School of Chemical Engineering, Sichuan University, Chengdu, China.
West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China.
J Fluoresc. 2017 May;27(3):883-894. doi: 10.1007/s10895-017-2024-3. Epub 2017 Jan 13.
It was studied that making conditions of a micro-volume fluorescence capillary biosensor for determining pyruvate (PA) and lactate (LA). The biosensor made under the optimized conditions could be used for sequential quantifications of LA in the range 0.10-1.2 mM and PA in 4-120 μM, and its recovery for PA and LA was in a satisfactory range 97-106% for human serum samples, with detection limits of 0.023 mM for LA (RSD < 1.89%, n = 11) and 0.87 μM for PA (RSD < 1.70%, n = 11). The new assay possessed these advantages that the LDH immobilizing on capillary realized the reuse of expensive enzyme in fluorospectrophotometry, and the consumption of serum samples or chemical reagents decreased to 9 μL in per assay, and the analytes no needed to preseparation, and it also are accurate and reliable. Consequently, the fluorescence capillary biosensor should have a good prospect in assaying PA and LA or LA/PA ratios for clinical medicines or biology field. The optimization conditions and parameters obtained in this study have also a certain guiding significance for the development of biochip based on glass substrate.
研究了一种用于测定丙酮酸（PA）和乳酸（LA）的微体积荧光毛细管生物传感器的制作条件。在优化条件下制作的生物传感器可用于连续定量测定 LA 的范围为 0.10-1.2 mM 和 PA 的范围为 4-120 μM，其对人血清样品中 PA 和 LA 的回收率在 97-106%的满意范围内，LA 的检测限为 0.023 mM（RSD < 1.89%，n = 11），PA 的检测限为 0.87 μM（RSD < 1.70%，n = 11）。新测定法具有以下优点：在荧光分光光度法中固定化 LDH 实现了昂贵酶的重复使用，每次测定的血清样品或化学试剂消耗量减少到 9 μL，且无需预分离分析物，同时具有准确性和可靠性。因此，荧光毛细管生物传感器在临床药物或生物学领域测定 PA 和 LA 或 LA/PA 比值方面具有良好的前景。本研究中获得的优化条件和参数对基于玻璃衬底的生物芯片的发展也具有一定的指导意义。
