• Title/Summary/Keyword: HPLC quantification

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Inhibitory Effect of Mixed Extracts Obtained from Astragali Radix and Lithospermi Radix on Matrix Metalloproteinases in IL-1β-induced SW1353 Cells and Quantitative Analysis of Active Compounds (황기, 지치 복합물의 연골세포에서의 Matrix Metalloproteinases 저해 효과 및 유효성분의 분석)

  • Choi, Doo Jin;Choi, Bo Ram;Lee, Dae Young;Choi, Soo Im;Lee, Young Seob;Kim, Geum Soog
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.4
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    • pp.247-258
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    • 2019
  • Background: Astragali radix (A) and Lithospermi radix (L) have long been used as traditional medicines due to their known anti-inflammatory effects. This study aimed at evaluating, their optimal mixing ratio and their functional compounds by investigating the inhibitory effects of mixed extracts of A and L and their active compounds on matrix metalloproteinases (MMPs). Methods and Results: A and L extracts were obtained by extraction at $80^{\circ}C$ using 50% and 70% fermented alcohol, respectively, and then mixed at a ratio of 5 : 5, 6 : 4, 7 : 3 and 8 : 2 (w/w). The activities of MMP-1, MMP-3, and MMP-13 were evaluated in interleukin-1beta ($IL-1{\beta}$)-induced SW1353 cells. The extract mixtures showed synergistic inhibitory effects on MMP-3 and MMP-13, higher than the effects of the individual A and L extracts. The 7 : 3 mixture (ALM16) showed the most effective MMPs inhibitory activity, while among the active ingredients, calycosin-7-O-${\beta}$-D-glucoside and lithospermic acid exhibited excellent MMPs inhibitory activity. Additionally, an HPLC method was established for simultaneous quantification of the effective components of the extract mixtures, and validated by measuring the linearity, precision and accuracy of the limit of detection (LOD) and limit of quantification (LOQ). Conclusions: ALM16 showed the most effective MMPs inhibitory activity. Calycosin-O-${\beta}$-D-glucoside, calycosin and lithospermic acid were identified as useful candidates, as they were the major functional compounds in the MMP inhibitory activity. Summarily, ALM16 might be a highly effective in osteoarthritis management, owing to its because it exhibits a protective effect on cartilage via excellent inhibition of MMPs.

Determination and Validation of Synthetic Antioxidants in Processed Foods Distributed in Korea

  • Park, Hyeon-Ju;Seo, Eunbin;Park, Jin-Wook;Yun, Choong-In;Kim, Young-Jun
    • Journal of Food Hygiene and Safety
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    • v.37 no.5
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    • pp.297-305
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    • 2022
  • Antioxidants are food additives that extend the shelf life of food products by preventing lipid rancidity caused by active oxygen. They can either be naturally-derived or manufactured synthetically via chemical synthesis. In this study, method validation of five synthetic antioxidants, namely butylated hydroxyanisole, butylated hydroxytoluene, tertiary butylhydroquinone, propyl gallate, and disodium ethylenediaminetetraacetic acid, was performed using a high performance liquid chromatography-ultraviolet visible detector, and the method applicability was evaluated by analyzing foods containing antioxidants. The coefficient of determination (R2) average was 0.9997, while the limit of detection and limit of quantification were 0.02-0.53 and 0.07-1.61 mg/kg, respectively. The intra and inter-day accuracies and precisions were 83.2±0.7%-98.7±2.1% and 0.1%-5.7% RSD, respectively. Inter-laboratory validation for accuracy and precision was conducted using the Food Analysis Performance Assessment Scheme quality control material. The results satisfied the guidelines presented by the AOAC International. In addition, the expanded uncertainty was less than 16%, as recommended by CODEX. Consequently, to enhance public health safety, the results of this study can be used as basis data for evaluating the intake of synthetic antioxidants and assessing their risks in Korea.

