Li Qiao-Jing, Li Yong-Sheng, Gao Xiu-Feng
School of Chemical Engineering, Sichuan University.
Anal Sci. 2015;31(5):413-9. doi: 10.2116/analsci.31.413.
Based on fluorescence capillary analysis technology, a method for quantitating lactate dehydrogenase (LDH) activity in a micro-volume sample was developed. Sample and reagent consumptions were merely 2 and 16 μL per time, respectively. The optimized test conditions were as follows. The reaction reagent consisted of 0.10 M phosphate buffer (pH 6.5), 0.30 mM NADH and 1.20 mM pyruvate. NADH standard was prepared with a phosphate buffer of pH 8.0, and its linear response was controlled in 0.05 - 0.30 mM. LDH standards containing 2.0 mM PEG could exhibit long-term stability. Under the optimized conditions, a linear response for LDH from 50 to 1200 U L(-1) and a detection limit of 31 U L(-1) were obtained with good precision (RSD: 2.1 - 2.2%, n = 10) and better recovery of 96 - 105%. The method's characteristics was high sensitivity, low consumptions, simple operations, good precision and reliability, lending itself to the miniaturization of fluorophotometer which transformed into a bedside instrument in the hospital.
基于荧光毛细管分析技术，开发了一种用于微量样品中乳酸脱氢酶（LDH）活性定量的方法。每次样品和试剂消耗量分别仅为2 μL和16 μL。优化后的测试条件如下。反应试剂由0.10 M磷酸盐缓冲液（pH 6.5）、0.30 mM NADH和1.20 mM丙酮酸组成。用pH 8.0的磷酸盐缓冲液制备NADH标准品，其线性响应控制在0.05 - 0.30 mM范围内。含有2.0 mM PEG的LDH标准品可表现出长期稳定性。在优化条件下，LDH在50至1200 U L⁻¹范围内呈线性响应，检测限为31 U L⁻¹，精密度良好（RSD：2.1 - 2.2%，n = 10），回收率较好，为96 - 105%。该方法具有灵敏度高、消耗低、操作简单、精密度和可靠性好的特点，有助于将荧光光度计小型化，转化为医院床边仪器。
