RRLC-TOF/MS鉴别防风血浆、尿液中成分及代谢产物

RRLC-TOF/MS in identification of constituents and metabolites of Radix Saposhnikoviae in rat plasma and urine

目的应用高分离度快速液相-飞行时间质谱(RRLC-TOF/MS)联用技术鉴别防风入血成分及尿液中的原型成分及代谢产物,以探索防风可能的药效成分及体内代谢过程。方法采用Angilent Zorbax Extend-C18柱(250mm×4.6mm,5μmid),甲醇-水流动相体系梯度洗脱,在飞行时间质谱正离子模式下扫描,利用TOF/MS得到的准确相对分子质量,对照化学成分数据库,对血浆、尿液中的防风成分及代谢物进行鉴别。结果鉴别出血浆中6个防风原型成分:蔗糖、升麻苷、升麻素、紫花前胡苷元、5-O-甲基维斯阿米醇苷和3'-O-i-丁酰亥茅酚;尿液中8个原型成分:升麻苷、divaricatacid、升麻素、5-O-甲基维斯阿米醇苷、(3S)-2,2-二甲基-3,5-二羟基-8-羟甲基-3,4-二氢-2H,6H-苯并-[1,2-b:5,4-b']双吡喃-6-酮、5-O-甲基维斯阿米醇、亥茅酚苷和汉黄芩素;及2个代谢产物:升麻素的葡萄糖醛酸苷和升麻素的同分异构体。结论该方法可靠、有效,可成功用于防风体内成分鉴别,并为进一步的药效实验提供参考和依据。

Objective To analyze the constituents and metabolites of Radix Saposhnikoviae（RS）in rat plasma and urine by rapid-resolution liquid chromatography-time of flight mass spectrometry（RRLC-TOF/MS）,so as to explore the active ingredients and metabolites of RS in vivo.Methods The separation was performed on a Angilent Zorbax Extend-C18（5 μm,250 mm×4.6 mm id）column,with a methanol-water mobile phase system used for gradient elution.Time-of-flight mass spectrometer（TOF/MS）was applied for qualitative analysis under positive ion mode.Based on the accurate molecular weight of TOF/MS detection and the compound list of RS established previously,the constituents and metabolites of RS in different matrix in vivo were identified.Results Six constituents of RS were identified in the plasma：sucrose,prim-O-glucosylcimifugin,cimifugin,nodakenetin,5-O-methylvisamminol,and 3＇-O-i-butyrylhammaudol.Eight constituents were identified in the urine：prim-O-glucosylcimifugin,divaricatacid,cimifugin,4＇-O-glucosyl-5-O-methylvisamminol,（3S）-2,2-dimethyl-3,5-dihydroxy-8-hydroxymethyl-3,4-dihydro-2H,6H-benzo-[1,2-b：5,4-b＇]dipyran-6-one,5-O-methylvisamminol,sec-O-β-D-glucosylhammaudol,and wogonin.Two metabolites were identified in the urine：glucuronide of cimifujin and an isomer of it.Conclusion The present method is reliable and effective for identifying compounds of RS in vivo,and it can provide a reference and evidence for the further pharmacodynamics experiments.