• Title/Summary/Keyword: Metabolites Analysis

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Isolation and quantitative analysis of metabolites from Scrophularia buergeriana and their hepatoprotective effects against HepG2 Cells (현삼 (Scrophularia buergeriana)에서 분리한 화합물의 함량분석 및 간세포 보호 효과)

  • Na, Hyeon Seon;Oh, Seon Min;Shin, Woo Cheol;Bo, Jeon Hwang;Kim, Hyoung-Geun;Yoon, Dahye;Yang, Seung Hwan;Lee, Young-Seob;Kim, Geum-Soog;Baek, Nam-In;Lee, Moon-Soon;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.399-406
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    • 2019
  • The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H2O fractions. The repeated silica gel or octadecyl SiO2column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

Applied Analysis for Metabolic Profiling of Trace-level Amino Acid in Biological Fluid (생체시료 중 미량 아미노산 대사 프로필을 위한 분석법 응용)

  • Nam, Hyung Wook;Park, Song-Ja;Pyo, Hee Soo;Paeng, Ki Jung
    • Analytical Science and Technology
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    • v.16 no.5
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    • pp.349-357
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    • 2003
  • The universality of low molecular weight metabolites (i.e. amino acids, steroid hormones) allows rapid and straightforward investigation of biochemistry of genetically un-characterized species. Thus in vivo metabolic profiling of amino acid in combination with multivariate data analysis (metabolomics) offers great potential in comparative biology. In this paper, amino acid profiles in biological fluid (media) were studied by using HPLC/FLD. HPLC procedure for amino acids require the formation of derivatives due to the low absorption of the free compounds. o-Phthalaldehyde (OPA) used in association with a thiol, such as 3-mercaptopropionic acid (3-MPA), is one of the most popular and sensitive reagents, which yield quickly fluorescent iso-indoles at room temperature. To improve unstability of OPA/3-MPA derivatization, we optimized injector programs for fixed injection times. Linear regressions for the standard curves were linear in the range 0.5 - 100.0 ppb, giving correlation coefficents above 0.99. The detection limit were 1.70 pmol(GLU) - 23.81 pmol(SER). It is practically useful when the amount of sample is very low on single cells.

Comparison of Soyasapogenol A, B Concentrations in Soybean Seeds and Sprouts

  • Kang, Eun-Young;Kim, Seung-Hyun;Kim, Sun-Lim;Seo, Su-Hyun;Kim, Eun-Hye;Song, Hong-Keun;Ahn, Joung-Kuk;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.2
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    • pp.165-176
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    • 2010
  • Soybean seeds contain many biologically active secondary metabolites, such as proteins, saponins, isoflavones, phytic acids, trypsin inhibitors and phytosterols. Among them, saponins in soybeans have attracted considerable interest because of their health benefits. Soyasaponin A and B are the most abundant types of saponins found in soybeans along with soyasapogenol (aglycone), which is a precursor of soyasaponin. The main purpose of this experiment was to determine the concentration of soyasapogenol in soybean seeds and sprouts as a function of seed size, usage, seed coat color and seed cotyledon color. The 79 Korean soybean varieties were cultivated at Yesan of Chungnam in 2006 for the analysis of soyasapogenol using HPLC with Evaporative Light Scattering Detection (ELSD). The total average concentration of soyasapogenol was $1313.52{\mu}g\;g^{-1}$ in soybean seeds and $1377.22{\mu}g\;g^{-1}$ in soybean sprouts. Soybean sprouts were about 5% higher than soybean seeds in average total soyasapogenol concentration. In the process of sprouting, the average soyasapogenol A content decreased by approximately 1.6%, but soyasapogenol B and total soyasapogenol increased by 8.31% and 4.88%, based on the content of soybean seeds. When classified according to the size of seeds, the total soyasapogenol concentration of soybean seeds were not significantly different (p<0.05) On average, small soybean seeds were increased by as much as $103.14{\mu}g\;g^{-1}$ in sprouting process. As a function of the use of the seeds, The total soyasapogenol in soybean seeds were significantly different (p<0.05). While, the soybean sprouts were not significant different (p<0.05). Altogether, sprout soybean seeds show the greatest change in content during the germination process. When seeds with different coat colors were compared, the total soyasapogenol concentration of soybean with yellow seed coats ($1357.30\mu g\;g^{1}$) was slightly higher than that of soybean with black ($1260.30{\mu}g\;g^{-1}$) or brown ($1263.62{\mu}g\;g^{-1}$) seed coats. For the color of the cotyledon, the total soyasapogenol concentration was significantly increased in green cotyledon during the germination and seedling process. The results of this study suggest the functional characteristics of soybeans through quantitative analysis of soyasapogenol. In addition, the concentration of soyasapogenol exhibited a change during the germination process, which was evaluated by the nutritional value of the soybean sprouts.

