Determination of L-carnitine, acetyl-L-carnitine and propionyl-L-carnitine in human plasma by high-performance liquid chromatography after pre-column derivatization with 1-aminoanthracene

被引:66
作者
Longo, A [1 ]
Bruno, G [1 ]
Curti, S [1 ]
Mancinelli, A [1 ]
Miotto, G [1 ]
机构
[1] SIGMA TAU IND FARMACEUT RIUNITE,DIV RES & DEV,DEPT BIOCHEM,I-00040 POMEZIA,ROME,ITALY
来源
JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS | 1996年 / 686卷 / 02期
关键词
carnitine; acetylcarnitine; propionylcarnitine; 1-aminoanthracene;
D O I
10.1016/S0378-4347(96)00219-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new sensitive high-performance liquid chromatographic procedure for the determination of L-carnitine (LC), acetyl-L-carnitine (ALC) and propionyl-L-carnitine (PLC) in human plasma has been developed. Precolumn derivatization with 1-aminoanthracene (1AA), performed in phosphate buffer in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) as catalyst, is involved. The fluorescent derivatives were isocratically separated on a reversed-phase column (C-18). The eluate was monitored with a fluorimetric detector set at 248 nm (excitation wavelength) and 418 nm (emission wavelength). Because of the presence of endogenous carnitines, the validation was performed using dialyzed plasma. The identity of the derivatized compounds was assessed by mass spectrometry and the purity of the chromatographic peaks was confirmed by HPLC-tandem mass spectrometry. The limits of quantitation were 5 nmol/ml for LC, 1 nmol/ml for ALC and 0.25 nmol/ml for PLC. The recovery of the extraction procedure was in the range 82.6%-95.4% for all 3 compounds. Good linearity (R approximate to 0.99) was observed within the calibration ranges studied: 5-160 nmol/ml for LC, 1-32 nmol/ml for ALC and 0.25-8 nmol/ml for PLC. Precision was in the range 0.3-16.8% and accuracy was always lower than 10.6%.
引用
收藏
页码:129 / 139
页数:11
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