PAHs 농도 분포에 따른 GC/MS와 HPLC의 분석특성에 관한 연구

홍좌령 ( Jwa Ryung Hong ),최광민 ( Kwang Min Choi ) 한국산업보건학회 2015 한국산업보건학회지 Vol.25 No.3

Objectives: The purpose of this study was to determine the best method to analyze PAHs at extremely low concentrations. To this end, 16 PAHswere analyzed simultaneously by GC/MS, HPLC/FLD and HPLC/UVD, and the analytical characteristics of HPLC and GC/MS were compared. Methods: This study was conducted by GC/MS and HPLC/FLD/UVD, and evaluated linearity, precision and detection limit. Standard solutions were prepared for 21 samples in the range of 0.00001~1.0 ㎍/mL and the samples were divided into four groups. All samples were made in three sets and analysis was replicated seven times. Results: Sixteen PAHs could be simultaneously separated by HPLC and GC/MS, and the adequate equipment was HPLC/FLD. The retention times by HPLC were shorter than GC/MS, and HPLC had better separation for most PAHs than GC/MS. The peaks of naphthalene and naphthalene-D8 partially overlapped for GC/MS. HPLC/FLD had a 20-2000 times lower limit of detection than GC/MS and UVD. However FLD was not adequate for analyzing acenaphthylene because it has too low a fluorescence quantum yield to be detected. The precision of HPLC/FLD/UVD and GC/MS showed less than 20% at 0.001 ㎍/mL PAHs and when the concentration was higher, the coefficient of variation was decreased. HPLC/FLD was better for the overall detection of limits. Conclusions: The results indicate that the HPLC/FLD method has good linear range, precision and a detection of limits from 0.00001~0.0001 ㎍/mL for all 16 PAHs. This study contributes to providing useful data for analysis technology and can be applied to occupational exposure measurement for PAHs in workplaces.

HPLC/UVD를 이용한 현미와 볏짚 중 MCPA의 잔류분석방법 확립

유기용(Ki-Yong Yoo),강대원(Dae-Won Kang),최용화(Yong-Hwa Choi),한성수(Seibg-Soo Han) 韓國雜草學會 2010 Weed&Turfgrass Science Vol.30 No.4

HPLC/UVD를 이용한 현미와 볏짚 중 MCPA의 잔류분석방법 확립하고자 용매별 추출효율과 충진제별 정제효율을 검토한 결과, 용매별 추출효율은 시료 20g에 acetone 200mL와 1N-HCl 100mL의 혼합용매(pH 3.6)를 사용하는 것이 87%로 높은 효율을 보였고, 충진제별 정제효율은 florisil(6mL, 1g) Sep-pak® cartridge column에서 1% methanol/ acetonitile 용출이 83%의 효율을 나타내었다. 이 결과를 토대로 현미와 볏짚 시료 중의 MCPA 잔류분석을 수행하였는바, 평균회수율은 현미의 경우 96.0%와 94.9%이었으며, 볏짚의 경우 92.5%와 88.2%로 높은 회수율이었고, 실험의 정밀도는 상대표준편차 1.5~5.7%로 매우 높았다. 벼 이앙 후 30일에 bentazone+MCPA(11+1.2%) 합제를 ha당 30kg과 60kg을 각각 처리하여 재배한 현미와 볏짚 중 MCPA 잔류량은 모든 처리에서 식품의약품안전청이 권고한 잔류허용량인 5㎍ mL-1 미만이었다. MCPA의 HPLC 분석조건과 잔류분석절차의 결과는 가수분해, 유도체화, saponification과 같은 복잡한 분석절차를 생략할 수 있고, Rt가 기존 16.5 min.에서 6.5 min.으로 빨라져 인체위해성과 시간소모성의 단점을 개선하였으며, 회수율을 높이고 정밀도가 높은 결과로써 현미와 볏짚 중 HPLC에 의한 MCPA 잔류분석에 유용하게 적용할 수 있을 것으로 판단된다. This study was carried out to establish the analytical method of MCPA residue in brown rice and rice straw by HPLC/UVD. When MCPA was extracted from sample under the pH 3.6 by adding acetone 200 mL and 1N-HCl 100 mL, the extraction efficiency was high by 87%. And purification efficiency was high by 83% when 5 mL of 1% methanol/acetonitrile was eluated by the florisil Sep-pak cartridge column. From spiking of 0.1㎍ mL-1 and 0.25㎍ mL-1 of MCPA to control sample, respectively, average recovery rate of MCPA in brown rice was 96.0% and 94.9% and that in rice straw was 92.5% and 88.2%, respectively. Precision of experiment was very high by relative standard deviation of 1.5% to 5.7%. In brown rice and rice straw treated with bentazone+MCPA (11+1.2%) of 30 kg and 60 kg per ha at 30 days after rice transplanting, respectively, maximum residue limit was under 0.05㎍-1 of the recommended rate of Korean Food and Drug Administration. From the above results, the analytical procedure of MCPA in plants such as hydrolysis, saponification and derivatization were ommited, and retention time was faster and recovery rate was higher than the existed results of HPLC/UVD. Therefore, these results were greatly improved and seemed to be usefully applied for residue analysis of MCPA in plants.

