• Title/Summary/Keyword: HPLC/PDA

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Development of HPLC-UV method for detection and quantification of seven organic acids in animal feed (사료 중 유기산 7종 동시분석법 개발 및 유통 사료의 모니터링)

  • Kim, Jin kug;Lee, Mi Jin;Lee, Ye Ji;Kim, Hye Jin;Jeong, Min Hee;Kim, Ho Jin
    • Analytical Science and Technology
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    • v.29 no.4
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    • pp.202-208
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    • 2016
  • 본 연구에서는 사료 첨가제로 이용되고 있는 유기산 7종(formic acid, malic acid, lactic acid, acetic acid, citric acid, fumaric acid, propionic acid)의 동시분석법 개발을 위한 연구를 실시하였다. 7종의 화합물은 표준물질의 Retention time과 UV spectra를 통해 구별하였고, 분석법 검증은 직선성, 민감성, 선택성, 정확성, 정밀성을 통하여 검증하였다. 그 결과로 LOD와 LOQ의 범위가 각각 43~26,755 μg/kg, 12-8,026 μg/kg으로 설정하였고, 평균 회수율이 79.3~95.2%로 우수하게 보였으며, intra-day, inter-day에 대한 전반적인 상대 표준 편차(%RSD)는 3.2% 미만으로 나타났다. 이와 같이 검증된 자료를 통해 유기산의 동시분석에 대한 직선성, 민감성, 선택성, 정확성 및 정밀성을 확인하였고, 높은 수준을 나타냄을 알 수 있었다. 이를 바탕으로 유기산이 검출되는 단미사료 46 가지를 분석에 적용하여 진행하였고, 정량과 동시분석 검출을 위한 방법은 RP-HPLC/UV 검출기를 이용하여 성공적으로 개발되었다. 따라서 본 연구결과를 바탕으로 하여 사료 중의 유기산의 분석이 신속하고 정확해졌을 뿐 아니라, 다른 종류의 사료 또한 이를 적용하여 효율적으로 이용할 수 있을 것으로 판단된다.

Isolation and Characterization of Surfactin-producing Bacillus amyloliquefaciens YJ07 from Spent Mushroom (Pleurotus eryngii) Substrates (새송이버섯 수확 후 배지로부터 surfactin 생성 Bacillus amyloliquefaciens YJ07의 분리 및 특성)

  • Shin, Pyung Gyun;Yoo, Young Bok;Cho, Yong Un;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.180-185
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    • 2011
  • Spent mushroom substrates (SMS) is a by-product remaining after a crop of mushrooms. About four surfactin-producing strains were isolated from SMS (Pleurotus eryngii). Among of them, one isolate, which designated to YJ07, potentially showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium. The biochemical characteristics of the strain YJ07 was similar with Bacillus subtilis and Bacillus amyloliquefaciens by Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis of the strain YJ07 also showed that the strain YJ07 was most closely related to Bacillus amyloliquefaciens with sequence similarity of 99.5%. On the basis of their biochemical characteristics and phylogenetic distinctiveness, the strain YJ07 was classified within the genus Bacillus as Bacillus amyloliquefaciens YJ07. The antifungal compound from B. amyloliquefaciens YJ07 was similar to lipopeptide surfactin from Bacillus subtilis by TLC and HPLC analysis.

Antioxidant Effects and Spectrophotometry Analysis of Alkaloids from Vinca minor (빈카 마이너 추출물에 함유된 알칼로이드들의 분광학적 분석 및 항산화 효능 연구)

  • Kim, Jun-Sub;Joo, Ji-Hye;Kang, Jo-Eun;Jang, Sun-Dong;Jung, Kyung-Hwan;Moon, Gi-Seong;Lee, Hyang-Yeol
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.1
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    • pp.16-22
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    • 2015
  • Alkaloids of Vinca minor L. were characterized by HPLC equipped with the photo diode array detector and compared their UV/vis absorption spectra with an index material such as vincamine. UV/vis absorption spectra of more than 9 compounds belong to various classes such as flavonoids were shown and characterized. Extracts with ethanol or hot water were tested against the DPPH (2,2-diphenyl-1-picrylhydrazyl) free-radical to investigate their antioxidant activity. Based on the results, those extracts show about 14-15% of antioxidant activity of quercetine and vitamin C used as standards.

Analysis of tetracyclines in shrimp samples based on a two-step extraction approach prior to high-performance liquid chromatography

