On-line capillary based quantitative analysis of a heterocyclic amine in human urine - Analytical Methods & Tools for Lyophilization

On-line capillary based quantitative analysis of a heterocyclic amine in human urine
Received 28 June 2006; revised 11 August 2006; revised 14 August 2006. Available online 8 September 2006.
Peter Viberga, b, , , Karl-Gustav Wahlundb and Kerstin Skoga
Journal of Chromatography A
Article in Press
ScienceDirect
Copyright ? 2006 Elsevier B.V. All rights reserved.
aDivision of Applied Nutrition and Food Chemistry, Department of Food Technology, Engineering, and Nutrition, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden
bDivision of Analytical Chemistry, Centre for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden
Abstract
A high through-put miniaturised separation-quantification method for the heterocyclic aromatic amine 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in urine was developed. The limit of detection was of 0.65 fmol (0.14 pg) injected or 65 pM. Heterocyclic aromatic amines are mutagenic and carcinogenic compounds formed at low levels in protein-rich food during cooking. Due to the low concentrations and the high complexity of the matrix in which they exist (food, blood, and urine), and the often small sample volumes (capillary blood; urine, blood and milk from small animals), very sensitive and selective analytical methods are required for their detection. Miniaturisation was accomplished by micro solid-phase extraction in capillaries with blue chitin as solid-phase, coupled on-line (in-capillary) to capillary electrophoresis with nanospray tandem mass spectrometric detection. This new technique provided a total analysis time of only 15 min, including extraction and separation, together with low sample and solvent consumption. Blue chitin showed high tolerance towards the urine matrix. Urine collected 12 h after consumption of fried chicken contained 1.8 nmol L-1 (0.40 pg ?L-1) of PhIP.
Keywords: On-line; In-capillary; Nanospray; Nanoelectrospray; Tandem mass spectrometry; Capillary electrophoresis; Blue chitin; PhIP; Heterocyclic amines; Food analysis

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