Establishment of reference intervals of plasma aminogram of healthy adults in Shanghai area based on high-performance liquid chromatography-tandem mass spectrometry technique

ZHU Xuan; ZHAN Yingying; ZHAO Yangyang; HU Yao; ZHANG Cuiping

doi:10.3969/j.issn.1674-3806.2025.02.09

Abstract：Objective To establish and validate the reference intervals of plasma aminogram of healthy adults in Shanghai area based on high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)technique.Methods A total of 341 apparently healthy individuals who underwent physical examination at the Physical Examination Center of Huashan Hospital Affiliated to Fudan University from January 2023 to October 2023 were recruited.Other 40 apparently healthy individuals were recruited from November 2023 to December 2023 for reference interval validation.HPLC-MS/MS technique was used to detect the levels of 28 kinds of amino acids in the plasma of the study subjects.The reference intervals of plasma aminogram of healthy adults in Shanghai area were established and validated.Results The methodological evaluation results of HPLC-MS/MS detection of adult plasma amino acids showed that the coefficient of variation of precision of the analytes was less than 15％,and the accuracy was between 86％and 115％,and the quantification lower limit deviation was less than 15％,and the coefficient of variation was less than 15％.The correlation coefficient(R2)of the linear range standard curve was greater than 0.99,and the validations of the carrying contamination rate and sample stability met the corresponding requirements.Based on this method,the reference intervals of 28 kinds of amino acids in the plasma of healthy adults in Shanghai area were established.The results of gender stratification analysis showed that the levels of most amino acids and metabolites in the plasma of healthy adult males were higher than those in the plasma of healthy adult females,and the differences were statistically significant(P＜0.05).The validation results showed that the reference intervals established in this study met the requirements of Define and Determine the Reference Intervals in Clinical Laboratory(WS/T 402-2012)standard.Conclusion In this study,the reference intervals of 28 kinds of plasma amino acids in healthy adults in Shanghai area are established and validated to provide a basis for clinical diagnosis and treatment.