LC-MS/MS를 이용한 농산물 중 제초제 Sulfonylurea계 36종 동시 시험법 개발

이수정,이정미,장귀현,김현경,김지영 한국환경농학회 2023 한국환경농학회지 Vol.42 No.2

Sulfonylurea herbicides are widely used in agriculture because they have a long residual period and high selectivity. An analytical method was developed using QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) technique for simultaneous determination of sulfonylurea herbicide residues in agricultural products by liquid chromatography tandem mass spectrometry and for establishment MRL (Maximum Residue Limit) of those herbicides in Korea. Extraction was performed using acetonitrile containing 0.1% formic acid with MgSO4 (anhydrous magnesium sulfate) and NaCl (sodium chloride) and the extract was cleaned up using MgSO4 and C18 (octadecyl). The matrix-matched calibration curves were composed of 7 concentration levels from 0.001 to 0.25 mg/kg and their coefficients of determination (R2 ) exceeded 0.99. The recoveries of three spiking levels (LOQ, 10LOQ, 50LOQ, n=5) were in the range of 71.7-114.9% with relative standard deviations of less than 20.0% for all the five agriculture products. All validation values met criteria of the European Union SANTE/11312/2021 guidelines and Food and Drug Safety Evaluation guidelines. Therefore, the proposed analytical method was accurate, effective, and sensitive for sulfonylurea herbicide residues determination in agricultural commodities. .

Identification and Management of Sulfonylurea-resistant Biotype of Scirpus planiculmis in Reclaimed Paddy Fields, Korea

Tae-Seon Park(박태선),Chung-Kil Kang(강충길),Jae-Eup Park(박재읍),Bon-Il Ku(구본일),Hong-Kyu Park(박홍규),Sita Ram Ghimire(기미어 시다람),Young-Doo Kim(김영두),Jae-Kwon Ko(고재권) 韓國雜草學會 2009 Weed&Turfgrass Science Vol.29 No.2

Sulfonylurea(SU)계 제초제에 대한 저항성 새섬매자기가 한국의 서산 간척지 논에서 발생한 것이 확인되었다. SU 제초제 저항성 새섬매자가 발생한 논은 1990년부터 13년 동안 연속적으로 SU계 혼합 제초제를 사용하였다. 온실 조건에서 서산과 무안에서 채취한 새섬매자기 구경을 이식 후 10일에 azimsulfuron, bensulfuron-methyl, cinosulfuron, ethoxysulfuron, imazosulfuron 그리고 pyrazosulfuron-ethyl을 처리하였을 때 무안의 새섬매자기는 각 제초제 추천량에서 방제가 되었으나 서산의 새섬매자기는 추천량의 5배량에서도 20-40% 생존되었다. 생체중 50%를 억제하는 각 제초제의 농도(GR50)는 두 지역에서 채취한 새섬매자기 간에 현저한 차이가 있었으며, 서산의 매자기에 대한 6 종류의 GR50은 무안의 새섬매자기 대한 GR50값 보다 47-100배 높게 나타났다. 또한 무안과 서산의 새섬매자기에서 추출한 acetolactate synthase(ALS)에 대한 pyrazosulfuron-ethyl의 50% 억제 농도(I50)각각 0.8 nM과 409 nM로 나타나, 서산의 새섬매자기가 무안의 새섬매자기 보다 511배 높은 저항성을 가지고 있음을 확인하였다. SU계 제초제 저항성 새섬매자기를 효과적으로 방제하기 위한 제초제 선발 결과 benzobicyclone이 효과적이었다. 간척지 담수직파논에서는 파종 전에 이행성 비선택성 제초제인 glyphosate SL를 처리한 다음, 파종 후 10일에 benzobicyclone+azimsufuron+metamifop GR를 처리하는 것이 매우 효과적이었다. A suspected biotype of Scripus planiculmis to be resistant to sulfonylurea (SU) herbicides was identified in Seosan reclaimed paddy fields in Korea. The fields have been cultivated for monocultural rice production with wet-direct seeding method and continuously treated with SU-based herbicide mixtures for thirteen years since 1990. In greenhouse studies, 6 different SU herbicides, such as azimsulfuron, bensulfuron-methyl, cinosulfuron, ethoxysulfuron imazosulfuron and pyrazosulfuron-ethyl, completely controlled the Muan assession of S. planiculmis at the recommended dose rate of each herbicide, however, the Seosan accession of S. planiculmis biotype was survived 20 to 45% even treated with 5 times higher dose of each recommended rate of all herbicides treated. The GR50 values of 6 SU herbicides for Seosan accession of S. planiculmis were 47 to 100 times higher than those for Muan accession of S. planiculmis. The I50 values pyrazosulfuron-ethyl to acetolactate synthase (ALS) extracted from Sean and Muan accession of S. planiculmis were 409 nM and 0.8 nM, respectively. The I50 value of Seosan was 511 times higher than that of Muan accession. These results suggested that the Seosan accession of S. planiculmis have strong resistant characteristics to 6 SU herbicides, respectively, indicating that resistance might be due to the alteration in the target site of ALS. In the pot experiment, benzobicyclon having different mechanisms of action from SU herbicides had excellent controlling effects on the resistant biotype when their plant height were applied below 5 cm. The management in reclaimed paddy fields was very effective under the treatment of systematic herbicides such as foliar applied glyphosate SL at 15 days before seeding followed by benzobicyclon-included herbicides at 10 days after seeding.

