Sadat CityUniversity, Department of Environmental Biotechnology, Genetic Engineering and Biotechnology Research Institute (GEBRI), Sadat City, Egypt.
Anal Chim Acta. 2013 Jul 17;787:226-32. doi: 10.1016/j.aca.2013.05.036. Epub 2013 May 27.
Reaction of formaldehyde with amino acids followed by oxidation with hydrogen peroxide to produce a fluorophore Norharman product is well known and was used for the spectrofluorimetric determination of l-tryptophan (Trp). This study aimed to use graphene oxide (GO) to enhance the selectivity and sensitivity of Trp in presence of other amino acids and possible interfering compounds. Different parameters such as pH, temperature, incubation time, and concentrations of formaldehyde, H2O2 and GO were studied to optimize the condition of determination. Experimental data showed that the maximum fluorescence intensity was achieved in pH 7.0-9.0 phosphate buffer mixed with 7-10% (v/v) formaldehyde and 1-2% (v/v) H2O2 as oxidizing agent at 60°C for 1h. On the basis of calibration curve of various concentrations of Trp in the presence of 20 μg mL(-1) GO, the lower limit of detection (LOD) of Trp was determined as 0.092 nmol mL(-1) and the lower limit of quantification (LOQ) was 0.3 nmol mL(-1). The selectivity of Trp in presence of other amino acids and possible interfering compounds were studied with and without GO. The data obtained after inner filter effect corrections revealed that the selectivity of Trp in presence of amino acids and other possible interfering agents was improved in the range of 76-96%, compared with that in absence of GO. The enhancement of selectivity in the presence of GO indicates that the Trp and other amino acid and possible interfering compounds were adsorbed by GO, and the selective uptaking of Trp-by the reaction with formaldehyde followed by oxidation with H2O2 at 60°C with high selectivity and sensitivity was achieved successfully.
甲醛与氨基酸反应,然后用过氧化氢氧化生成荧光团诺拉曼产物,这是众所周知的,用于测定 l-色氨酸(Trp)的荧光分光光度法。本研究旨在使用氧化石墨烯(GO)来提高 Trp 在存在其他氨基酸和可能的干扰化合物时的选择性和灵敏度。研究了不同的参数,如 pH 值、温度、孵育时间,以及甲醛、H2O2 和 GO 的浓度,以优化测定条件。实验数据表明,在 pH 7.0-9.0 的磷酸盐缓冲液中,加入 7-10%(v/v)的甲醛和 1-2%(v/v)的 H2O2 作为氧化剂,在 60°C 下孵育 1 小时,可获得最大的荧光强度。在存在 20μg mL(-1) GO 的情况下,各种浓度 Trp 的校准曲线的基础上,确定 Trp 的检测限(LOD)为 0.092 nmol mL(-1),定量限(LOQ)为 0.3 nmol mL(-1)。研究了在存在和不存在 GO 的情况下,其他氨基酸和可能的干扰化合物存在时 Trp 的选择性。在内滤效应校正后获得的数据表明,与不存在 GO 相比,Trp 在存在氨基酸和其他可能的干扰剂时的选择性提高了 76-96%。在存在 GO 的情况下选择性增强表明,Trp 和其他氨基酸和可能的干扰化合物被 GO 吸附,并且通过在 60°C 下与 H2O2 反应进行选择性吸收,成功地实现了高选择性和灵敏度。