• Title/Summary/Keyword: HPLC-RI

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Establishment of Analytical Method for Fenhexamid Residue in Korean Cabbage, Apple, Mandarin and Green Pepper (HPLC를 이용한 배추, 사과, 감귤, 고추 중 살균제 Fenhexamid의 정밀 분석법 확립)

  • Lee, Hye-Ri;Riu, Myoung-Joo;Park, Hee-Won;Na, Ye-Rim;Song, Hyuk-Hwan;Keum, Young-Soo;Zhu, Yongzhe;Kim, Jeong-Han
    • The Korean Journal of Pesticide Science
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    • v.13 no.4
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    • pp.223-231
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    • 2009
  • This study was performed to develop a precise single residue analytical method of fungicide fenhexamid in representative crops for general residue analytical method which could be applied to most of crops. Korean cabbage, mandarin, apple and green pepper were selected, macerated, extracted with acetone, concentrated and partitioned with dichloromethane. Then the extracts were concentrated and cleaned-up through Florisil column with ethyl acetate/0.1% acetic acid in hexane [15:85, (v/v)] before concentration and analysis with HPLC. LOQ (Limit of Quantitation) of fenhexamid was 1 ng (S/N>10) and MQL (Method Quantitative Limit) was 0.01 mg/kg. Recoveries were measured at two fortification levels (10 MQL and 50 MQL) on crop samples and ranged from 85.2% to 94.8% (mean recoveries) and coefficients of variation were <10% regardless of sample type.

Lipase Inhibitory Mode of Dieckol Isolated from Eisenia bicyclis Ethanol Extract (Eisenia bicyclis 에탄올 추출물로부터 분리한 Dieckol의 Lipase 저해 Mode)

  • Jung, Seul-A;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;Cho, Ji-Young;Kim, Tae-Wan;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.112-118
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    • 2013
  • This study was performed to investigate the possible use of Eisenia bicyclis (EB) ethanol extract to inhibit activity against lipase. In tests, the lipase inhibitory activity of EB ethanol extract was noted as being 43, 27, and 24% at concentrations of 5, 2.5, and 1 mg/ml, respectively. Isolation was carried out by liquid and liquid extraction, silica-gel column chromatography, and HPLC. The results showed that the lipase inhibitory activity of the ethyl acetate (EA) fraction from EB ethanol extract exhibited the strongest lipase inhibitory activity with an $IC_{50}$ value of 1.31 mg/ml. The EA fraction was separated using silica-gel column chromatography and we obtained 22 sub-fractions. Amongst them, the EA1 fraction showed the highest lipase inhibitory activity with an $IC_{50}$ value of 0.54 mg/ml. Eight peaks were obtained from the EA1 fraction by HPLC. Fraction 5 also showed a strong lipase inhibitory activity with an $IC_{50}$ value of 0.37 mg/ml. The fraction 5 was identified as dieckol and the inhibition pattern analyzed from Lineweaver-Burk plots revealed a non-competitive inhibitor. These results suggest that EB has potential as a natural anti-obesity agent.

Anti-atopic dermatitis effects of Parasenecio auriculatus via simultaneous inhibition of multiple inflammatory pathways

  • Kwon, Yujin;Cho, Su-Yeon;Kwon, Jaeyoung;Hwang, Min;Hwang, Hoseong;Kang, Yoon Jin;Lee, Hyeon-Seong;Kim, Jiyoon;Kim, Won Kyu
    • BMB Reports
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    • v.55 no.6
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    • pp.275-280
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    • 2022
  • The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.

Isolation and characterization of antifungal violacein producing bacterium Collimonas sp. DEC-B5 (항진균활성 violacein 색소를 생산하는 Collimonas sp. DEC-B5 균주의 분리 및 특성)

  • Lee, Ye-Rim;Mitchell, Robert J.;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.212-219
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    • 2016
  • Forty-nine pigments were extracted from the collections of 106 pigment producing bacteria from the plant rhizosphere soil. Antibacterial activity test was performed in the subjects of the extracted pigments with plant pathogenic bacteria including Xanthomonas axonopodis and Xanthomonas campestris, and with plant pathogenic fungi including Botrytis cinerea, Colletotrichum acutatum, and Fusarium oxysporum. The yellow pigment by Chryseobacterium sp. RBR9 and the red pigment by of Methylobacterium sp. RI13 showed the antibacterial activities against Xanthomonas axonopodis and Xanthomonas campestris. The violet pigment by Collimonas sp. DEC-B5 showed the antibacterial activity as well as the antifungal activities against Botrytis cinerea and Fusarium oxysporum. Especially, the violet pigment inhibited the growth of Botrytis cinerea more than 65% at MIC $20{\mu}M$. Upon the HPLC analysis result for the isolation of pigment with antifungal activity, violacein (91.6%) and deoxyviolacein (8.4%) were isolated for the pigment by Collimonas sp. DEC-B5. The production amount of the pigment was increased more than 10 times higher when D-mannitol 1.5% and yeast extract 0.2% were added as the nitrogen source to SCB medium. This study suggests that produced violacein by Collimonas sp. DEC-B5 will be effective to control strawberry gray-mold rot fungi by its preventive activity.

