Determination of amino acids in rat serum based on an UPLC method with pre-column derivatization of PITC
10.13200/j.cnki.cjb.004248
- VernacularTitle:基于异硫氰酸苯酯柱前衍生化-超高效液相色谱法检测大鼠血清中氨基酸含量
- Author:
ZHANG Ruihu
- Publication Type:Journal Article
- Keywords:
Serum;
Phenyl isothiocyanate(PITC);
Pre-column derivatization;
Ultra-high performance liquid chromatography(UPLC);
Amino acids
- From:
Chinese Journal of Biologicals
2024;37(6):723-730
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop an ultra performance liquid chromatography(UPLC)method for simultaneous determination of 19 amino acids in rat serum by employing the pre-column derivatization of phenyl isothiocyanate(PITC),verify and preliminarily apply the method.Methods The analysis was performed on an ACQUITY UPLC~?BEH C_(18) column(2.1mm×100 mm,1.7 μm) by using 0.1 mol/L sodium acetate buffer solution-acetonitrile as mobile phase at the flow rate of 0.3 mL/min in gradient elution.The column temperature was fixed at 52 ℃,and the detection wavelength was 254 nm.The specificity,linearity,precision,stability and accuracy of the method were verified based on the optimized chromatographic conditions and preparation method of sample solution,and the contents of 19 amino acids in rat serum were determined by using the developed method.Results All 19 amino acids in rat serum were separated well within 10 min,and the blank derivatization solvent and internal standard compounds had no interference with the analysis of amino acids.All the components exhibited good linear correlation in the given range(r ≥ 0.998 0),the limit of detection was 0.004 6-0.186 5 μg/mL,and the limit of quantitation was 0.013 9-0.559 6 μg/mL.The RSDs of precision and stability tests were all less than 4%.The average spike recoveries ranged from 90% to 110% and the RSDs were less than 5%.The contents of amino acids in rat serum ranged from 4.837 to 110.233 μg/mL.Conclusion The developed method is sensitive,simple and feasible,which is suitable for the analysis of amino acids in rat serum.
