LC-MS/MS를 이용한 설사성패류독소의 분석조건 확립

이가정(Ka Jeong Lee),스즈키 도시유키(Toshiyuki Suzuki),김풍호(Poong Ho Kim),오은경(Eun Gyoung Oh),송기철(Ki Cheol Song),김지회(Ji Hoe Kim) 한국식품과학회 2009 한국식품과학회지 Vol.41 No.4

설사성패류독의 신속정밀 분석조건 확립을 위하여 LC-MS/MS를 사용하여 이동상, 분석용 column 및 collision energy 등을 변화시키면서 시험하였다. 50 mM formic acid와 2 mM ammonium formate가 함유된 acetonitrile 수용액을 이동상으로 사용하였을 때 OA와 DTX1이 검출되었다. Collision energy는 독소 성분에 따라 달리하는 것이 다성분 동시분석에 적합하였으며 OA와 DTX1 고유의 fragment ion들은 48 V 정도에서 최적의 intensity로 확인되었다. Column의 종류에 따라서는 C8 column의 경우 OA, DTX1, DTX3, PTX2 및 YTX 모두 검출 가능하였으나 실제 검출 대상이 OA와 DTX1인 경우에는 일반적으로 사용되는 Csub{18} column도 적합한 것으로 확인되었다. 본 연구에서 확립한 LC-MS/MS 분석 조건의 검출한계는 OA와 DTX1 모두 1 ng/g, 정량한계는 각각 3ng/g이었고, 표준독 성분을 첨가한 시료에서 process efficiency는 굴의 경우 91-118%, 진주담치에서는 96-117%이었고, matrix의 영향은 거의 없었다. 마우스 시험에서 양성을 나타낸 시료를 LCMS/MS법으로 분석한 결과, 일부 시료에서만 OA 및 DTX1이 검출되어 두 시험법의 독성은 일치하지 않았으며 LC-MS/MS법은 마우스 시험법보다 하루 이상 분석시간을 단축할 수 있었다. To establish and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and accurate quantitation of diarrhetic shellfish poisoning (DSP) toxins, we compared the results from different mobile phases and columns used for their analysis and collision energies for MS/MS experiments. Clear peaks of okadaic acid (OA) and dinophysistoxin-1 (DTX1) were obtained by using a mobile phase comprising aqueous acetonitrile containing 2 mM ammonium formate and 50 mM formic acid. The collision energies were optimized to facilitate the most sensitive detection for each toxin, namely, OA, DTX1, pectenotoxin-2 (PTX2), or yessotoxin (YTX). Further, the maximum ion response was obtained at a collision energy of 48 V for OA and DTX1. We compared the analytical performance of Csub{8} and Csub{18} columns. A wide range of toxins namely, OA, DTX1, PTX2, and YTX, except DTX3, were detected by both the columns. Although DTX3 was only detected by the Csub{8} column, we found that the Csub{18} column was also suitable for the quantitation of OA and DTX1 the toxins responsible for inducing diarrhea. The limit of detection of OA and DTX1 by the established LC-MS/MS conditions was 1 ng/g, and the limit of quantitation of the toxins under the same conditions was 3 ng/g. The process efficiencies were 91-118% for oysters (Crassostrea gigas) and 96-117% for mussels (Mytilus galloprovincialis) further, we observed no significant effect of matrix during the ionization process in LC-MS/MS. The comparison between mouse bioassay (MBA) and LC-MS/MS yielded varying results because low concentrations of OA and DTX1 were detected by LC-MS/MS in some shellfish samples, which provided positive results on MBA for DSP. The analysis time required by MBA for DSP analysis can be reduced by LC-MS/MS.

ELISA-LC/MS/MS 병행에 의한 식품 중 aflatoxins 분석

김경열,남민지,남보람,류희정,송정언,심원보,이수형,정덕화,Kim, Kyeong-Yeol,Nam, Min-Ji,Nam, Bo-Ram,Ryu, Hee-Jung,Song, Jeong-Eon,Shim, Won-Bo,Lee, Soo-Hyung,Chung, Duck-Hwa 한국환경보건학회 2010 한국환경보건학회지 Vol.36 No.1

High performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS) have been widely used to quantify aflatoxins in food, but these methods are expensive, time-consuming, unsuitable for analysis of the routine screening of large sample numbers and require derivatization and high level techniques to perform. The objective of this study is to detect aflatoxins in a large number of foods by a high efficient analytical system of combined enzyme linked immunosorbent assay (ELISA) for screening and LC/MS/MS for confirmation. The samples spiked individually with aflatoxin $B_1$ (0.5 and 1.0 ng/g) and total aflatoxins (10 ng/g) were analyzed by ELISA and LC/MS/ MS, and the recoveries for ELISA and LC/MS/MS were 71.8~119.2% and 70.8~135.3%, respectively. A total of 378 samples (grains, nuts, soybean and fermented soybean foods, pepper and fermented pepper foods) were purchased from the six major cities in Korea and analyzed by ELISA-LC/MS/MS system. Twenty two (5.8%; peanut: 11, pistachio: 2, walnut: 6, almond: 1, pepper powder: 1, pepper paste: 1) out of 378 samples were screened as aflatoxin B1 positive by ELISA, but, 4 (1.1%; peanut: 2, pistachio:1, pepper powder: 1) out of the 22 samples screened were confirmed as aflatoxins positive at levels of 1.02~52.79 ng/g by LC/MS/MS. ELISA-LC/MS/MS system provides a more rapid, accurate and cost-effective method for the detection of aflatoxins in large number of samples.

식품에서 당살초 판별을 위한 LC-ESI-MS/MS 분석법과 KASP 마커 개발

박보름,이선희,엄권용,노은영,한경문,황진우,김형일,백선영 한국식품위생안전성학회 2022 한국식품위생안전성학회지 Vol.37 No.2

Known for its effectiveness in weight loss and diabetes prevention, Gymnema sylvestre products can be found in the US, Japanese, and Indian markets. However, the recommended dosage or safety of these products has not yet been proven. Therefore, development of an analytical method for detecting the content of Gymnema sylvestre in food products is required. Accordingly, this study proposes an analysis method that can examine Gymnema sylvestre in food using LC-ESI-MS/MS and KASP (Kompetitive Allele-Specific PCR) markers. In LC-ESI-MS/MS, a simultaneous analysis method for gymnemic acid and deacylgymnemic acid was optimized using negative ionization mode, and its validation test was completed for solid and liquid samples. In addition, KASP markers were prepared by finding the specific SNP of G. sylvestre in ITS2 and matK through DNA barcodes. The two KASP markers returned positive FAM fluorescence result when combined with G. sylvestre, and this aspect was confirmed in raw G. sylvestre as well. The applicability of the method was tested on 21 different food and healthy functional products containing G. sylvestre purchased on the internet. As a result, although there was a difference in the ratios of gymnemic acid and deacylgymnemic acid in LC-ESI-MS/MS, the index component was detected in all 21 products samples. In the KASP analysis, 9 products returned positive FAM result, and the rest of the products were found to be containing G. sylvestre extract. This study is the first study to use the dual system of LC-ESI-MS/MS and KASP for the analysis of G. sylvestre. The study has confirmed that these two methods are applicable to the examine G. sylvestre content in food products.

LC-MS/MS를 이용한 Homocysteine 측정과 그 유용성 평가

전선희 ( Sun Hee Jun ),임미숙 ( Mi Suk Lim ),정영순 ( Yong Sun Jung ),송정한 ( Jung Han Song ) 대한임상검사과학회 2005 대한임상검사과학회지(KJCLS) Vol.37 No.1

Total homocysteine is now considered a risk factor for cardiovascular diseases. I increased interest has led to a multitude of studies requiring the determination of total homocysteine in conjunction with other factor. There are various methods for measuring total homocysteine, including HPLC, FPIA, GC-MS and LC-MS/MS. The most recent method for measuring total homocysteine uses a deuterium-labelled internal standard and tandem mass spectrometry. This development requires no derivatization and therefore leads to an increase in sample throughput compared to other techniques. We have evaluated the method for homocysteine by the LC-MS/MS method, and the correlation between the FPIA method and the LC-MS/MS method. The standard curve (0, 5, 10, 20, 50, 100 uM) was linear over the range examined (up to 100 uM). The lower limit of quantification (CV < 10 %) was 0.5 uM/L and the lower limit of detection (S/N >3) was 0.1 uM/L. Intra-assay variation and inter-assay variation were both <6 %. The comparision study for homocysteine concentration showed good correlation (r=0.9684) between the FPIA method and LC-MS/MS methods. Our conclusion is that the method showed relatively good precision, and was rapid and accurate.

