液相层析串联质谱法检测脑脊液食欲素诊断1型发作性睡病的初步探索OA

Objective Narcolepsy type 1(NT1)is known to be associated with low levels of hypocretin-1(Hcrt-1)in cerebrospinal fluid(CSF).The standard method for Hcrt-1 measurement is radioimmunoassay(RIA)with imported re-agents,but this antibody-dependent method is limited to radiation safety-certified lab,gradual radioactivity degradation,and slow turn-around time.The purpose of this study is to explore a non-radioactive,faster,and antibody independent do-mestic method in China for Hcrt-1 detection.Methods Repeated testing of cerebrospinal fluid from 14 clinically diag-nosed NT1 patients and 10 non-narcolepsy patients was performed using liquid chromatography-tandem mass spectrometry(LC-MS/MS)technology,including the establishment and optimization of fundamental methodological procedures.The main steps involved the addition of non-radioactive isotope-labeled internal standards to the cerebrospinal fluid,followed by solid-phase extraction,mass spectrometry signal acquisition,and quantitative analysis.The results were then com-pared with the corresponding radioimmunoassay(RIA)findings.Results The LC-MS/MS method showed faster speed,and good linearity across a wider range of synthesized standard(5～2 500 pg/ml),and good repeatability.Although this ab-solute-quantitation-based LC-MS/MS method and RIA method have different reading values in Hcrt-1 quantitation,they both can segregate NT1 group from non-NT1 group well.Conclusion Although larger cohorts are needed to set up a stan-dard method in China,LC-MS/MS method is proved to be an easier,safer,faster,and possibly more accurate method for Hcrt-1 quantitation and detection for NT1 diagnosis.