콩잎 열수추출물의 지표성분인 camelliaside A의 정량분석을 위한 HPLC-UVD 분석법 밸리데이션

김정호,리승환,문시원,박기훈 한국응용생명화학회 2022 Journal of Applied Biological Chemistry (J. Appl. Vol.65 No.3

콩잎의 기능성식품소재연구가 활발히 진행되고 있지만 기능성식품소재로서 콩잎을 관리할 수 있는 밸리데이션 방법은 제시되지 않고있다. 본 연구에서는 콩잎에서 7개의 kaempferol 유도체를 UHPLC-ESI-Q-TOF-MS로 동정하고 이들 중 camelliaside A를 지표물질로 선정하였다. 콩잎열수추출물에서 camelliaside A를 정량할 수 있는 HPLC-UVD 밸리데이션 방법을 식품의약품안전처의 건강기능성식품에 대한 가이드라인에 따라 구축하였다. 개발된 HPLC-UVD 밸리데이션 방법은 특이성, 정확도, 일내정밀도, 일간정밀도, 검출한계, 정량한계 및 직선성에서 가이드라인을 만족하는 결과값으로 검증되었다. 또한 camelliaside A 분석법을 콩잎열수추출물에 적용하여 적합한 수준의 평가값을 얻었다.

HPLC-UVD/MS를 이용한 농산물 중 Oxycarboxin의 잔류분석

정명근 ( Myoung-gun Choung ),주하은 ( Haeun Joo ),권희주 ( Heeju Kwon ),정주영 ( Juyoung Jeong ),김기쁨 ( Gi-ppeum Kim ),황영선 ( Young-sun Hwang ),이영득 ( Young Deuk Lee ) 한국환경농학회 2017 한국환경농학회 학술대회집 Vol.2017 No.-

Oxycarboxin(5,6-dihydro-2-methyl-N-phenyl-1,4-oxathiin-3-carboxamide-4,4-dioxide) as oxanthiin fungicide is a systemic fungicide commonly used as a seed treatment to control various fungi that cause seed and seeding diseases in agronomic and horticultural crops. An analytical method was developed using HPLC-UVD/MS to precisely determine the residue of oxycarboxin. The oxycarboxin residue was extracted with acetone from representative samples of five raw products which comprised hulled rice, soybean, Kimchi cabbage, green pepper, and apple. The extract was diluted with saline water, and dichloromethane partition was followed to recover oxycarboxin from the aqueous phase. Florisil column chromatography was additionally employed for final purification of the extract. Oxycarboxin was separated and quantitated by HPLC with UVD using a Zorbax SB-AQ C₁₈ column. Accuracy of the proposed method was validated by the recovery from crop samples fortified with oxycarboxin at 3 levels per crop in each triplication. Mean recoveries ranged from 78.3% to 96.1% in five representative agricultural commodities. The coefficients of variation were less than 10%. Limit of quantitation of oxycarboxin was 0.04 mg/kg as verified by the recovery experiment. A confirmatory technique using LC/MS with selected-ion monitoring was also provided to clearly identify the suspected residue. Therefore, this analytical method was reproducible and sensitive enough to determine the residue of oxycarboxin in agricultural commodities.

