浙江中医药大学学报2025，Vol.49Issue(8)：968-982,15.DOI:10.16466/j.issn1005-5509.2025.08.003
基于钌络合物荧光试剂的雷公藤甲素柱前衍生HPLC-FLD检测方法初探
Preliminary Study on HPLC-FLD Method for Pre-Column Derivatization of Triptolide by Using Ruthenium Complex Fluores-cent Reagent
摘要
Abstract
[Objective]To investigate a simple method for the rapid detection and precise analysis of trace triptolide(TP)in vitro.[Methods]Initially,4-carboxy-4'-methyl-2,2'-bipyridine(Me-bpy-COOH)and cis-bis(2,2'-bipyridine)dichlororuthenium(Ⅱ)(cis-Ru(bpy)2CL2)were used as raw materials to synthesize tripyridine ruthenium monocarboxylic acid fluorescent reagent,namely bis(2,2'-bipyridyl)(4-methyl-4'-carboxy-2,2'-bipyridyl)ruthenium(Ⅱ)bishexafluorophosphate[Ru(bpy)2(bpy-COOH)2PF6].Subsequently,the pre-column derivatization reaction conditions of Ru(bpy)2(bpy-COOH)2PF6 with TP was optimized through orthogonal test and a methodological evaluation of the high performance liquid chromatography-fluorescence detection(HPLC-FLD)of triptolide-terpyridine ruthenium derivative(TTRD)was conducted.Taking commercially available Kunxian Capsules as object,the newly established evaluation method was adopted for detection.[Results]A fluorescent derivative of TP,TTRD was obtained and was characterized by proton nuclear magnetic resonance(1H-NMR)and ultra high performance liquid chromatography-mass spectrometry(UPLC-MS).The optimized pre-column derivatization reaction conditions were a 4:1 molar ratio of Ru(bpy)2(bpy-COOH)2PF6 to TP,a reaction time of 2 hours and a reaction temperature of 45℃.This study presented a HPLC-FLD method for the pre-column derivatization of TP.The method demonstrated a good linear relationship within the range of 50 to 1 200 μg·L-1.The TP content in Kunxian Capsules was measured by using this method,yielding a result of 78.78 μg·g-1,which fell within the prescribed range of national drug standards.[Conclusion]The pre-column-derived HPLC-FLD method for TP exhibited good sensitivity,accuracy and reliability,expanding the HPLC detection range for TP.关键词
三联吡啶钌/雷公藤甲素/柱前衍生/正交试验/高效液相色谱-荧光检测法/昆仙胶囊/结构表征/含量测定Key words
tripyridine ruthenium/triptolide/pre-column derivatization/orthogonal test/high performance liquid chromatography-fluorescence detection/Kunxian Capsules/structural characterization/content determination分类
医药卫生引用本文复制引用
汤梦洁,徐蓓华,张慧敏,周晓祺,丁中鹏,石森林..基于钌络合物荧光试剂的雷公藤甲素柱前衍生HPLC-FLD检测方法初探[J].浙江中医药大学学报,2025,49(8):968-982,15.基金项目
浙江省重点科技创新团队项目(2011R09042-06)Zhejiang Provincial Key Science and Technology Innovation Team Project(2011R09042-06) （2011R09042-06）