한국 논에서 제초제 저항성잡초의 발생 현황, 메카니즘 및 방제

박태선(Tae Seon Park),성기영(Ki Yeong Seong),조현숙(Hyun Suk Cho),서명철(Myung Chul Seo),강항원(Hang Won Kang),박기웅(Kee Woong Park) 충남대학교 농업과학연구소 2014 농업과학연구 Vol.41 No.2

Sulfonylurea (SU)-resistant weeds of eight annual weeds, Monochoria vaginalis, Scirpus juncoides and Cyperus difformis, etc., and four perennial weeds, Scirpus planiculmis, Sagittaria pigmaea, Eleocharis acicularis and Sagittaria trifolia as of 2013 since identification Monochoria korsakowii in the reclaimed rice field in 1998. And the resistant Echinochloa oryzoides to ACCase and ALS inhibitors has been confirmed in rice fields of the southern province, Korea in 2009. In the beginning, the M. vaginalis, S. juncoides and C. difformis of these SU-resistant weeds were rapidly and individually spreaded in different fields, however, these resistant weeds have been occurring simultaneously in the same filed recently. The resistant biotype by weed species demonstrated about 10- to 1,000-fold resistance, based on GR50 values of the SU herbicides tested. And the resistant biotype of E. oryzoides to cyhalofop-butyl, pyriminobac-methyl, and penoxsulam was about 14, 8, and 11 times more resistant than the susceptible biotype base on GR50 values. The products and applied area of SU-included herbicides have been increased rapidly, and have accounted for about 69% and 96% in Korea, respectively. In Korea, the main cause of SU-resistant weed is extensive use of these herbicides. The top ten herbicides by applied area were composed of all SU-included herbicides by 2003. The concentrated and successive treatment of ACCase and ALS inhibitors for control of barnyardgrass in rice led up to the resistance of E. oryzoides. Also, SU-herbicides like pyrazosulfuron-ethyl and imazosulfuron which effective to barnyardgrass can be bound up with the resistance of E. oryzoides. The ALS activity isolated from the resistant biotype of M. korsakowii to SU-herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity (I50) of the SU-resistant M. korsakowii was 14- to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of [<SUP>14</SUP>C]bensulfuron uptake and translocation. Acetolactate synthase (ALS) genes from M. vaginalis resistant and susceptible biotypes against SU-herbicides revealed a single amino acid substitution of proline (CCT), at 197th position based on the M. korsakowii ALS sequence numbering, to serin (TCT) in conserved domain A of the gene. Carfentrazone-ethyl and pyrazolate were used mainly to control SU-resistant M. vaginalis by 2006 in Korea. However, the alternative herbicides such as benzobicyclone, to be possible to control simultaneously the several resistant weeds, have been developing and using broadly, because the several resistant weeds have been occurring simultaneously in the same fieled. The top ten herbicides by applied area in Korea have been occupied by products of 3-way mixture type including herbicides with alternative mode of action for the herbicide resistant weeds. Mefenacet, fentrazamide and cafenstrole had excellent controlling effects on the ACCase and ALS inhibitors resistant when they were applied within 2nd leaf stage.

