Rapid UPLC-MS/MS method for routine analysis of plasma pristanic, phytanic, and very long chain fatty acid markers of peroxisomal disorders
Rapid UPLC-MS/MS method for routine analysis of plasma pristanic, phytanic, and very long chain fatty acid markers of peroxisomal disorders
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DOI：
10.1194/jlr.d800019-jlr200
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发表时间：
2008-08-01
影响因子：
6.5
通讯作者：
Al-Mokhadab, Manhal
中科院分区：
文献类型：
--
作者：
Al-Dirbashi, Osama Y.;Santa, Tomofumi;Al-Mokhadab, Manhal
定量测定血浆中的丁酸、植酸和长链脂肪酸（即六糖酸、四糖酸和二十二糖酸）是研究多种过氧化物酶体疾病（pd）的主要方法。现有的方法通常基于GC-MS，耗时且费力。本文以4-[2-（N，N-二甲氨基）乙基氨基磺酰基]-7-（2-氨基乙基氨基）-2,1,3-苯并恶二唑（DAABD-AE）为原料，建立了一种快速、特异的液相色谱串联质谱分析方法。衍生化是为了改善这些脂肪酸的质谱特性。血浆中（20 μ l）的分析物在大约2小时内用DAABD-AE进行水解、提取和衍生化。衍生物在反相柱上分离，用正离子电喷雾串联质谱法检测，注射至注射时间为5 min。校正图在覆盖生理和病理浓度的范围内呈线性。低、高浓度的日内（n = 12）和日内（n = 10）变化均小于9.2%。在正常血浆（n = 250）中为每种化合物建立参考区间，并与文献一致。使用确诊患者的标本（n = 20），可以可靠地检测到各种pd。总之，该方法允许在5分钟的分析运行中检测至少9个pd。此外，这种衍生化方法可能适用于其他携带羧基的疾病标志物。
Quantification of pristanic acid, phytanic acid, and very long chain fatty acids (i.e., hexacosanoic, tetracosanoic, and docosanoic acids) in plasma is the primary method for investigateing a multitude of peroxisomal disorders (PDs). Typically based on GC-MS, existing methods are time-consuming and laborious. In this paper, we present a rapid and specific liquid chromatography tandem mass spectrometric method based on derivatization with 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole (DAABD-AE). Derivatization was undertaken to improve the poor mass spectrometric properties of these fatty acids. Analytes in plasma (20 mu l) were hydrolyzed, extracted, and derivatized with DAABD-AE in similar to 2 h. Derivatives were separated on a reverse-phase column and detected by positive-ion electrospray ionization tandem mass spectrometry with a 5 min injection-to-injection time. Calibration plots were linear over ranges that cover physiological and pathological concentrations. Intraday (n = 12) and interday (n = 10) variations at low and high concentrations were less than 9.2%. Reference intervals in normal plasma (n = 250) were established for each compound and were in agreement with the literature. Using specimens from patients with established diagnosis (n = 20), various PDs were reliably detected. In conclusion, this method allows for the detection of at least nine PDs in a 5 min analytical run. Furthermore, this derivatization approach is potentially applicable to other disease markers carrying the carboxylic group.