• 한국식품저장유통학회지
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• 제30권1호
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• pp.28-41
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• 2023

Non-aflatoxigenic Aspergillus oryzae and aflatoxigenic A. flavus cannot be clearly identified by partial sequencing of the internal transcribed spacer (ITS) and 18S ribosomal ribonucleic acid (18S rRNA) regions. This study aimed to compare the accuracy among three aflatoxin detection methods using ultra-performance liquid chromatography (UPLC), high-performance liquid chromatography (HPLC), and an enzyme-linked immunosorbent assay (ELISA) kit and to select the non-aflatoxigenic Aspergillus sp. isolated from soybean paste. All analytical methods were suitable according to the international standards of Codex Alimentarius FAO-WHO (CODEX) or the Ministry of Food and Drug Safety (MFDS). UPLC exhibited the best of limit of detection (LOD) and limit of quantification (LOQ). Based on UPLC, HPLC, and the ELISA kit assay, the P5 and P7 strains isolated from soybean paste had 1,663.49, 1,468.12, and >20 ㎍/kg and 1,470.08, 1,056.73, and >20 ㎍/kg, respectively, detected and re-identified as A. flavus. In contrast, the P3 and P4 strains (A. oryzae), which were detected below the MFDS standards in all assays, were confirmed as non-aflatoxigenic fungi. Among the methods evaluated for quantitative analysis of aflatoxin, UPLC and HPLC are superior in terms of accuracy, and the ELISA kit rapidly detects low concentrations of aflatoxin. Furthermore, this study demonstrates that any Aspergillus sp. isolated for use as a fermentation starter should be analyzed for potential aflatoxin production using UPLC and HPLC for accurate quantitative analysis or ELISA for the rapid detection of low-level concentrations of aflatoxin.

• Food Science and Biotechnology
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• 제17권3호
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• pp.679-682
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• 2008

A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

• 한국식품저장유통학회지
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• 제19권3호
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• pp.438-442
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• 2012

칼슘보충제로 칼슘 결합 펩타이드를 만들기 위해, 보리 단백질에 Flavourzyme을 사용하여 18시간 동안 가수분해 하였고, 가수분해 된 보리 단백질은 3 kDa 이하로 한외여과 하였다. 한외여과 된펩타이드는 ion exchange chromatography와 normal-phase HPLC를 사용하여 칼슘 결합 펩타이드를 분리하였고, 분리된 각 분획의 칼슘결합력을 측정하였다. 그 결과 가장 높은 칼슘 결합력을 보인 분획을 칼슘 chelation을 위한 소재로 얻었고, 따라서 본 연구 결과 얻어진 보리 단백질 가수분해물은 칼슘보충제로의 사용이 가능하다고 판단된다.

• 분석과학
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• 제32권1호
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• pp.24-34
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• 2019

The safety of food is occasionally questionable, as there have been some reports of products contaminated with illegal adulterants. In this study, the presence of 16 sedative-hypnotics and sleep inducers in dietary supplements was determined by ultra-high-performance liquid chromatography with UV detection (UPLC-UV) and quadruple Orbitrap mass spectrometry (Q-Orbitrap-MS). The UPLC method was validated, providing a linearity (R2) of more than 0.999, and LODs and LOQs that ranged from 0.2 to 0.5 and 0.6 to $1.5{\mu}g\;mL^{-1}$, respectively. The repeatabilities were 0.2-8.4 % (intra-day) and 0.3-4.5 % (inter-day), and the accuracies were 89.0-117.0 % (intra-day) and 87.8-111.9 % (inter-day). The mean recoveries of the spiked samples ranged from 98.7 to 107.3 %. The relative standard deviation (%RSD) of the stability was less than 2.4 %. Using the developed method, one sedative-hypnotic compound, phenobarbital, was detected in one of the nineteen samples tested. In addition, the major characteristic fragment ions of each target compound were confirmed using Q-Orbitrap-MS for higher accuracy. Monitoring the presence of these 16 sedative-hypnotics and sleep inducers in dietary supplements should be pursued in the interest of human health, and the results of this study confirmed that the developed method has value for this application.