Differential Herbicide Response of Sulfonylurea-Resistant Scirpus juncoides Roxb. Accessions to Sulfonylurea Herbicides

Tae-Seon Park(박태선),Hong-Kyu Park(박홍규),In-Yong Lee(이인용),Byung-Chul Moon(문병철),Bon-Il Ku(구본일),Chung-Kil Kang(강충길),Young-Doo Kim(김영두),Jae-Kwon Ko(고재권) 韓國雜草學會 2009 Weed&Turfgrass Science Vol.29 No.3

국내 4개 지역의 수집종간 sulfonylurea(SU)계 제초제 저항성을 올챙이고랭이의 저항성 정도 차이를 구명하기 위하여 SU계 제초제들에 대한 식물체 및 acetolactate synthase(ALS) 반응 차이를 감수성 계통과 비교 분석하였다. 김제, 나주 지역에서 채집된 저항성 올챙이고랭들은 SU계 제초제들의 기준량에서 거의 생육에 영향을 받지 않았다. 사용된 SU계 제초체들의 기준량에 대한 김제와 나주지역계통의 생체중은 각각 95~109% 및 90~98%를 보였다. 그란 김해 채집계통의 처리된 설포닐우레아계 제초제들의 기준량에 대한 생체중은 무처리 대비 41~58% 나타내어 중간정도의 저항성을 보였다. 김제와 나주 채집종에 대한 ALS 50% 억제 제초제 농도인 I50은 감수성 채집계통에 비해 각각 283~1,074배 및 157~870배 높게 나타났다. 그러나 서산 채집계통의 I50은 감수성인 수언 채집계통에 비해 5~11배 높게 나타났다. 설포닐우레아계 제초제 저항성 올챙이고랭이에 대한 비설포닝우레아계통 제초제들 중 13종류의 제초제를 시험한 결과 벤조비싸이크론과 부로모부타이드 입제가 매우 효과적이었다. Three sulfonylurea (SU)-resistant Scirpus juncoides Roxb. accessions were tested for levels of resistance to four SU herbicides which have been widely using in paddy fields of Korea since 1990, based on whole plant response and sensitivity of the target enzyme, acetolactate synthase (ALS). Gimje and Naju accessions were not affected to the survival by treatment with recommended doses of all SU herbicides tested. The fresh weight of Gimje and Naju accessions were 95% (ethoxysulfuron) to 109% (bensulfuron-methyl) and 90% (ethoxysulfuron) to 98% (imazosulfuron) to the recommended doses of all SU herbicides tested than the susceptible Suwon accession, respectively. However, the fresh weight for Seosan accession displayed an intermediate response and was only 41% (ethoxysulfuron) to 58% (imazosulfuron) more resistant than the susceptible accession. I50 values the ALS for the Gimje and Naju accessions were 283 (parzosulfuron ethyl)- to 1,074 (bebsulfuron-mthyl)-fold and 157 (bensulfuron-methyl)- to 870 (prazosulfuron-ethyl)-fold higher to the recommended doses of all SU herbicides tested than the susceptible Suwon accession, respectively. However, the I50 value for Seosan accession was 4- to 9-fold more resistant than the susceptible accession, as determined by I50 values of ALS. Benzobicyclone SC and bromobutide GR of alternative herbicides tested showed the very high efficacy to S. juncoides seedlings without rice injury.

