分析测试学报2024，Vol.43Issue(5)：703-713,11.DOI:10.12452/j.fxcsxb.24020301
柱前衍生/高效液相色谱-串联质谱法测定氨基酸工业品中26种游离手性氨基酸含量
Determination of 26 Free Chiral Amino Acids in Industrial Products by Pre-column Derivatization and HPLC-MS/MS
摘要
Abstract
A highly sensitive and selective pre-column derivatization coupled with high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)assay was developed for the deter-mination of the D,L configurations of 26 free chiral amino acids in industrial products.The samples were dissolved and extracted in 0.1 mol/L HCl solution,and then derivatized by Marfey reagent un-der alkaline condition at 37℃for 16 hours.Baseline separation of the derivatives was achieved on a Phenomenex Kinetex F5(250 mm×5 mm,4.6 µm)column.The mobile phase consisted of 10 mmol/L ammonium acetate and acetonitrile by gradient elution.Reverse phase chromatographic reso-lution of the enantiomers was obtained within 27 min.The derived analytes were sensitively detected using an electrospray ionization source in negative ion mode(ESI-)and multiple reaction monitoring(MRM)mode.The results showed that the derivatized products of 26 chiral amino acids could be ef-fectively separated.The mass concentrations of 26 chiral amino acids showed a good linear relation-ship with their corresponding peak areas in a corresponding range.The limits of quantification were determined to be 0.5-12.5 ng/mL and correlation coefficients(r2)not less than 0.9940.The extrac-tion recoveries for all analysts were in the ranges of 80.3%-118%and the relative standard deviations were in the ranges of 0.47%-9.8%.The method was applied to determine chiral amino acids in 180 industrial samples.The results indicated that D-amino acids were detected in L-type pure amino acid reagent,L-type stable isotope-labeled amino acid internal standard reagent,single L-amino acid raw material,and composite amino acid raw material powder.It is the first time to evaluate the quality of chiral amino acids in relevant industrial products in China,and it was found that the monitoring and management of chiral configurations need to be strengthened for amino acid complex powders.It also thoroughly investigates the potential impact on the accuracy of test results in the food,environmental,chemical,and pharmaceutical industries sectors when the amino acid purity reagents are used as standards or isotopic internal standards for the detection of relevant samples.With the advantages of rapid,accurate,high sensitivity,and simple sample pretreatment,this method can be an effective technical tool for quality management and health assessment in the fields of research reagents,bio-pharmaceuticals,chemical food and nutritional health.关键词
手性氨基酸/手性衍生化/高效液相色谱-串联质谱/工业品Key words
chiral amino acids/chiral derivatization/HPLC-MS/MS/industrial products分类
化学化工引用本文复制引用
匡慧颖,李子樱,朱波,张玉鑫,倪一平,肖伟敏,叶新玉,杨国武,杨燕..柱前衍生/高效液相色谱-串联质谱法测定氨基酸工业品中26种游离手性氨基酸含量[J].分析测试学报,2024,43(5):703-713,11.基金项目
国家重点研发计划科研项目(2021YFF0701903) （2021YFF0701903）
