柱前衍生-荧光检测-高效液相色谱-质谱法测定尿中游离多胺
Pre-column Derivatization-HPLC-MS Determination of Free Polyamines in Urine with Fluorescence Detection
摘 要
用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙基氯甲酸酯作为柱前衍生化试剂,在乙腈中,以pH 9的硼酸钠缓冲溶液为催化剂,40 ℃下衍生反应10 min后获得稳定的荧光产物.在Eclipse XDB-C8色谱柱上,通过梯度洗脱对5种多胺进行了分离和在线质谱定性.激发波长(λex)和发射波长(λem)分别为333 nm和390 nm.采用大气压化学电离源(APCI)正离子模式,实现了人尿中游离多胺的质谱定性及荧光定量测定.检出限(3S/N)在4.98~9.31 fmol.此方法应用于尿样中多胺的测定,并测得回收率在94%~98%之间.
柱前衍生
荧光检测
多胺
HPLC-MS
Pre-column derivatization
Fluorescence detection
Polyamines
Abstract
A new fluorescence reagent,1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate was used as pre-column derivatizing agent in an acetonitrile solution,catalyzed by the borate buffer of pH 9 to react with polyamines at 40 ℃ for 10 min,to give stable fluorescence products.Five polyamines were then separated on the Eclipse XDB-C8 chromatographic column by gradient elution with acetonitrile-H2O solution of different concentrations,and on-line analyzed qualitatively with APCI mode by MS and quantitatively by HPLC with fluorescence detection at the wavelengths of (λex) 333 nm and (λem) 390 nm.Detection limits of the 5 polyamines (3S/N) found were in the range of 4.98-9.31 fmol.Test for recovery was made by addition of standard polyamine solutions to a urine sample,and the values of recovery found were in the range of 94%-98%.
中图分类号 O657.31
所属栏目 工作简报
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收稿日期 2008/12/18
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备注陈向明(1978-),男,山东招远人,讲师,硕士,主要从事色谱分析研究.
引用该论文: CHEN Xiang-ming,DONG Xiu-li,LI Hong-juan,LIU De-sheng. Pre-column Derivatization-HPLC-MS Determination of Free Polyamines in Urine with Fluorescence Detection[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2009, 45(3): 278～280
陈向明,董秀丽,李洪娟,刘德胜. 柱前衍生-荧光检测-高效液相色谱-质谱法测定尿中游离多胺[J]. 理化检验－化学分册, 2009, 45(3): 278～280
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参考文献
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【2】Price J R,Metz P A.HPLC of 9-fluorenylmethylchloroformate-polyamine derivatives with fluorescence detection[J].Chromatographia,1987,24:795．
【3】陈冠华,刘云惠.氯甲酸芴甲酯柱前衍生高效液相色谱法测定植物中多胺[J].分析化学,1998,26(8):945．
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【5】吴筱丹,楼健.高效液相色谱法分析柿子中多胺[J].理化检验-化学分册,2004,40(2):72．
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