Ogony Joshua, Mare Suneetha, Wu Wei, Ercal Nuran
Department of Chemistry, University of Missouri-Rolla, 1870 Miner Circle, Rolla, MO 65409, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Oct 20;843(1):57-62. doi: 10.1016/j.jchromb.2006.05.027. Epub 2006 Jun 21.
2-Mercaptoethylamine (cysteamine) is an aminothiol compound used as a drug for the treatment of cystinosis, an autosomal recessive lysosomal storage disorder. Because of cysteamine's important role in clinical settings, its analysis by sensitive techniques has become pivotal. Unfortunately, the available methods are either complex or labor intensive. Therefore, we have developed a new rapid, sensitive, and simple method for determining cysteamine in biological samples (brain, kidney, liver, and plasma), using N-(1-pyrenyl) maleimide (NPM) as the derivatizing agent and reversed-phase high performance liquid chromatography (HPLC) with a fluorescence detection method (lambda(ex)=330 nm, lambda(em)=376 nm). The mobile phase was acetonitrile and water (70:30) with acetic acid and o-phosphoric acid (1 mL/L). The calibration curve for cysteamine in serine borate buffer (SBB) was found to be linear over a range of 0-1200 nM (r(2)=0.9993), and in plasma and liver matrix, the r(2) values were 0.9968 and 0.9965, respectively. The coefficients of the variation for the within-run and between-run precisions ranged from 0.68 to 9.90% and 0.63 to 4.17%, respectively. The percentage of relative recovery ranged from 94.1 to 98.6%.
2-巯基乙胺（半胱胺）是一种氨基硫醇化合物，用作治疗胱氨酸病的药物，胱氨酸病是一种常染色体隐性溶酶体贮积症。由于半胱胺在临床环境中的重要作用，通过灵敏技术对其进行分析已变得至关重要。不幸的是，现有的方法要么复杂，要么劳动强度大。因此，我们开发了一种新的快速、灵敏且简单的方法，用于测定生物样品（脑、肾、肝和血浆）中的半胱胺，该方法使用N-(1-芘基)马来酰亚胺（NPM）作为衍生剂，并采用反相高效液相色谱（HPLC）结合荧光检测法（激发波长λ(ex)=330 nm，发射波长λ(em)=376 nm）。流动相为乙腈和水（70:30），含有乙酸和正磷酸（1 mL/L）。发现半胱胺在丝氨酸硼酸盐缓冲液（SBB）中的校准曲线在0 - 1200 nM范围内呈线性（r(2)=0.9993），在血浆和肝基质中，r(2)值分别为0.9968和0.9965。批内精密度和批间精密度的变异系数分别在0.68%至9.90%和0.63%至4.17%之间。相对回收率的百分比在94.1%至98.6%之间。
