Liu Qing-ye, Liang Yue-yuan, Liang Ai-hui, Jiang Zhi-liang
School of Environment and Resource, Guangxi Normal University, Guilin 541004, China.
Guang Pu Xue Yu Guang Pu Fen Xi. 2009 Sep;29(9):2535-8.
In acetate buffer solution and in the presence of glucose oxidase (GOD), glucose reduced the dissolved oxygen to form H2O2 that oxidized catalytically the excess KI to from I3- by horseradish peroxidase (HRP). The I3- combines respectively with rhodamine S (RhS), rhodamine 6G(Rh6G), butyl-rhodamine B(b-RhB) and rhodamine B(RhB) to form RhS-I3, Rh6G-I3, b-RhB-I3 and RhB-I3 associated particles that result in fluorescence quenching at 556, 556, 584 and 584 nm, respectively. Under the optimal conditions, the concentration of glucose in the range of 0.083-9.99, 0.17-8.33, 0.33-8.33 and 0.33-9.99 micromol x L(-1) is linear with their fluorescence quenching at 556, 556, 584 and 584 nm, with detection limits of 0.059, 0.17, 0.21 and 0.16 micromol x L(-1) glucose. And the regression equation was deltaF = 40.0c + 3.0, deltaF = 23.9c + 8.1, deltaF = 25.6c + 4.2, and deltaAF = 18.4c + 0.8, respectively. The RhS system was the most sensitive and stable, and was chosen for use. Influence of some foreign substances on the RhS fluorescence quenching determination of 6.67 micromol x L(-1) glucose was examined, with a relative error of +/- 10%. Results showed that 1000-fold Mg2+ and Cu2+, 300-fold Mn2+, 100-fold Zn2+, Al3+ and Co2+, 60-fold L-tyrosine, urea and nicotinic acid, 50-fold Fe3+, HSA and BSA, 10-fold sucrose, vitamin B2, L-lysine, L-glutamic acid and L-cystine did not interfere with the determination. This RhS fluorescence quenching assay was applied to the determination of glucose in the serum samples with satisfactory results.
在醋酸盐缓冲溶液中，在葡萄糖氧化酶（GOD）存在的情况下，葡萄糖将溶解氧还原形成H₂O₂，H₂O₂在辣根过氧化物酶（HRP）的作用下将过量的KI催化氧化形成I₃⁻。I₃⁻分别与若丹明S（RhS）、若丹明6G（Rh6G）、丁基罗丹明B（b-RhB）和罗丹明B（RhB）结合形成RhS-I₃、Rh6G-I₃、b-RhB-I₃和RhB-I₃缔合颗粒，分别导致在556、556、584和584nm处的荧光猝灭。在最佳条件下，葡萄糖浓度在0.083 - 9.99、0.17 - 8.33、0.33 - 8.33和0.33 - 9.99 μmol·L⁻¹范围内，其在556、556、584和584nm处的荧光猝灭呈线性关系，葡萄糖的检测限分别为0.059、0.17、0.21和0.16 μmol·L⁻¹。回归方程分别为ΔF = 40.0c + 3.0、ΔF = 23.9c + 8.1、ΔF = 25.6c + 4.2和ΔF = 18.4c + 0.8。RhS体系最灵敏且稳定，被选用。考察了一些外来物质对6.67 μmol·L⁻¹葡萄糖的RhS荧光猝灭测定的影响，相对误差为±10%。结果表明，1000倍的Mg²⁺和Cu²⁺、300倍的Mn²⁺、100倍的Zn²⁺、Al³⁺和Co²⁺、60倍的L-酪氨酸、尿素和烟酸、50倍的Fe³⁺、HSA和BSA、10倍的蔗糖、维生素B₂、L-赖氨酸、L-谷氨酸和L-胱氨酸不干扰测定。该RhS荧光猝灭法用于血清样品中葡萄糖的测定，结果令人满意。
