Validation of a Rapid Quantitative Method for the Residues of Nitrofuran Metabolites in Loach by Accelerated Solvent Extraction and HPLC Triple Quadrupole Mass Spectrometry

Ryu, Eun Chae,Han, Yun-jeong,Park, Seong-soo,Lim, Chul-joo,Choi, Sunok,Park, Se Chang The Korean Society of Food Hygiene and Safety 2016 한국식품위생안전성학회지 Vol.31 No.2

미꾸라지에서의 Nitrofuran계 대사물질인3-amino-2-oxazolidone(AOZ), 5-morpholinomethyl-3-amino-2-oxazolidinone(AMOZ), 1-ammino-hydantoin (AHD)와 semicarbazide(SEM)의 잔류량을 검사하기 위해HPLC-MS/MS를 이용한 신속한 정량법이 개발되었다. 2-nitrobenzaldehyde (2-NBA)를 이용해 $50^{\circ}C$에서 1시간 동안 산 가수분해와 유도체화 과정을 거친 뒤에, 액-액 분배로 정제와 추출을 하였다. 회수율은 음성시료에 3가지 농도 0.5, 1.0, $2.0{\mu}g/kg$의 표준액을 첨가하여 평가하였고 평균 회수율은 75.1-108.1% 이었다. 정밀성(%RSD)은 일내 8.7% 이하, 일간 8.5% 이하였다. 직선성은 NBAOZ는 $0.2-20{\mu}g/Kg$, NBAMOZ는 $0.8-20{\mu}g/Kg$, NBAHD는 $0.2-20{\mu}g/Kg$, NBSEM 는 $0.1-20{\mu}g/Kg$ 범위에서 모두 상관계수 0.99이상이었다. 검출한계(LOD)는 NBAOZ $0.06{\mu}g/Kg$, NBAMOZ $0.24{\mu}g/Kg$, NBAHD $0.06{\mu}g/Kg$, NBSEM $0.03{\mu}g/Kg$이었고, 정량한계(LOQ)는 NBAOZ $0.2{\mu}g/Kg$, NBAMOZ $0.8{\mu}g/Kg$, NBAHD $0.2{\mu}g/Kg$, NBSEM $0.1{\mu}g/Kg$ 이었다. 가수분해 및 유도체화 소요시간을 1시간으로 줄여 만든 신속 간편한 이 시험법이 미꾸라지 중 nitrofuran metabolites잔류량 분석에 적합함을 확인할 수 있었다. A rapid method using HPLC-MS/MS has been developed for quantitative determination of the metabolites of nitrofurans, namely 3-amino-2-oxazolidone (AOZ), 5-morpholinomethyl-3-amino-2-oxazolidinone (AMOZ), 1-ammino-hydantoin (AHD) and semicarbazide (SEM) in loach. The extraction procedure was founded on simultaneous acidic hydrolysis and derivatization using 2-nitrobenzaldehyde (2-NBA) for 1 hour at $50^{\circ}C$, followed by purification with liquid-liquid extraction. Recovery was evaluated by spiking standards into blank samples at three levels (0.5, 1.0 and $2.0{\mu}g/kg$), and the mean recovery was 75.1-108.1%. Precision values expressed as the relative standard deviation (%RSD) were ${\leq}8.7%$ and ${\leq}8.5%$ for intra-day and inter-day precision, respectively. Linearity was studied in the range of $0.2-20{\mu}g/Kg$ for NBAOZ, $0.8-20{\mu}g/Kg$ for NBAMOZ, $0.2-20{\mu}g/Kg$ for NBAHD, and $0.1-20{\mu}g/Kg$ for NBSEM, and the obtained coefficient correlations (r) were ${\geq}0.99$ for all compounds. Limits of detection (LODs) for the derivatized nitrofuran metabolites were established at $0.06{\mu}g/Kg$ for NBAOZ, $0.24{\mu}g/Kg$ for NBAMOZ, $0.06{\mu}g/Kg$ for NBAHD, and $0.03{\mu}g/Kg$ for NBSEM. Limits of quantification (LOQs) were established at $0.2{\mu}g/Kg$ for NBAOZ, $0.8{\mu}g/Kg$ for NBAMOZ, $0.2{\mu}g/Kg$ for NBAHD, and $0.1{\mu}g/Kg$ for NBSEM. This simplified rapid method for reducing the derivatization and hydrolysis times can be applied to the determination of nitro-furan residues in loach.

