In this work, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for screening and confirmation of 64 illicit compounds in dietary supplements. The target ...compounds were illegally used pharmaceutical drugs, prohibited compounds, and not authorized ingredients for different therapeutics (sexual enhancement, weight loss, muscular strengthening, and relaxing products). The validation procedure was performed to evaluate selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, and precision according to the Association of Official Analytical Chemists guidelines. The linearity was >0.98 in the range of 0.5-200 µg L
. The LOQs were in the range 1-10 µg kg
for all target compounds. The accuracy (expressed as recovery) was 78.5-114%. The precision (expressed as the relative standard deviation) was below 9.15%. The developed method was applied for the determination of illicit compounds in dietary supplements collected from websites. As a result, the total detection rate was 13.5% (27 samples detected in 200 samples). The concentrations of detected samples ranged from 0.51 to 226 mg g
. The proposed methodology is suitable for monitoring the adulteration of illicit compounds in dietary supplements.
In this study, an analysis method was established for the quantification of residues of halquinol and its metabolites in livestock and fishery products using liquid chromatography–tandem mass ...spectrometry (LC–MS/MS). We selected beef, pork (muscle and fat), chicken, egg, milk, flat fish, eel, and shrimp as target samples for validation of the method owing to them being typical livestock and fishery products. Validation of the developed analysis method was performed using liquid chromatography–tandem mass spectrometry (LC–MS/MS) at three concentration levels (0.5, 1, and 2 × the maximum residue limits) following the Codex Alimentarius (CODEX) guidelines (CAC/GL 71–2009). For all samples, correlation coefficients (R
2
) exceeded 0.99, recoveries ranged between 75.59 and 119.36%, and coefficients of variation (CV) ranged between 1.39 and 28.66%, thus satisfying CODEX guidelines. In addition, inter-laboratory validation was conducted, and the resulting recoveries and CVs satisfied the CODEX guidelines; LOQ was established as 10 μg kg
–1
for pig muscle and 5 μg kg
–1
for the other samples. Therefore, the analysis method developed in this study can accurately and precisely screen for and quantify halquinol and its metabolites in livestock and fishery products.
As veterinary drugs available for fish is very restricted, there is growing trials for repurposing livestock drugs as aquatic animal drugs. Tylosin is one of the most effective antibiotics to treat ...bacterial infections approved for livestock, and would be used in fish. Hence, we investigated the toxicological and microbiological aspects of tylosin to establish health-based guidance value (HBGV) and maximum residue limit (MRL) in fishes, and reevaluated the microbiological acceptable daily intake (mADI) based on updated relevant data and international guildeline. Lastly, exposure assessment was performed to confirm the appropriateness of MRL. By investigating available microbiologcial studies on tylosin, the microbiological point of departure was determined as 0.308 μg/mL, which was mean 50% minimum inhibitory concentration (MIC
50
), obtained from the Food Safety Committee of Japan (FSCJ) evaluation report. Furthermore, as a factor for the derivation of mADI, the volume of colon content was recently changed to 500 mL in compliance with the International Cooperation on Harmonization of Technical Requirements for Registration of Veterinary Medicinal Products (VICH) guidelines. This was previously defined as the mass of colon content (220 g). We applied correction factor 0.224 to the mean MIC
50
for tylosin in the equation of mADI, since the drug is transformed to metabolites with reduced activity prior to entering the colon and bound to fecal materials within the colon of human. The mADI was evaluated as 0.01 mg/kg bw/day. Finally, the hazard index, calculated by dividing the estimated chronic dietary exposure by mADI, did not exceed 100%, suggesting that chronic dietary exposure to tylosin residues from veterinary use was unlikely to be a public health concern. Overall, this study contributes significantly in updating HBGV by application of the concept of mADI for the first time in Korea based on the revised microbiological risk assessment guidelines and in providing scientific rationale for the risk management of veterinary drug residues in food.
