Preliminary Study on HPLC-FLD Method for Pre-Column Derivatization of Triptolide by Using Ruthenium Complex Fluores-cent Reagent
10.16466/j.issn1005-5509.2025.08.003
- VernacularTitle: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
- Author:
Mengjie TANG
1
;
Beihua XU
1
;
Huimin ZHANG
1
Author Information
1. 浙江中医药大学药学院 杭州 311400
- Publication Type:Journal Article
- Keywords:
tripyridine ruthenium;
triptolide;
pre-column derivatization;
orthogonal test;
high performance liquid chromatography-fluorescence detection;
Kunxian Capsules;
structural characterization;
content determination
- From:
Journal of Zhejiang Chinese Medical University
2025;49(8):968-982
- CountryChina
- Language:Chinese
-
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.
