• Title/Summary/Keyword: HPLC/PDA

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Analytical Method Validation of Quercetin in Changnyeong Onion Extract as a Functional Ingredient for Functional Health Food (건강기능식품 기능성원료로서 창녕양파추출액의 지표성분 Quercetin 분석법)

  • Jeon, Seon-Young;Jeong, Eun-Jeong;Baek, Jeong-Hwa;Cha, Yong-Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.4
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    • pp.565-569
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    • 2011
  • Validation of quercetin as a marker compound in the standardization of Changnyeong onion extract developed for functional health food was attempted by analytical method. The specificity was satisfied with retention time and photo diode array (PDA) spectrum by analysis of quercetin using HPLC and comparison with standard compound. It showed a high linearity in the calibration curve as coefficient of correlation ($R^2$) of 0.9986, and the limit of detection (LOD) and limit of quantitation (LOQ) were 0.2 mg/L and 0.5 mg/L, respectively. Recovery rate test with quercetin concentration of 0.05, 0.075 and 0.1 mg/mL was revealed in the high range of 82.36~95.26%, 82.70~98.24% and 87.91~95.11%, respectively. The intra-day and inter-day precision in quercetin for Changnyeong onion extracts was 0.10~3.28% and 0.96~5.79%, respectively. Therefore, application of quercetin was validated in analytical method as a marker compound in Changnyeong onion extracts.

Isolation and Characterization of Thermophilic Bacillus sp. UJ03 from Spent Mushroom (Flammulina velvtipes) Substrates (팽이버섯 수확 후 배지로부터 고온성 Bacillus sp. UJ03의 분리 및 특성)

  • Gal, Sang-Wan;Cho, Soo-Jeong
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1481-1486
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    • 2011
  • Spent mushroom substrate (SMS) is a by-product remained after a crop of mushrooms. About seven thermophilic strains were isolated from SMS (Flammulina velvtipes). Among them, one isolate, designated UJ03, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain UJ03 produced cellulase and xylanase as extracellular hydrolases. The strain UJ03 was identified as a member of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain UJ03 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain UJ03 was classified within the genus Bacillus, for which the name Bacillus sp. UJ03 is proposed. The antifungal compound from Bacillus sp. UJ03 was similar to lipopeptide iturin A of Bacillus sp.

Structural Identification of $Siderophore_{AH18}$ from Bacillus subtilis AH18, a Biocontrol agent of Phytophthora Blight Disease in Red-pepper (Bacillus subtilis AH18의 고추역병 방제능과 $Siderophore_{AH18}$의 구조분석)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.326-335
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    • 2008
  • The siderophore ($siderophore_{AH18}$) of Bacillus subtilis AR18 was determined to be one of catechol type and purified by using Amberlite XAD-2, Sephadex LR-20 chromatography, and reversed-phase RPLC. The $Siderophore_{AH18}$ was identified bacillibactin with its structure by GC-MS, $^1H$-NMR, and $^{13}C$-NMR. $Siderophore_{AH18}$ (bacillibactin) had been confirmed its molecular weight of 883 and chemical structure of $(2,3-dihydroxybenzoate-glycine-threonine)_3$. Purified $siderophore_{AH18}$ showed strong biocontrol ability towards the spore of Phytophthora capsici on PDA and able to effectively suppress (55%) P. capsici causing red-pepper blight in the pot in vivo test.

Enhanced Lovastatin Production by Solid State Fermentation of Monascus ruber

  • Xu Bao-Jun;Wang Qi-Jun;Jia Xiao-Qin;Sung Chang-Keun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.78-84
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    • 2005
  • The purpose of this study was to optimize the solid state cultivation of Monascus ruber on sterile rice. A single-level-multiple-factor and a single-factor-multiple-level experimental design were employed to determine the optimal medium constituents and to optimize carbon and nitrogen source concentrations for lovastatin production. Simultaneous quantitative analyses of the ${\beta}$-hydroxyacid form and ${\beta}$-hydroxylactone for of lovastatin were performed by the high performance liquid chromatography (HPLC) method with a UV photodiode-array (PDA) detector. The total lovastatin yield ($4{\sim}6\;mg/g$, average of five repeats) was achieved by adding soybean powder, glycerol, sodium nitrate, and acetic acid at optimized levels after 14 days of fermentation. The maximal yield of lovastatin under the optimal composition of the medium increased by almost 2 times the yield observed prior to optimization. The experimental results also indicated that the ${\beta}$-hydroxylactone form of lovastatin (LFL) and the ${\beta}$-hydroxyacid form of lovastatin (AFL) simultaneously existed in solid state cultures of Monascus ruber. while the latter was the dominant form in the middle-late stage of continued fermentation. These results indicate that optimized culture conditions can be used for industrial production of lovastatin to obtain high yields.