The globalization of food distribution has made necessary to secure safe products to the general consumers through the rapid detection of harmful additives on the field. For this purpose, we ...developed a cuvette-type localized surface plasmon resonance (LSPR) sensor that can be easily used by consumers with conventional ultraviolet-visible light spectrophotometer for
measurements. Gold nanoparticles were uniformly deposited on a transparent substrate via a self-assembly method to obtain a plasmonically active chip, and the chemical receptor p-nitroaniline (
-NA) was functionalized to stabilize the device sensitivity under external temperature and pH conditions. The fabricated chip was fixed onto a support and combined with a cuvette-type LSPR sensor. To evaluate the applicability of this sensor on the field, sensitivity and quantitative analysis experiments were conducted onto melamine as a model sample from harmful food additives. Under optimal reaction condition (2 mM
-NA for 20 min), we achieved an excellent detection limit (0.01 ppb) and a dynamic range allowing quantitative analysis over a wide concentration range (0.1-1000 ppb) from commercially available milk powder samples.
The mi-iuy croaker
Miichthys miiuy
has immense commercial value in the Republic of Korea. The red drum
Sciaenops ocellatus
is widely produced by aquaculture, although its price is approximately 25% ...that of
M. miiuy
.
S. ocellatus
has black spots on its tail, enabling it to be distinguished from
M. miiuy
based on appearance. However, identifying
S. ocellatus
after simple processing steps, such as skin removal and dicing, is difficult. Certain traders misrepresent and sell
S. ocellatus
as
M. miiuy
or cultured
M. miiuy
for illegal economical gain. Therefore, an accurate and rapid identification method is required to distinguish between
M. miiuy
and
S. ocellatus
in the field. Here, a method for rapid field identification was developed based on species-specific primers using a portable ultra-fast PCR instrument. The ultra-fast real-time PCR method can complete the entire analytical procedure, including DNA isolation, amplification, and detection, within 30 min, thus maintaining the accuracy of identifying
M. miiuy
and
S. ocellatus
products on site. Forty-nine commercial products were tested, and all samples were successfully identified. Thus, the developed method is rapid, efficient tool for ensuring consumer protection.
Bisphenol A (BPA) is considered to be an endocrine disruptor, but the mechanisms by which it disrupts endocrine functions are poorly understood. Here, we have shown that BPA binds both estrogen ...receptor (ER)-α and ER-beta (ER-β) using a fluorescence polarization competitive binding assay. In addition, we found that BPA induced cell proliferation by modulating cell cycle-related genes in the MCF-7 human mammary cancer cell line. Moreover, using a BG1 luciferase ER transactivation assay, we found that BPA has estrogenic activity. Modulating the MAPK pathway by using an ERK inhibitor (PD98059) or a JNK inhibitor (SP600125) had no effect on the ability of BPA to induce estrogenic activity. However, the antiestrogen, ICI 182,780, and the p38 inhibitor, PD 169316 successfully blocked BPA-induced estrogenic activity. Our findings suggest that BPA mimics ER-dependent estrogenic activity by targeting proteins that regulate the cell cycle and p38 MAPK.
Bisphenol A exerts estrogen receptor-dependent estrogenic activity by targeting proteins that regulate the cell cycle and p38 MAPK.
BACKGROUND: Spiropidion and its metabolite are tetramic acid insecticide and require the establishment of an official analysis method for the safety management because they are newly registered in ...Korea. Therefore, this study was to determine the analysis method of residual spiropidion and its metabolite for the five representative agricultural products.
METHODS AND RESULTS: Three QuEChERS methods (original, AOAC, and EN method) were applied to optimize the extraction method, and the EN method was finally selected by comparing the recovery test and matrix effect results. Various adsorbent agents were applied to establish the clean up method. As a result, the recovery of spiropidion was reduced when using the dispersive-SPE method with MgSO 4 , primary secondary amine (PSA), graphitized carbon black (GCB) and octadecyl (C18) in soybean. Color interference was minimized by selecting the case including GCB and C18 in addition to MgSO 4 . This method was established as the final analysis method. LC-MS/MS was used for the analysis by considering the selectivity and sensitivity of the target pesticide and the analysis was performed in MRM mode. The results of the recovery test using the established analysis method and inter laboratory validation showed a valid range of 79.4-108.4%, with relative standard deviation and coefficient of variation were less than 7.2% and 14.4%, respectively.
