目的研究还原型谷胱甘肽(GSH)、半胱氨酸(Cys)、维生素C(VC)和亚铁离子(Fe2+)等常见还原性物质对五价无机砷[As(Ⅴ)]的还原作用及特点。方法模拟体内环境建立还原体系[37℃、pH7.0的磷酸盐缓冲液、3μmol/L As(Ⅴ)、60mmol/L巯基化合物(GSH和Cys)、0-30mmol/L VC、0-40mmol/LFe^2+],采用高效液相色谱-氢化物发生-原子荧光法(HPLC-HG-AFS)检测各种反应过程中的三价无机砷[As(Ⅲ)]和As(Ⅴ)。结果As(Ⅴ)还原率和As(Ⅲ)生成率随GSH浓度增加(0~60mmol/L)而增加;当GSH浓度为60mmol/L时,As(Ⅴ)还原率和As(Ⅲ)生成率达到峰值;随着GSH浓度继续增加(60~80mmol/L),As(Ⅴ)还原率和As(Ⅲ)生成率反而下降;巯基化合物中的巯基(-SH)在还原As(Ⅴ)过程中起关键作用,60mmol/L的-SH反应30分钟后As(Ⅴ)还原率达到60%,As(Ⅴ)还原率与-SH的具体存在形式(GSH或Cys)无关;单独VC或Fe^2+不能还原As(Ⅴ),但是当VC浓度大于10mmol/L或Fe2+浓度大于20mmol/L时,可以增加60mmol/L的-SH的还原作用(P〈0.01)。结论巯基化合物中的-SH可以非酶促还原As(Ⅴ),VC和Fe^2+可以增强-SH的还原作用。
Objective To study the role of glutathione(GSH), cysteine(Cys), vitamin C(VC) and ferrous ion(Fe^2+) in the reduction of arsenate [ As ( Ⅴ ) ] and the characteristic of reduction.Methods The reaction conditions [ ( 37 ℃, pH7.0 phosphate buffer) and the concentrations of As ( V ) ( 3μmol/L), GSH ( 60mmol/L), Cys ( 60mmol/L), VC (0 - 30mmol/L) and Fe^2+ (0 - 40mmol/L) ] were simulated in vitro. Arsenite[As( Ⅲ ) ] and [As( Ⅴ ) ] were determined by the high performance liquid chromatography-hydride generation-atomic fluorescence spectrometry(HPLC-HG-AFS). Results The reduction, of As( Ⅴ ) and As( Ⅲ) formation, increased with the concentration, of GSH(0 - 60mmol/L), and reached a maximum values at the concentration of 60mmol/L GSH, then decreased with the concentration of(60- 80mmol/L) GSH. -SH in suffhydryl compound, could play an important role in the reduction of As( Ⅴ ), and at the concentration of 60mmol/ L -SH(both GSH and Cys) could reduce 60% of As( Ⅴ ) to As( Ⅲ) for 30 min. VC or Fe^2+ could not reduce As( Ⅴ ) alone, but when the concentration of VC or Fe^2+ reached to 10mmol/L or 20mmol/L, they could have a significantly synergistic effect with 60mmol/L -SH on the reduction of As(Ⅴ ) ( P 〈 0.01 ). Conclusion -SH could reduce As( Ⅴ ) alone without any reductase. VC and Fe^2+ could have a synergistic effect with -SH on the reduction of As( V ).