Validation of a Rapid Quantitative Method for the Residues of Nitrofuran Metabolites in Loach by Accelerated Solvent Extraction and HPLC Triple Quadrupole Mass Spectrometry

Eun Chae Ryu,Yun-jeong Han,Seong-soo Park,Chul-joo Lim,Sunok Choi,Se Chang Park 한국식품위생안전성학회 2016 한국식품위생안전성학회지 Vol.31 No.2

미꾸라지에서의 Nitrofuran계 대사물질인3-amino-2-oxazolidone (AOZ), 5-morpholinomethyl-3-amino-2-oxazolidinone (AMOZ), 1-ammino-hydantoin (AHD)와 semicarbazide (SEM)의 잔류량을 검사하기 위해HPLC-MS/MS를 이용한 신속한 정량법이 개발되었다. 2-nitrobenzaldehyde (2-NBA)를 이용해 50℃에서 1시간 동안 산 가수분해와 유도체화 과정을 거친 뒤에, 액-액 분배로 정제와 추출을 하였다. 회수율은 음성시료에 3가지 농도 0.5, 1.0, 2.0 μg/kg의 표준액을 첨가하여 평가하였고 평균 회수율은 75.1-108.1% 이었다. 정밀성(%RSD)은 일내 8.7% 이하, 일간 8.5% 이하였다. 직선성은 NBAOZ는 0.2-20 μg/Kg, NBAMOZ는 0.8-20 μg/Kg, NBAHD는 0.2-20 μg/Kg, NBSEM 는 0.1-20 μg/Kg 범위에서 모두 상관계수 0.99이상이었다. 검출한계(LOD)는 NBAOZ 0.06 μg/Kg, NBAMOZ 0.24 μg/Kg, NBAHD 0.06 μg/Kg, NBSEM 0.03 μg/Kg이었고, 정량한계(LOQ)는 NBAOZ 0.2 μg/Kg, NBAMOZ 0.8 μg/Kg, NBAHD 0.2 μg/Kg, NBSEM 0.1 μg/Kg 이었다. 가수분해 및 유도체화 소요시간을 1시간으로 줄여 만든 신속 간편한 이 시험법이미꾸라지 중 nitrofuran metabolites잔류량 분석에 적합함을 확인할 수 있었다. A rapid method using HPLC-MS/MS has been developed for quantitative determination of the metabolites of nitrofurans, namely 3-amino-2-oxazolidone (AOZ), 5-morpholinomethyl-3-amino-2-oxazolidinone (AMOZ), 1-ammino-hydantoin (AHD) and semicarbazide (SEM) in loach. The extraction procedure was founded on simultaneous acidic hydrolysis and derivatization using 2-nitrobenzaldehyde (2-NBA) for 1 hour at 50℃, followed by purification with liquid-liquid extraction. Recovery was evaluated by spiking standards into blank samples at three levels (0.5, 1.0 and 2.0 μg/kg), and the mean recovery was 75.1-108.1%. Precision values expressed as the relative standard deviation (%RSD) were ≤ 8.7% and ≤ 8.5% for intra-day and inter-day precision, respectively. Linearity was studied in the range of 0.2-20 μg/Kg for NBAOZ, 0.8-20 μg/Kg for NBAMOZ, 0.2-20 μg/Kg for NBAHD, and 0.1- 20 μg/Kg for NBSEM, and the obtained coefficient correlations (r) were ≥ 0.99 for all compounds. Limits of detection (LODs) for the derivatized nitrofuran metabolites were established at 0.06 μg/Kg for NBAOZ, 0.24 μg/Kg for NBAMOZ, 0.06 μg/Kg for NBAHD, and 0.03 μg/Kg for NBSEM. Limits of quantification (LOQs) were established at 0.2 μg/Kg for NBAOZ, 0.8 μg/Kg for NBAMOZ, 0.2 μg/Kg for NBAHD, and 0.1 μg/Kg for NBSEM. This simplified rapid method for reducing the derivatization and hydrolysis times can be applied to the determination of nitrofuran residues in loach.