설사성 패류독의 LC-MS/MS에 의한 분석

윤소미,장준호,신일식,이종옥,이종수,Yun, So-Mi,Jang, Jun-Ho,Shin, Il-Shik,Lee, Jong-Ok,Lee, Jong-Soo 한국식품영양과학회 2007 한국식품영양과학회지 Vol.36 No.7

LC-MS/MS법에 의하여 설사성 패류독소 7종(okadaic acid, dinophysistoxin-1, -3, pectebotoxin-1, -2, -6, yessotoxin)의 동시 분석 조건을 확립하였고, 2006년 3월부터 9월까지 매월 5종의 이매패(굴, 진주담치, 반지락, 가리비, 개조개, 6월부터 9월까지는 가리비 대신 피조개)를 정기적으로 통영시장에서 구입하여 분석하였으며, 기존의 마우스 검사법과 비교하였다. 마우스 검사법으로는 총 35종 시료 중 3월의 굴과 진주담치에서 0.05${\sim}$0.1 MU/g의 독성을 나타내었으나, 이미 알려진 설사성 패류독소는 전혀 검출되지 않아 다른 성분에 의한 것으로 판단되었다. 한편, LC-MS/MS법에 의하여서는 6월 진주담치에서 dinophysistoxin-1(DTX1)이 0.05 ${\mu}g/g$ 검출되었으며, 8월의 모든 시료에서 okadaic acid(OA)가 0.01${\sim}$0.02 ${\mu}g/g$ 검출되었다. 이들은 외국의 기준치[0.05 MU/g(일본), OAs(OA+DTX1) 0.16 ${\mu}g/g$(EU)]보다 매우 낮은 함량으로 식품 위생상 안전하였다. Diarrhetic shellfish poisons (DSP) such as okadaic acid (OA), dinophysistoxin-1 (DTX1), pectenotoxin-1(PTX1), PTX2, PTX6 and yessotoxin (YTX) were determined simultaneously by LC-MS/MS and mouse bioassay in the shellfishes (oyster, mussel, Washington purple clam, ark shell, scallop and short necked clam) collected at Tongyeong, from March to September, 2006. Oyster and mussel were found to contain DSP (0.05${\sim}$0.1 MU/g) in March by mouse bioassay; however, no DSP components were detected on the LC-MS/MS. Also, a small amount of DTX1 (0.05 ${\mu}g/g$) in mussel (June) and OA (0.01${\sim}$0.02 ${\mu}g/g$) in 5 species of shellfishes(August) were determined by LC-MS/MS.

LC-MS/MS 및 GC-MS/MS를 활용한 수산물 중 디아제팜의 정량분석법 개발

신다솜,강희승,김주혜,정지윤,이규식,Shin, Dasom,Kang, Hui-Seung,Kim, Joohye,Jeong, Jiyoon,Rhee, Gyu-Seek 한국식품위생안전성학회 2018 한국식품위생안전성학회지 Vol.33 No.2

본 연구는 국내 생산 및 수입 양식 수산물에 대해 잔류할 수 있는 향정신성 의약품인 디아제팜 대한 안전관리강화기반을 위해 마련되었다. 중국인민공화국 국가 표준시험법(GB 29697-2013)을 기반으로 전처리 방법을 개선하여 GC-MS/MS 시험법을 확립하였으며, LC-MS/MS 방법과의 기기간 검증을 통해 확립된 시험법의 선택성, 정량한계 및 회수율에 대한 검증을 통해 디아제팜 시험법으로서의 유효성을 확인하였다. LC-MS/MS의 경우 아세토니트릴로 추출 후 PSA를 이용해 정제하였고, GC-MS/MS의 경우 아세토니트릴로 추출후 $C_{18}$카트리지를 이용해 정제하였다. 디아제팜은 표준용액을 정량한계를 포함한 농도에 따라 검량선을 작성한 결과 두 기기 모두 $r^2$> 0.99 이상의 직선성을 확인하였다. 본 실험에서의 검출한계와 정량한계는 LC-MS/MS 및 GC-MS/MS 모두 0.0004 mg/kg, 0.001 mg/kg 수준이었으며, 평균 회수율은 각각 99.8~106%, 109~124%이었다. 또한, 분석오차는 모두 15% 이하로 정확성 및 재현성이 우수하였으며, CODEX 가이드라인 규정에 만족하는 수준이었다. 따라서 개발된 시험법은 안전한 수산물의 국내 유통과 잔류실태조사를 위해 활용될 것으로 기대한다. The aim of this study was to develop an analytical method for the quantification of diazepam residues in fishery products, using liquid and gas chromatography-tandem mass spectrometry (LC-MS/MS and GC-MS/MS). The sample utilized in the study was extracted from the fish sample (crucian carp) using 0.1% formic acid in acetonitrile. For the utilization of the purification process, the dispersive solid phase extraction (dSPE) was used for LC-MS/MS, dSPE and SPE was used for GC-MS/MS, respectively. To be sure, the standard calibration curves showed a good linearity as the noted correlation coefficients, $r^2$ was > 0.99. The average recoveries for accuracy ranged in 99.8~124% for the samples which were fortified at three different levels (0.001, 0.002 and 0.010 mg/kg). The correlation coefficient for the precision effect was measured at a range of 4.01~11.8%. The limit of detection (LOD) for the diazepam analysis was 0.0004 mg/kg, and the limit of the quantification (LOQ) was 0.001 mg/kg. The proposed analytical method was characterized with a high accuracy and acceptable sensitivity to meet the established Codex Alimentarius Commission (CAC/GL71-2009) guideline requirements. We therefore established the optimal analysis method for the determination of diazepam in the fishery products using LC-MS/MS and GC-MS/MS. It would be applicable to analyze the diazepam residues in fishery products in further studies on this subject.

