天泰1号多类别多成分质量控制方法研究
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摘要
目的建立同时快速测定新天泰1号制剂中19种药效成分(人参皂苷Rg1、Rb1、Rd,小檗碱、表小檗碱、药根碱、巴马汀、非洲防己碱、黄连碱、吴茱萸碱、吴茱萸次碱、去氢吴茱萸碱、柠檬苦素、金丝桃苷、姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素、松果菊苷、毛蕊花糖苷)的超高效液相色谱-电喷雾三重四极杆质谱(UHPLC-ESI-Qq Q-MS/MS)新方法,为产品提供全面高效的质量控制方法。方法采用Agilent 1290II高效液相色谱系统,Agilent Zorbax Eclipse Plus C18色谱柱(150 mm×4.6 mm,3.5μm),乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱;采用正负离子多反应监测模式对各成分特定离子对进行监测,测定新天泰1号样品中19种活性成分含量。结果在15 min内完成19种活性成分的准确测定,极大地提高了分析效率;19种活性成分在线性范围内线性关系良好(r2>0.999 0);19种活性成分人参皂苷Rg1、Rb1、Rd及小檗碱、表小檗碱、药根碱、巴马汀、非洲防己碱、黄连碱、吴茱萸碱、吴茱萸次碱、去氢吴茱萸碱、柠檬苦素、金丝桃苷、姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素、松果菊苷、毛蕊花糖苷的回收率分别为94.80%、96.78%、95.59%、96.88%、97.74%、100.08%、96.27%、100.25%、98.32%、97.16%、95.60%、95.28%、96.81%、95.22%、96.85%、95.31%、93.86%、94.79%、95.20%;19种成分在3批样品中的定量测定结果分别为2.28~2.49 mg/g、0.82~0.90 mg/g、1.22~1.32 mg/g、14.44~15.50 mg/g、3.71~3.99 mg/g、3.26~3.49 mg/g、3.09~3.33 mg/g、4.39~4.72 mg/g、4.56~4.92 mg/g、0.52~0.57 mg/g、0.30~0.33 mg/g、4.46~4.76 mg/g、3.02~3.24 mg/g、2.59~2.76 mg/g、6.03~6.38 mg/g、1.47~1.58 mg/g、1.90~2.08 mg/g、3.40~3.88 mg/g、1.53~1.74 mg/g。结论 UHPLC-ESI-MS/MS多反应监测法可同时快速对新天泰1号中多种不同类型的活性成分进行监测,该方法快速,灵敏度高,重复性好,可用于本品质量控制,亦为中药质量研究提供参考和借鉴。
Objective To establish an effective UHPLC-ESI-MS/MS method for the simultaneous determination of 19 active components(ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside) of different types in Xintiantai I(XI), and provide a comprehensive and efficient quality control method for traditional Chinese medicine(TCM). Methods The analysis was performed on an Agilent 1290 system with a Agilent Zorbax Eclipse Plus C18 column(150 mm × 4.6 mm, 3.5 μm) at a flow rate of 0.4 m L/min using acetonitrile and 0.1% formic acid aqueous solution as mobile phase. Mass spectrometric detection was performed on multiple reaction monitoring(MRM) in positive and negative ionization mode. The contents of 19 active components in XI were determined by monitoring the specific ions of each component. Results The 19 active components were accurately determined in 15 min and had the good linearity(r2 > 0.999) within the linear ranges. The average recovery rates of ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside were 94.80%, 96.78%, 95.59%, 96.88%, 97.74%, 100.08%, 96.27%, 100.25%, 98.32%, 97.16%, 95.60%, 95.28%, 96.81%, 95.22%, 96.85%, 95.31%, 93.86%, 94.79%, and 95.20%, respectively; The contents of three batches XI of the 19 components were in the ranges of 2.28—2.49, 0.82—0.90, 1.22—1.32, 14.44—15.50, 3.71—3.99, 3.26—3.49, 3.09—3.33, 4.39—4.72, 4.56—4.92, 0.52—0.57, 0.30—0.33, 4.46—4.76, 3.02—3.24, 2.59—2.76, 6.03—6.38, 1.47—1.58, 1.90—2.08, 3.40—3.88, and 1.53—1.74 mg/g, respectively. Conclusion The developed UHPLC-ESI-MS/MS-MRM method is fast, sensitive, and reproducible for TCM quality control. It can be used for the quality control of XI, which also provides reference for TCM quality research.