Effective Herbicides for Control of Sulfonylurea-Resistant Monochoriα vaginαlis in Paddy Field

Yong In Kuk,Oh Do Kwon 韓國作物學會 2003 Korean journal of crop science Vol.48 No.4

Monochoria vaginalis is one of the most troublesome resistant weeds in Korean rice culture. Thus, the objectives of this study were to evaluate the response of M. vaginalis resistant to sulfonylurea(SU) herbicides and to determine alternative herbicides for the control of resistant M. vaginalis in direct seeded and transplanted rice culture in Korea. In greenhouse studies, the resistant biotype was 31-, 38-, 3172-, and 7-fold more resistant to ben-sulfuron-methyl, cyclosulfamuron, imazosulfuron, and pyrazosulfuron-ethyl, respectively, than the susceptible biotype, indicating cross-resistance to the SU herbicides used in this study. Non-SU herbicides, butachlor, carfentrazone-ethyl, mefenacet, pretilachlor, pyrazolate, and thiobencarb, several SU herbicide-based mixtures, ethoxysulfuron plus fentrazamide, pyrazosulfuron-ethyl plus pyrazolate plus simetryn, and non-SU herbicide-based mixtures, pyrazolate plus butachlor, pyrazolate plus pretilachlor, simetryn plus molinate, carfentrazone-ethyl plus butachlor, and carfentrazone-ethyl plus thiobencarb can be used to control both the resistant and susceptible biotypes of M. vaginalis when applied before the second leaf stage. In the field experiment, the resistant biotype of M. vaginalis that survived from the paddy fields treated with a SU herbicide-based mixture could effectively be controlled by using mixtures of bentazone plus MCPA, bentazone plus mecoprop-P, and bentazone plus 2,4-D when applied at 2 or 4 main leaves. Our results suggest that the SU-resistant M. vaginalis had not developed multiple resistances to herbicides with different modes of action. In particular, bentazone plus MCPA and bentazone plus mecoprop-P were effective control measures after failure to control resistant M. vaginalis in Korean rice culture.

Effective Herbicides for Control of Sulfonylurea-Resistant Monochoria vaginalis in Paddy Field

Kuk, Yong-In,Kwon, Oh-Do The Korean Society of Crop Science 2003 Korean journal of crop science Vol.48 No.4

Monochoria vaginalis is one of the most troublesome resistant weeds in Korean rice culture. Thus, the objectives of this study were to evaluate the response of M. vaginalis resistant to sulfonylurea(SU) herbicides and to determine alternative herbicides for the control of resistant M. vaginalis in direct seeded and transplanted rice culture in Korea. In greenhouse studies, the resistant biotype was 31-, 38-, 3172-, and 7-fold more resistant to ben-sulfuron-methyl, cyclosulfamuron, imazosulfuron, and pyrazosulfuron-ethyl, respectively, than the susceptible biotype, indicating cross-resistance to the SU herbicides used in this study. Non-SU herbicides, butachlor, carfentrazone-ethyl, mefenacet, pretilachlor, pyrazolate, and thiobencarb, several SU herbicide-based mixtures, ethoxysulfuron plus fentrazamide, pyrazosulfuron-ethyl plus pyrazolate plus simetryn, and non-SU herbicide-based mixtures, pyrazolate plus butachlor, pyrazolate plus pretilachlor, simetryn plus molinate, carfentrazone-ethyl plus butachlor, and carfentrazone-ethyl plus thiobencarb can be used to control both the resistant and susceptible biotypes of M. vaginalis when applied before the second leaf stage. In the field experiment, the resistant biotype of M. vaginalis that survived from the paddy fields treated with a SU herbicide-based mixture could effectively be controlled by using mixtures of bentazone plus MCPA, bentazone plus mecoprop-P, and bentazone plus 2,4-D when applied at 2 or 4 main leaves. Our results suggest that the SU-resistant M. vaginalis had not developed multiple resistances to herbicides with different modes of action. In particular, bentazone plus MCPA and bentazone plus mecoprop-P were effective control measures after failure to control resistant M. vaginalis in Korean rice culture.

