|
KMID : 0606920070150040266
|
|
Biomolecules & Therapeutics
2007 Volume.15 No. 4 p.266 ~ p.272
|
|
|
Simultaneous Determination of C22-C26 Very Long - Chain Fatty Acids Following tert-Butyldimethylsilyl Derivatization by Stable Isotope GC- MS for the Screening of Adrenoleucodystrophy
|
|
Yoon Hye-Ran
|
|
|
Abstract
|
|
|
A rapid analytical method was developed to quantify very long-chain fatty acids (VLCFAs, C22:0, C24:0, C26:0) in human plasma with good sensitivity and specificity using tert-butyldimethylsilyl (TBDMS) derivatization and stable isotope GC-MS selective ion monitoring (GC-MS/SIM). Two-hundred and fifty μL of plasma was fortified with deuterated stable isotope internal standards (d3-C22:0, d3-C24:0, d3-C26:0) and standard mixtures of chloroform and methanol, and then extracted with hexane and acetonitrile. To upper layer of liquid-liquid-extraction, N-(t-Butyldimethylsilyl)-N-methyltrifluoroacetamide was added and then heated to 60 oC for 30 min to produce the TBDMS derivatives. Derivatives of VLCFAs were analyzed by GC-MS/SIM. Calibration curves showed a linear relationship for the target compounds in the concentration range of 10-4 ~ 2× 103 μg/mL with the correlation coefficient ranging from 0.996 to 0.999. The limit of quantification for the plasma was 10-4 ~2×10-4 μg/mL (S/N=3). When applied to the plasma specimens of patients with peroxisomal disorder, X-linked adrenoleucodystropy (ALD, Mckusick 202370), the method clearly differentiated normal subjects from ALD patients. The C24:0/C22:0 and C26:0/C22:0 ratios were significantly elevated in the plasma of patients with X-linked ALD compared to normal subjects. The new developed method might be useful for a rapid and sensitive
diagnosis of X-linked ALD and other peroxisomal disorders.
|
|
|
KEYWORD
|
|
|
GC-MS/SIM, TBDMS derivative, docosanoic, tetracosanoic, hexacosanoic, peroxisomal disorder, X-linked Adrenoleucodystropy
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|
|