Biodegradation of Ochratoxin A by Aspergillus tubingensis Isolated from Meju

  • Cho, Sung Min;Jeong, Seong Eun;Lee, Kyu Ri;Sudhani, Hemanth P.K.;Kim, Myunghee;Hong, Sung-Yong;Chung, Soo Hyun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1687-1695
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    • 2016
  • Ochratoxin A (OTA), a mycotoxin, contaminates agricultural products and poses a serious threat to public health worldwide. Microbiological methods are known to be a promising approach for OTA biodegradation because physical and chemical methods have practical limitations. In the present study, a total of 130 fungal isolates obtained from 65 traditional Korean meju (a fermented starter for fermentation of soybeans) samples were examined for OTA-biodegradation activity using thin-layer chromatography. Two fungal isolates were selected for OTA-biodegradation activity and were identified as Aspergillus tubingensis M036 and M074 through sequence analysis of the beta-tubulin gene. After culturing both A. tubingensis isolates in Soytone-Czapek medium containing OTA (40 ng/ml), OTA-biodegradation activity was analyzed using high-performance liquid chromatography (HPLC). Both A. tubingensis strains degraded OTA by more than 95.0% after 14 days, and the HPLC analysis showed that the OTA biodegradation by the A. tubingensis strains led to the production of ochratoxin α, which is much less toxic than OTA. Moreover, crude enzymes from the cultures of A. tubingensis M036 and M074 led to OTA biodegradation of 97.5% and 91.3% at pH 5, and 80.3% and 75.3% at pH 7, respectively, in a buffer solution containing OTA (40 ng/ml) after 24 h. In addition, the OTA-biodegrading fungi did not exhibit OTA production activity. Our data suggest that A. tubingensis isolates and their enzymes have the potential for practical application to reduce levels of OTA in food and feed.

Assay of $\beta$-Galactosidase Using High Performance Liquid Chromatography (고속액체크로마토그래피를 이용한 유당분해효소의 활성도 측정)

  • Shin, Myung Gon;Chang, Pahn Shick;Min, Bong Kee;Kim, Sun Chang
    • Analytical Science and Technology
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    • v.5 no.4
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    • pp.465-469
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    • 1992
  • An analytical procedure is presented for the quantitative determination of lactose, glucose, and galactose in the hydrolyzate of lactose by ${\beta}$-galactosidase with high-performance liquid chromatography. An Aminex HPX-87C column at $85^{\circ}C$ and refractive index detector were used to resolve lactose, glucose, and galactose in only 12 minutes with distilled and deionized water as a mobile phase. The validity of high-performance liquid chromatography as a method for the assay of ${\beta}$-galactosidase was supported by recovery experiments and comparision of results with those by ONPG method, a spectrophotometric assay. The procedure was appropriate for determination of sugars in the enzyme reaction mixture and could by applied to analysis of ${\beta}$-galactosidase activity.

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Structure-Guided Identification of Novel Phenolic and Phenolic Amide Allosides from the Rhizomes of Cimicifuga heracleifolia

  • Yim, Soon-Ho;Kim, Hyun-Jung;Jeong, Na-Ri;Park, Ki-Deok;Lee, Young-Ju;Cho, Sung-Dong;Lee, Ik-Soo
    • Bulletin of the Korean Chemical Society
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    • v.33 no.4
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    • pp.1253-1258
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    • 2012
  • Two phenolic allopyranosides and two phenolic amide allopyranosides, along with eight known phenolic compounds, including cimicifugic acids, shomaside B, fukiic acid, isoferulic acid, and piscidic acid, were isolated from the n-butanolic extract of rhizomes of Cimicifuga heracleifolia. On-line spectroscopic data for UV, NMR, and MS from a combination of LC-NMR and LC-MS techniques directly and rapidly provided sufficient structural information to identify and confirm all the structures of major phenolic compounds in the extract, in addition to their HPLC profiles. This combined analytic information was then used as a dereplication tool for structure-guided screening in order to isolate unknown phenolic compounds in the extract. Successive fractionation and purification using semi-preparative HPLC acquired four unknown allopyranosides, and their structures were identified as cis-ferulic acid 4-O-${\beta}$-D-allopyranoside, trans-ferulic acid 4-O-${\beta}$-D-allopyranoside, trans-feruloyltyramine 4-O-${\beta}$-D-allopyranoside, and trans-feruloyl-(3-O-methyl)dopamine 4-O-${\beta}$-D-allopyranoside, based on a subsequent spectroscopic interpretation.