Biocontrol of Anthracnose in Pepper Using Chitinase, ${\beta}$-1,3 Glucanase, and 2-Furancarboxaldehyde Produced by Streptomyces cavourensis SY224

  • Lee, So Youn;Tindwa, Hamisi;Lee, Yong Seong;Naing, Kyaw Wai;Hong, Seong Hyun;Nam, Yi;Kim, Kil Yong
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1359-1366
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    • 2012
  • A strain of Streptomyces cavourensis subsp. cavourensis (coded as SY224) antagonistic to Colletotrichum gloeosporioides infecting pepper plants was isolated. SY224 produced lytic enzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase, and protease in respective assays. To examine for antifungal activity, the treatments amended with the nonsterilized supernatant resulted in the highest growth inhibition rate of about 92.9% and 87.4% at concentrations of 30% and 10%, respectively. However, the sterilized treatments (autoclaved or chloroform treated) gave a lowered but significant inhibitory effect of about 63.4% and 62.6% for the 10% supernatant concentration, and 75.2% and 74.8% for the of 30% supernatant concentration in the PDA agar medium, respectively, indicative of the role of a non-protein, heat stable compound on the overall effect. This antifungal compound, which inhibited spore germination and altered hyphal morphology, was extracted by EtOAc and purified by ODS, silica gel, Sephadex LH-20 column, and HPLC, where an active fraction was confirmed to be 2-furancarboxaldehyde by GS-CI MS techniques. These results suggested that SY224 had a high potential in the biocontrol of anthracnose in pepper, mainly due to a combined effect of lytic enzymes and a non-protein, heat-stable antifungal compound, 2-furancarboxaldehyde.

Validation of Simultaneous Analysis Method of Standard Compounds in Fermented Kalopanax pictus Nakai by Bioconversion (생물전환을 통한 음나무발효물의 지표성분 설정 및 동시분석법 검증)

  • Jang, Won Hui;Lee, Wha Young;Lee, Bong Jin;Kim, Jean Man;Park, Seon Ju
    • The Korean Journal of Food And Nutrition
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    • v.32 no.2
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    • pp.148-154
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    • 2019
  • The aim of this study was to select compounds for the standardization of fermented Kalopanax pictus Nakai (KP-F), to develop the analysis method using HPLC-PDA and to perform method validation. KP-F is a fermented powder developed to improve the original physiological activities and create a new functionality. Eleutheroside E, Acanthoside B, and Syringaresinol were selected as the standard compounds and developed our own method for simultaneous analysis. The analyte was isolated using C18 column with a gradient elution of 0.05 M phosphoric acid in water and methanol as the mobile phase at a flow rate of 1 mL/min and detected at 210 nm. As a result, all standard compounds showed good linearity with an $R^2$ (coefficient of correlation) of 1.000 and for the limit of detection range of $0.710{\sim}0.831{\mu}g/mL$, and the limit of quantification as $2.150{\sim}2.520{\mu}g/mL$. The precision was RSD (%) of less than 4.80%, while the accuracy was 4.70%>RSD (%) for the range 102.44~110.48%. In conclusion, the developed analysis method is suitable for the detection of Eleutheroside E, Acanthoside B, and Syringaresinol in KP-F.

Whitening Activity of Abeliophyllum distichum Nakai Leaves According to the Ratio of Prethanol A in the Extracts

  • Jang, Tae-Won;Choi, Ji-Soo;Kim, Hoi-Ki;Lee, Eun-Ja;Han, Man-Wook;Lee, Ki-Beom;Kim, Do-Wan;Park, Jae-Ho
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.667-674
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    • 2018
  • In this study, we evaluated the whitening activity of prethanol A and water extracts from Abeliophyllum distichum Nakai. The extracts were prepared using 0, 50, 70, and 100% prethanol A at $121^{\circ}C$, 1.2 atm for 15 minutes. To confirm effective extraction, the acteoside content of each extract was analyzed with the HPLC-PDA method. The antioxidant activity was evaluated using DPPH and ABTS scavenging activity assays, and the whitening activity was evaluated based on inhibitory activities on the protein and mRNA expression of tyrosinase, tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), and microphthalmia-associated transcription factor (MITF) in B16 F10 cells. Each extract showed strong antioxidant and whitening activity. $IC_{50}$ values of antioxidant activity from each extract were in order of 100%, 70%, 50%, and 0%. In addition, whitening activity inhibited the protein and mRNA expression of melanin synthesis factor, following the same pattern as antioxidant activity. In conclusion, water and prethanol A extracts of A. distichum showed effective antioxidant and whitening activity and are thus considered to be valuable materials for whitening cosmetics. The results of this study will also provide basic data for the safe and efficient production of A. distichum as a cosmetic material.

