Validation and further optimisation of a cyclodextrin-modified micellar electrokinetic capillary chromatography method for urine profiling

被引:6
作者
Guillo, C
Perrett, D
Hanna-Brown, M [1 ]
机构
[1] Kings Coll London, Dept Pharm, London SE1 9NN, England
[2] Barts & London Queen Marys Sch Med & Dent, Dept Med, London EC1A, England
关键词
capillary electrophoresis; sulphated beta-cyclodextrin-modified MECC; metabonomics; urine profiling;
D O I
10.1365/s10337-004-0218-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Metabonomic studies require efficient and high-resolution analytical probes to monitor changes in the 'metabolic fingerprint'. The advantageous characteristics of Capillary Electrophoresis, enabling highly efficient separations of diverse components present in minute sample volumes, may therefore prove a useful tool in biofluid analysis. This paper describes the optimisation and validation of a sulphated beta-cyclodextrin-modified MECC method for urine profiling. Cyclodextrin substitution, experimental conditions including capillary length, injection mode and time, applied voltage, temperature and capillary pre-conditioning procedures were investigated and optimised. Precision, linearity, sensitivity and robustness of the method were assessed, as well as urine stability. The validated sulphated beta-cyclodextrin-modified MECC method allows for the separation of over 80 urinary analytes in under 25 min, using a sodium borate/SDS/sulfated beta-cyclodextrin (25/75/6.25 mM) electrolyte and an 18 kV applied voltage in a 40 cm (effective length), 50 mum i.d. fused silica capillary at 20 degreesC, pre-conditioned with HCl for 5 min and BGE for 1 min, and UV diode array detection (190-600 nm). Suck methodology should prove invaluable in the rapid comparison of urine profiles and indication of metabolic disorders or abnormalities.
引用
收藏
页码:S157 / S164
页数:8
相关论文
共 57 条
[1]  
Adam T, 1999, CLIN CHEM, V45, P2086
[2]   Screening method for inherited disorders of purine and pyrimidine metabolism by capillary electrophoresis with reversed electroosmotic flow [J].
Adam, T ;
Lochman, P ;
Friedecky, D .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2002, 767 (02) :333-340
[3]  
Alfazema LN, 1998, ADV EXP MED BIOL, V431, P171
[4]  
Alfazema LN, 2000, ELECTROPHORESIS, V21, P2503
[5]  
[Anonymous], 2001, Guidance for industry, bioanalytical method validation
[6]   Rapid identification of the bacterial pathogens responsible for urinary tract infections using direct injection CE [J].
Armstrong, DW ;
Schneiderheinze, JM .
ANALYTICAL CHEMISTRY, 2000, 72 (18) :4474-4476
[7]  
Barbas C, 1998, CLIN CHEM, V44, P1340
[8]  
BOOMSMA F, 1993, CLIN CHEM, V39, P2503
[9]   Comparison of capillary electrophoretic and liquid chromatographic determination of hypoxanthine and xanthine for the diagnosis of xanthinuria [J].
Bory, C ;
Chantin, C ;
Boulieu, R .
JOURNAL OF CHROMATOGRAPHY A, 1996, 730 (1-2) :329-331
[10]   Sensitive and high-throughput analyses of purine metabolites by dynamic pH junction multiplexed capillary electrophoresis: A new tool for metabolomic studies [J].
Britz-McKibbin, P ;
Nishioka, T ;
Terabe, S .
ANALYTICAL SCIENCES, 2003, 19 (01) :99-104