곡류 중 T-2 및 HT-2 독소 동시 정량분석의 유효성 검증 및 실태조사
Survey and method validation of simultaneous quantitative analysis of T-2 and HT-2 toxins in cereals
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본 연구에서는 곡류 중 트리코테센류 곰팡이독소인 T-2 독소 및 HT-2 독소의 LC-MS/MS 분석방법을 검증하고 국내 유통 곡류 중 T-2 독소 및 HT-2 독소의 오염실태를 파악하였다. 곡류 중의 T-2 독소 및 HT-2 독소를 분석하기 위해, 염화나트륨을 포함한 90% 메탄올 용액으로 추출, 원심분리, 여과, 4% 염화나트륨용액으로 희석하고, 원심분리한 후, 여과한 후 면역친화성칼럼에 의해 정제한 시료를 LC-MS/MS 동시정량 분석하였다. T-2 독소 및 HT-2 독소의 검출한계 및 정량한계는 각각 0.5 μg/kg 및 1.5 μg/kg 얻었다. matrix-matched 표준 검량식에서 상관계수 0.99 이상의 직선성을 얻었으며, T-2독소와 HT-2 독소 2배에서 10배의 정량한계로 표준용액을 첨가한 시료에서 회수율은 T-2독소와 HT-2 독소 각각 100.6±7.2 %, 96.8 ± 9.4 %로 EU 가이드라인에서 제시하는 유효성 기준을 만족하였다. LC-MS/MS 정량법을 이용하여 국내 곡류 9품목 115건에 대해 T-2 독소와 HT-2 독소의 오염도를 조사하여 본 결과, 전체 곡류 115건 중에서 T-2 독소는 83건, HT-2 독소는 93건 검출되었으며 오염도는 T-2 독소는 N.D~37.1 ug/kg, HT-2 독소는 N.D~5.4 ug/kg 으로 낮은 수준이었으며, 오염도는 유럽기준치(100 μg/kg)이내 이었다. 본 연구에서 개발된 곡류 중 T-2 독소 및 HT-2 독소에 대한 분석법은 향후 우리나라 곡류 중 곰팡이독소 안전관리를 위한 시험법으로 활용가능하며, 오염도 자료는 안전성 평가의 기초자료로 활용이 가능할 것으로 사료된다.
Abstract
The aim of this study was to develop an analytical method for determination of T-2 toxin and HT-2 toxin level in cereals and to survey their levels using LC-MS/MS. The T-2 and HT-2 toxins were simultaneously analyzed by electrospray ionization with a positive ion mode and multiple reaction monitoring (MRM) after filteration and immuno-affinity column clean-up. A matrix-matched standard calibration used for quantification and recoveries of T-2 and HT-3 toxins were in the range of 100.6±7.2% and 96.8±9.4%, respectively. Limits of detection and quantification of T-2 and HT-2 toxins were estimated to be 0.5 and 1.5 μg/kg, respectively. Each repeatability (RSRr) of T-2 and HT-2 toxins was determined to be 0.9~6.0%, and 4.9~6.1%, respectively. Total 115 samples cereals were collected from 9 types of cereals for analysis. The positive percentages of T-2 and HT-2 toxins obtained from collected samples were found to be 72% and 80%, respectively. The contamination level of T-2 toxin and HT-2 toxin in cereals were 37.1 μg/kg, and 5.4 μg/kg, respectively. Therefore, this study suggests that the developed method could be an useful analytical method to determine the T-2 and HT-2 toxin level in cereals and the present data could be used as a reference to estimate the risk assessment.
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