전남지역에서 발생한 Sulfonylurea계 제초제 저항성 쇠털골 방제

권오도(Oh-Do Kwon),국용인(Yong-In Kuk),조승현(Seong-Hyeon Cho),신해룡(Hae-Ryoung Shin) 韓國雜草學會 2009 Weed&Turfgrass Science Vol.29 No.3

전남지방 간척지 벼 담수직파재배 농가포장에서 쇠털골의 발생이 2006년 이래 점차 증가되고 있다. 이러한 잡초가 발생한 곳은 벼 단작 재배지로 잡초방제를 위해 6~8여년간 설포닐우레아(sulfonylurea, SU)계 혼합제초제를 연속 사용해 온 지역이었다. 따라서 본 실험은 이러한 쇠털골에 대해 SU계 제초제 저항성여부와 처리시기별 토양처리제초제의 방제효과를 알아보고자 수행하였다. 쇠털골의 수집종중 감수성 생태형(전북 익산)은 pyrazosulfuron-ethyl과 bensulfuron-methyl의 추천량(21, 51g ha-1)과 배량(42, 102g ha-1) 처리에 의해 처리시기(써레질 후 7일, 14일)나 약량에 관계없이 100% 방제되었으나, 저항성 생태형(전남 신안)은 이들 제초제의 표준량 및 배량에서도 각각 2.4~16.4%와 40.6~67.1%의 낮은 방제효과를 보여 SU계 제초제에 저항성을 판단된다. 이러한 SU계 제초제 저항성 쇠털골의 방제는 써레질 직후에 thiobencarb, bromobutide, benzobicyclon, oxndiazon 처리에서 99~100%, 써레질 8일후에 thiobencarb, bromobutide, benzobicyclon 단제와 benfuresate·bifenox, piperophos·dimethametryn 혼합체 처리에서 98.3~100%, 써레질 15일후에 benfuresate·bifenox, molinate·simetryn·MCPB, thiobencarb·simetryn 처리에서 94.4~98.0%의 방제효과를 보였다. 본 연구 결과는 한국에서 SU계 제초제 저항성 쇠털골을 확인한 첫 보고이며, 추후 저항성 기작 등에 관한 연구가 병행되어야 할 것이다. The Eleocaris acicularis has been increased in reclaimed paddy fields in Jeonnam, Korea since 2006. These paddy fields were in monoculture rice production using sulfonylurea (SU) herbicide-based mixtures for 6-8 consecutive years. This research was conducted to investigate whether E. acicularis collected in Jeonnam, Korea has resistance to SU herbicides and determine alternative application times. The susceptible biotype of E. acicularis was completely controlled with the respective recommended rates and 2 times of the recommended rates of the bensulfuron-methyl and pyrazosulfuron-ethyl regardless of herbicide application times (7 and 14 days after harrowing), but the resistant biotype was only 2.4~16.4% and 40.6~67.1% controlled with the respective recommended rates and 2 times of the recommended rates of the herbicides. The resistant biotype could be controlled by other herbicides having different modes of action from SU herbicides, such as thiobencarb, bromobutide, benzobicyclon, and oxadiazon when they were applied at 0 day after harrowing (DAH) and thiobencarb, bromobutide, benzobicyclon, benfuresate·bifenox, and piperophos·dimethametryn when they were applied at 8 DAH. On the other hand, benfuresate·bifenox, molinate·simetryn·MCPB, and thiobebcarb·simetryn when they were applied at 15 DAH gave acceptable control (94.4~98.0%) of the resistant biotype. Our results are the first report of E. acicularis resistant to SU herbicides to in Korea, and thus further works are required to investigate resistance mechanism.

