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超高效液相色谱-串联四极杆质谱法比较新鲜与炮制后蟾酥的化学成分

[Comparison of chemical composition between fresh and processed Bufonis Venenum by UPLC-TQ-MS].

作者信息

Wang Zi-yue, Wang Hong-lan, Zhou Jing, Ma Hong-yue, Gong Yan, Yan Wen-li, Qian Da-wei

出版信息

Zhongguo Zhong Yao Za Zhi. 2015 Oct;40(20):3967-73.

Abstract

Toad venom is the Bufo bufo gargarizans or B. melanostictus after the ears of the gland secretion, used in the treatment of various cancers in recent years. Research shows that the main anti-tumor components in bufadienolide. Bufadienolide have free type structure and conjunct type structure. To identify and clarify the difference between bufogenin and bufotoxin contained in Bufonis Venenum, which was from B. bufo gargarizans, an UPLC-TQ-MS method has been established. UPLC-TQ-MS method was used to identify and quantify the major bufadienolides in Bufonis Venenum. UPLC-TQ-MS assay with positive ion mode was performed on a Waters ACQUITY UPLC BEH C, (2.1 mm x 100 mm, 1.7 µm) with the mobile phase consisting of 0. 1% aqueous formic and acidacetonitrile in gradient elution at a flow rate of 0.4 mL · min⁻¹ and the column temperature was set at 35 °C. By comparing their retention time and high resolution mass data of Bufonis Venenum extracts, 37 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were free-type bufadienolides (bufogenin), 11 of them were conjugated bufadienolides. There were significant differences in the main composition between fresh and processed Bufonis Venenum. The study found that the chemical composition of toad venom through great changes after processing, conjunct type content is much less, free type content as well change.

摘要

蟾酥是中华大蟾蜍或黑眶蟾蜍耳后腺的分泌物,近年来用于治疗各种癌症。研究表明,蟾毒配基是主要的抗肿瘤成分。蟾毒配基有游离型结构和结合型结构。为了鉴别和阐明中华大蟾蜍蟾酥中所含蟾毒配基和蟾毒素之间的差异,建立了超高效液相色谱-串联四极杆质谱法(UPLC-TQ-MS)。采用UPLC-TQ-MS法对蟾酥中的主要蟾毒配基进行鉴别和定量。在Waters ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7 µm)色谱柱上以正离子模式进行UPLC-TQ-MS分析,流动相为0.1%甲酸水溶液和乙腈,梯度洗脱,流速为0.4 mL·min⁻¹,柱温设定为35℃。通过比较蟾酥提取物的保留时间和高分辨率质谱数据,在正离子模式下通过MS/MS分析初步鉴定出37种有效成分。其中26种为游离型蟾毒配基(蟾毒配基),11种为结合型蟾毒配基。鲜蟾酥和炮制后蟾酥的主要成分存在显著差异。研究发现,蟾酥经炮制后化学成分发生了很大变化,结合型含量减少很多,游离型含量也有所变化。

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