| 摘要: | 本研究的目的為國內首次利用液相層析- 電灑游離/質譜/質譜技術來大量及專一性地檢測新生兒採血濾紙樣品中 (約含3.6 .mu.l 血樣) 是否帶有過氧化脢體失調疾病 (如: 腦白質腎上腺營養不良症 (Adrenoleukodystrophy, ALD), 髓磷質腎上腺神經病變(adrenomyelinoneuropathy, AMN), 過氧化脢體生成缺陷 (peroxisomal biogenesis defects, PBD), 雷弗素姆氏病變 (Refsum disease))。血液中某些極長鏈脂肪酸的濃度是診斷先天性過氧化脢體失調症重要的參數，如: 二十、二十二、二十四及二十六烷酸 (eicosanoic, docosanoic, tetracosanoic and hexacosanoic acids) 等等。目前過氧化脢體失調的的檢測方式大都利用氣相層析或氣相層析質譜連線技術偵測極長鏈脂肪酸，雖然具有靈敏度及專一性高之優點，但常會有步驟緩慢及自動化困難之情形。大量分析、高專一性、低偵測極限與富含分析物結構物訊息是液相層析-電灑游離/質譜/質譜連線技術的優點，因此選擇此技術開發應用於新生兒極長鏈脂肪酸的偵測，而不同型式的先天性過氧化脢體失調症也將可在一次分析中同時篩檢出來。因此相對於傳統方法，新的先天性過氧化脢體失調症篩檢方法將會更準確,快速,經濟及簡便。目前本實驗室發展液相層析/質譜/質譜連線偵測氨基酸、醯基肉毒鹼及17 羥孕酮已成功應用於新生兒遺傳代謝疾病之偵測。
In this study, our most important goal was to provide a high throughput and specific method with potential to screen for many of the peroxisomal disorders (ex. adrenleukodystrophy (ALD), adrenomyelinoneuropathy (AMN), peroxisomal biogenesis defects (PBD), Refsum disease and etc.) with a 3-mm blood spot (~3.6 .mu.l blood impregnated on filter paper) from newborns using high performance liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Several very long chain fatty acids (VLCFA)-eicosanoic (C20:0), docosanoic (C22:0), tetracosanoic (C24:0) and hexacosanoic (C26:0) acids are the most important plasma parameters for the diagnosis and monitoring of the peroximal disorders in newborns. Currently, neonatal screening procedures for the peroximal disorders are based on its determination by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). Analyses of VLCFA by GC and GC/MS methods, while highly sensitive and specific, is still slow and difficult to automate.LC-ESI/MS/MS is known for its high throughput, high specificity, low detection limit and abundant structure information and is applied to the etermination of VLCFA of infants in this study. It is practicable to detect all VLCFA at a time related to peroxisomal disorders in blood using LC/MS/MS. Compare with traditional screening method, this new method should be fast, reliable, reproducible and relatively cheap. Currently, the amino acids, acylcarnitine and 17-hydroxyprogesterone profiling of dry blood specimens using LC/MS/MS has been developed and recognized as a useful tool for screening inherited metabolic defects of newborns in our laboratory. |