후기 중년 여성의 노화불안에 미치는 영향 요인

남은채(Eun-Chae Nam),김향수(Hyang-Soo Kim),김구민(Doe-Yeon Kang),김나은(Ku-Min),강도연(Kim․,Na-Eun Kim),류승민(Seung-Min Ryu),박예원(Ye-Won Park),정다은(Da-Eun Jung) 한국응용과학기술학회 (구.한국유화학회) 2021 한국응용과학기술학회지 Vol.38 No.1

본 연구는 후기 중년 여성의 자아존중감;폐경 증상;배우자지지;노화불안의 정도를 파악하고 노화불안에 미치는 영향 요인을 알아보고자 하는 서술적 조사연구로 2020년 5월 23일부터 6월 13일까지 3주 동안 대한민국에 거주하는 기혼자 중 후기 중년 여성을 대상으로 온라인 설문지를 이용하여 총 110명의 자료를 수집하였다. 자료 분석 결과;후기 중년 여성의 노화불안에 미치는 영향 요인은 폐경 증상(β =.515;p<.001);자아존중감(β=-.268;p=.001) 순으로 회귀모형의 설명력은 41.4%로 나타났다. 따라서 노화불안을 낮추기 위해서는 후기 중년 여성의 폐경 증상에 대한 적극적인 중재가 필요하며 자아존중감을 높이기 위한 방안이 마련되어야 할 것이다. This study is a descriptive research to determine the degree of self-esteem;menopausal symptom;spousal support of middle-aged women and investigate the factors affecting aging anxiety. For 3 weeks from May 23 to June 13;2020;data total of 110 people were collected using online questionnaires for late middle-aged women. As a result of date analysis;factors affecting aging anxiety in late middle-aged women were menopausal symptoms(B=17.943;p<.001);self-esteem(B=-.585;p=.001) and the explanatory power of the regression model was 41.4%. Therefore;in order to lower againg anxiety;active intervention for menopausal symptoms of late middle-aged women is required;and measures should be taken to increase self-esteem.

LC-MS/MS를 이용한 채소류 및 서류의 클로로젠산 함량 조사

류지은(Ji-Eun Ryu),김운호(Woon-Ho Kim),배호정(Ho-Jeong Bae),정유정(You-Jung Jung),최영주(Young-Ju Choi),문경은(Kyeong-Eun Moon),최종철(Jong-Chul Choi),채경석(Kyung-Suk Chae),이진희(Jin-Hee Lee),도영숙(Young-Sook Do),최옥경(Ok-Kyung C 한국식품영양과학회 2020 한국식품영양과학회지 Vol.49 No.8