Monitoring of Sugar and Low-Calorie Sweetener Content in Alcoholic Beverages (유통 주류의 당류 함량 및 저칼로리 감미료 사용 실태 조사)

  • Young-Sun Cho;Jin-A Jeong;Hye-Jung Kwon;Han-Taek Kim;Ji-Yeon Lee;Hye-Won Lim;Eun-Bin Lee;Hye-Jin Kim;Won-Joo Lee;Myung-Jin Lee
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.281-287
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    • 2024
  • This study investigated the total sugar and low-calorie sweetener content in 72 alcoholic beverages. The samples included 10 takjus, 6 yakjus, 5 cheongjus, 5 beers, 12 fruit wines, 5 sojus, 5 general distilled alcoholic beverages, 9 liqueurs, and 15 other alcoholic beverages. Sugar and allulose content were analyzed using HPLC-RI, and the content of five sweeteners was analyzed using HPLC-UV and LC-MSMS. The average sugar content in the alcoholic beverages was 4.13±5.16 g/100 g. When categorized by type, the sugar content ranged from 0.00 to 8.92 g/100 g, 0.00 to 30.55 g/100 g, and 0.14 to 17.02 g/100 g in fermented (takju, yakju, cheongju, beer, and fruit wine), distilled (soju, general distilled alcoholic beverages, and liqueur), and other alcoholic beverages, respectively. Sugar content was the highest in liqueur, with the average content being 12.41±9.66 g/100 g. Among low-calorie sweeteners, acesulfame potassium, sodium saccharin, aspartame, and sucralose were detected in concentrations ranging from 28.6-121.5, 42.3, 34.1-141.5, and 23.3-88.1 mg/kg, respectively. Cyclamate and allulose were not detected in any of the alcoholic beverages. Sweeteners were detected in 24 out of the 72 alcoholic beverages, and their content complied with the standards stipulated for food additives.

Variation of Saponin Content in the Decoctions of Platycodi Radix (II) (길경(桔梗)의 추출조건에 따른 사포닌함량의 변화 (제2보))

  • Yoo, Dae-Seok;Choi, Yeon-Hee;Cha, Mi-Ran;Choi, Chun-Whan;Kim, Mi-Ri;Yon, Gyu-Hwan;Hong, Kyung-Sik;Lee, Byung-Hoe;Kim, Eun-Ju;Cho, Sang-Wook;Kim, Young-Sup;Ryu, Shi-Yong;Kang, Jong-Seong
    • Korean Journal of Pharmacognosy
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    • v.41 no.2
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    • pp.147-152
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    • 2010
  • The capability of the solvents for extracting the bioactive saponins from the roots of Platycodon grandiflorum (Campanulaceae) was investigated to obtain an ideal extract which contained bioactive saponins with high quality and high quantity. The content of eight representative saponins in extracts, such as deapioplatycoside E, platycoside E, platyconic acid A and platycodin D, platycodin $D_3$, platycodin $D_2$, polygalacin $D_2$, polygalacin D were analyzed simultaneousely by the modified HPLC analytical method. The validation test of the modified qualitative and quantitative analytical method employing the ELSD equipped HPLC for eight representative saponins in the roots extract of P. grandiflorum showed a good linearity, precision and accuracy. the correlation coefficient ($r^2$) values of the calbration curves for each saponins were observed to be over 0.9990. LOD and LOQ of each saponin was calculated as $0.10{\mu}g{\sim}0.40{\mu}g$ (LOD) and $0.40{\mu}g{\sim}0.80{\mu}g$ (LOQ), respectively. Recovery rates of each saponin were also calculated as over 98%, respectively. With exception of two saponins, platyconic acid A and platycodin D, The content of eight saponins in extracts was decreased proportionally to the increment of the water ratio of solvent for extraction. Consequently, as aquous alcohol was used as a solvent for extracting the saponin components from powdered roots of P. grandiflorum, the water content in the aquous alcohol was seemed to be a critical factor for extracting efficacy. The 60-80% ratio of alcohol in the aquous alcohol were deduced to be suitable and recommendable for the preparation of roots extract of P. grandiflorum which contained saponins with high quality and high quantity.

Analyses of Capsaicinoids and Ascorbic Acid in Pepper (Capsicum annum L.) Breeding Lines (고추의 육성계통에 따른 Capsaicinoids와 Ascorbic Acid 분석)

  • Jung, Mi-Ri;Hwang, Young;Kim, Hae-Young;Jeong, Heong-Sang;Park, Ji-Sung;Park, Dong-Bok;Lee, Jun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.11
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    • pp.1705-1709
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    • 2010
  • Peppers (Capsicum annuum L.) are a rich source of phytochemicals including polyphenolics, flavonoids, capsaicinoids and ascorbic acid. Capsaicinoids are a group of 12 or more related alkaloids responsible for the pungent sensation in the fruits of the genus Capsicum. Ascorbic acid is another functional and nutritional constituent of peppers. In this study, the contents of two major capsaicinoids (capsaicin and dihydrocapsaicin) and ascorbic acid in 131 pepper breeding lines were quantified by HPLC. In 131 pepper breeding lines, capsaicin and dihydrocapsaicin contents were in the range of 0.0 to 219.6 and 0.0 to 110.8 mg/100 g, respectively. The breeding lines with higher capsaicin content contained higher dihydrocapsaicin content as well. Ascorbic acid contents were 264.9 to 1695.5 mg/100 g for the 131 pepper breeding lines. The analytical method validation parameters including accuracy, repeatability, and reproducibility were calculated to ensure the method's validity. This study provides basic information to plant breeders and biotechnologists who are planning to breed genotypes with high content of phytochemicals.