高效液相色谱法测定喹噁林类药物对刺参组织DNA的氧化损伤

为探索喹噁林类药物在刺参养殖中使用的安全性问题，采用高效液相色谱-紫外法对喹噁林类药物引起刺参组织DNA氧化损伤效应进行研究。采集经过不同添加剂量、不同时间喹噁林药物处理过的刺参组织，提取DNA，酶解后用高效液相色谱-紫外法对DNA氧化损伤标志物8-羟基脱氧鸟苷（8-hydroxy-2’-deoxyguanosine，8-OHdG）进行测定分析。结果表明：高效液相色谱-紫外检测法，操作简单，具有广泛的适用性、稳定性和准确性，灵敏度高，并且样品用量少，分析速度快。8-OHdG的含量随着乙酰甲喹质量浓度的增加和处理时间的延长而显著增加（P＜0.05），存在质量浓度-反应、时间-反应关系，并且是一个持续性的过程。喹烯酮对DNA的氧化损伤也是随着添加剂量的增加而显著加重（P＜0.05）。说明乙酰甲喹和喹烯酮能引起刺参组织DNA的氧化损伤，对其遗传物质具有一定的遗传毒性，因此需要规范喹噁林类药物在刺参养殖中的使用。

Determination of Oxidative DNA Damage Induced by Quinoxaline in Apostichopus japonicas Tissue Using High Performance Liquid Chromatography

8-Hydroxy-2’-deoxyguanosine (8-OHdG) is one of the main modified products from oxidative DNA damage
when deoxyguanosine (dG) is oxidized by a large amount of active oxygen species (ROS). Once 8-OhdG has avoided to
be repaired within the body, it maybe the initiation factor causing mutagenic, teratogenic and carcinogenic effect. 8-OHdG
is a metabolic end product that is stable within the body. It is not formed by non-DNA oxidation pathway outside the cell.
The content of 8-OHdG can reflect the degree of oxidative damage in DNA. Thus, it is important to determine the content
of 8-OHdG in order to evaluate the interaction between oxidative damage, oxidative stress and DNA damage in the body.
The quinoxaline-induced DNA oxidative damage in Apostichopus japonicas tissues was determined by high performance liquid
chromatography (HPLC) in this study. Sea cucumber tissue treated with quinoxalines at different concentrations and for different
time periods were collected. DNA were extracted and treated enzymatically, and then 8-OHdG as an indicator of DNA oxidative
damage was measured and analyzed by HPLC. The results showed that the HPLC-UV method displayed a wide applicability,
stability and accuracy, high sensitivity, less samples and fast analysis. The 8-OHdG content remarkably increased along with
increasing concentration and processing time of mequindox. The dose- and time-dependent response relationship existed,
which were a continual process. The oxidative damage to DNA caused by quinocetones significantly increased with increasing
concentration (P < 0.05). It was indicated that there was obvious genetic toxicity resulting from oxidative DNA damage caused by
mequindox and quinocetone in sea cucumber. Thus, it is necessary to control strictly quinoxalines applied in sea cucumber.