Application of LC-MS/MS method in detection of 10 kinds of amino acids in serum of patients with unexplained recurrent spontaneous abortion

XU Hong; HAN Yanjie; ZHANG Yao; LIU Li

doi:10.3969/j.issn.1002-266X.2017.44.005

Abstract：Objective To detect the changes of 10 kinds of amino acid levels in patients with unexplained recurrent spontaneous abortion (URSA) by high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method,then to evaluate the performance of this method.Methods Forty URSA patients were used as the observation group,40 normal early pregnancy women (＜ 20 weeks) were arranged as the control group.LC-MS/MS method was used to detect ornithine,citrulline,glycine,alanine,valine,arginine,leucine,methionine,phenylalanine,and tyrosine levels in the two groups.The specificity,precision,recovery rate,and stability of LC-MS/MS method were also analyzed.Results The serum ornithine levels in the observation group and control group were (89.3 ± 8.9) and (33.6 ± 5.4)μ mol/L,respectively;citrulline levels were separately (18.6 ± 3.2) and (14.3 ± 2.4) μmol/L,and glycine levels were separately (230.2 ± 27.3) and (202.2 ± 19.9) μmol/L(all P ＜ 0.01).No significant difference was found in the levels of serum alanine,valine,arginine,leucine,methionine,phenylalanine',and tyrosine between the two groups (all P ＞0.05).The specificity analysis showed that the amino acid m/z was observed in the serum of the observation group,and there was only internal standard m/z in methanol blank solution,no m/z of the corresponding amino acid.The precision and recovery rate analysis showed that the recovery rate of the method was 82.1％-93.6％,the within-day relative standard deviation (RSD) was 1.5％-5.1％ and the between-days RSD was 2.5％-7.1％.The stability analysis showed that the average recovery rate was 95.3％-103.2％ for 2-7 days.Conclusions The levels of serum ornithine,citrulline,and glycine increase in URSA patients,but no significant changes are found in the serum alanine,valine,arginine,leucine,methionine,phenylalanine and tyrosine.The LC-MS/MS method has high recovery rate,repeatability,and stability in detection of serum amino acids.