LC-MS/MS를 이용한 벌꿀 중 grayanotoxin 분석법 연구 및 실태조사

이숙연(Sook-Yeon Lee),최윤주(Youn-Ju Choi),이강봉(Kang-Bong Lee),조태용(Tae-Yong Cho),김진숙(Jin-Sook Kim),손영욱(Young-Wook Son),박재석(Jae-Seok Park),임성임(Sung-Im Im),최희정(Hee-Jung Choi),이동하(Dong-Ha Lee) 한국식품과학회 2008 한국식품과학회지 Vol.40 No.1

본 연구는 야생꿀을 비롯한 벌꿀, 벌집채꿀 등 국내산 및 수입산 꿀의 안전관리를 위한 grayanotoxin(GTX)의 시험분석법 확립 및 실태조사를 위하여 수행하였다. GTX 표준품 Ⅰ, Ⅱ, Ⅲ는 LC-MS/MS로 분자량을 확인한 후, 시판품인 III를 제외한 Ⅰ과 Ⅱ는 NMR을 이용하여 구조를 확인하였다. 총 111건(국내산 벌꿀 25건, 국내산 야생꿀 21건, 벌집채꿀 13건, 수입산 벌꿀 44건, 수입산 야생꿀 8건)의 벌꿀시료는 메탄올을 사용하여 벌꿀-메탄올 용액을 만들어 tC18 cartridge에 loading 한 후, 여과된 액을 동량의 증류수로 희석하여, 이온화장치로 ESI를 장착한 triplequadrupole LC-MS/MS를 이용하여 분석하였다. LC의 용리액은 1% 포름산이 첨가된 “메탄올-물”을 사용하는 것이 10분 이내의 분석시간대에 나타나는 피크의 모양과 감도가 우수한 경향을 나타내었다. 본 방법을 이용하여 검체 중의 GTX Ⅰ, Ⅱ, Ⅲ의 함유량을 조사한 결과 총 111건 중 수입산 야생꿀 3건(2.7%)에서 GTX Ⅰ, Ⅱ, Ⅲ가 검출되었고, 수입산 야생꿀 1건에서 GTX Ⅰ, Ⅲ가 검출되었다. GTX Ⅰ의 검출량은 최소 3.13 ± 0.00 mg/kg에서 최고 12.93 ± 0.01 mg/kg으로 나타났고 GTX Ⅱ는 0.84 ± 0.01 mg/kg, 0.92 ± 0.00 mg/kg, 1.08 ± 0.01 mg/kg의 함량을 나타내어 GTX Ⅰ에 비해 낮은 수치를 나타내었다. GTX Ⅲ는 최소 0.25 ± 0.01 mg/kg에서 최고 3.29 ± 0.74 mg/kg으로 함량에 큰 차이를 보였다. 본 방법을 이용한 총 111건의 벌꿀 시료의 GTX분석시 수입산 야생꿀 4건에서만 GTX가 검출됨을 알 수 있었다. 본 연구에서는 비휘발성 또는 극성 때문에 GTX 분석시 GC 및 GC-MS에서 분석이 어려운 벌꿀시료를 대상으로 전처리 시간의 단축을 모색함과 동시에 LC-MS/MS를 이용한 시험분석법을 개발할 수 있었고, 모니터링을 통하여 네팔, 터키 등 특정 지역의 야생꿀의 섭취를 제한하는 과학적 근거를 마련할 수 있었다. This study was performed to establish analysis methods, and evaluated for grayanotoxin in domestic/foreign honey and wild honey. The molecular weight of grayanotoxins Ⅰ, Ⅱ and Ⅲ, excluding grayanotoxin Ⅲ that has been commercialized, were analyzed by LC-MS/MS. Then, the molecular structure of grayanotoxins Ⅰ and Ⅱ were analyzed by NMR. A total 111 samples (25 Korean honey, 21 Korean wild honey, 13 Korean honeycomb honey, 44 foreign honey, 8 foreign wild honey) were examined to determined whether or not each sample contained grayanotoxins Ⅰ, Ⅱ, and Ⅲ. The honey samples were mixed with methanol and loaded into a tC18 cartridge, the filtrate was diluted with water, and the mixture was then analyzed by ESI triple-quadrupole LC-MS/MS. Grayanotoxins were only found in the foreign wild honey and were not detected in Korean honey, Korean honeycomb honey, or Korean wild honey. Three of the samples contained grayanotoxin Ⅰ, Ⅱ, and Ⅲ, and one sample contained only grayanotoxins Ⅰ and Ⅲ. The lowest level for grayanotoxin I was 3.13 ± 0.00 mg/kg, and the highest level was 12.93 ± 0.01 mg/kg. The levels of grayanotoxin Ⅱ were 0.84 ± 0.01 mg/kg, 0.92 ± 0.00 mg/kg and 1.08 ± 0.01 mg/kg, respectively. The lowest level of grayanotoxin Ⅲ was 0.25 ± 0.01 mg/kg and the highest level was 3.29 ± 0.74 mg/kg. Through this study, safety management for foreign wild honey has been enabled.