HPLC-UVD를 이용한 축산식품 중 Novobiocin의 시험법 확립

박희라(Hee-ra Park),권찬혁(Chan-Hyeok Kwon),이종구(Jong-Goo Lee),김형수(Hyung-Soo Kim),채영식(Young-Sik Chae),오재호(Jae-Ho Oh),권기성(Kisung Kwon) 한국식품과학회 2012 한국식품과학회지 Vol.44 No.3

노보비오신은 동물용의약품 중 약산성 항생제이며, 우리나라의 경우 선진국에 비해 항생제의 사용이 높은 만큼 동물용의약품의 잔류 사례는 계속해서 증가하고 있다. 이에 따라 식품 중 잔류동물용의약품의 안전관리를 위해 축산 식품 중 닭고기, 소고기 및 우유에서 노보비오신에 대한 HPLC-UVD를 이용한 잔류 분석법을 확립하였다. 각 시료 일정량에 메탄올을 가하고 균질화하여 대상 성분을 추출하고 액-액분배법으로 지방성분을 제거한다. 대상성분이 추출된 여액을 실리카 카트리지를 이용하여 정제한 다음 HPLC-UVD로 분석하였다. 확립된 시험법의 정량한계와 직선성은 CODEX 기준에 적합한 수준으로 각각 0.02 ㎎/㎏과 0.999(r²)이었으며 회수율은 닭고기, 소고기 및 우유에서 각각 88.8±5.6-100.3±4.4, 88.8±7.2-97.0±3.2 및 88.1±4.3-92.8±3.6%이었다. 본 연구에서 확립된 노보비오신의 시험법은 동물용의약품의 안전관리를 위한 공정시험법으로 활용될 것으로 기대된다. Novobiocin is a coumarin-containing antibiotic, and has a longer half-life in various animals than other veterinary medicines. A simple and rapid high-performance liquid chromatography assay for the determination of residual novobiocin levels in chicken, beef and milk has been developed and validated. The separation condition for HPLC/UVD was optimized by a MG Ⅱ C₁? (4.6 ㎜ ID×250 ㎜, 5 ㎛) column with 0.1% formic acid in H₂O/0.1% formic acid in Acetonitrile (40/60, v/v) as the mobile phase at a flow rate of 1.0 mL/min and the detection wavelength was set at 340 nm. Residues were extracted from tissue by blending with methanol. After liquid-liquid partitioning, lipid materials were removed with n-hexane and purification as Silica (1 g, 6 mL) cartridge with 10 mL acetone/dichloromethane (10/90, v/v). Limit of quantification and linearity performed by the analytical method were 0.02 ㎎/㎏ and 0.999 (r²), and the recovery range was 88.8±5.6-100.3±4.4, 88.8±7.2-97.0±3.2 and 88.1±4.3-92.8±3.6%. It is expected that this analytical method with regards to novobiocin in chicken, beef and milk could be applied as an official method to administer food safety on veterinary medicines.

HPLC-UVD/MS를 이용한 농산물 중 fenoxycarb, pyriproxyfen 및 methoprene의 분석법 확립

이수진(Su-Jin Lee),김영학(Young-Hak Kim),송이슬(Lee-Seul Song),황영선(Young-Sun Hwang),임정대(Jung-Dae Lim),손은화(Eun-Hwan Sohn),임무혁(Moo Hyeog Im),도정아(Jung-Ah Do),오재호(Jae-Ho Oh),권기성(Kisung Kwon),이중근(Joong-Keun Lee),이영 한국농약과학회 2011 농약과학회지 Vol.15 No.3

