利用UPLC-TQ-MS比较蟾酥鲜品和蟾酥商品化学成分

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

蟾酥为蟾蜍科动物中华大蟾蜍Bufo bufo gargarizans或黑框蟾蜍B.melanostictus等近缘种的耳后腺分泌的白色浆汁加工而成,近年来多用于治疗各种癌症,有一定的攻毒抗癌、消肿止痛作用。研究表明其主要抗肿瘤成分为蟾蜍二烯内酯类。蟾蜍二烯内酯有游离型和结合型2种结构。该文通过超高效液相色谱质谱联用技术(UPLC-TQ-MS)对中华大蟾蜍的蟾酥鲜品中蟾蜍二烯酸内酯类成分进行研究。采用UPLC-TQ-MS的正离子多反应监测(MRM)模式。色谱条件:ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),柱温35.0℃,流速0.4 m L·min-1,流动相0.1%甲酸超纯水溶液-乙腈溶液。最终结合相关文献推导出蟾酥鲜品的37种蟾蜍二烯内酯成分,其中26种为游离型结构,11种为结合型结构。与蟾酥商品比较,二者之间有18种化合物存在显著差异,研究发现蟾酥的部分化学成分经加工处理后有很大的变化,7种结合型蟾蜍二烯内酯大量减少,11种游离型蟾蜍二烯酸内酯含量也随之明显改变,可能是在加工处理过程中蟾蜍二烯酸内酯部分结合型转化成了游离型。

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 C18（ 2. 1 mm × 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 m L·min-1and the column temperature was set at 35 ℃. 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.