경북 지역 Sulfonylurea계 저항성 논잡초 발생양상과 분포

김상국,김학윤 한국잔디학회 2013 Weed & Turfgrass Science Vol.2 No.2

The study was carried out to obtain the basic information on distribution and occurrence of weeds including resistant weeds to sulfonylurea herbicides in paddy fields of Gyeongbuk province. In weed distribution on life cycle, annual weed was occupied by 91.8% and perennials were 8.2%, respectively. In morphological distribution of weeds, broad leaf weeds were 85.7%, sedges was 9.1%, and grass weed was 3.8%, respectively. Sulfonylurea-resistant weeds in paddy fields of twelve regions were widely occurred except for Yecheon region. The six sulfonylurea resistant biotypes occurred in paddy fields of Gyeongbuk province were Lindernia dubia, Eleocharis kuroguwai, Monochoria vaginalis and Ludwigia prostrata. It revealed that occurrence rate and area of sulfonylurea-resistant weeds were about 15.9 % and 22,420 ha in Gyeongbuk province. 본 연구는 경상북도 지역의 논에 발생하는 잡초종의 발생과 분포 및 sulfonylurea계 제초제 저항성잡초 발생비율을 조사한 것으로 생활사별 논잡초 분포는 일년생이 91.8%로서 다년생 8.2%보다 많았으며, 형태적 분포는 광엽잡초85.7%, 사초과 9.1%, 화본과 3.8%였다. 잡초종은 물달개비, 미국외풀, 피, 여뀌바늘, 올챙이고랭이, 미국가막사리이었고, 물달개비가 우점하는 지역과 발생률은 포항, 의성,상주, 경주 순으로 각각 85.1%, 76.3%, 73.5%, 58.9%였으며, 미국외풀은 영덕, 고령, 군위, 영양, 안동, 예천, 구미,영천 순으로 발생률은 각각 96.3%, 93.8%, 92.7%, 90.7%,86.8%, 70.6%, 66.7%, 56.2%로 나타났다. 지역별로 주요논잡초는 물달개비, 미국외풀, 너도방동사니, 피, 올챙이고랭이 순이었고, 우점도를 1981년, 1992년 및 2012년도와비교하면 미국외풀(42.8%), 물달개비(37.5%), 너도방동사니(7.9%)순이었다. 논에 발생한 sulfonylurea계 제초제 저항성 잡초는 물달개비, 여뀌바늘, 미국외풀, 올챙이고랭이로 총 4종이었고, 제초제 저항성잡초 가운데 가장 많은 발생면적을 보인 초종은 미국외풀로 11.1%를 차지하였다. 2012년 조사된 지역에 대한 제초제 저항성 잡초 발생률은15.9%정도로 경북지역의 벼 재배면적(141,005ha)을 고려하면 제초제 저항성잡초 발생면적은 22,420ha정도로 추정되었다.

Research Paper : Difference in ultraweak photon emissions between sulfonylurea-resistant and sulfonylurea-susceptible biotypes of Scirpus juncoides following the application of a sulfonylurea herbicide

( Hidehiro Inagaki ),( Yoshiki Ishida ),( Akira Uchino ),( Kimihiko Kato ),( Chizuko Kageyama ),( Hiroyuki Iyozumi ),( Hideki Nukui ) 한국잡초학회 2008 Weed Biology and Management Vol.8 No.2

농산물 중 Sulfonylurea계 제초제의 공정 잔류분석법 개발

이영득 ( Young Deuk Lee ) 한국환경농학회 2018 한국환경농학회 학술대회집 Vol.2018 No.-

An analytical method was developed to simultaneously determine residues of four sulfonylurea herbicides including chlorimuron-ethyl, ethametsulfuron-methyl, rimsulfuron and tribenuron-methyl in agricultural products for the official purpose. All of target analytes were extracted with acetonitrile containing 0.5 M ammonium chloride from raw or moistened samples. The extract was diluted with saline water, and subjected to n-hexane washing to remove nonpolar co-extractives while the analyte remained in the aqueous phase. Enhancing ionic strength of the aqueous phase, dichloromethane partition was then followed to recover all the analytes. Weak anion-exchange column chromatography using SPE-aminopropyl cartridge was employed to further purify the extract prior to determination by high-performance liquid chromatography (HPLC) with detection of ultraviolet absorption at 234 nm. Linear gradient elution of acetonitrile/phosphate buffer (pH 3.0) mixture on an octadecylsilyl column was successfully applied to separate each residue of the analyte. Total of five representative types of agricultural commodities which comprised brown rice, mandarine, green pepper, potato and soybean were selected for validation of the method. A confirmatory and/or alternative quantitative method were also provided using LC coupled with the mass spectrometric technique.