Quantification of biogenic amines in human plasma based on the derivatization with N-hydroxy-succinimidyl fluorescein-O-acetate by high-performance liquid chromatography.
An HPLC method for the determination of biogenic amines based on the precolumn derivatization with N-hydroxysuccinimidyl fluorescein-O-acetate (SIFA) has been developed. The derivatization was performed at 45 degrees C for 30 min in borate buffer (pH 8.0). The derivatives were separated on a ZORBAX Eclipse XDB-C8 column (150 mm x 4.6 mm id; 5 mum) and monitored by fluorescence detection (excitation, 469 nm; emission, 512 nm). The LODs (S/N = 3) for spermine, spermidine, putrescine, cadaverine, and phenethylamine were 0.4, 0.2, 0.3, 0.5, and 0.4 nM, respectively. The developed method has been successfully applied to the determination of biogenic amines in human plasma of three healthy volunteers and four cancer patients. Average recoveries for plasma samples ranged from 94 to 106% and coefficients of variation ranged from 1.8 to 4.6%. Deproteinization of plasma was accomplished with ACN to precipitate interfering substances and the centrifuged supernatant was used directly for analysis.
Deng
Ying-Hua
YH
Department of Chemistry, Wuhan University, Wuhan, P. R. China.
Zhang
Hua-Shan
HS
Du
Xiao-Lan
XL
Wang
Hong
H
D000588:Amines / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D015394:Molecular Structure; D010777:Photochemistry; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013388:Succinimides
D000588:Amines; C411986:N-hydroxysuccinimidyl fluorescein-O-acetate; D013388:Succinimides