본 연구는 LC-MS/MS를 이용해 다소비 채소류 및 서류에 함유된 9종의 chlorogenic acid 이성질체를 분석하기 위하여 시료 추출 후 QuEChERS kit으로 정제하는 전처리법을 확립하였고, Syncronis C18 컬럼을 사용하여 30분 동안 분석하는 이동상 용매 조성과 9종의 이성질체의 정량 및 정성 이온을 선택하여 negative 모드에서 분석하는 기기 분석법을 확립하였다. 확립한 분석법은 AOAC 가이드라인에 따라 특이성, 직선성, 검출한계, 정량한계, 정확성, 정밀성을 확인하여 유효성을 검증하였다. 다소비 채소류 및 서류 43품목(48품종)의 chlorogenic acids 함량을 분석한 결과, 24품목(29품종)에 함유되었음을 확인하였다. 채소류의 경우 39품목(42품종) 중 22품목(25품종)에 chlorogenic acids가 함유되었으며(0.01±0.01~38.31±8.73 ㎎/100 g FW), 취나물, 케일, 미나리, 우엉, 상추, 브로콜리, 콜리플라워, 가지, 치커리, 도라지 순으로 함량이 높게 나타났다. 서류는 고구마(호박고구마, 밤고구마), 감자(로즈밸리, 수미), 마, 토란 등 4품목(6품종)을 분석하였는데, 고구마의 경우 밤고구마(1.31±0.52 ㎎/100 g FW)보다 호박고구마(1.42±0.39 ㎎/100 g FW)에, 감자의 경우 수미(0.09±0.03 ㎎/100 g FW)보다 로즈밸리(0.37±0.07 ㎎/100 g FW)에 chlorogenic acids 함량이 높게 나타났다. 또한 취나물, 미나리, 고구마, 감자를 대상으로 조리시간에 따른 chlorogenic acids 함량 변화를 확인한 결과, 취나물과 미나리는 30초 삶았을 때 가장 높은 함량을 나타내었고, 감자는 30분, 고구마는 20분쪘을 때 chlorogenic acids 함량이 높게 나타났다. 본 연구에서는 다소비 채소류 및 서류의 chlorogenic acids 함량과 조리시간에 따른 chlorogenic acids 함량 변화를 확인하여 식품 내 chlorogenic acids 함량에 관한 기초자료로 제공하고자 하였다. In this study, a sample preparation method and simultaneous determination method were developed to determine the chlorogenic acids content of frequently consumed vegetables, root and tuber crops in Korea by liquid chromatography coupled to mass spectrometry. The developed method was validated for its specificity, linearity, limit of detection, limit of quantification, recovery, accuracy, and precision based on the AOAC guidelines. Among the 39 items of vegetables, 22 items (0.01±0.01∼38.31±8.73 ㎎/100 g fresh weight (FW)) were quantified. The chlorogenic acids content of the vegetables was in the following order: chwinamul (38.31±8.73 ㎎/100 g FW), kale (22.93±0.75 ㎎/100 g FW), water dropwort (10.67±4.46 ㎎/100 g FW), burdock root (9.72±2.52 ㎎/100 g FW), red lettuce (leaf) (8.66±5.37 ㎎/100 g FW), broccoli (2.45±0.50 ㎎/100 g FW), and cauliflower (2.35±0.36 ㎎/100 g FW). Among the four items of root and tuber crops, sweet potato (1.42±0.39 ㎎/100 g FW) and potato (0.37±0.07 ㎎/100 g FW) were quantified. In addition, the chlorogenic acids content of chwinamul and water dropwort was highest when boiled for 30 seconds. Sweet potato steamed for 20 minutes and potato steamed for 30 minutes showed high chlorogenic acids contents.

Excitatory neuron–specific SHP2-ERK signaling network regulates synaptic plasticity and memory

Ryu, Hyun-Hee,Kim, TaeHyun,Kim, Jung-Woong,Kang, Minkyung,Park, Pojeong,Kim, Yong Gyu,Kim, Hyopil,Ha, Jiyeon,Choi, Ja Eun,Lee, Jisu,Lim, Chae-Seok,Kim, Chul-Hong,Kim, Sang Jeong,Silva, Alcino J.,Kaang AAAS 2019 Science signaling Vol.12 No.571

