High Performance Liquid Chromatography Analysis of 2-mercaptoethylamine (Cysteamine) in Biological Samples by Derivatization with N-(1-pyrenyl) Maleimide (NPM) Using Fluorescence Detection
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 (λex = 330 nm, λem = 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 (r2 = 0.9993), and in plasma and liver matrix, the r2 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%.
J. Ogony et al., "High Performance Liquid Chromatography Analysis of 2-mercaptoethylamine (Cysteamine) in Biological Samples by Derivatization with N-(1-pyrenyl) Maleimide (NPM) Using Fluorescence Detection," Journal of Chromatography B, Elsevier, Jan 2006.
The definitive version is available at http://dx.doi.org/10.1016/j.jchromb.2006.05.027
Keywords and Phrases
Cystinosis; High performance liquid chromatography; Oxidative stress; Thiols
Article - Journal
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