Tacrolimus의 혈중농도측정법 비교 및 간이식환자에서의 집단 약동학

김은영,강원구,곽혜선 한국임상약학회 2008 한국임상약학회지 Vol.18 No.1

This study aimed to compare a microparticle enzyme immunoassay (MEIA) with a liquid chromatography-tandem mass spectrometry (LC/MS/MS) technique for the measurement of tacrolimus concentrations in adult liver transplant recipients, to investigate how the assay choice influenced the population pharmacokinetics of tacrolimus and to identify patient characteristics that affected pharmacokinetic parameters in each assay. Tacrolimus concentrations from 29 liver (n=52 paired-samples) transplant recipients measured by both MEIA and LC/MS/MS were used to evaluate the performance of these methods in the clinical setting. Tacrolimus pharmacokinetics was studied independently using MEIA and LC/MS/MS data in 70 adult patients using a population approach performed with NONMEM. Patient characteristics which influenced pharmacokinetic parameters in each assay were compared. The relation between LC/MS/MS and MEIA measurements was best described by the regression equation MEIA=1.465*LC/MS/MS- 1.336 (r=0.91). Multiple linear regression analysis showed significant inverse relationships between assay difference and hematocrit (Hct) (p<0.025) in liver graft recipients. In MEIA, the population estimate of tacrolimus CL/F and apparent volume of distribution (Vd/F) were found to be 10.1 L/h and 226 L, and in LC/MS/MS, 13 L/h and 305 L respectively. Neither patient's age, weight, gender, grafted hepatic weight, albumin concentration, nor markers of liver function influenced tacrolimus CL/F. The final model of CL/F was found to be 10.1+(Hct/Hct mean)12.0 in MEIA and 13+(1+Hct/578) in LC/MS/MS indicating that CL/F was influenced by hematocrit.

Analysis of Myosin Heavy Chain Isoforms from Longissimus Thoracis Muscle of Hanwoo Steer by Electrophoresis and LC-MS/MS

