日本血吸虫中间宿主钉螺浸杀法给药后螺体内氯硝柳胺含量的变化研究

doi:10.11853/j.issn.1003.8280.2021.04.011

摘要/Abstract

摘要： 目的建立高效液相色谱法（HPLC）检测钉螺体内氯硝柳胺含量，探讨日本血吸虫中间宿主钉螺浸杀法给药后螺体内氯硝柳胺含量的变化。方法空白钉螺滴加氯硝柳胺标准品溶液，制备成含量为4.00、0.40和0.04 μg/g的样品。样品冷冻干燥后，置于冷冻粉碎机中，液氮粉碎。用含有4%乙酸的二氯化碳溶液超声萃取。萃取液氮气吹干、复溶，HPLC法检测氯硝柳胺含量，计算精密度和回收率。将钉螺浸泡于8.00、2.00、0.50和0.12 mg/L的氯硝柳胺水溶液中，分别在2、20、40、60、120、180、300、420和540 min取出钉螺、清洗。给药后的钉螺分为2组，第1组钉螺按照上述样品前处理方法，HPLC法检测氯硝柳胺含量；第2组经过清洗，恢复饲养48 h后，敲击法判断钉螺死亡数。结果建立的HPLC测定方法回收率为99.64%～110.36%，最低检出限为0.01 mg/L，最低定量限0.05 mg/L，相对标准偏差<10.00%。钉螺在接触浓度为8.00、2.00、0.50和0.12 mg/L的药液2 min时，体内药物含量达到最高，分别为1.96、1.67、1.08和0.98 μg/g；死亡率分别为33.33%、36.67%、16.67%和0；各浓度组在60 min后降至最低点，分别为1.32、0.76、0.75和0.62 μg/g，然后均有一定程度的升高。结论建立的样品前处理方法和HPLC法可满足测定要求，钉螺体内的氯硝柳胺含量呈先降低后升高的现象，产生这一现象的原因有待进一步研究。

关键词: 氯硝柳胺含量, 高效液相色谱, 杀螺活性, 钉螺

Abstract: Objective To establish a high-performance liquid chromatography (HPLC) method for measuring the content of niclosamide in Oncomelania hupensis snails, the intermediate host of Schistosoma japonicum, and to investigate the change in the content of niclosamide in snails after treatment with the immersion killing method. Methods The niclosamide standard solution was added to blank O. hupensis snails to prepare the samples with concentrations of 4.00, 0.40, and 0.04 μg/g, and after freeze drying, the samples were crushed with liquid nitrogen in a cryogenic sample crusher. Ultrasonic extraction was performed using carbon dichloride solution containing 4% acetic acid, and then the extracted liquid was blow-dried with nitrogen and redissolved. HPLC was used to measure the content of niclosamide, and precision and recovery were calculated. The snails were immersed in niclosamide solution at concentrations of 8.00, 2.00, 0.50, and 0.12 mg/L and were taken out and washed after 2, 20, 40, 60, 120, 180, 300, 420, and 540 minutes. The snails were then divided into groups 1 and 2; the snails in group 1 were treated with the above sample pretreatment method, and HPLC was used to measure the content of niclosamide; the snails in group 2 were washed and fed for 48 hours, and the knocking method was used to measure the number of dead snails. Results The HPLC method established had a recovery rate of 99.64%-110.36%, with a limit of detection of 0.01 mg/L, a limit of quantification of 0.05 mg/L, and a relative standard deviation of <10%. The concentrations of niclosamide in snails were 1.96, 1.67, 1.08, and 0.98 μg/g, respectively, after exposure to the drug solution at a concentration of 8.00, 2.00, 0.50, and 0.12 mg/L for 2 minutes, with mortality rates of 33.33%, 36.67%, 16.67%, and 0 respectively. After 60 minutes of exposure, the concentration decreased to the lowest level, i.e., 1.32, 0.76, 0.75, and 0.62 μg/g, respectively, followed by a certain degree of increase. Conclusion The established sample pretreatment method and HPLC method can be used for measurement, and the content of niclosamide in snails first decreases and then increases, which requires further studies for explanation.

Key words: Niclosamide content, High-performance liquid chromatography, Molluscicidal activity, Oncomelania hupensis

中图分类号:

R184.38

邢云天, 戴建荣, 曲国立, 梁幼生, 张苏阳, 冯柏年. 日本血吸虫中间宿主钉螺浸杀法给药后螺体内氯硝柳胺含量的变化研究[J]. 中国媒介生物学及控制杂志, 2021, 32(4): 441-446.

XING Yun-tian, DAI Jian-rong, QU Guo-li, LIANG You-sheng, ZHANG Su-yang, FENG Bai-nian. Change in the content of niclosamide in Oncomelania hupensis, the intermediate host of Schistosoma japonicum, after treatment with the immersion killing method[J]. Chines Journal of Vector Biology and Control, 2021, 32(4): 441-446.

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