Quantification of the Bioactive Components of the Rhizomes of Curcuma wenyujin and Assessment of Its Anti-inflammatory Effect in Benign Prostatic Hyperplasia-1 Cells

  • Seo, Chang-Seob;Park, Eunsook;Lee, Mee-Young;Shin, Hyeun-Kyoo
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.238-243
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    • 2019
  • In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini $C_{18}$ columns ($250mm{\times}4.6mm$, $5{\mu}m$) at $40^{\circ}C$ by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin $E_2$ ($PGE_2$) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were $0.10-0.32{\mu}g/mL$ and $0.30-0.98{\mu}g/mL$, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 - 5.92 mg/g, 4.72 - 4.86 mg/g, and 1.06 - 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed $PGE_2$ levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit antiinflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.

Comparison of Vitamin B5 Content and True Retention in Commonly Consumed Vegetables by Different Cooking Methods (국내 다소비 채소류의 조리에 따른 비타민 B5 함량 및 잔존율 비교)

  • Jin Ju, Park;Arin, Park;Eunji, Park;Youngmin, Choi
    • Journal of the Korean Society of Food Culture
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    • v.37 no.6
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    • pp.540-546
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    • 2022
  • This study aimed to determine the changes in the vitamin B5 content of raw and cooked vegetables. The nineteen vegetables were subjected to different cooking methods, viz. blanching, boiling, pan-broiling, and steaming. Vitamin B5 was quantified by reversed-phase high-performance liquid chromatography (HPLC) using photodiode-array (PDA) detection (200 nm). The standard reference materials (SRM) were used to validate the accuracy of vitamin B5 measurement method used in this study. The cooking yields ranged from 82.63 to 107.62% and decreased in most of the vegetables except bitter melon, curled mallow, and eggplant. The raw kabocha squash, Danhobak, had the highest vitamin B5 content (0.671 mg/100 g) among the samples. All cooked vegetables showed lower vitamin B5 content compared to the raw samples. The true retention ranged from 0% (crown daisy, blanching) to 84.49% (kabocha squash, steaming). These results indicate that vitamin B5 is degraded after cooking. Pan-broiling and steaming are better cooking methods than the others for retaining vitamin B5. The true retention of vitamin B5 in the samples markedly depends on the cooking method and food matrix. These results can be used as important basic data for nutritional evaluation of meals.

Chondroprotective and Anti-inflammatory Effects of ChondroT, A New Complex Herbal Medication

  • Jung Up Park;WonWoo Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.103-103
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    • 2022
  • Ganghwaljetongyeum (GHJTY) is a complex herbal decoction comprising 18 plants; it is used to treat arthritis. In order to develop a new anti-arthritic herbal medication, we selected 5 out of 18 GHJTY plants by using bioinformatics analysis. The new medication, called ChondroT, comprised water extracts of Osterici Radix, Lonicerae Folium, Angelicae Gigantis Radix, Clematidis Radix, and Phellodendri Cortex. This study was designed to investigate its chondroprotective and anti-inflammatory effects to develop an anti-arthritic herb medicine. ChondroT was validated using a convenient and accurate high-performance liquid chromatography. photodiode array (HPLC-PDA) detection method for simultaneous determination of its seven reference components. The concentrations of the seven marker constituents were in the range of 0.81-5.46 mg/g. The chondroprotective effects were evaluated based on SW1353 chondrocytes and matrix metalloproteinase 1 (MMP1) expression. In addition, the anti-inflammatory effects of ChondroT were studied by Western blotting of pro-inflammatory enzymes and by enzyme-linked immunosorbent assay (ELISA) of inflammatory mediators in lipopolysaccharides (LPS)-induced RAW264.7 cells. ChondroT enhanced the growth of SW1353 chondrocytes and also significantly inhibited IL-1β-induced MMP-1 expression. However, ChondroT did not show any effects on the growth of HeLa and RAW264.7 cells. The expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was induced by LPS in RAW264.7 cells, which was significantly decreased by pre-treatment with ChondroT. In addition, ChondroT reduced the activation of NF-κB and production of inflammatory mediators, such as IL-1β, IL-6, PGE2, and nitric oxide (NO) in LPS-induced RAW264.7 cells. These results show that ChondroT exerted a chondroprotective effect and demonstrated multi-target mechanisms related to inflammation and arthritis. In addition, the suppressive effect was greater than that exhibited by GHJTY, suggesting that ChondroT, a new complex herbal medication, has therapeutic potential for the treatment of arthritis.

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