用高效液相色谱─电化学检测器法检测Fenton反应中的羟自由基

用高效液相色谱─电化学检测器（ＨＰＬＣ－ＥＣ）法建立了体外测定羟自由基（ＯＨ）的方法。在Ｆｅｎｔｏｎ反应中，Ｆｅ２＋（０．３３ｍｍｏｌ．Ｌ－１－８．０ｍｍｏｌ·Ｌ－１）可剂量依赖地增加ｏＨ生成量。而Ｈ２Ｏ２（０．０００２５％—０．３％）同ＯＨ发生无明显的量效关系，高浓反的Ｈ２Ｏ２反而不利于ＯＨ的生成。反应体系中加入ＡＴＰ或ＡＤＰ均可显著地增加ＯＨ的生成量并有较好的量效关系．ＡＴＰ的另一个代谢产物ＡＭＰ则对ＯＨ的发生无影响。卡托普利可剂量依赖地体外淬灭ＯＨ。

Determination of hydroxyl radical in Fenton reactionby using high performance liquid chromatograph connected with electrochemical detector

A method for detecting hydroxyl radi-cal produced in Fenton reaction by usinghigh perform-ance liquid chromatogragh connected withelectrochemical detector(HPLC-EC)was established.and the effects of Fe2+,H2O2 as well as ATP,ADPand AMP on hydroxyl radical production in Fenton re-action were investigated.By using this method,effectof captopril to eliminate hydroxyl radical in vitro wasalso studied.In the assay,salicylic acid(SA)capturedhydroxyl radical in Fenton reaction and was oxidizedto 2,3-dihydroxybenzoic acid and 2,5-dihydroxy-benzoic acid.Which could be determined by captoprilHPLC-EC with a detecting limit of 0.4ng In ourstudy,it was found that Fe2+(0.33 mmol·L -1 -8.0mmol·L-1)dose-dependently produced hydroxyl rad-ical,while H2O2(0.00025%-0.3 %)didn't ATP andADP,but not AMP,could significantly increase theproduction of hydroxyl radical.Captopril coulddose-dependently eliminate hydroxyl radical producedin Fenton reaction.