Survey of Aflatoxin B1 and Ochratoxin A on Commercial Dried Red Pepper and Red Pepper Powder (유통 건고추 및 고춧가루의 아플라톡신 B1과 오크라톡신 A 오염도 조사)

  • Jegal, Seung;Kim, Ji-Hyeung;Joo, Gwang-Sig;Jung, Se-Jin;Na, Hyeon-Ju;Jo, Nam-Gyu;Lee, Jea-Man;Kim, Yong-Hee
    • Journal of Food Hygiene and Safety
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    • v.28 no.3
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    • pp.267-271
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    • 2013
  • A survey of aflatoxin $B_1$ and ochratoxin A was conducted on dried red pepper and red pepper powder. Total number of 193 samples were collected from local markets in Incheon. The presence of aflatoxin $B_1$ and ochratoxin A was determined by high performance liquid chromatography (HPLC) with fluorescence detector using immunoaffinity column clean-up. The recovery rate of aflatoxin $B_1$ and ochratoxin A were more than 80% and the limits of quantification were 0.13 ${\mu}g/kg$ for aflatoxin $B_1$ and 0.30 ${\mu}g/kg$ for ochratoxin A. Aflatoxin $B_1$ was detected in 33 samples (17.1%) with a range of 0.14~9.67 ${\mu}g/kg$ and ochratoxin A was detected in 40 samples (20.7%) with a range of 0.31~3.31 ${\mu}g/kg$. These results show that the occurrence of aflatoxin $B_1$ and ochratoxin A in dried red pepper and red pepper powder tested in this study is low compared with the standard in Korea Food Code (10 ${\mu}g/kg$ as aflatoxin $B_1$ and 7 ${\mu}g/kg$ as ochratoxin A).

Anti-cancer Activity of Styrax japonica Bark Extrats (때죽나무(Styrax japonica) 수피 추출물의 항암 활성)

  • Kwon, Oh-Woong;Kim, Woo-Jin;Lee, Hak-Ju
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.1
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    • pp.68-77
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    • 2014
  • A compound has been isolated from the methanol extract of Styrax japonica bark using conventional chromatographic methods including silica gel chromatography, TLC and HPLC. The molecular formula of Styraxlignolide F analyzed by spectrometric analyses using FAB-MS, NMR was found to be $C_{27}H_{34}O_{11}Na$. The cytotoxicity of the styralignolide F was showed 15.2% in $1.0mg/m{\ell}$ on human kidney cell (HEK 293). As anticancer activity of $CH_2Cl_2$ fraction, over 60% of AGS and MCF-7 cells were inhibited in concentration of $1.0mg/m{\ell}$. In the results of anticancer test using quantification of Bcl-2, $CH_2Cl_2$ fraction showed lower Bcl-2 and p53 expression than those of styraxlignolide F and other fractions. In apoptosis of human lung carcunoma cancer cell (A549), $CH_2Cl_2$ fraction showed the highest inhibition rate (46.9%) and styralignolide F was the next (43.5%). The $CH_2Cl_2$ fraction showed higher anti-cancer activities than isolated substance (styraxlignolide F), probably due to the crude extract showing synergic effects by other components.

Residual Characteristics of Bistrifluron and Fluopicolide in Korean Cabbage for Establishing Pre-Harvest Residue Limit (생산단계 잔류허용기준 설정을 위한 엇갈이배추 중 bistrifluron과 fluopicolide의 잔류 특성 연구)

  • Lee, Dong Yeol;Jeong, Dong Kyu;Choi, Geun-Hyoung;Lee, Deuk-Yeong;Kang, Kyu Young;Kim, Jin Hyo
    • The Korean Journal of Pesticide Science
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    • v.19 no.4
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    • pp.361-369
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    • 2015
  • This study investigated residual characteristics of bistrifluron and fluopicolide in Korean cabbage, and suggested the pre-harvest residue limits (PHRLs) based on their dissipation patterns and biological half-lives. The pesticides were sprayed on Korean cabbage in two different region under greenhouse conditions at the recommended dose, respectively. The samples for residue analysis were harvested at 0 (2 hr), 1, 2, 4, 6, 8 and 10 days after treatment, and analyzed by HPLC after clean-up with Florisil SPE. The limit of quantification (LOQ) was $0.03mg\;kg^{-1}$ for bistrifluron and fluopicolide, and the recoveries ranged from 87.2-110.6% with below 5% of RSD. The biological half-lives of field I and field II were 3.9 and 4.2 days for bistrifluron and 4.9 and 4.2 days for fluopicolide, respectively. The PHRL of bistrifluron and fluopicolide were recommended as 3.83 and $3.23mg\;kg^{-1}$ for 10 days before harvest, respectively.