<P><B>Cell type–specific RASopathy</B></P><P>The neurodevelopmental disorder Noonan syndrome is often caused by activating mutations in the phosphatase SHP2 that enhance RAS signaling. However, SHP2 is present in multiple neuron types as well as glia; thus, where the mutant protein has its pathological effects is unclear. Ryu <I>et al.</I> examined one NS-associated SHP2 mutation in isolated cell types from mice and determined that its presence in only excitatory neurons resulted in electrophysiological and cognitive effects. This was because certain adaptor proteins that interact with SHP2 to mediate RAS signaling are abundant in excitatory but not inhibitory neurons. These findings reveal that cell type–specific variations within the RAS signaling network underlie the phenotypes of NS and possibly other “RASopathies”.</P><P>Mutations in RAS signaling pathway components cause diverse neurodevelopmental disorders, collectively called RASopathies. Previous studies have suggested that dysregulation in RAS–extracellular signal–regulated kinase (ERK) activation is restricted to distinct cell types in different RASopathies. Some cases of Noonan syndrome (NS) are associated with gain-of-function mutations in the phosphatase SHP2 (encoded by <I>PTPN11</I>); however, SHP2 is abundant in multiple cell types, so it is unclear which cell type(s) contribute to NS phenotypes. Here, we found that expressing the NS-associated mutant SHP2<SUP>D61G</SUP> in excitatory, but not inhibitory, hippocampal neurons increased ERK signaling and impaired both long-term potentiation (LTP) and spatial memory in mice, although endogenous SHP2 was expressed in both neuronal types. Transcriptomic analyses revealed that the genes encoding SHP2-interacting proteins that are critical for ERK activation, such as GAB1 and GRB2, were enriched in excitatory neurons. Accordingly, expressing a dominant-negative mutant of GAB1, which reduced its interaction with SHP2<SUP>D61G</SUP>, selectively in excitatory neurons, reversed SHP2<SUP>D61G</SUP>-mediated deficits. Moreover, ectopic expression of GAB1 and GRB2 together with SHP2<SUP>D61G</SUP> in inhibitory neurons resulted in ERK activation. These results demonstrate that RAS-ERK signaling networks are notably different between excitatory and inhibitory neurons, accounting for the cell type–specific pathophysiology of NS and perhaps other RASopathies.</P>

Synthesis and Antifungal Activity of 2,5-Disubstituted-6-arylamino-4,7-benzimidazolediones

Ryu, Chung-Kyu,Song, Eun-Ha,Shim, Ju-Yeon,You, Hea-Jung,Choi, Ko Un,Choi, Ik Hwa,Lee, Eun Young,Chae, Mi Jin 梨花女子大學校 藥學硏究所 2002 藥學硏究論文集 Vol.- No.11

2,5-Disubstituted-6-arylamino-4,7-benzimidazolediones were synthesized and tested for in vitro antifungal activity against pathogenic fungi. Among them, 6-arylamino-5-chloro-2-(2-pyridyl)-4,7-benzimidazolediones exhibited potent antifungal activity against Candida specis and Aspergillus niger.

Synthesis and Antifungal Activity of 2,5-Disubstituted-6-arylamino-4,7-benzimidazolediones

Ryu, Chung-Kyu,Song, Eun-Ha,Shim, Ju-Yeon,You, Hea-Jung,Choi, Ko Un,Choi, Ik Hwa,Lee, Eun Young,Chae, Mi Jin 梨花女子大學校 藥學硏究所 2003 藥學硏究論文集 Vol.- No.12

2,5-Disubstituted-6-arylamino-4,7-benzimidazolediones were synthesized and tested for in vitro antifungal activity against pathogenic fungi. Among them, 6-arylamino-5-chloro-2-(2-pyridyl)-4,7-benzimidazolediones exhibited potent antifungal activity against Candida species and Aspergillus niger.

복분자 착즙박을 이용한 식초의 이화학적 특성 및 항산화 활성

류은혜(Eun-Hye Ryu),채규서(Kyu-Seo Chae),김성웅(Sung-Woong Gim),김용석(Yong-Suk Kim),김기덕(Ki-Deok Kim),권지웅(Ji-Wung Kwon) 한국식품과학회 2021 한국식품과학회지 Vol.53 No.1