  • Thinnakorn Sukkhunthod;Thanakorn Pluangklang;Sumita Boonnab;Sira Sansuk;Phitchan Sricharoen;Maliwan Subsadsana
    • Analytical Science and Technology
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    • v.37 no.4
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    • pp.211-219
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    • 2024
  • This study presents a sensitive and reliable method for determining tetracycline (TC), oxytetracycline (OTC), and chlortetracycline (CTC) residues in shrimp samples. A two-step process involving liquid-liquid extraction (LLE) followed by solid-phase extraction (SPE) was developed prior to HPLC analysis. The target analytes were effectively extracted using EDTA/McIlvaine buffer (pH 4.0): methanol (80:20, %v/v), with subsequent clean-up using a C18 SPE cartridge. HPLC separation was conducted on a C18 column (250 mm × 4.6 mm i.d., 5 ㎛) at 30 ℃, using 0.01 % trifluoroacetic acid (A) and acetonitrile (B) as the mobile phase. A gradient elution protocol was applied, transitioning from 85(A):15(B) %v/v to 70(A):30(B) %v/v at 7 min, with a 5 min hold, followed by adjustment to 85(A):15(B) %v/v for 13-14 min. The detection was performed using photodiode array (PDA) at 365 nm with a flow rate of 1.0 mL/min. The calibration curves exhibited good linearity within a concentration range of 0.4-6.0 ㎍/mL (R2 > 0.995). The limits of detection (LOD) for TC, OTC, and CTC in shrimp were 0.034, 0.029, and 0.021 ㎍/mL, respectively. The limits of quantitation (LOQ) for TC, OTC, and CTC were found to be 0.114, 0.097, and 0.071 ㎍/mL, respectively. Recoveries of TC, OTC, and CTC from spiked shrimp samples ranged from 91.0 % to 95.5 %, 92.4 % to 97.2 %, and 93.3 % to 96.6 %, respectively. This method was successfully applied to the determination of TC, OTC, and CTC residues in shrimp samples sourced from various local markets.

Determination of amitraz by high-performance Liquid chromatography with photodiode array detection and method validation (HPLC-PDA를 이용한 Amitraz의 분석법 확립 및 검증)

  • Yun, Hyeong-Jun;Yun, So-Mi;Lee, Myoung-Heon;Son, Seong-Wan
    • Korean Journal of Veterinary Research
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    • v.48 no.1
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    • pp.33-38
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    • 2008
  • In veterinary medicine amitraz has been used as an insecticide to eliminates mites, lice, and ticks in dogs, cats, goats, swine and cattle. The objective of present study was to developed an analytical method using one-step extraction and determination of the amitraz in veterinary drugs by liquid chromatography (LC). The amitraz was analyzed by LC equipped with Waters XTerra RP18 ($4.8{\times}250mm;\;5{\mu}m;\;Waters,\;USA$) analytical column, using 75% acetonitrile (acetonitrile/D.W; 75/25) at 1.0 ml/ min. The UV-VIS detection of amitraz was made at 290 nm. Calibration graphs were linear with very good correlation coefficients ($r^2>0.9999$) from $80{\sim}120{\mu}g/ml$. The limit of detection was $0.09{\mu}g/ml $ and limit of quantification was $0.27{\mu}g/ml $. The method showed good intra-day precision (CV 0.05~0.09%) and inter-day precision (CV 0.06~0.18%).

Comparison of Valerenic Acids and Valepotriates Production According to the Culture Conditions for Cultured Roots of Valeriana fauriei var. dasycarpa Hara

  • Li, Mei-Yang;Ahn, Jun-Cheul;Kim, Kwang-Soo;Kim, Ok-Tae;Park, Yoon-Jung;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.2
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    • pp.101-106
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    • 2006
  • We established a practical method for rapid and large-scale production of Valeriana fauriei var. dasycarpa Hara roots by bioreactor culture and confirmed valerenic acids and valepotriates production. We also compared valerenic acids and valepotriates production patterns according to various media conditions. Among the media tested, B5 medium gave the maximum biomass production of 101 g fresh weight, which was a 5.03-fold multiplication rate obtained 4 weeks after inoculation of 20 g of fresh weight. The best production of total valerenic acids $(7.86\;mg/l)$ and valepotriates $(8.96\;mg/l)$ was B5 medium.

Isolation and Purification of Several Substances Produced by Fusarium graminearum and Their Antimicrobial Activities (Fusarium graminearum이 생산하는 몇가지 물질의 분리정제 및 항균 활성)

  • 김병섭;김건우;이종규;이인원;조광연
    • Korean Journal Plant Pathology
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    • v.11 no.2
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    • pp.158-164
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    • 1995
  • 토마토의 엽권에서 분리한 Fusariym graminearum이 분비하는 물질은 벼 도열병균(Pyricularia oryzae)의 여러 종의 식물병원 진균에 대한 항균활성을 나타내었으며, 이러한 활성물질을 PDA에서 본 균을 배양 한 후 chloroform으로 추출하여 분리정제 하였다. HPLC에 의하여 5종류의 활성 물질을 분획하였으며, 그중 1번(F402) 화합물을 벼 도열병균(P. oryzae)을 포함한 22개 식물 병원 진균에 대하여 살균 활성범위를 조사한 결과, 이 화합물은 50$\mu\textrm{g}$/ml 농도에서 Pythium ultimum, Rhizoctonia solani, Sclerotinia sclerotiorum은 전혀 억제하지 못하였으며, Phytophthora spp., Cladosporium fulvum, Fusarium spp., Corynespora cassicola에는 어느 정도의 활성이 있었지만 낮게 나타났고, P. oryzae, Cochliobolus miyabeanus, Alternaria solani는 100% 억제하여 활성이 높게 나타났다. 또한 장내 세균에 대한 활성을 MIC로 비교할 때 Streptococcus pyogenes, Streptococcus faecium에 대하여는 각각 12.5, 25 $\mu\textrm{g}$/ml였고 Staphylococcus aureus는 25-50$\mu\textrm{g}$/ml으로 나타났으며, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella aerogenes, Enterobacter cloacae에서는 100$\mu\textrm{g}$/ml 이상으로 활성이 나타나지 않았다. F402를 200$\mu\textrm{g}$/ml의 농도로 직접 살포한 식물체에서의 방제효과는 벼도열병, 벼 깨씨무늬병, 보리 흰 가루병에 대하여는 80%이상이었으나, 벼 잎집무늬마름병, 오이 잿빛곰팡이병, 토마토 역병, 밀 녹병에서는 낮았다.