Metabolomic analysis of perilla seeds harvested from Korea and China (국내산 및 중국산 들깨 종자의 대사체 분석)

  • Gu, Suyeon;Choi, Nayoung;Son, Yejin;Park, Ji Yeong;Choi, Sung-Gil;Lee, Myoung Hee;Kim, Hyun-Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.411-419
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    • 2019
  • A comprehensive comparison of fatty acid and metabolite profiles in 8 species of perilla seeds harvested from Korea and China was carried out to understand the correlation between cultivation conditions and the quality of perilla seeds. Metabolomic studies revealed that the perilla seeds were distinct from each other, based on the partial least squares -discriminant analysis (PLS-DA) score plots; 23 compounds including metabolites, fatty acids, and phytosterols that predominantly contributed to this interspecies difference were identified. Changes in the growth environment affected these compounds. In particular, phenolic compounds were affected by rainfall and temperature. Moreover, temperature and altitude showed negative and positive correlations, respectively, to oleic acid. Although more studies on the effect of various environmental factors on cultivation of perilla plants as well as their seeds are needed, the present results provide information that would be useful in identifying perilla seeds produced in specific regions and in the cultivation of good quality perilla.

The co-injection of antioxidants with foot-and-mouth disease vaccination altered growth performance and blood parameters of finishing Holstein steers

  • Seo, Jakyeom;Song, Minho;Jo, Namchul;Kim, Woonsu;Jeong, Sinyong;Kim, Jongnam;Lee, Seyoung;Seo, Seongwon
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.792-799
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    • 2019
  • Objective: This study was conducted to evaluate whether the co-injection of antioxidants together with foot-and-mouth disease (FMD) vaccination has the potential to attenuate the negative effects caused by vaccination in Holstein finishing steers. Methods: A total of 36 finishing Holstein steers (body weight [BW]: $608{\pm}45.6kg$, 17 months old) were randomly allocated to one of three treatments: i) control (CON, only FMD vaccination without any co-injection), ii) co-injection of commercial non-steroidal anti-inflammatory drugs (NSAID) with FMD vaccination at a ratio of 10:1 (NSAID vol/FMD vaccine vol) as a positive control (PCON), iii) co-injection of commercial mixture of vitamin E and selenium with FMD vaccination (VITESEL) (1 mL of FMD vaccine+1 mL of antioxidants per 90 kg of BW). Changes in growth performance and blood parameters because of treatments were determined. Results: No significant difference in BW, average daily gain, and dry matter intake of the steers was observed among the treatments. The FMD vaccination significantly increased white blood cells (WBC), neutrophils, platelets, and mean platelet volume (p<0.01) in blood analysis. The count of lymphocyte tended to increase after vaccination (p = 0.08). In blood analysis, steers in VITESEL tended to have higher numbers of WBC, neutrophils, and platelets compared to that of other treatments (p = 0.09, 0.06, and 0.09, respectively). Eosinophils in VITESEL were higher than those in PCON (p<0.01). Among blood metabolites, blood urea nitrogen and aspartate transaminase were significantly increased, but cholesterol, alanine transferase, inorganic phosphorus, Mg, and albumin were decreased after FMD vaccination (p<0.01). Conclusion: The use of antioxidants in FMD vaccination did not attenuate growth disturbance because of FMD vaccination. The metabolic changes induced by vaccination were not controlled by the administration of antioxidants. The protective function of antioxidants was effective mainly on the cell counts of leukocytes.

Knowledge Modeling and Database Construction for Human Biomonitoring Data (인체 바이오모니터링 지식 모델링 및 데이터베이스 구축)

  • Lee, Jangwoo;Yang, Sehee;Lee, Hunjoo
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.607-617
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    • 2020
  • Human bio-monitoring (HBM) data is a very important resource for tracking total exposure and concentrations of a parent chemical or its metabolites in human biomarkers. However, until now, it was difficult to execute the integration of different types of HBM data due to incompatibility problems caused by gaps in study design, chemical description and coding system between different sources in Korea. In this study, we presented a standardized code system and HBM knowledge model (KM) based on relational database modeling methodology. For this purpose, we used 11 raw datasets collected from the Ministry of Food and Drug Safety (MFDS) between 2006 and 2018. We then constructed the HBM database (DB) using a total of 205,491 concentration-related data points for 18,870 participants and 86 chemicals. In addition, we developed a summary report-type statistical analysis program to verify the inputted HBM datasets. This study will contribute to promoting the sustainable creation and versatile utilization of big-data for HBM results at the MFDS.

