Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1291, Issue -, Pages 10-18Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2013.03.061
Keywords
Amino acids; Ultrasound assisted dispersive; liquid-liquid micro extraction; Ethyl chloroformate derivatization; Gas chromatography-mass spectrometry
Funding
- Council of Scientific and Industrial Research
- CSIR, New Delhi [OLP-0004, BSC0111]
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A rapid and economical method for the simultaneous determination of 20 amino acids in complex biological and food matrices (hair, urine and soybean seed samples) has been developed using ultrasound assisted dispersive liquid-liquid micro extraction (UA-DLLME). The method involves simultaneous derivatization and extraction followed by gas chromatography-mass spectrometric (GC-MS) analysis of amino acids. The parameters of UA-DLLME were optimized with the aid of design of experiments approach. The procedure involves the rapid injection of mixture of acetonitrile (disperser solvent), trichloroethylene (TCE) (extraction solvent) and ethylchloroformate (derivatization reagent) into the aqueous phase of sample extract containing pyridine. The Plackett-Burman design has indicated that, the factors such as volume of disperser and extraction solvents and pH were found to be significantly affects the extraction efficiency of the method. The optimum conditions of these factors based on central composite design were found to be 250 mu L of acetonitrile, 80 mu L of TCE and pH of 10. The limit of detection and limit of quantification were found to be in the range of 0.36-3.68 mu g L-1 and 1.26-12.01 mu g L-1 respectively. This is the first application of DLLME for the analysis of amino acids in any matrices. The advantages like (i) in situ derivatization and extraction of amino acids without any prior lyophilization and cleanup of sample, (ii) low consumption of extraction solvent, (iii) fast and simple, (iv) cost-effective and (iv) good repeatability make the method amenable for the routine analysis of amino acids in clinical, toxicological, nutritional and quality control laboratories. (C) 2013 Elsevier B.V. All rights reserved.
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