• Journal of Applied Biological Chemistry
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• 제65권4호
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• pp.343-348
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• 2022

백지는 산형과(Umbelliferae)에 속하는 구릿대(Angelica dahurica)의 뿌리를 건조한 것으로, 다양한 약리작용을 하는 것으로 알려져 예로부터 한약재로 사용되어 왔다. 백지의 대표적인 활성 성분인 coumarin을 포함하여 다양한 성분이 알려져 있으며, 최근 백지의 다양한 생리활성이 보고되고 있다. 본 연구는 백지를 건강기능식품의 기능성 원료로 활용하고자, 백지의 지표성분 설정 및 이에 대한 UPLC 분석법을 확립하여 분석법 유효성 검증을 실시하였다. 백지의 지표성분으로는 oxypeucedanin hydrate, byakangelicol, oxypeucedanin, imperatoin, phellopterin, isoimperatorin의6종을 설정하였다. 분석법 유효성 검증은 특이성, 직선성, 검출한계, 정량한계, 정확성, 정밀성 등의 항목을 이용하여 확인하였다. 본 시험법에서 지표성분 6종은 서로 간섭없이 분리되어 특이성을 확인하였고, 결정계수의 값도 0.999 이상으로 나타나 높은 직선성을 나타내었다. 분석법 유효성 검증 결과, 지표 성분 6종의 RSD는 5% 이하로 나타났으며 90-110%의 범위의 회수율을 나타내 높은 정확성과 정밀성이 있는 것을 확인할 수있었다. 이러한 결과는 UPLC를 이용한 백지의 지표성분 분석법이, 새롭게 제시한 6종에 대해 적합한 동시분석 분석법임을 나타낸다. 따라서, 본 연구를 통하여 확립된 백지의 지표성분 분석법이 향후 백지(구릿대)의 건강기능식품 기능성 원료 개발을 위한 기초자료로 활용될 것으로 기대된다.

• 분석과학
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• 제34권4호
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• pp.160-171
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• 2021

Synthetic azo dyes are used extensively in herbal medicines to render the medicines more visually attractive to consumers. This study developed and validated a rapid high-performance liquid chromatography (HPLC) method to determine whether synthetic colorants such as Tartrazine, Auramine O, Metanil yellow, Sunset yellow, and Orange II are used extensively in Typha orientalis. To increase the recovery of the synthetic dyes, this method employed containing 50 mM ammonium acetate in 70 % methanol at first extraction and 100 mM HCl in 70 % methanol at second extraction. Five synthetic pigments in Typha orientalis were separated by gradient elution with a mobile phase consisting of acetonitrile and 50 mM ammonium acetate in distilled water at ultra-violet (UV) detection 428 nm or 500 nm. Additionally, this study established the liquid chromatography tandem mass spectrometry (LC-MS/MS) method to confirm positive samples suspected by HPLC results. The HPLC-UV method had good linearity, indicating r²> 0.999. The recoveries of the samples spiked with three different concentration ranged from 73.8~91.5 %, and relative standard deviation values indicated 0.2~5.2 %. The established LC-MS/MS could successfully identify the synthetic pigments in herbal medicine samples. The study demonstrates that Typha orientalis adulterated by yellowish synthetic dyes can be successfully distinguished when using the HPLC-UV method.

• 분석과학
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• 제30권5호
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• pp.221-225
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• 2017

Thioctic acid is a vitamin-like antioxidant which is prepared as tablets and injection. The Korean Pharmacopoeia (KP XI) contains monograph for the quality control of raw thioctic acid using ultra-violet visible spectrophotometry and its formulations using high performance liquid chromatography (HPLC). In British Pharmacopoeia 2013 (BP2013), another HPLC method is used for the assay test of thioctic acid material. For the international harmonization, we present an HPLC method for quantitation of thioctic acid in both raw material and tablets. Method validation was performed to determine linearity, precision, accuracy, system suitability, and robustness. The linearity of calibration curves in the desired concentration range was high ($r^2=0.9995$), while the RSDs for intra- and inter-day precision were 0.93 ~ 1.26 % and 1.40 ~ 1.76 %, respectively. Accuracies ranged from 98.13-100.00 %. Since the system suitability, intermediate-precision and robustness of the assay were satisfactory, this method will be a valuable addition to the Korean Pharmacopoeia (KP XI).

