Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
J Sep Sci. 2014 Mar;37(5):465-575. doi: 10.1002/jssc.201301188. Epub 2014 Jan 29.
Sarcosine has been identified as a potential prostate cancer marker. To provide determination of this compound, a number of methods are developing. In this study, we optimized a method for its separation by hydrophilic interaction LC with electrochemical detection (ED). Due to the fact that mobile phases commonly used for this type of separation altered the LODs measured by electrochemical detectors, we applied postcolumn dosing of buffer suitable for ED. The optimized conditions were mobile phase A acetonitrile, mobile phase B water in the ratio A/B 70:30, with postcolumn addition of mobile phase C (200 mM phosphate buffer pH 9). The optimal mixing ratio was A + B/C 1:1 with a flow rate of 0.80 mL/min (0.40 + 0.40 mL/min) and detection potential of 1000 mV. Due to the optimization of the parameters for effective separation, which had to meet the optimal parameters of ED, we reached a good resolution for separation also with a good LOD (100 nM). In addition, we successfully carried out sarcosine analysis bound on our modified paramagnetic microparticles with the ability to preconcentrate sarcosine isolated from artificial urine.
肌氨酸已被确定为一种潜在的前列腺癌标志物。为了对这种化合物进行定量，人们正在开发许多方法。在这项研究中，我们通过亲水相互作用 LC 与电化学检测 (ED) 对其分离方法进行了优化。由于常用于此类分离的流动相改变了电化学检测器测量的 LO D 值，因此我们采用了适合 ED 的缓冲液的柱后添加。优化条件为流动相 A 乙腈，流动相 B 水的比例为 A/B 70:30，柱后添加流动相 C（200 mM 磷酸盐缓冲液，pH 值 9）。最佳混合比为 A + B/C 1:1，流速为 0.80 mL/min（0.40 + 0.40 mL/min），检测电位为 1000 mV。由于必须满足 ED 的最佳参数，因此我们对有效分离的参数进行了优化，从而实现了良好的分离度和良好的 LO D（100 nM）。此外，我们还成功地在经过修饰的顺磁微球上进行了肌氨酸分析，该微球具有从人工尿液中浓缩肌氨酸的能力。
