Dietzen Dennis J, Weindel Annette L
Department of Pediatrics, Washington University School of Medicine, St. Louis, MO, USA.
Methods Mol Biol. 2010;603:27-36. doi: 10.1007/978-1-60761-459-3_3.
Analysis of clinically relevant amino acids using ion-exchange chromatography coupled to photometric detection has been an indispensable component in the detection of inborn errors of metabolism for six decades. Detection of amino acids using mass spectrometry offers advantages in speed and analytic specificity. Employing methanol extraction and controlled butylation, C8 reversed-phase chromatography, and MS/MS detection, 32 amino acids are quantified in 20 min with clinically appropriate imprecision in plasma, urine, and CSF. Quantitation is linear to 1,000 micromol/L and limits of detection are at least 1.0 micromol/L. Important isobaric amino acids are distinguished by chromatography or by unique patterns of fragmentation following collision-induced dissociation. The technique employs commercially available reagents and may be expanded and customized for specific clinical or research settings.
六十年来，使用离子交换色谱法结合光度检测对临床相关氨基酸进行分析一直是检测先天性代谢缺陷不可或缺的组成部分。使用质谱法检测氨基酸在速度和分析特异性方面具有优势。采用甲醇萃取和可控丁基化、C8反相色谱法以及MS/MS检测，可在20分钟内对血浆、尿液和脑脊液中的32种氨基酸进行定量，其不精密度在临床可接受范围内。定量线性范围至1000微摩尔/升，检测限至少为1.0微摩尔/升。重要的等压氨基酸通过色谱法或碰撞诱导解离后的独特碎片模式来区分。该技术使用市售试剂，可针对特定临床或研究环境进行扩展和定制。