Fenoxycarb, pyriproxyfen and methoprene are juvenile hormone mimic insecticide, These insecticides have been widely used for mosquito, fly, scale insects, and Lepidoptera. The purpose of this study was to develop a simultaneous determination procedure of fenoxycarb, pyriproxyfen and methoprene residues in crops using HPLC-UVD/MS. These insecticide residues were extracted with acetone from representative samples of four raw products which comprised brown rice, apple, green pepper, and Chinese cabbage. The extract was diluted with saline water, and then n-hexane/dichloromethane partition was followed to recover these insecticides from the aqueous phase. Florisil column chromatography was additionally employed for final clean up of the extract. The analytes were quantitated by HPLC-UVD/MS, using a C₁? column. The crops were fortified with each insecticide at 3 levels per crop. Mean recovery ratios were ranged from 80.0 to 104.3% in four representative agricultural commodities. The coefficients of variation were less than 4.8%. Quantitative limit of fenoxycarb, pyriproxyfen, and methoprene was 0.04 ㎎/㎏ in crop samples. A HPLC-UVD/MS with selected-ion monitoring was also provided to confirm the suspected residues. The proposed simultaneous analysis method was reproducible and sensitive enough to determine the residues of fenoxycarb, pyriproxyfen and methoprene in the agricultural commodities.

Valinamide carbamate계 살균제 iprovalicarb의 잔류 분석법 개발

정명근 한국국제농업개발학회 2019 韓國國際農業開發學會誌 Vol.31 No.1

This experiment was conducted to establish an analytical method for residues of iprovalicarb, as recently developed a valinamide carbamate fungicide showing effect on pathogens of the oomycetes, in crops using HPLC-UVD/MS for the official purpose. Iprovalicarb residue was extracted with acetonitrile from representative samples of five raw products which comprised apple, green pepper, Kimchi cabbage, hulled rice and soybean. The extract was diluted with large volume of saline water and directly partitioned into n-hexane/dichloromethane(20/80, v/v) to remove polar co-extractives in the aqueous phase. For the hulled rice and soybean samples, n-hexane/acetonitrile partition was additionally employed to remove non-polar lipids. The extract was finally purified by optimized florisil column chromatography. On an octadecylsilyl column in HPLC, iprovalicarb was successfully separated from coextractives of sample, and sensitively quantitated by ultraviolet absorption at 210 nm with no interference. Accuracy and precision of the proposed method was validated by the recovery experiment on every crop samples fortified with iprovalicarb at 3 concentration levels per crop in each triplication. Mean recoveries ranged from 87.5 to 101.6% in five representative agricultural commodities. The coefficients of variation were all less than 10%, irrespective of sample types and fortification levels. Limit of quantitation( LOQ) of iprovalicarb was 0.04 mg/kg as verified by the recovery experiment. A confirmatory method using LC/MS with selected-ion monitoring technique was also provided to clearly identify the suspected residue. 본 연구에서는 HPLC-UVD/MS를 이용하여 농산물 중valinamide carbamate계 살균제 iprovalicarb의 잔류 분석법을확립하였다. 대표 농산물은 사과, 고추, 배추, 현미 및 콩을 선정하였고, acetonitrile를 이용하여 추출한 iprovalicarb는 nhexane/ dichloromethane(20/80, v/v) 혼합용매를 이용한 액-액분배법과 florisil 흡착크로마토그래피법으로 정제하여 HPLCUVD/ MS의 분석시료로 사용하였다. Iprovalicarb의 정량분석을위한 최적 HPLC-UVD 분석 조건을 확립하였으며, 정량한계(LOQ)는 0.04 mg/kg 이었다. 각 대표 농산물에 대한 정량한계, 정량한계의 10배 및 50배 수준에서 회수율을 검토한 결과, 모든 처리농도에서 87.5~101.6% 수준의 회수율을 나타내었고, 반복 간 변이계수(CV)는 최대 7.7%를 나타내어 잔류분석 기준인 회수율 70~120% 및 분석오차 10% 이내를 충족시키는만족한 결과를 도출하였으며, LC/MS/SIM을 이용하여 실제 농산물 시료에 적용하여 재확인 하였다. 이상의 결과로 신규iprovalicarb의 HPLC-UVD/MS 분석법은 국내·외 농산물 잔류농약 검사 시 검출한계, 회수율 및 분석오차 면에서 국제적분석기준을 만족하는 신뢰성이 확보된 정량 분석법으로 사용가능할 것이다.

