• Title/Summary/Keyword: HPLC-MS-MS

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The VHR Dual-Specificity Protein Tyrosine Phosphatase (DS-PTPase) Inhibitor Isolated from Gastrodia elata (천마(Gastrodia elata)로부터 분리한 VHR DS-PTPase 저해 물질)

  • Lee, Myung-Sun;Oh, Won-Keun;Bae, Eun-Young;Ahn, Soon-Cheol;Sohn, Cheon-Bae;Osada, Hiroyuki;Ahn, Jong-Seog
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.505-509
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    • 2002
  • In the course of screening program for VHR DS-PTPase (dual-specificity protein tyrosine phosphatase) from natural sources, Gastrodia elata was selected. One compound showing potent inhibitory activity was isolated by the solvent extraction and column chromatography including silica gel, ODS RP-18, Sephades LH-20, and HPLC. This compound was identified as baicalein by several NMR techniques such as $^1H-NMR$, $^{13}C-NMR$, and DEPT. Baicalein showed selective inhibitory activity against VHR DS-PTPase with $IC_{50}=2.4\;{\mu}M$, and showed cytotoxicity against several human cancer cell lines with an $GI_{50}$ of $5.26{\sim}12.93\;{\mu}g/mL$ range, including, melanoma (LOX-IMVI), lung cancer (NCI H23 and A549), colon cancer (HCT 116 and SW 620), prostate cancer (PC-3), and leukemia (MOLT 4F).

Evaluation of the Physiological Activity and Identification of the Active Ingredients of Crab Apple (Malus prunifolia Borkh.) Extracts (꽃사과(Malus prunifolia Borkh.) 추출물의 생리활성 평가 및 활성 성분의 규명)

  • Shin, Hyun Young;Kim, Hoon;Jeong, Eun-Jin;Kim, Hyun-Gyeong;Lee, Kyung-Haeng;Bae, Yun-Jung;Kim, Woo Jung;Lee, Sanghyun;Yu, Kwang-Won
    • The Korean Journal of Food And Nutrition
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    • v.34 no.5
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    • pp.477-486
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    • 2021
  • To utilize Malus pruniforia Borkh. as a functional material, cold-water (CW), hot-water (HW), and 70% ethanol (EtOH) extracts were prepared, and their antioxidant and anti-inflammatory activities were compared. The antioxidant activity of the HW extract evaluated by ABTS and DPPH radical scavenging and FRAP activity was significantly effective. The total polyphenol content of the HW extract was also higher by 15.5±0.7 mg GAE/g extract compared to other extracts. The EtOH extract showed significantly decreased TNF-α (39.8%), IL-6 (65.5%), and NO (34.9%) levels in RAW 264.7 cells compared to the LPS-induced control group. The levels of IL-6 (21.1%) and IL-8 (19.3%) were significantly decreased by treatment of EtOH extract in HaCaT keratinocytes induced with TNF-α and IFN-γ. The UHPLC-MS results indicated that the EtOH extract might have chlorogenic acid and phlorizin as the major compounds. This was validated using HPLC-DAD, which showed that the EtOH extract had higher levels of chlorogenic acid and phlorizin (1,185±58 and 470±10 ㎍/g extract, respectively). In conclusion, the present study suggested that the anti-inflammatory activity of the EtOH extract was more effective than the CW and HW extracts, and chlorogenic acid and phlorizin could be used as indicator compounds and functional substances.

Simultaneous Analysis of the Compounds of Natural Cosmetic Resources Containing Chrysanthemum zawadskii, Perilla frutescens, Rosa multiflora and their Anti-oxidative Activity (산구절초, 자소엽, 찔레를 함유한 천연 화장품소재의 다성분 동시분석과 항산화활성)

