HPLC-UV검출기를 이용한 논토양 중 oxolinic acid 분석

Determination of Oxolinic Acid in Paddy Soil by HPLC Coupled with UV Detector

논토양 중 quinolone계의 항균성물질인 oxolinic acid에 대하여 HPLC-UV검출기를 이용하여 정량하는 잔류분석방법을 설정하였다. 논토양 시료는 지역과 토성이 다른 2토양을 공시하였으며, 토양 중 oxolinic acid는 4 M-KOH : MeOH(1 : 3, v/v) 혼합용매로 추출하여 추출액을 산성화 한 다음 dichloromethane으로 액 액분배하여 dichloromethane층을 탈수, 농축 한 후 HPLC로 분석하였다. Oxolinic acid의 머무름 시간은 10.2 min이었으며, 검출파장은 262 nm였다. 표준검량선은 $1{\sim}40$ ng에서 결정계수($r^2$)가 $0.999^{**}$로 직선성을 보였으며, 직선회귀식은 y=378.99x+135.08이었다. 논토양 무처리시료에 oxolinic acid를 0.2 mg/kg과 1.0 mg/kg 수준으로 표준품을 첨가하여 회수율을 측정한 결과 토양 1의 평균 회수율은 각각 $90.9{\pm}4.52%$ (n=3)와 $95.0{\pm}0.23%$ (n=3)였으며, 각각의 변이계수는 4.97%와 0.24%였다 토양 2에서는 $92.2{\pm}1.15%$ (n=3)와 $93.1{\pm}0.31%$ (n=3)로 각각의 변이계수는 1.25%와 0.33%이었다. 검출한계는 두 토양 공히 0.05 ppm(50 ppb)이었다.

This study was performed to examine analytical method of a quinolone compound, oxolinic acid in paddy soil by HPLC coupled with UV detector. Two types of soil texture in different regions were used for this experiment. Oxolinic acid was extracted by a 4 M-KOH : MeOH(1 : 3, v/v) mixtures and acidified followed by liquid-liquid partitioning in dichloromethane. Dichlormethane layer was dehydrated, evaporated and analyzed by HPLC (262 nm). Retention time was 10.2 min. The standard calibration curve of oxolinic acid showed linearity ($r^2>0.999^{**}$, y=378.99x+135.08) in the range of $1{\sim}40$ ng. The mean recoveries, evaluated from fortified soil samples at two concentration levels of 0.2 mg/kg and 1.0 mg/kg, were $90.9{\pm}4.52%$(C.V. 4.97%) and $95.0{\pm}0.23%$(C.V. 0.24%) for soil 1 and $92.2{\pm}1.15%$(C.V. 1.25%) and $93.1{\pm}0.31%$ (C.V. 0.33%) for soil 2, respectively The detection limits of two types of soils were same as 0.05 ppm. Overall, the present analytical method of oxolinic acid by HPLC coupled with UV detector seems to be used reasonably.