含多酸化合物新型洗手液抗甲型流感病毒效果及毒性评价

目的：配制含有多酸化合物的新型洗手液，并探讨其在人手部抗流感病毒的效果和安全性。方法：采用MDCK细胞
流感病毒模型研究2种多酸化合物K10Na［(VO)3(SbW9O33)2］?26H2O(PM-1001)和K15［Pr(BW11O39)2］?28H2O(Pr-B)
的体外抗病毒作用，选择3个适宜的多酸剂量配制成含高、中、低剂量多酸洗手液；依据美国标准方法进行洗手液抗病毒效果评价，含2种多酸化合物的洗手液各有5名志愿者参与实验，将每名志愿者的10个手指随机分成5组，在手指上涂0.02 mL组织培养半数感染剂量（TCID50）为107.5/0.1 mL的病毒悬液后，分别在加病毒后、病毒液晾干后和用不同洗手液处理后收集病毒，采用MDCK细胞培养法测定病毒滴度；检测流感病毒在手部晾干30和60 min后的浓度和活力变化；选择含高浓度PM-1001的
洗手液，按照《消毒技术规范》（2008版）利用昆明小鼠和白色家兔进行毒性评价。结果：PM-1001和Pr-B抗流感病毒的治疗指数(TI)分别为82.68和69.13；对照组的病毒浓度为104.33±0.66TCID50/0.1 mL，各洗手液处理组组织培养法均未检测到病毒，各洗手液处理组病毒浓度均低于对照组（P<0.001），30和60 min收集的病毒浓度分别为103.98 ±0.42TCID50/0.1 mL和103.83±0.35TCID50/0.1 mL，与对照组比较差异无统计学意义（P>0.05）；含高浓度PM-1001的洗手
液对昆明小鼠的经口毒性实验半数致死浓度（LD50）大于5　000　mg/kg^-1（体质量），对家兔完整皮肤刺激指数为0.33，对3只家兔眼刺激的平均评分分别为0、0.67和0.33。结论：含多酸化合物的新型洗手液可有效抑制手部甲型流感病毒，属实际无毒级的物质，对家兔完整皮肤和眼无刺激性。

Evaluation on effect of anti-A-influenza virus and toxicity of new hand-washing agent with heterpolyoxometalates

Objective To prepare a new hand-washing agent with heterpolyoxometalates and to study its anti-Ainfluenza virus effect and toxicity.Methods The anti-virus effects of the two compounds K10 Na[(VO) 3(SbW9 O33) 2 ]·26H2 O(PM-1001)and K15 [Pr(BW11 O39) 2 ]·28H2 O(Pr-B)were detected on MDCK cells. Three levels of heterpolyoxometalates(high,middle,and low doses)were chosen to add into hand-washing agents;the anti-A-influenza virus effect test was performed according to the American Society for Testing and Materials(ASTM)standard assessment protocols.In this test,10volunteers were randomly divided into twogroups for testing hand-washing agents with PM-1001and Pr-B respectively.Every volunteer's ten fingers were divided into 5groups randomly.After all the finger taps were contaminated with 0.02mL of 107.5 tissue culture infectious dose(TCID50)/0.1mL live human influenza A virus,the H1N1titers were assessed with MDCK cell culture method after contaminating,air drying and treatment with different hand-washing agents. The concentrations and viabilities of H1N1on hands 30and 60min after hand drying were also assessed.The handwashing agent with high dose of PM-1001 was chosen for toxicity assessment using Kunming mice and rabbits according to Disinfection Technical Specifications(2008).Results The therapeutic indexes(TI)of PM-1001and Pr-B were 82.68and 69.13.The H1N1concentration in control group was 104.33±0.66 TCID50 /0.1mL.The fingers in treatment groups had no viable H1N1recovered.The virus concentrations in treatment groups had significant differences compared with control group(P<0.001).The H1N1virus concentrations in 30and 60 min groups were 103.98 ±0.42 TCID50 /0.1 mL and 103.83±0.35 TCID50 /0.1 mL and had no significant differences compared with control group(P>0.05).The LD50 in mouse acute toxicity test was>5 000mg·kg-1,the rabbit skin irritation index was 0.33,and the eye stimulation indexes of three rabbits were 0,0.67,and 0.33.Conclusion The new hand-washing agents including PM-1001and Pr-B are highly effective in reducing influenza A virus on human hands. It has no toxicity and no stimulation to skin and eyes of rabbits.