  • Ham, Ha Neul;Shrestha, Abinash Chandra;Kim, Ju Eun;Lee, Tae Bum;Yoo, Byoung Wan;Kim, Min Sook;Kim, Kwang Sang;Cha, Joon-Seok;Lee, Yong Mun;Kim, Jeong Yeob;Leem, Jae Yoon
    • Korean Journal of Pharmacognosy
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    • v.49 no.4
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    • pp.312-321
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    • 2018
  • Recently, consumer demand for functional cosmetics containing natural ingredients has been greatly expanded. To develop the natural cosmetic materials, we selected 3 plants, Chrysanthemum zawadskii Herbich (CZ), Perilla frutescens (L.) Britton var. acuta Kudo (PF), and Rosa multiflora Thunberg (RM) which showed high total flavonoid contents (TFC), total polyphenol contents (TPC), and strong DPPH radical scavenging effect. We determined astragalin, chlorogenic acid, and rosmarinic acid as a marker compound for quantitative analysis of the content of each material and standardization of the quality standards and manufacturing standards through LC/MS analysis. HPLC-DAD was used to simultaneously analyze these marker components of three natural product complexes (Mix) and to validate the analytical method through experiments such as linearity, accuracy and precision. The detection wavelengths were set at 210, 265, and 330 nm. The detected 3 compounds from extract of CZ, PF, RM showed significant linearity ($R^2${\geq_-}$0.9947). The limit of detection (LOD) of chlorogenic acid, astragalin and rosmarinic acid were $8.29{\mu}g/ml$, $2.28{\mu}g/ml$, and $27.00{\mu}g/ml$, respectively. The limit of quantification (LOQ) of chlorogenic acid, astragalin and rosmarinic acid were $25.11{\mu}g/ml$, $6.92{\mu}g/ml$, and $81.83{\mu}g/ml$, respectively. The contents of the three indicators of Mix were 19.82-24.71 mg/g of chlorogenic acid, 43.80-46.02 mg/g of astragalin, and 46.33-48.57 mg/g of rosmarinic acid.

Arsenic species in husked and polished rice grains grown at the non-contaminated paddy soils in Korea (국내 비오염 논토양에서 재배한 현미와 백미 중 비소화학종 함량)

  • Kim, Da-Young;Kim, Ji-Young;Kim, Kye-Hoon;Kim, Kwon-Rae;Kim, Hyuck-Soo;Kim, Jeong-Gyu;Kim, Won-Il
    • Journal of Applied Biological Chemistry
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    • v.61 no.4
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    • pp.391-395
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    • 2018
  • There is an increasing concern over arsenic (As) contamination of paddy soil and rice with regard to food safety. This study was conducted to investigate total and inorganic As concentration in one hundred husked and polished rice samples collected at the non-contaminated paddy soil in Korea. Arsenic species in rice samples were extracted using 1% nitric acid ($HNO_3$) with a microwave oven and were measured using high performance liquid chromatography coupled with inductively coupled plasma-mass spectrometry. Mean concentrations of total As in husked rice and polished rice were 0.18 and $0.11mg\;kg^{-1}$, respectively. Also, average inorganic As concentrations in husked rice and polished rice were 0.11 and $0.07mg\;kg^{-1}$, respectively. These levels are lower than the standard guideline value 0.35 and $0.2mg\;kg^{-1}$ for inorganic As in husked and polished rice recommended by Codex Committee on Contaminants in Foods and Ministry of Food and Drug Safety, respectively. The mean of the inorganic As ratio for the total amount of As was 0.65 and 0.67 for husked rice and polished rice, respectively, and the range was from 0.08 to 1.0. For health risk assessment, the average value of cancer risk probability was $9.24{\times}10^{-5}$ and ranged from $2.30{\times}10^{-5}$ to $1.90{\times}10^{-5}$. Therefore, human exposure to As through dietary intake of surveyed rice samples might considered to be a low health risk.

Dammarane-type triterpene oligoglycosides from the leaves and stems of Panax notoginseng and their antiinflammatory activities