본 연구에서는 복분자 가공 후 버려지는 부산물의 활용성을 높이기 위하여 착즙박의 발효를 통해 식초를 제조한 후, 전통 발효식초의 소재로서의 이용 가능성을 살펴보고자 발효 특성 및 항산화 활성을 포함한 이화학적 특성을 분석하였다. 알코올 발효후 이화학적 특성 분석 결과, 총 산도와 알코올 함량은 착즙액보다 착즙박에서 각각 0.49, 1.32% 만큼 낮게 나타났으며, 초산발효 후에는 알코올 함량과 당도가 착즙액에서 각각 0.72, 0.6 °Brix 만큼 높게 나타났다. 착즙박에서 pH는 0.32 낮았고 총 산도가 0.38% 높게 나타났다. 이러한 결과에 비추어 볼 때 착즙박 또한 원활한 발효가 진행됨이 확인되었다. 유기산 분석 결과 착즙박에서는 acetic acid가 착즙액에서는 citric acid가 더 높게 나타났다. 복분자 착즙박을 이용한 식초의 총 폴리페놀, 총 플라보노이드, 총 안토시아닌 함량을 측정한 결과 각각 51.58 TAE mg/mL, 9.55RUE mg/mL, 6.05 CYE mg/mL로 대조구인 복분자 착즙액 식초보다 높은 함량을 보였다. 복분자 착즙박 식초의 산화 방지 활성측정을 위해 DPPH 및 ABTS 라디칼 소거능의 IC<SUB>50</SUB> 값을 측정한 결과 각각 10.23, 18.30 μL/mL로 착즙액 식초보다 높은 활성을 나타내었으며, 환원력 및 FRAP 또한 착즙박을 이용한 식초에서 높게 나타났다. 이러한 결과를 종합해볼 때 복분자 가공 부산물로 버려지는 착즙박은 초산발효 결과가 우수하고, 높은 산화 방지 활성을 나타내어 기능성 발효식초로 활용이 가능할 것으로 판단된다. In this study, vinegar was prepared using black raspberry pomace to increase its utilization capacity. As a result of alcohol fermentation, the final alcohol content was 8.90% and the sugar content was 7.03°Brix. The total acid content after acetic acid fermentation was 4.44%, which was 0.38% higher than that of the juice. Among the organic acid content, the acetic acid content was higher in the pomace than in the juice. The total polyphenol (51.58 TAE mg/mL), total flavonoid (9.55 RUE mg/mL), and total anthocyanin (6.05 CYE mg/mL) contents of vinegar produced using black raspberry residue were higher than those of the juice. In addition, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity, reducing power, and ferric reducing antioxidant power (FRAP) activity were higher than those of the juice, indicating strong antioxidant properties. Thus, black raspberry pomace has excellent acetic acid fermentation ability and high antioxidant activity, indicating that it can be used as a functional fermented vinegar.

식물유래 추출물의 주요 학교운동장 잡초 발아억제 활성 조사

이은지(Eun Ji Lee),강동민(Dong Min Kang),김민채(Min Chae Kim),류주현(Ju Hyun Ryu) 한국원예학회 2022 한국원예학회 학술발표요지 Vol.2022 No.11

shRNA-based Gene Therapy System Using Functional Peptides for the Treatment of Psoriasis

Yoo Eun LEE,Yeong Chae RYU,Byeong Hee HWANG 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10

Recently, the Interleukin-23 p19 subunit (IL-23p19) activating interleukin 23 only has been found to play an important role in the inflammatory response of psoriasis. This study aimed to deliver the shRNA silencing IL-23p19 with high specificity via self-assembled nanocomplexes for psoriasis treatment delivery. Nanocomplexes were formed by electrostatic attraction between fusion peptides and shRNAs and were confirmed by gel retardation assay. Moreover, the size and positive charge of nanocomplex were measured by DLS analysis. In addition, the stability of shRNAs in nanocomplex was tested under serum presence. Fluorescence imaging and flow cytometry analysis confirmed the cell permeability of the nanocomplex containing Cy3-labeled siRNAs into RAW 264.7 macrophage cells. Also, shRNAs via nanocomplexes effectively inhibited 34% of IL23-p19 mRNA expression in the cells. The fusion peptide did not reduce cell viability even at high concentrations in the cells. Thus, gene therapy based on nanocomplexes with IL-23p19 shRNAs could be effectively used to treat various related diseases, including psoriasis.