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Assay of bradykinin metabolites in human body fluids by CE‐LIF coupled with transient ITP preconcentration
Author(s) -
Chen Ying,
Zhang Lan,
Xu Liangjun,
Lin JinMing,
Chen Guonan
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200800477
Subject(s) - tris , chemistry , chromatography , derivatization , electrolyte , bradykinin , detection limit , molar concentration , tetrafluoroborate , buffer solution , ionic liquid , high performance liquid chromatography , biochemistry , organic chemistry , receptor , electrode , catalysis
A CE with LIF detection was developed for the separation and determination of three bradykinin (BK) metabolites: BK2‐9, BK1‐7 and BK1‐5. BK fragments were derivatized with 5‐(4, 6‐dichloro‐s‐triazin‐2‐ylamino) fluorescein before CE‐LIF analysis. Eighty millimolar of Tris‐H 3 PO 4 (pH 9.0) was selected as the derivatization media. Three BK fragments were baseline separated within 10 min by using 0.2 M TAPS‐Tris buffer (pH 8.5) as the running buffer. Meanwhile we have also developed a simple, quick, and sensitive on‐column transient ITP preconcentration for CE‐LIF detection of three BK fragments. Ten millimolar of Tris‐HCl (pH 9.0) was chosen as the leading electrolyte and 0.2 M TAPS‐Tris (pH 8.5) containing 10 mM 1‐butyl‐3‐methylimidazolium tetrafluoroborate as the terminating electrolyte and also served as the running buffer during CZE separation. By using this transient ITP coupled with CE‐LIF, concentration detection limits ( S / N =3) for BK2‐9, BK1‐7 and BK1‐5 were 0.3, 0.1 and 0.1 pmol/L, respectively. This method has been applied to the assay of human saliva and plasma samples with satisfactory results.