  • Li, Juan;Wang, Ru-Feng;Zhou, Yue;Hu, Hai-Jun;Yang, Ying-Bo;Yang, Li;Wang, Zheng-Tao
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.377-384
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    • 2019
  • Background: Inflammation is widespread in the clinical pathology and closely associated to the progress of many diseases. Triterpenoid saponins as a key group of active ingredients in Panax notoginseng (Burk.) F.H. Chen were demonstrated to show antiinflammatory effects. However, the chemical structures of saponins in the leaves and stems of Panax notoginseng (PNLS) are still not fully clear. Herein, the isolation, purification and further evaluation of the antiinflammatory activity of dammarane-type triterpenoid saponins from PNLS were conducted. Methods: Silica gel and reversed-phase C8 column chromatography were used. Furthermore, preparative HPLC was used as a final purification technique to obtain minor saponins with high purities. MS, NMR experiments, and chemical methods were used in the structural identifications. The antiinflammatory activities of the isolated saponins were assessed by measuring the nitric oxide production in RAW 264.7 cells stimulated by lipopolysaccharides. Real-time reverse transcription polymerase chain reaction was used to measure the gene expressions of inflammation-related gene. Results: Eight new minor dammarane-type triterpene oligoglycosides, namely notoginsenosides LK1-LK8 (1-8) were obtained from PNLS, along with seven known ones. Among the isolated saponins, gypenoside IX significantly suppressed the nitric oxide production and inflammatory cytokines including tumor necrosis $factor-{\alpha}$, interleukin 10, interferon-inducible protein 10 and $interleukin-1{\beta}$. Conclusion: The eight saponins may enrich and expand the chemical library of saponins in Panax genus. Moreover, it is reported for the first time that gypenoside IX showed moderate antiinflammatory activity.

MTHFR, As3MT and GSTO1 Polymorphisms Influencing Arsenic Metabolism in Residents Near Abandoned Metal Mines in South Korea (폐금속광산 지역 주민의 비소 대사에 영향을 미치는 MTHFR, As3MT, GSTO1 유전자 다형성)

  • Surenbaatar, Ulziikhishig;Kim, Byoung-Gwon;Son, Hyun-Jin;Cho, Seong-Sik;Kim, Gwon-Min;Lim, Hyoun-Ju;Kwon, Jung-Yeon;Kim, Ki-Hwan;Hong, Young-Seoub
    • Journal of Environmental Health Sciences
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    • v.47 no.6
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    • pp.530-539
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    • 2021
  • Background: In South Korea, areas around abandoned metal mines are designated as regions with high arsenic (As) contamination. However, studies assessing urinary As exposure, As metabolism, and relevant genetic polymorphisms in residents of these metal mine areas are lacking. Objectives: To identify factors associated with As exposure and evaluate the effects of MTHFR, As3MT, and GSTO1 genetic polymorphisms on As metabolism in residents of abandoned metal mine areas by measuring urinary As species. Methods: Urinary As species (arsenite [As3+], arsenate [As5+], monomethyl arsonic acid, and dimethylarsinic acid) were isolated using high-performance liquid chromatography in combination with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). Four genetic polymorphisms (MTHFR A222V, MTHFR E429A, GSTO1 A140D, As3MT M287T) were analyzed in 144 residents of four areas around abandoned metal mines. Results: The study sample was comprised of 34.7% men and 65.3% women, with a mean age of 70.7±10.9 years. The urinary inorganic As concentration was higher among those consuming more than half locally produced rice (0.31 ㎍/L) than those consuming less than half such rice (0.18 ㎍/L). The urinary dimethylarsinic acid concentration was higher in the group that had consumed seafood in the past day (31.68 ㎍/L) than in those who had not (22.37 ㎍/L). Furthermore, individuals heterozygous in the MTHFR A222V and GSTO1 A140D polymorphism had higher urinary arsenic species concentrations than did individuals with a wild type or homozygous for the variant allele. Conclusions: Consumption of locally produced rice was associated with inorganic As exposure, whereas seafood consumption was associated with organic As exposure among residents of abandoned metal mine areas. There was no clear association between MTHFR A222V and GSTO1 A140D polymorphisms and As metabolism.

Development and characterization of hyaluronic acid-based orally disintegrating film containing vitamin D (히알루론산 기반 비타민 D 함유 구강용해필름의 제조 및 특성평가)

  • Kang, Seo-Yeon;An, Da-Yeon;Han, Jung-Ah
    • Korean Journal of Food Science and Technology
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    • v.54 no.3
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    • pp.327-333
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    • 2022
  • An orally disintegrating film (ODF) based on hyaluronic acid (HA) containing vitamin D was developed. The vitamin D content in the ODF was set based on the adequate intake (AI) of vitamin D from 0 to 10 AI (0, 1, 4, 7, and 10AI). The control (0AI) had the highest thickness and showed the longest disintegration time among the samples. The moisture content of the ODFs was significantly lower in those with vitamin D compared to the control. As the amount of vitamin D increased, the water vapor permeability (WVP) of the ODFs decreased, and the opacity significantly increased. The tensile strength was higher in the films containing vitamin D compared to the control films. However, the elongation at the break showed no significant difference among the films. The vitamin D content in the film was reduced by 25.7-44.2% during processing compared to the amount that was originally added. Based on the above results, a new and convenient vitamin D delivery system, an ODF, could be successfully produced.

