Devall A J, Blake R, Langman N, Smith C G S, Richards D A, Whitehead K J
Pain Signalling Group, Department of Pharmacology, Division of Neuroscience, The Medical School, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Apr 1;848(2):323-8. doi: 10.1016/j.jchromb.2006.10.049. Epub 2006 Nov 13.
The development of a HPLC method using a monolithic C18 column is described using fluorescence detection for the assay of 21 amino acids and related substances with derivatisation using ortho-phthaldialdehyde (OPA) in the presence of 3-mercaptopropionic acid (3-MPA). The method employs a tertiary gradient and has a run time of 24 min. Linearity (r2) for each amino acid was found to be greater than 0.99 up to a 10 microM concentration; reproducibility across all analyses (relative standard deviation (R.S.D.)) was between 0.97 and 6.7% and limit of detection (LOD) between 30 and 300 fmol on column. This method has been applied to the analysis of amino acids in both spinal microdialysis and cerebral spinal fluid samples.
本文描述了一种使用整体式C18柱的高效液相色谱（HPLC）方法，该方法采用荧光检测，通过在3-巯基丙酸（3-MPA）存在下用邻苯二甲醛（OPA）衍生化来测定21种氨基酸及其相关物质。该方法采用三元梯度，运行时间为24分钟。发现每种氨基酸在浓度高达10微摩尔时的线性（r2）大于0.99；所有分析的重现性（相对标准偏差（R.S.D.））在0.97%至6.7%之间，柱上检测限（LOD）在30至300飞摩尔之间。该方法已应用于脊髓微透析和脑脊液样品中氨基酸的分析。
