백출과 지황 중 Fosthiazate 및 기장 중 Tricyclazole과 Tebuconazole의 LC-MS/MS와 GC-MS/MS 분석법 개발

Abstract

Fosthiazate is an organophosphorus nematicide and insecticide. Atractylodes rhizome white and Rehmannia root is herbal medicine listed in the Korean Pharmacopeia (KP). This study develops a method for analyzing fosthiazate using GC-NPD. Cleanup using an SPE Cartridge was performed twice. This method using GC-NPD was complex to extract, partition and cleanup. After that, the method was developed by QuEChERS method using LC-MS/MS to improve unnecessary preparation procedures. LC-MS/MS on electrospray mode was used for the analysis of fosthiazate. In multiple reaction monitoring, m/z=228 and m/z=104.15 were engaged as the quantification ion and qualification ion, respectively. Method limit of quantitation (MLOQ) was 0.01 mg/kg. The linearity of matrix-matched calibration curve (r2) was ≥0.99 at the calibration range of 0.001-0.05 mg/kg. For the recovery test, 5 g of macerated Atractylodes rhizome white or Rehmannia root was treated with standard solutions at MLOQ and 10MLOQ levels. Recovery rates were in the range of 70-120% (RSD≤20%) at two spiked levels (MLOQ and 10MLOQ) for Atractylodes rhizome white or Rehmannia root. Proso millet (Panicum miliaceum L.) is the longest used summer cereal in human in addition to wheat and barley. Tricyclazole and tebuconazole is a fungicide which has been used for various crops including proso millet. LC-MS/MS on electrospray mode was used for the analysis of tricyclazole. In multiple reaction monitoring, m/z=136 and m/z=163.15 were engaged as the quantification ion and qualification ion, respectively. GC-MS/MS on electron ionization mode was used for the analysis of tebuconazole. In multiple reaction monitoring, m/z=125 and m/z=99.1 were engaged as the quantification ion and qualification ion, respectively. Method limit of quantitation (MLOQ) was 0.01 mg/kg. The linearity of matrix-matched calibration curve (r2) was ≥0.99 at the calibration range of 0.0025-0.25 mg/kg. For the recovery test, 10 g of macerated grain or straw of proso millet was treated with standard solutions at 10MLOQ and 50MLOQ levels. Recovery rates were in the range of 70-120% (RSD≤20%) at two spiked levels (10MLOQ and 50MLOQ) for grain and straw. In field study, tricyclazole 20 % suspension concentrate formulation was treated on each 4 plots (50/40/30 days, 40/30/21 days, 30/21/14 days and 21/14/7 days before harvest) with three replicates. After harvest, grain and straw were analyzed to show that the residue levels of trycyclazole decreased according to treatment time. Maximum levels of residue were 5.68 mg/kg and 1.13 mg/kg for grain and straw in treatment plot of 21/14/7 days before harvest. In field study of tebuconazole, tebuconazole 20 % suspension concentrate formulation was treated on each 4 plots (50/40/30 days, 40/30/21 days, 30/21/14 days and 21/14/7 days before harvest) with three replicates. After harvest, grain and straw were analyzed to show that the residue levels of tebuconazole decreased according to treatment time. Maximum levels of residue were 1.66 mg/kg and 1.20 mg/kg for grain and straw in treatment plot of 21/14/7 days before harvest.

Fosthiazate는 유기인계 살충제이고 Atractylodes rhizome white 및 Rehmannia root는 대한민국약전에 등재 된 약초이다. 이 연구는 GC-NPD를 사용하여 해당 생약에서 fosthiazate 분석법을 개발하였다. SPE 카트리지를 사용한 정제가 두 번 수행되었으며 GC-NPD를 사용하는 이 분석법은 추출, 분배, 정제 모두 복잡했다. 이 분석법을 개선하기 위해 LC-MS/MS를 사용하는 QuEChERS 법으로 분석법을 확립하여 불필요한 준비 절차를 개선하였다. 회수율 시험을 위해, 5 g의 Atractylodes rhizome white 또는 Rehmannia root는 MLOQ 및 10MLOQ 수준에서 표준 용액으로 처리하였고. 2 개의 처리수준 (MLOQ 및 10MLOQ)에서 회수율은 70-120 % (RSD≤20 %)였다. 기장은 밀과 보리 외에 인간에서 가장 오래 사용되는 여름 곡물이다. Tricyclazole과 tebuconazole은 기장을 포함한 다양한 작물에 사용되는 살균제이다. 이 연구에서 정량한계(MLOQ)는 0.01 mg/kg이었다. 회수율 시험을 위해, 10g의 알곡 또는 짚의 기장을 10MLOQ 및 50MLOQ 수준의 표준 용액으로 처리 하였다. 알곡 및 짚에 대한 2 개의 처리수준 (10MLOQ 및 50MLOQ)에서 회수율은 70-120 % (RSD≤20 %) 범위였다. 살균제 tricyclazole 20 % 액상수화제를 3 회 반복하여 각각의 4 개의 구역 (50/40/30 일, 40/30/21 일, 30/21/14 일 및 21/14/7 일)에서 3 회 반복 처리 하였다. 수확 후, 알곡 및 짚을 분석하여 tricyclazole의 잔류 수준이 처리 시간에 따라 감소 함을 보여 주었다. 수확 전 21/14/7 일의 처리구역에서 알곡 및 짚에 대한 최대 잔류 물 수준은 5.68 mg/kg 및 1.13 mg/kg이었다. 살균제 tebuconazole 20 % 액상수화제를 각각의 4 개의 구역 (수거 전 50/40/30 일, 40/30/21 일, 30/21/14 일 및 21/14/7 일)으로 처리 하였다. 수확 후, 알곡 및 짚을 분석하여 처리 시간에 따라 tebuconazole의 잔류 수준이 감소 함을 보여 주었다. 수확 전 21/14/7 일의 처리구역에서 알곡 및 짚에 대한 최대 잔류 물 수준은 1.66 mg/kg 및 1.20 mg/kg이었다.