Comparison of the bioactive compounds and anti-inflammatory effects found in different flower colors from Abeliophyllum distichum Nakai (미선나무 꽃 색에 따른 생리활성 화합물 및 항염증 활성 비교)

  • Jang, Tae-Won;Choi, Ji-Soo;Han, So-Yeon;Park, Hye-Jeong;Lee, Da-Yoon;Min, Young-Sil;Park, Jae-Ho
    • Journal of Applied Biological Chemistry
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    • v.65 no.3
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    • pp.203-213
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    • 2022
  • Abeliophyllum distichum (A. distichum, Korean endemic plant) is one genus and one species in the Oleaceae family. According to the color variation of petals and calyx, A. distichum is classified as A. distichum (white flower), A. distichum for. lilacinum (pink flowers), A. distichum for. eburneum (ivory flowers), and Okhwang 1 (golden flowers). In previous studies, bioactivities (antioxidant, anti-inflammatory, and anti-cancer) of A. distichum have been reported. We conducted a comparison of the differences in bioactive compounds and the anti-inflammatory effects on macrophages among four flowers of A. distichum (FAD). The identification and quantification of glycosides were analyzed by HPLC/PDA and LCMS. These results were shown FAD has rutin, hirsutrin, and acteoside. Antioxidant activity of FAD significantly decreased reactive oxygen species. In addition, FAD reduced the expression of pro-inflammatory mediators (nitric oxide, iNOS, and COX-2) in lipopolysaccharide-induced RAW 264.7 cells. For further study, we investigated the regulation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. In conclusion, FAD may exert anti-inflammatory effects by suppressing inflammatory mediators via regulations of NF-κB and MAPK signaling pathways. Therefore, these findings suggest that FAD is a potential resource as a preventative or therapeutic agent for inflammation.

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.

Mechanism of Panax notoginseng saponins modulation of miR-214-3p/NR1I3 affecting the pharmacodynamics and pharmacokinetics of warfarin

  • Yuting Yang;Zhenyu Zhai;Huiming Yao;Ling He;Jun Shao;Zirong Xia;Juxiang Li
    • Journal of Ginseng Research
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    • v.48 no.5
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    • pp.494-503
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    • 2024
  • Background: With the prevalence of dietary supplements, the use of combinations of herbs and drugs is gradually increasing, together with the risk of drug interactions. In our clinical work, we unexpectedly found that the combination of Panax notoginseng and warfarin, which are herbs that activate blood circulation and remove blood stasis, showed antagonistic effects instead. The purpose of this study was to evaluate the drug interaction between Panax notoginseng saponins (PNS) and warfarin, the main active ingredient of Panax notoginseng, and to explore the interaction mechanism. Methods: The effects and mechanisms of PNS on the pharmacodynamics and pharmacokinetics of warfarin were explored mainly in Sprague-Dawley rats and HepG2 cells. Elisa was used to detect the concentrations of coagulation factors, HPLC-MS to detect the blood concentrations of warfarin in rats, immunoblotting was employed to examine protein levels, qRT-PCR to detect mRNA levels, cellular immunofluorescence to detect the localization of NR1I3, and dual luciferase to verify the binding of miR-214-3p and NR1I3. Results: PNS significantly accelerated warfarin metabolism and reduced its efficacy, accompanied by increased expression of NR1I3 and CYP2C9. Interference with NR1I3 rescued the accelerated metabolism of warfarin induce by PNS co-administration. In addition, we demonstrated that PNS significantly reduced miR-214-3p expression, whereas miR-214-3p overexpression reduced NR1I3 and CYP2C9 expression, resulting in a weakened antagonistic effect of PNS on warfarin. Additionally, we found that miR-214-3p bound directly to NR1I3 3'-UTR and significantly downregulated NR1I3 expression. Conclusion: Our study demonstrated that PNS accelerates warfarin metabolism and reduces its pharmacodynamics by downregulating miR-214-3p, leading to increased expression of its target gene NR1I3, these findings provide new insights for clinical drug applications to avoid adverse effects.