UPLC-Q-TOF/MS分析不同比例丹参配伍藜芦毒性生物碱变化规律

doi:10.6023/A12080606

化学学报 ›› 2012, Vol. 70 ›› Issue (21): 2257-2264.DOI: 10.6023/A12080606 上一篇下一篇

研究论文

UPLC-Q-TOF/MS分析不同比例丹参配伍藜芦毒性生物碱变化规律

李飞^a,b, 王宇光^b, 杨亮^b, 朱冠秀^a,b, 梁乾德^b, 马增春^b, 肖成荣^b, 谭洪玲^b, 汤响林^b, 高月^b

a 中南大学湘雅医学院长沙 570228;
b 军事医学科学院放射与辐射医学研究所北京 100850

投稿日期:2012-08-30 发布日期:2012-09-22
通讯作者: 王宇光, 高月 E-mail:gaoyue@nic.bmi.ac.cn; wang_yuguang@139.com
基金资助:
项目受国家重点基础研究发展计划(Nos. 2011CB505304, 2012CB518402)、国家自然科学基金(No. 81073149)和北京市自然科学基金(No. 7112110)资助.

Regularity of Toxic Alkaloids during the Combination of Veratrum nigrum and Salvia miltiorrhiza by UPLC-Q-TOF/MS

Li Fei^a,b, Wang Yuguang^b, Yang Liang^b, Zhu Guanxiu^a,b, Liang Qiande^b, Ma Zengchun^b, Xiao Chengrong^b, Tan Hongling^b, Tang Xianglin^b, Gao Yue^b

a Xiangya Medical College Central Sourth University, Changsha 570228;
b Beijing Institute of Radiation Medicine, Beijing 100850

Received:2012-08-30 Published:2012-09-22
Supported by:
Project supported by National Basic Research Program of China (Nos. 2011CB505304, 2012CB518402), the National Natural Science Foundation of China (No. 81073149) and the Natural Science Foundation of Beijing (No. 7112110).

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摘要/Abstract

利用超高效液相色谱-飞行时间质谱联用技术研究不同配伍比例的丹参藜芦的化学指纹图谱, 并分析毒性生物碱在不同比例配伍混煎前后溶出量的变化. 同时利用小鼠的急性毒性实验来考察配伍后的毒性变化规律. 结果显示在正离子模式下, 各配伍组(A～H)样本之间的差异能得到明显区分, 并结合Loadings图谱和数据库确定了配伍化学指纹图谱中的17种藜芦生物碱毒性成分, 当固定藜芦用量为LD₅₀且藜芦比例大于丹参时, 藜芦定、伪介芬胺等生物碱溶出高于或与藜芦单煎液相当, 但随着丹参比例增加, 上述生物碱溶出呈逐渐下降趋势, 当丹参比例大于藜芦时, 生物碱溶出低于藜芦单煎液. 不同比例丹参藜芦配伍急性毒性实验毒性变化规律与上述生物碱含量的变化趋势吻合, 表明上述毒性生物碱可能是藜芦与丹参特定比例配伍后毒性增强主要化学标志物.

关键词: 十八反, 丹参, 藜芦, 急性毒性实验, 液质联用

In traditional chinese medicine, the combination of veratrum nigrum and salvia miltiorrhiza is traditional incompatibility. In order to investigate the intrinsic relevance of acute toxicity experiment and main toxic alkaloids variation in different ratio, ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to reveal the chemical profile of decoctions of veratrum nigrum and salvia miltiorrhiza in different ratio (10∶0, 10∶1, 5∶1, 2∶1, 1∶1, 1∶2, 1∶5, 1∶10). The analyses were performed on a reversed-phase C₁₈ column using a binary eluent under gradient conditions. At the same time, the trend of toxicity diversity between the veratrum nigrum and salvia miltiorrhiza was explored through acute toxicity experiment of mice. Seventeen alkaloids of veratrum nigrum can be detected, The t_R-m/z pairs and mean mass can be obtained by selected ion chromatogram, after extracted by Markerlynx XS, the dataset were subjected to principal component analysis (PCA) and orthogonal partial least squared discriminant analysis (OPLS-DA) to holistically compare the difference between samples, and variant changes of veratrum nigrum were observed. After fixed the dosage of veratrum nigrum as LD₅₀, when the ratio of veratrum nigrum was greater than salvia miltiorrhiza, the dissolution of some alkaloids was greater than or equal to the single decoction group of veratrum nigrum, such as verdine, pseudojervine, zygacine, rubijervine, neogermidine, germidine, 3-angeloylzygadenine and so on, with the increase proportion of salvia miltiorrhiza, these alkaloids dissolution showed a declining trend. When the ratio of veratrum nigrum was less than salvia miltiorrhiza, the dissolution of the alkaloids was less than the single decoction group of veratrum nigrum. The result of acute toxicity experiment is matched with the trend of these alkaloids, which showed us these alkaloids may be the main chemical markers of enhanced toxicity during the combination of veratrum nigrum and salvia miltiorrhiza in different ratio.

Key words: eighteen incompatible pairs, salvia miltiorrhiza, veratrum nigrum, acute toxicity experiment, ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF/MS)

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李飞, 王宇光, 杨亮, 朱冠秀, 梁乾德, 马增春, 肖成荣, 谭洪玲, 汤响林, 高月. UPLC-Q-TOF/MS分析不同比例丹参配伍藜芦毒性生物碱变化规律[J]. 化学学报, 2012, 70(21): 2257-2264.

Li Fei, Wang Yuguang, Yang Liang, Zhu Guanxiu, Liang Qiande, Ma Zengchun, Xiao Chengrong, Tan Hongling, Tang Xianglin, Gao Yue. Regularity of Toxic Alkaloids during the Combination of Veratrum nigrum and Salvia miltiorrhiza by UPLC-Q-TOF/MS[J]. Acta Chimica Sinica, 2012, 70(21): 2257-2264.

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UPLC-Q-TOF/MS分析不同比例丹参配伍藜芦毒性生物碱变化规律

Regularity of Toxic Alkaloids during the Combination of Veratrum nigrum and Salvia miltiorrhiza by UPLC-Q-TOF/MS

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