HPLC-UVD를 이용한 살균제 Fenpyrazamine의 시험법 개발 및 검증

도정아 ( Jung Ah Do ),박혜진 ( Hyejin Park ),권지은 ( Ji Eun Kwon ),이지영 ( Ji Young Lee ),이미영 ( Mi Young Lee ),조윤제 ( Yoon Je Cho ),이상목 ( Sang Mok Lee ),김명애 ( Myung Ae Kim ),최시원 ( Si Won Choi ),김희정 ( Hee Jeong K 한국환경농학회 2013 한국환경농학회 학술대회집 Vol.2013 No.-

Fenpyrazamine은 일본 Sumitomo Chemical에서 개발한 pyrazole계 살균제로 작물에 빠르게 침 투하여 곰팡이의 발아관과 균사체 신장을 억제하는 작용기작으로 강력한 살균작용을 나타낸다. 채소류 등의 잿빛곰팡이병, 잿빛무늬병 등의 방제에 효과적이며, 또한 포유동물에 고도의 안전성과 낮은 환경지속성으로 인해 수확 전까지 다양한 작물에 사용이 가능하다. 미국 환경보호청(US Environmental Protection Agency)에서는 2013년에 미국 연방 살충제, 살균제, 살서제법(Federal Insecticide, Fungicide, and Rodenticide Act, FIFRA)에 따라 아몬드와 상추 등에 fenpyrazamine의 사용을 허가하고 0.02 ppm (아몬드와 피스타치오), 0.7 ppm (인삼), 2.0 ppm (잎상추), 5.0 ppm (산딸기류 열매) 등의 잔류허용기준을 설정하였다(78 Fed. Reg. 14,461 (3/6/13)). 국내에서는 2013년에 복숭아(2.0 mg/kg), 포도(5.0 mg/kg) 및 감귤(2.0 mg/kg)에 잔류허용기준 설정이 추진 중인 신규농약으로, 이에 따라 본 연구에서는 국내 생산 및 향후 수입되는 다양한 농산물 중 잔류 할 수 있는 fenpyrazamine에 대한 안전성을 확보하기 위해 잔류량을 신속하고 정확하게 측정할 수 있는 효율적인 공정 시험법을 확립하였다. Fenpyrazamine은 증기압이 2.89 x 10-8 Pa(25℃)로 휘 발성이 낮고 분자 구조상에서 pyrazole 구조가 있어 단파장을 흡수하는 특성을 가지고 있기 때문 에 HPLC-UVD를 분석기기로 선택하였으며, 243 nm에서 최대흡광도를 나타내어 검출파장으로 선택하 였다. 추출용매는 물질의 용해도와 추출효율을 고려하여 acetonitrile로 선정하였으며, n-octanol/water 분배계수(logPow, 25℃)가 3.52의 중간 비극성의 물리화학적 특징을 고려하여 액-액 분배 단계에 서 dichloromethane과 물의 분배로 극성불순물을 제거하였고, 정제단계에서는 silica cartridge를 이 용하여 hexane/acetone(95/5, v/v) 10 mL로 카트리지를 세척한 다음 hexane/acetone(85/15, v/v) 10 mL로 용출해 주었을 때 다양한 매트릭스 간섭 물질로부터 fenpyrazamine을 효과적으로 정제할 수 있었다. 확립된 시험법으로 복숭아, 포도, 감귤을 대상으로 처리농도 0.05, 0.5, 5.0 mg/kg으로 각각 5반복의 회수율 실험을 시행한 결과 회수율 범위가80.5∼102.7%이었고, 분석오차가 10% 미 만으로 시험법의 정확성을 확인하였으며, LC-MS를 통한 재확인 과정을 수행함으로써 시험법의 신 뢰성과 선택성을 확보할 수 있었다. 따라서 본 연구에서 개발한 시험법은 농산물에 잔류하는 fenpyrazamine을 분석하기 위한 공정 시험법으로 활용할 수 있을 것으로 판단된다.

