2-(α-羟基戊基)苯甲酸钾盐的急性毒性和致突变作用研究

Studies on acute toxicity and mutagenicity of potassium 2-(1-hydroxypentyl)-benzoate

目的：研究2-（a-羟基戊基）苯甲酸钾盐（dl—PHPB）的急性毒性及致突变性。方法：急性毒性试验采用Bliss法，昆明小鼠分为5组，每组10只，雌雄各半，分别按256．0、286．3、320．0、357．8、400．0、447．2mg／kg静脉注射dl—PHPB，观察注射后至给药后14d动物的毒性反应性质及程度，计算半数致死剂量（LD50）及95％可信限；Ames试验采用平板掺入预培养法，选用鼠伤寒沙门氏菌株TA97、TA98、TA100和TA102，每皿10．5～5000．0μg浓度范围内，检测加和不加S9条件下对Ames菌的致突变性；微核试验采用雄性昆明小鼠，分为3组，每组6只，分别按23．3、70、210mg／kg单次静脉注射dl—PHPB，注射后24h计数骨髓嗜多染红细胞微核率（MNPCEs）；体外染色体畸变试验在加和不加S9条件下，检测11．0～88．0μg／mL dl—PHPB对CHL细胞染色体畸变率的影响。结果：小鼠静脉注射dl—PHPB后，各剂量动物出现--过性呼吸急促、自发活动减少、俯卧少动等症状，随剂量增加至≥320．0mg／kg时，部分动物出现濒死症状，并在给药后2～30min内死亡，死亡率随剂量的增加而增加，256．0～447．2mg／kg6个剂量组死亡率依次为0、0、10％、30％、70％、100％。未死亡动物在给药30min后逐渐恢复正常。14d观察期期间动物未见其他异常。其LD50为373．3mg／kg，95％可信限为355．6～392．0mg／kg；Ames试验结果显示，在每皿0．5—5000．0μg浓度范围内未引起4种测试菌株回复突变数增加；微核试验结果显示，dl—PHPB在23．3～210．0mg／kg范围内，各组动物MNPCEs均低于4‰；CHL细胞体外染色体畸变试验中，dl—PHPB在11．0～88．0μg／mL浓度范围内致CHL细胞染色体畸变率〈5％。结论：在本实验条件下，小鼠静脉注射dl—PHPB的LD50为373．3mg／kg，致突变性3项试验均为阴性结果。

OBJECTIVE： To evaluate the acute toxicity and mutagenicity of potassium 2-（1-hydroxypentyl）- benzoate（dl-PHPB）. METHODS： KM mice were intravenously treated with dl-PHPB at the doses of 256.0, 286.3, 320.0, 357.8, 400.0 and 447.2 mg/kg. We observed the clinical signs of toxicity and calculated the median lethal dose （LDs0） and LDs0 95% confidence interval. Mutagenicity of dl-PHPB was studied by Ames test, bone marrow micronucleus study in male mice and in vitro CHL cell mammalian chromosome aberration assay. In Ames test, the tester strains used were Salmonella typhimurium TA97, TA98, TA100 and TA102, mutagenicity was evaluated with the plate incorporation method beginning at 0.5, 5, 50, 500 and 5 000 μg per plate in the absence or presence of S9 metabolic activation. In bone marrow micronucleus study, KM male mice were intravenously treated with dl-PHPB at the dosse of 210, 70 and 23.3 mg/kg. Bone marrow cells were harvested from mice at 24 hours after dosing. 1 000 PCEs per animal were examined microscopically for the presence of micronucleated polychromatic erythrocytes（MNPCEs）. The in vitro CHL cell mammalian chromosome aberration study on dl-PHPB consisted of the premilinary toxicity assay and chromosome aberration assay, with and without S9 metabolic activation system, and chromosomal aberration percentage per dose were analyzed. RESULTS ： In the acute toxicity study, all mice after dosing developed transient toxic symptoms such as tachypnea, sautonomic activity decreased, prostration, then the dying state were observed at dosage of ≥ 320.0 mg/kg , mice appeared respiratory depression, cyanosis, loss of righting reflex, and died during 2 to 30 minute after administration. The mortality rate of mice treated with dl-PHPB from 256.0 to 447.2 mg/kg were 0, 0, 10%, 30%, 70%, 100%, respectively. The survival mice returned to normal after 30 min. In theM-days observation period, there were no abnormal clinical signs in the surviving animals and the general autopsy showed no abnormality in major organs. The intravenous median lethal dose of dl-PHPB was 373.3 mg/kg in mice, LDs0 95% confidence interval was 355.6 to 392.0 mg/kg. In the Ames assay, no positive mutagenic response was observed. In bone marrow micronucleus study, a single bolus intravenous administration of dl-PHPB at dosages of 23.3, 70 and 210 mg/kg didn＇ t induce a significant increase in the number of MNPCEs in the bone marrow, MNPCEs were less than 4%0 at three dosages. In CHL chromosome aberration assay, dl-PHPB achieved a chromosome aberration of less than 5 % in the presence and absence of S9 mixture. CONCLUSION： The intravenous median lethal dose of dl-PHPB was 373.3 mg/kg in mice, and dl- PHPB was not mutagenic in these 3 screening mutation assays.