• Mycobiology
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• 제42권4호
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• pp.353-360
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• 2014

Makgeolli is a traditional Korean alcoholic beverage. The flavor of makgeolli is primarily determined by metabolic products such as free sugars, amino acids, organic acids, and aromatic compounds, which are produced during the fermentation of raw materials by molds and yeasts present in nuruk, a Korean fermentation starter. In this study, makgeolli was brewed using the wild yeast strain Saccharomyces cerevisiae Y98-5, and temporal changes in the metabolites during fermentation were analyzed by ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. The resultant data were analyzed by partial least squares-discriminant analysis (PLS-DA). Various metabolites, including amino acids, organic acids, sugar alcohols, small peptides, and nucleosides, were obviously altered by increasing the fermentation period. Changes in these metabolites allowed us to distinguish among makgeolli samples with different fermentation periods (1, 2, 3, 6, 7, and 8 days) on a PLS-DA score plot. In the makgeolli brewed in this study, the amounts of tyrosine ($463.13{\mu}g/mL$) and leucine ($362.77{\mu}g/mL$) were high. Therefore, our results indicate that monitoring the changes in metabolites during makgeolli fermentation might be important for brewing makgeolli with good nutritional quality.

• Journal of Microbiology and Biotechnology
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• 제26권6호
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• pp.1124-1131
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• 2016

In this study, the global proteome of the IPEC-J2 cell line was evaluated using ultra-high performance liquid chromatography coupled to a quadrupole Q Exactive^™ Orbitrap mass spectrometer. Proteins were isolated from highly confluent IPEC-J2 cells in biological replicates and analyzed by label-free mass spectrometry prior to matching against a porcine genomic dataset. The results identified 1,517 proteins, accounting for 7.35% of all genes in the porcine genome. The highly abundant proteins detected, such as actin, annexin A2, and AHNAK nucleoprotein, are involved in structural integrity, signaling mechanisms, and cellular homeostasis. The high abundance of heat shock proteins indicated their significance in cellular defenses, barrier function, and gut homeostasis. Pathway analysis and annotation using the Kyoto Encyclopedia of Genes and Genomes database resulted in a putative protein network map of the regulation of immunological responses and structural integrity in the cell line. The comprehensive proteome analysis of IPEC-J2 cells provides fundamental insights into overall protein expression and pathway dynamics that might be useful in cell adhesion studies and immunological applications.

• 한국입자에어로졸학회지
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• 제18권2호
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• pp.37-50
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• 2022

Due to the syndrome coronavirus 2 (SARS-CoV-2) pandemic, the subway passenger cabin should be continuously sterilized. However, a disinfectant such as chlorine is toxic and can lead to different issues to human health. In this paper, we introduced a novel disinfectant based on natural product (Dendropanax morbifera extract). Via ultra-high performance liquid chromatography - mass spectrometer (UHPLC-MS), different compounds from Dendropanax morbifera extract showed antivirus potentials. Antimicrobial experiments confirmed that the air-disinfectant containing Dendropanax morbifera can eliminate harmful microorganisms including Gram (-), Gram (+), and yeast within 5 mins. The as-prepared air-disinfectant also showed high antivirus activity against H1N1, HRV, and EV71. Deodorization test also indicates that the as-prepared air-disinfectant can lower the harmful gas such as ammonia and trimethylamine in the atmosphere. To evaluate the potential of air-disinfectant containing Dendropanax morbifera in practical applications, different safety tests including acute oral toxicity, acute skin irritation, and eye irritation were conducted. Results showed that the as-prepared disinfectant did not negatively affect tested animals during these safety investigations.