CONCLUSION(S): Spiropidion and its metabolite could be analyzed with a modified QuEChERS method, and the established method would be widely available to ensure the safety of residual insecticides in Korea.
플루옥사스트로빈은 Strobilurus 속의 버섯에서 추출한 천 연물을 기반으로 개발된 살균제로서 곰팡이성 질병 방제에 효과적이다. 잔류물의 정의는 유럽(EU), 미국(EPA), 일본 (JFCRF)에서는 플루옥사스트로빈과 플루옥사스트로빈 Z 이 성질체의 합으로 정의하고 있으며, 감자, 대두 등 90품목 에 대하여 0.01-60 mg/kg으로 잔류허용기준이 ...설정되어 있 다. 코덱스(CODEX)와 국내에는 잔류허용기준이 설정되어 있지 않음에 따라 본 연구에서는 추후 국내·외 수입 및 재 배 농산물 중 플루옥사스트로빈에 대한 잔류허용기준 준수 여부 확인을 위한 시험법을 개발하고자 하였다. 전처리 과 정은 플루옥사스트로빈의 물리·화학적 특성을 고려하여 QuEChERS법을 이용한 추출 및 정제방법으로 최적화하였 으며, LC-MS/MS를 이용하여 시험법을 개발하였다. 추출 용매는 아세토니트릴로 하고, MgSO4 및 PSA를 이용하여 정제과정을 확립하였다. 대표 농산물 5종에 대해 0.01, 0.1 및 0.5 mg/kg의 처리농도로 실험을 진행한 결과, 플루옥사 스트로빈 및 플루옥사스트로빈 Z 이성질체의 결정계수(R2) 는 0.998 이상이고 플루옥사스트로빈의 평균 회수율(n=5) 은 75.5-100.3%, 플루옥사스트로빈 Z 이성질체는 75.0- 103.9%이었다. 상대표준편차는 플루옥사스트로빈이 5.5% 이하, 플루옥사스트로빈 Z 이성질체가 4.3% 이하로 확인 되었다. 또한 시험법의 유효성을 확인하기 위해 외부 실험 기관인 광주지방식품의약품안전청과의 실험실간 검증을 진 행하였으며, 검증 결과 두 실험실간의 회수율은 플루옥사 스트로빈의 경우 80.3-101.4%, 플루옥사스트로빈 Z 이성질 체는 80.2-105.0%이었고, 상대표준편차는 모두 18.1% 이하 로 정확성 및 재현성이 우수함을 확인할 수 있었다. 따라 서 본 연구 결과는 CODEX 가이드라인(CAC/GL 40-1993, 2003) 및 식품의약품안전평가원의 가이드라인(MFDS, 2016) 에 만족함에 따라 공정시험법으로 활용 가능할 것이다.
Fluoxastrobin a fungicide developed from Strobilurus species mushroom extracts, can be used as an effective pesticide to control fungal diseases. In this study, we optimized the extraction and purification of fluoxastrobin according to its physical and chemical properties using the QuEChERS method and developed an LC-MS/MS-based analysis method. For extraction, we used acetonitrile as the extraction solvent, along with MgSO4 and PSA. The limit of quantitation of fluoxastrobin was 0.01 mg/kg. We used 0.01, 0.1, and 0.5 mg/kg of five representative agricultural products and treated them with fluoxastrobin. The coefficients of determination (R2) of fluoxastrobin and fluoxastrobin Z isomer were > 0.998. The average recovery rates of fluoxastrobin (n=5) and fluoxastrobin Z isomer were 75.5–100.3% and 75.0–103.9%, respectively. The relative standard deviations (RSDs) were < 5.5% and < 4.3% for fluoxastrobin and fluoxastrobin Z isomer, respectively. We also performed an interlaboratory validation at Gwangju Regional Food and Drug Administration and compared the recovery rates and RSDs obtained for fluoxastrobin and fluoxastrobin Z isomer at the external lab with our results to validate our analysis method. In the external lab, the average recovery rates and RSDs of fluoxastrobin and fluoxastrobin Z isomer at each concentration were 79.5–100.5% and 78.8–104.7% and < 18.1% and < 10.2%, respectively. In all treatment groups, the concentrations were less than those described by the ‘Codex Alimentarius Commission’ and the ‘Standard procedure for preparing test methods for food, etc.’. Therefore, fluoxastrobin is safe for use as a pesticide.
