Al Lawati Haider A J, Al-Nadabi Mira M, Varma Gouri B, Suliman Fakhr Eldin O, Al-Abri Hasnaa
Department of Chemistry, College of Science, Sultan Qaboos University, PO Box 36, Al-Khod, 123, Oman.
Luminescence. 2014 Dec;29(8):1148-53. doi: 10.1002/bio.2675. Epub 2014 Apr 30.
A highly sensitive, rapid and economical method for the determination of amlodipine (AM) in biological fluids was developed using a peroxyoxalate chemiluminescence (CL) system in a lab-on-a-chip device. Peroxyoxalate-CL is an indirect type of CL that allows the detection of native fluorophores or compounds derivatized with fluorescent labels. Here, fluorescamine was reacted with AM, and the derivatization product was used in a bis-(2,4,6-trichlorophenyl)oxalate-CL system. Fluorescamine reacts selectively with aliphatic primary amine at neutral or basic pH. As most of the calcium channel blocker and many cardiovascular drugs do not contain primary amine, the developed method is highly selective. The parameters that influenced the CL signal intensity were studied carefully. These included the chip geometry, pH, concentration of reagents used and flow rates. Moreover, we confirmed our previous observation about the effects of imidazole, which is commonly used in the bis-(2,4,6-trichlorophenyl)oxalate-CL system as a catalyst, and found that the signal was significantly improved when imidazole was absent. Under optimized conditions, a calibration curve was obtained with a linear range (10-100 µg/L). The limit of detection was 3 µg/L, while the limit of quantification was 10 µg/L. Finally the method was applied for the determination of AM in biological fluids successfully.
利用芯片实验室装置中的过氧草酸酯化学发光(CL)系统,开发了一种用于测定生物流体中氨氯地平(AM)的高灵敏度、快速且经济的方法。过氧草酸酯 - CL是一种间接型化学发光,可用于检测天然荧光团或用荧光标记衍生化的化合物。在此,荧光胺与AM反应,衍生化产物用于双 -(2,4,6 - 三氯苯基)草酸酯 - CL系统。荧光胺在中性或碱性pH下与脂肪族伯胺选择性反应。由于大多数钙通道阻滞剂和许多心血管药物不含伯胺,因此所开发的方法具有高度选择性。仔细研究了影响CL信号强度的参数。这些参数包括芯片几何形状、pH值、所用试剂的浓度和流速。此外,我们证实了我们之前关于咪唑影响的观察结果,咪唑在双 -(2,4,6 - 三氯苯基)草酸酯 - CL系统中常用作催化剂,发现不存在咪唑时信号显著改善。在优化条件下,获得了线性范围为(10 - 100μg/L)的校准曲线。检测限为3μg/L,定量限为10μg/L。最后,该方法成功应用于生物流体中AM的测定。