농식품 환경 분야(PF) : HPLC-UVD를 이용한 농산물 중 Quinmerac의 시험법 개발

이현숙 ( Hyun Sook Lee ),윤지영 ( Ji Young Yoon ),이한진 ( Han Jin Lee ),이상목 ( Sang Mok Lee ),김희정 ( Heejung Kim ),조윤제 ( Yoon Jae Cho ),천소영 ( So Young Chun ),정지윤 ( Jiyoon Jeong ),장문익 ( Moon Ik Chang ),이은주 ( Eun 한국환경농학회 2014 한국환경농학회 학술대회집 Vol.2014 No.-

Quinmerac은 독일의 BASF사가 개발한 quinnoline carboxylic acid계 선택성 제초제 또는 생장 조절제인데, 세포신장, 세포부피 증가를 촉진하고 세포분열을 도와주는 auxin의 역할을 하여 국내에서는 복숭아 등 과실류에 과실비대 및 숙성 촉진을 목적으로 생장조절제로서 사용되고 있다. Quinmerac은 분자 내에 carboxylic acid를 함유하고 있는 pKa가 4.32인 약산성 화합물로 현행 분석법은 pH 조절에 의한 세척 및 추출과정으로 구성된 ion-associated partition법을 이용하여 정제한 후 carboxylic acid기를 ester화하여 중성의 화합물로 변환하여 추가 액-액 분배를 한 후GC-NPD를 이용하여 분석하게 되어있다. 이는 여러 단계를 거쳐 분석방법이 복잡하고 전처리 시간이 오래 걸리며, 간섭효과 영향 등의 문제점이 있어 이를 개선하여 보다 쉽고, 간편한 분석법 개선이 요구되었다. 개선된 분석법은 검체에 NaOH 수용액을 가하여 알칼리성으로 조절한 후 acetone 으로 추출하고 quinmerac의 해리성을 이용하여 액-액 분배는 ion-associated partition을 적용하였고 C18 SPE 카트리지를 이용하여 정제한 후 유도체화 없이 HPLC-UVD로 기기분석 하였다. 정량을 위한 검량선은 농도범위 0.02~0.1 ng에서 측정하였으며, 직선성( R 2 )은 0.999 이상이었고, 정량한계는 0.04 mg/kg이었다. 분석법의 유효성 확인을 위해 감귤, 고추 및 현미에 첨가농도 0.04, 0.08 및 0.4 mg/kg 수준 (LOQ, LOQ의 2배 및 LOQ의 10배)에서 위의 분석법을 적용하여 회수율 시험을 한 결과 73.4~91.3% 범위 이었고, 분석오차는 10% 미만으로 코덱스 가이드라인(CAC/GL 40)에 적합하였다. 본 연구에서 개선된 시험법은 향후 국내 유통 농산물을 대상으로 한 잔류실태 조사연구에 활용될 계획이다.

HPLC-UVD/MS를 이용한 농산물 중 bentazone의 분석법 확립

김영학(Young-Hak Kim),이수진(Su-Jin Lee),송이슬(Lee-Seul Song),황영선(Young-Sun Hwang),이영득(Young Deuk Lee),정명근(Myoung-Gun Choung) 한국농약과학회 2011 농약과학회지 Vol.15 No.2

Bentazone is benzothiadiazole group herbicide, and used to foliage treatment. This herbicide have already been widely used for cereals and vegetables planting in worldwide. This experiment was conducted to establish a determination method for bentazone residue in crops using HPLC-UVD/MS. Bentazone residue was extracted with acetone (adjusted pH 1 with phosphoric acid) from representative samples of five raw products which comprised hulled rice, soybean, apple, green pepper, and Chinese cabbage. The extract was diluted with saline water, and dichloromethane partition was followed to recover bentazone from the aqueous phase. Florisil column chromatography was additionally employed for final clean up of the extract. The bentazone was quantitated by HPLC with UVD, using a YMC ODS AM 303 (4.6 × 250 ㎜) column. The crops were fortified with bentazone at 3 levels per crop. Mean recovery ratio were ranged from 82.0% for a 0.2 ㎎/㎏ in apple to 97.9% for a 0.02 ㎎/㎏ in Chinese cabbage. The coefficients of variation were ranged from 0.5% for a 0.02 ㎎/㎏ in soybean to 9.7% for a 0.02 ㎎/㎏ in Chinese cabbage. Quantitative limit of bentazone was 0.02 ㎎/㎏ in representative five crop samples. A LC/MS with selected-ion monitoring was also provided to confirm the suspected residue. Therefore, this analytical method was reproducible and sensitive enough to determine the residue of bentazone ill agricultural commodities.