In this study, 27 chemicals found in household products, which became an issue in Korea were screened for the agonistoc and antagonistic effects against human estrogen receptor using official ...Organization for Economic Cooperation and Development (OECD) in vitro assays, STTA assay using ERα-HeLa-9903 cell line and BG1Luc ER TA assay. In the case of human ER agonist screening by two assays, all tested chemicals did not show agonist effect against ER. In ER antagonist test by BG1Luc ER TA assay, five surfactants α-dodecyl-ω-hydroxypoly(oxyethylene), alcohols C16-18 ethoxylated, nonylphenol, ethoxylated, 3,6,9,12,15,18,21-heptaoxatritriacontan-1-ol, and α-dodecyl-ω-hydroxypoly(oxy-1,2-ethanediyl)) were found to exhibit weak antagonistic activities. The agonist/antagonist effects against human estrogen receptor of various chemicals, used in Korea by OECD test guideline are reported in this study. These results indicated that two OECD in vitro assays will can be applied in Korea by screening of agonistic/antagonistic effects against human ER of various chemicals.
•OECD in vitro assays applied for screening of the agonistic/antagonistic effect against human ER on 27 chemicals.•No chemicals exhibited the agonistic effect against human ER.•Five surfactants have been found week antagonistic activities mediated by binding affinity against ER.
DNA 바코드를 이용한 가정간편식 제품의 원재료 모니터링 연구 유연철; 홍예원; 김정주 ...
Han'gug sigpum wi'saeng anjeonseong haghoeji,
06/2020, Volume:
35, Issue:
3
Journal Article
Peer reviewed
Open access
In this study, we monitored the raw materials in home-meal replacement (HMR) products, which have shown more than 63% growth in market size for two years. A total of 89 HMR products were purchased ...and the DNA barcodes of 112 raw materials in the product samples were analyzed. In order to identify the raw material species, a primer set specific for the 16S ribosomal RNA region of each raw material species was amplified. The amplicon was purified and sequenced, and then used to perform a BLAST search provided by the National Institutes of Health (NIH). The species of the raw material was determined by comparing the nucleotide sequences of the species registered in GenBank with identity and match score. Twenty-four species and three genera were identified from 112 raw materials. Three genera were identified at the genus level because a large number of species belonging to the same genus exist within 98% of the identity criteria. The results of the determination were compared with the available raw materials suggested in the Korea Food Code to determine the Korean name and availability of the foods. Six non-listed species were determined to be edible according to information provided by influential domestic and foreign organizations. 본 연구는 최근 소비가 크게 증가하고 있는 가정간편식의 원료에 대한 모니터링을 수행하였다. 다양한 유형의 가정간편식 제품을 구입하여 112개 원료의 DNA 바코드를 분석하였다. 원재료의 종을 동정하기 위하여 DNA 바코드 증폭에 주로 이용되는 미토콘드리아의 16S ribosomal RNA 유전자 부위를 증폭하는 프라이머 세트를 이용하였다. PCR 산물은 정제하여 염기서열을 분석한 후, 이를 이용하여 미국국립보건원에서 제공하는 BLAST search를 수행하였다. GenBank에 등록되어 있는 종의 염기서열과 유사도(Identity)와 매치 점수(Match score)를 비교하여 원료의 종을 판별하였다. 112개의 원료에서 24개의 종(Species)과 3개의 속(Genus)를 동정하였다. 3개의 속은 Identity의 기준이 되는 98% 이내에 해당하는 종이 다수 존재하여 속 수준에서 판별하였다. 판별 결과를 「식품의 기준 및 규격(제2019-57호)」 중 '(별표 1) 사용할 수 있는 원료 목록'에서 제시하는 사용 가능한 원료와 비교하여 국명 및 섭취 가능 여부를 판단하였으며, 등재되어 있지 않은 6개 종은 국제적으로 공인된 기구에서 어획량에 대한 정보를 확인하고, 식용 근거, 학명·이명 등을 확인하여 식용 가능 여부를 판단하였다.
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