김갑돈 한국축산식품학회 2014 한국축산식품학회지 Vol.34 No.5

The purpose of this study was to analyze myosin heavy chain (MHC) isoforms in bovine longissimus thoracis (LT) muscleby liquid chromatography (LC) and mass spectrometry (MS). LT muscles taken from Hanwoo (Korean native cattle) steer(n=3) used to separate myosin bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The peptide querieswere obtained from the myosin bands by LC-MS/MS analysis following in-gel digestion with trypsin. A total of 33 and 43queries were identified as common and unique peptides, respectively, of MHC isoforms (individual ions scores >43 indicateidentity or extensive homology, <0.05). MHC-1 (IIx), -2 (IIa), -4 (IIb), and -7 (slow/I) were identified based on the Mowsescore (5118, 3951, 2526, and 2541 for MHC-1, -2, -4, and -7, respectively). However, more analysis is needed to confirmthe expression of MHC-4 in bovine LT muscle because any query identified as a unique peptide of MHC-4 was not found. The queries that were identified as unique peptides could be used as peptide markers to confirm MHC-1 (14 queries), -2 (8queries), and -7 (21 queries) in bovine LT muscle; no query identified as a unique peptide of MHC-4 was found. LC-MS/MS analysis is a useful approach to study MHC isoforms at the protein level. The purpose of this study was to analyze myosin heavy chain (MHC) isoforms in bovine longissimus thoracis (LT) muscleby liquid chromatography (LC) and mass spectrometry (MS). LT muscles taken from Hanwoo (Korean native cattle) steer(n=3) used to separate myosin bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The peptide querieswere obtained from the myosin bands by LC-MS/MS analysis following in-gel digestion with trypsin. A total of 33 and 43queries were identified as common and unique peptides, respectively, of MHC isoforms (individual ions scores >43 indicateidentity or extensive homology, <0.05). MHC-1 (IIx), -2 (IIa), -4 (IIb), and -7 (slow/I) were identified based on the Mowsescore (5118, 3951, 2526, and 2541 for MHC-1, -2, -4, and -7, respectively). However, more analysis is needed to confirmthe expression of MHC-4 in bovine LT muscle because any query identified as a unique peptide of MHC-4 was not found. The queries that were identified as unique peptides could be used as peptide markers to confirm MHC-1 (14 queries), -2 (8queries), and -7 (21 queries) in bovine LT muscle; no query identified as a unique peptide of MHC-4 was found. LC-MS/MS analysis is a useful approach to study MHC isoforms at the protein level.

LC-MS/MS 와 GC-MS/MS 를 이용한 에센셜 오일 중 320종 잔류농약 분석법 개발

오가향 ( Ka Hyang Oh ),박성막 ( Sung Mak Park ),이소민 ( So Min Lee ),정소영 ( So Young Jung ),곽병문 ( Byeong-mun Kwak ),이미기 ( Mi-gi Lee ),이미애 ( Mi Ae Lee ),최성민 ( Sung Min Choi ),빈범호 ( Bum-ho Bin ) 대한화장품학회 2021 대한화장품학회지 Vol.47 No.4

에센셜 오일(essential oil)은 1 개의 단일 식물형 및 식물종이 만들어낸 향이 나는 식물 재료를 물리적인 방법으로 얻어낸 휘발성 물질로 방부, 살균, 항균 효과가 뛰어나 화장품, 향료, 아로마 테라피 치료 등의 목적으로 폭 넓게 사용되고 있다. 에센셜오일은 추출 및 농축과정을 거치게 되는데 이때 재배 과정 중 살포된 농약 또한 추출 및 농축이 됨으로써 인체에 유해할 수 있다. 본 연구에서는 에센셜 오일 중 320 종의 잔류농약을 분석하기 위하여 LC-MS/MS와 GC-MS/MS를 이용하였으며 기존의 정제과정인 hexane 대신 freezing 과정을 이용하여 전처리방법을 개선하였다. 에센셜 오일을 분석한 결과 기준치가 설정되어 있지 않은 chlorpyrifos, piperonyl butoxide, silafluofen 성분이 basil oil 및 clove leaf oil에서 검출되었다. 따라서 에센셜 오일에 대한 잔류농약 모니터링이 지속적으로 필요할 것으로 판단된다. Essential oil is a volatile substance obtained by physically obtaining fragrant plant materials made by one single plant and plant species, and is widely used for cosmetics, fragrances, and aroma therapy due to its excellent preservation, sterilization, and antibacterial effects. When essential oil would undergo the extraction and concentration processes, the agricultural chemicals thereof would be extracted and concentrated only to be harmful to the human body. This study analyzes 320 residual agricultural chemicals concentrated in the essential oil, and to this end, LC-MS/MS and GC-MS/MS are used, while the freezing process is applied instead of the conventional refining process hexane, to improve the preprocessing method. As a result of analyzing the essential oil, such ingredients as chlorpyrifos, piperonyl butoxide and silafluofen have been detected in Basil oil and Clove leaf oil. Hence, it is perceived that the residual agricultural chemicals should continue to be monitored for the essential oil.