Toxicity Assessment of Silver Ions Compared to Silver Nanoparticles in Aqueous Solutions and Soils Using Microtox Bioassay

Microtox 생물검정법을 이용한 은 이온과 은 나노입자의 수용액과 토양에서의 독성 비교 평가

This study was conducted to assess the microbial toxicity of ionic silver solution ($Ag^+N$) and silver nanoparticle suspension ($Ag^0NP$) based on the Microtox bioassay. In this test, the light inhibition of luminescent bacteria was measured after 15 and 30 min exposure to aqueous solutions and soils spiked with a dilution series of $Ag^+N$ and $Ag^0NP$. The resulting dose-response curves were used to derive effective concentration (EC25, $EC_{50}$, EC75) and effective dose ($ED_{25}$, $ED_{50}$, $ED_{75}$) that caused a 25, 50 and 75% inhibition of luminescence. In aqueous solutions, $EC_{50}$ value of $Ag^+N$ after 15 min exposure was determined to be < $2mg\;L^{-1}$ and remarkably lower than $EC_{50}$ value of $Ag^0NP$ with $251mg\;L^{-1}$. This revealed that $Ag^+N$ was more toxic to luminescent bacteria than $Ag^0NP$. In soil extracts, however, $ED_{50}$ value of $Ag^+N$ with 196 mg kg-1 was higher than $ED_{50}$ value of $Ag^0NP$ with $104mg\;kg^{-1}$, indicating less toxicity of $Ag^+N$ in soils. The reduced toxicity of $Ag^+N$ in soils can be attributed to a partial adsorption of ionic $Ag^+$ on soil colloids and humic acid as well as a partial formation of insoluble AgCl with NaCl of Microtox diluent. This resulted in lower concentration of active Ag in soil extracts obtained after 1 hour shaking with $Ag^+N$ than that spiked with $Ag^0NP$. With longer exposure time, EC and ED values of both $Ag^+N$ and $Ag^0NP$ decreased, so their toxicity increased. The toxic characteristics of silver nanomaterials were different depending on existing form of Ag ($Ag^+$, $Ag^0$), reaction medium (aqueous solution, soil), and exposure time.

$Ag^+$ 이온을 주성분으로 하는 $Ag^+N$과 $Ag^0$ 나노입자를 주성분으로 하는 $Ag^0NP$의 미생물학적 독성을 Microtox 생물검정법을 이용하여 수용액과 토양에서 용량-반응관계를 이용하여 비교, 평가하였다. 수용액 실험에서 Vibrio fisheri의 50% 발광 저해율을 보여주는 $EC_{50}$ 값은 $Ag^+N$이 $Ag^0NP$ 보다 현저히 낮게 나타나, 이온상태의 $Ag^+N$이 독성이 훨씬 높음을 알 수 있었다. 노출시간이 15분에서 30분으로 증가하면 독성 또한 증가했다. 반대로 토양 추출액 실험에서는 $Ag^+N$의 $ED_{50}$ 값이 $Ag^0NP$의 값 보다 높아, $Ag^+N$의 독성이 더 낮게 나타났다. 이것은 $Ag^+N$의 $Ag^+$가 토양 입자 또는 부식산에 강하게 흡착 되거나, Microtox 희석제 NaCl과 반응하여 난용해성 AgCl 침전물을 형성하여, 토양 추출액 중의 활성 Ag 농도가 감소한 것에 기인하는 것으로 판단되었다. Microtox 분석에 의한 Ag 나노용액의 생물학적 독성은 Ag의 존재형태 ($Ag^+$, $Ag^0$), 반응매질 (수용액, 토양), 노출시간에 따라 서로 상이한 결과를 보여 주었다.