Determination of free amino acids in pig plasma by precolumn derivatization with 6-N-aminoquinolyl-N-hydroxysuccinimidyl carbamate and high-performance liquid chromatography.
A high-performance liquid chromatographic method for measuring amino acids in pig plasma has been developed by using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate as a precolumn derivatization reagent. With the method presented here it was possible to separate 24 amino acids in pig plasma within one single run in 45 min, while only 18 amino acids were properly separated with the original AccQ.Tag conditions used for analysis of hydrolysate samples. The recovery was above 90% for most amino acids, and the mean coefficient of variation of the retention times below 0.30%. The within- and between-assay reproducibility for the determination of plasma amino acids showed C.V.s below 2.2% and 4.5%, respectively, for most amino acids analyzed. In the present work, most of the plasma amino acids were assayed with high sensitivity, accuracy and good reproducibility in a relatively short time and on very small amounts of sample.
Reverter
M
M
Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, Uppsala, Sweden. mariona.reverter@huv.slu.se
Lundh
T
T
Lindberg
J E
JE
D000596:Amino Acids / Q000097:blood* / Q000737:chemistry; D000634:Aminoquinolines / Q000737:chemistry*; D000818:Animals; D002219:Carbamates / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013552:Swine
C078178:6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; D000596:Amino Acids; D000634:Aminoquinolines; D002219:Carbamates