Determination of Pyribencarb and Its Metabolite KIE-9749 in Agricultural Commodities by High-Performance Liquid Chromatography (액체크로마토그래피를 이용한 농산물중 pyribencarb와 대사물 KIE-9749의 잔류 분석법)

  • Kwon, Hyeyoung;Hong, Su-Myeong;Kim, Taek-Kyum;Kim, Hyung-Jin;Moon, Byeong-Chul;Kyung, Kee-Sung
    • The Korean Journal of Pesticide Science
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    • v.20 no.3
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    • pp.236-246
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    • 2016
  • The aim of this study was to develop an analytical method for the determination of pyribencarb and its metabolite KIE-9749 in agricultural commodities. The experiment was performed with a range of concentrations $0.05{\sim}2.5{\mu}g/g$ in apple, green pepper, potato, hulled rice, soybean, pear, peach, grape and cucumber. Each samples were extracted with acetone and cleaned by dichloromethane/saline water partition and purified with Florisil solid phase extraction (SPE) cartridge and aminopropyl SPE cartridge. Pyribencarb and KIE-9749 were separated and quantified by HPLC/UVD at 265nm using acetonitrile and water as mobile phase. The recoveries of pyribencarb and KIE-9749 were within 78.3~108.4% and 73.9~113.7% with RSD below 12.2% and 15.0%, respectively. The limits of quantification (LOQ) were both $0.05{\mu}g/g$. LC/ESI-MS/MS was optimized for confirmation of residue identity.

Establishment of Pre-Harvest Residue Limits (PHRL) of Flubendiamide and Pyriofenone on Strawberry (Fragaria ananassa Duch.) (생산단계 잔류허용기준설정을 위한 Flubendiamide 및 Pyriofenone의 딸기(Fragaria ananassa Duch.) 중 경시적 잔류특성 연구)

  • Kim, Hee-Gon;Kim, Ji-Yoon;Hur, Kyung-Jin;Kwon, Chan-Hyeok;Hur, Jang Hyun
    • The Korean Journal of Pesticide Science
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    • v.21 no.1
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    • pp.62-67
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    • 2017
  • In this study, the residue patterns of flubendiamide and pyriofenone registered in the strawberry were investigated to predict pre-harvest residue limit (PHRL). The samples were harvested at 0, 1, 2, 3, 5, 7, 10 days after treatment and the pesticide residues were analyzed by HPLC/UVD. The limit of quantification (LOQ) was $0.01mg\;kg^{-1}$ for flubendiamide and pyriofenone. The recovery levels of flubendiamide and pyriofenone were $90.9{\pm}2.2%$ and $81.9{\pm}0.8%$, $87.7{\pm}2.1%$ and $85.3{\pm}1.1%$ for spiked levels of 0.01 and $0.1mg\;kg^{-1}$, respectively. The values of biological half-lives for field 1 and field 2 were 8.1 and 7.2 days for flubendiamide, 7.0 and 6.9 days for pyriofenone. According to these results, we recommends the level of PHRL on strawberry for flubendiamide and pyriofenone as 1.87 and $3.76mg\;kg^{-1}$ at 10 days before harvest, respectively.

Dissipation Pattern of Fungicides Boscalid and Pyraclostrobin in Jujube (대추 중 살균제 boscalid와 pyraclostrobin의 잔류 소실 특성)

  • Jo, Hyeong-Wook;Sohn, Sang-Hyun;Kim, Kyoung-Jin;Hwang, Gi-Jun;Jo, Beom-Haeng;Gil, Seok-Ju;Kwon, Chan-Hyeok;Moon, Joon-Kwan
    • The Korean Journal of Pesticide Science
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    • v.21 no.1
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    • pp.17-25
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    • 2017
  • Dissipation pattern and biological half-lives of fungicides boscalid and pyraclostrobin were calculated on jujube. The pesticides were sprayed on jujube in two different field at the standard rate, respectively. The raw agricultural commodities were harvested at 0 (2 hr), 1, 3, 5, 7, 10 and 14 days after treatment, and analyzed by HPLC/DAD. The method limit of quantification (MLOQ) was $0.02mg\;kg^{-1}$ for boscalid and pyraclostrobin. The recovery ranged 101.8~109.3% with below 5% of CV (Coefficient of variation) for boscalid and 104.2~115.4% with below 5% of CV for pyraclostrobin. An average initial deposit at field 1 and field 2 samples were observed 0.40 and $0.48mg\;kg^{-1}$ for boscalid and, 0.76 and $0.57mg\;kg^{-1}$ for pyraclostrobin, respectively. The biological half-lives of field 1 and field 2 were 11.0 and 13.2 day for boscalid, and 6.1 and 12.7 days for pyraclostrobin.