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Comparison of the antioxidant properties and flavonols in various parts of Korean red onions by multivariate data analysis

  • Park, Mi Jin;Ryu, Da Hye;Cho, Jwa Yeong;Ha, In Jong;Moon, Jin Seong;Kang, Young-Hwa
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.919-927
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    • 2018
  • To compare the antioxidant properties and flavonols in various parts; dry skin (DS) and edible portion (EP), of 8 red onions (Allium cepa L, ROs), total content of phenolics (TPC), flavonoids (TFC), and anthocyanins (TAC) and DPPH radical scavenging properties were estimated and the content of six flavonols were quantified by HPLC-PDA analysis. The major component of DS and EP of RO was quercetin and quercetin-4'-glucoside, respectively. Score plots of the PCA and PLS-DA were segregated by flavonols content and antioxidant properties according to the EP and DS of ROs. Loading plot of the PCA showed that the quercetin and sum of flavonol content were highly correlated with antioxidant activity of ROs. Therefore, flavonol content and antioxidant activity can be used as markers for distinct parts of ROs.

Antimicrobial polyhydroxybutyrate submicron fiber mat loaded with extract of Hypericum perforatum

  • Beran, Milos;Horna, Ales;Vorisek, Viktor;Berkova, Eliska;Korinkova, Radka;Trousil, Vojtech;Hrubanova, Marketa
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.257-270
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    • 2022
  • The aim of this work was to prepare a new biodegradable polyhydroxybutyrate (PHB) submicron fiber mat loaded with hypericin-rich Hypericum perforatum raw extract by centrifugal spinning technology, an alternative approach to the traditional method of electrospinning to fabricate nanofibers or microfibers from solutions at high speed and low cost. Hypericins in methanol/acetone extract of H. perforatum were determined by UHPLC-MS/MS and HPLC/PDA. Submicron fiber mats composed of pure PHB or PHB enriched with H. perforatum extract were prepared using a pilot plant demonstrator for the centrifugal spinning technology and characterized by SEM. Singlet oxygen production was quantified by the 1,3-diphenylisobenzofuran (DPIBF) method in hexane. The results proved a significant production of singlet oxygen by the prepared submicron fiber mat. We also found a significant antibacterial activity against the bacterial strain Escherichia coli CCM 5417 by a method in accordance with JIS Z 2801/ISO 22196 standards. The H. perforatum extract-enriched PHB submicron fiber mats showed potential for the development of self-cleaning and antimicrobial air filters.

Methyl Jasmonate-mediated Enhancement of Phenylethanoid Glycoside in Callus from Abeliophyllum distichum (cultivar Okhwang1)

  • Tae-Won Jang;So-Yeon Han;Da-Yoon Lee;Seo-Yoon Park;Woo-Jin Oh;Jae-Ho Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2023.04a
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    • pp.53-53
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    • 2023
  • Abeliophyllum distichum, one of the Korean endemic plant, is a significant pharmaceutical plant resource. A. distichum with phenylethanoid glycoside can use to regulate the development of cancer, DNA damage with radicals, and the generation of inflammatory mediators. In this study, we investigated whether the biomass, content of phenylethanoid glycoside, and growth rate of callus derived from A. distichum (cultivar Okhwang1, CAD) change in the absence or presence of plant hormones (2,4-Dichlorophenoxyacetic acid; 2, 4-D and 1-Naphthaleneacetic acid; NAA). The results showed that the best biomass, the growth rate of callus, and the contents of phenylethanoid glycoside were cultivated on Murashige and Skoog (MS) growth medium fortified with 1 ppm 2,4-D + 2 ppm NAA after 4 weeks. In a further study, CAD was cultivated on MS growth medium fortified with an elicitor (Methyl Jasmonate, MeJA). The results showed that CAD turned to brown color and fragile form with the elicitor. HPLC-PDA analysis revealed that the contents of phenylethanoid glycoside in the elicitor-treated group were higher than in the elicitor-non-treated group. These results are consistent with the findings of Arano-Varela H et al.,'s study which is that acteoside production can increase after the treatment of MeJA. Therefore, this study can be used to develop an effective and sustainable production of useful substances as an alternative to plant cultivation.

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