Objective To establish an effective UHPLC-ESI-MS/MS method for the simultaneous determination of 19 active components(ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside) of different types in Xintiantai I(XI), and provide a comprehensive and efficient quality control method for traditional Chinese medicine(TCM). Methods The analysis was performed on an Agilent 1290 system with a Agilent Zorbax Eclipse Plus C18 column(150 mm × 4.6 mm, 3.5 μm) at a flow rate of 0.4 m L/min using acetonitrile and 0.1% formic acid aqueous solution as mobile phase. Mass spectrometric detection was performed on multiple reaction monitoring(MRM) in positive and negative ionization mode. The contents of 19 active components in XI were determined by monitoring the specific ions of each component. Results The 19 active components were accurately determined in 15 min and had the good linearity(r2 > 0.999) within the linear ranges. The average recovery rates of ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside were 94.80%, 96.78%, 95.59%, 96.88%, 97.74%, 100.08%, 96.27%, 100.25%, 98.32%, 97.16%, 95.60%, 95.28%, 96.81%, 95.22%, 96.85%, 95.31%, 93.86%, 94.79%, and 95.20%, respectively; The contents of three batches XI of the 19 components were in the ranges of 2.28—2.49, 0.82—0.90, 1.22—1.32, 14.44—15.50, 3.71—3.99, 3.26—3.49, 3.09—3.33, 4.39—4.72, 4.56—4.92, 0.52—0.57, 0.30—0.33, 4.46—4.76, 3.02—3.24, 2.59—2.76, 6.03—6.38, 1.47—1.58, 1.90—2.08, 3.40—3.88, and 1.53—1.74 mg/g, respectively. Conclusion The developed UHPLC-ESI-MS/MS-MRM method is fast, sensitive, and reproducible for TCM quality control. It can be used for the quality control of XI, which also provides reference for TCM quality research.
引文
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[11]康文静,梁冰,李淑芳,等.去氢吴茱萸碱及盐酸小檗碱对D-半乳糖致衰老模型小鼠学习记忆的影响[J].中国药科大学学报,2010,41(4):372-374.
[12]刘春丽,陈虹,姜勇,等.松果菊苷对血管性痴呆大鼠行为学、氧自由基以及胆碱能神经递质代谢速率的影响[J].中国药理学通报,2013,29(7):1035-1036.
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[14]佘一鸣,胡永慧,韩立云,等.中药质量控制的研究进展[J].中草药,2017,48(12):2557-2563.
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[17]刘昌孝,陈士林,肖小河,等.中药质量标志物(Q-Marker):中药产品质量控制的新概念[J].中草药,2016,47(9):1443-1457.
[2]Li L,Liu Z R,Liu J F,et al.Ginsenoside Rd attenuates beta-amyloid-induced tau phosphorylation by altering the functional balance of glycogen synthase kinase 3beta and protein phosphatase 2A[J].Neurobiol Dis,2013,54:320-328.
[3]Jung H A,Min B S,Yokozawa T,et al.Anti-Alzheimer and antioxidant activities of Coptidis Rhizoma alkaloids[J].Biol Pharm Bull,2009,32(8):1433-1438.
[4]Huang M,Jiang X,Liang Y B,et al.Berberine improves cognitive impairment by promoting autophagic clearance and inhibiting production ofβ-amyloid in APP/tau/PS1mouse model of Alzheimer’s disease[J].Exp Gerontol,2017,91:25-33.
[5]Luo T,Shen X,Li S,et al.The protective effect of jatrorrhizine against oxidative stress in primary rat cortical neurons[J].CNS Neurol Disord Drug Targets,2016,16(5):617-623.
[6]Yuan S M,Gao K,Wang D M,et al.Evodiamine improves congnitive abilities in SAMP8 and APPswe/PS1ΔE9 transgenic mouse models of Alzheimer’s disease[J].Acta Pharm Sin,2011,32(3):295-302.
[7]Peng J H,Zhang C E,Wei W,et al.Dehydroevodiamine attenuates tau hyperphosphorylation and spatial memory deficit induced by activation of glycogen synthase kinase-3 in rats[J].Neuropharmacology,2007,52(7):1521-1527.
[8]刘安昌,娄红祥,赵丽霞.姜黄素药理活性及体内代谢[J].国外医药:植物药分册,2004,19(1):1-5.
[9]李锦松,陈剑鸿,孟民杰.金丝桃苷药理作用及其作用机制的研究进展[J].广东药学院学报,2015,31(2):269-272.
[10]石慧,王喜军.柠檬苦素类化合物的药理作用研究进展[J].中医药学报,2014,42(4):128-129.
[11]康文静,梁冰,李淑芳,等.去氢吴茱萸碱及盐酸小檗碱对D-半乳糖致衰老模型小鼠学习记忆的影响[J].中国药科大学学报,2010,41(4):372-374.
[12]刘春丽,陈虹,姜勇,等.松果菊苷对血管性痴呆大鼠行为学、氧自由基以及胆碱能神经递质代谢速率的影响[J].中国药理学通报,2013,29(7):1035-1036.
[13]李洋,洪莉,刘成,等.松果菊苷药理作用研究进展[J].医药导报,2017,36(1):56-60.
[14]佘一鸣,胡永慧,韩立云,等.中药质量控制的研究进展[J].中草药,2017,48(12):2557-2563.
[15]中国药典[S].四部.2015.
[16]尤娇娇,彭志红,韩凤梅.HPLC-MS/MS法同时测定平卧菊三七水提物中9种主要有效成分[J].中草药,2017,48(2):294-298.
[17]刘昌孝,陈士林,肖小河,等.中药质量标志物(Q-Marker):中药产品质量控制的新概念[J].中草药,2016,47(9):1443-1457.