Simultaneous Determination of B Group Vitamins in Supplemented Food Products by High Performance Liquid Chromatography-Diode Array Detection

  • Suh, Joon-Hyuk;Yang, Dong-Hyug;Lee, Byung-Kyu;Eom, Han-Young;Kim, Un-Yong;Kim, Jung-Hyun;Lee, Hye-Yeon;Han, Sang-Beom
    • Bulletin of the Korean Chemical Society
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    • v.32 no.8
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    • pp.2648-2656
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    • 2011
  • A simple HPLC-DAD method was developed and validated to determine B group vitamin content (thiamine, riboflavin, nicotinamide, pantothenic acid, pyridoxine and folic acid) in supplemented food samples, i.e., infant formula, cereal, low-calorie food, a multi-vitamin pill and a vitamin drink. In this study, the most significant advantages were simultaneous determination of the six B group vitamins in various food matrices and a small number of sample treatment steps that required only an organic solvent, acetonitrile. Moreover, this method prevents reduction of column durability, because the mobile phase does not contain ion-pairing reagents. Analytes were separated on a Develosil RPAQUEOUS $C_{30}$ (4.6 mm ${\times}$ 250 mm, 5 ${\mu}M$ particle size) column with a gradient elution of acetonitrile and 20 mM phosphate buffer (pH 3.0) at a flow rate between 0.8 and 1.0 mL/min. Detection was performed at 275 nm, except for that of pantothenic acid (205 nm). The calibration curves for all six vitamins showed good linearity with correlation coefficients ($r^2$) higher than 0.995. The developed method was validated with respect to linearity, intra- and inter-day accuracy and precision, limit of quantification (LOQ), recovery and stability. The method showed good precision and accuracy, with intra- and inter-assay coefficients of variation less than 15% at all concentrations. The recovery was carried out according to the standard addition procedure, with yields ranging from 89.8 to 104.4%. This method was successfully applied to the determination of vitamin B groups in supplemented food products.

Residue Dissipation Patterns of Indoxacarb and Pymetrozine in Broccoli under Greenhouse Conditions (시설재배 브로콜리 중 Indoxacarb 및 Pymetrozine의 잔류 소실특성)

  • Yang, Seung-Hyun;Lee, Jae-In;Choi, Hoon
    • Korean Journal of Environmental Agriculture
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    • v.39 no.1
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    • pp.75-82
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    • 2020
  • BACKGROUND: This study was carried out to establish pre-harvest residue limits (PHRLs) of indoxacarb and pymetrozine in broccoli under greenhouse conditions, based on dissipation patterns and biological half-lives of pesticides during 10 days after application. METHODS AND RESULTS: The field studies were conducted in two different greenhouse, located in Chungju-si (Field 1) and Gunsan-si (Field 2). Samples were collected at 0, 1, 2, 3, 5, 7 and 10 days after spraying pesticide suspension. The analytical methods for indoxacarb and pymetrozine using HPLC-DAD were validated by recoveries ranging of 94.3-105.4% and 81.8-96.0%, respectively, and MLOQ (Method Limit of Quantification) of 0.05 mg/kg. Biological half-lives of indoxacarb and pymetrozine were 2.9 and 3.2-3.8 days in broccoli, respectively. The lower 95% confidence intervals of dissipation rate constant of indoxacarb were determined as 0.1508 (Field 1) and 0.2017 (Field 2), whereas those of pymetrozine were calculated as 0.1489 (Field 1) and 0.1577 (Field 2). CONCLUSION: The significant differences were not observed between the dissipation rates of indoxacarb and pymetrozine in broccoli. The major factor affecting residue dissipation was the dilution effect by fast growth. The PHRLs for 10 days prior to harvest were recommended as 30.06 (Field 1) and 18.07 (Field 2) mg/kg for indoxacarb, and 4.84 (Field 1) and 4.43 (Field 2) mg/kg for pymetrozine, respectively.