A possible mechanism to the antidepressant-like effects of 20 (S)-protopanaxadiol based on its target protein 14-3-3 ζ

  • Chen, Lin;Li, Ruimei;Chen, Feiyan;Zhang, Hantao;Zhu, Zhu;Xu, Shuyi;Cheng, Yao;Zhao, Yunan
    • Journal of Ginseng Research
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    • v.46 no.5
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    • pp.666-674
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    • 2022
  • Background: Ginsenosides and their metabolites have antidepressant-like effects, but the underlying mechanisms remain unclear. We previously identified 14-3-3 ζ as one of the target proteins of 20 (S)-protopanaxadiol (PPD), a fully deglycosylated ginsenoside metabolite. Methods: Corticosterone (CORT) was administered repeatedly to induce the depression model, and PPD was given concurrently. The tail suspension test (TST) and the forced swimming test (FST) were used for behavioral evaluation. All mice were sacrificed. Golgi-cox staining, GSK 3β activity assay, and Western blot analysis were performed. In vitro, the kinetic binding analysis with the Biolayer Interferometry (BLI) was used to determine the molecular interactions. Results: TST and FST both revealed that PPD reversed CORT-induced behavioral deficits. PPD also ameliorated the CORT-induced expression alterations of hippocampal Ser9 phosphorylated glycogen synthase kinase 3β (p-Ser9 GSK 3β), Ser133 phosphorylated cAMP response element-binding protein (p-Ser133 CREB), and brain-derived neurotrophic factor (BDNF). Moreover, PPD attenuated the CORT-induced increase in GSK 3β activity and decrease in dendritic spine density in the hippocampus. In vitro, 14-3-3 ζ protein specifically bound to p-Ser9 GSK 3β polypeptide. PPD promoted the binding and subsequently decreased GSK 3β activity. Conclusion: These findings demonstrated the antidepressant-like effects of PPD on the CORT-induced mouse depression model and indicated a possible target-based mechanism. The combination of PPD with the 14-3-3 ζ protein may promote the binding of 14-3-3 ζ to p-GSK 3β (Ser9) and enhance the inhibition of Ser9 phosphorylation on GSK 3β kinase activity, thereby activating the plasticity-related CREBeBDNF signaling pathway.

Comparative transcriptome and metabolome analyses of four Panax species explore the dynamics of metabolite biosynthesis

  • Hyunjin, Koo;Yun Sun, Lee;Van Binh, Nguyen;Vo Ngoc Linh, Giang;Hyun Jo, Koo;Hyun-Seung, Park;Padmanaban, Mohanan;Young Hun, Song;Byeol, Ryu;Kyo Bin, Kang;Sang Hyun, Sung;Tae-Jin, Yang
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.44-53
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    • 2023
  • Background: The genus Panax in the Araliaceae family has been used as traditional medicinal plants worldwide and is known to biosynthesize ginsenosides and phytosterols. However, genetic variation between Panax species has influenced their biosynthetic pathways is not fully understood. Methods: Simultaneous analysis of transcriptomes and metabolomes obtained from adventitious roots of two tetraploid species (Panax ginseng and P. quinquefolius) and two diploid species (P. notoginseng and P. vietnamensis) revealed the diversity of their metabolites and related gene expression profiles. Results: The transcriptome analysis showed that 2,3-OXIDOSQUALENE CYCLASEs (OSCs) involved in phytosterol biosynthesis are upregulated in the diploid species, while the expression of OSCs contributing to ginsenoside biosynthesis is higher in the tetraploid species. In agreement with these results, the contents of dammarenediol-type ginsenosides were higher in the tetraploid species relative to the diploid species. Conclusion: These results suggest that a whole-genome duplication event has influenced the triterpene biosynthesis pathway in tetraploid Panax species during their evolution or ecological adaptation. This study provides a basis for further efforts to explore the genetic variation of the Panax genus.

Analytical method of aflatoxins in edible oil and infant-children foods (식용유지와 영유아식품 중 아플라톡신 분석방법)

  • Hu, Soo-Jung;Park, Seung-Young;Kim, Soon-Sun;Lee, Joon-Goo;Song, Ji-Young;Kang, Eun-Gi;Lee, Hyun-Sook;Cho, Dae-Hyun
    • Analytical Science and Technology
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    • v.24 no.2
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    • pp.150-157
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    • 2011
  • Aflatoxins are secondary metabolites of the molds of Aspergillus flavus and Aspergillus parasiticus. They are highly carcinogenic compounds and can affect a wide range of vegetable commodities such as cereals (especially corn), nuts, peanuts, fruits and oil seeds, in the field and during storage. In fact, oilseeds are often stored for weeks in conditions that promote the mould growth, and the possible consequent presence of aflatoxins in oilseeds can lead to their transfer in oil. In addition, aflatoxins can be found as a natural contaminant in multi-cereals and beans making baby food for infants and young-children. The objective of this study was to validate the liquid extraction method or develop an analytical method for edible oil and infant-children foods. Therefore, this study developed condition of extract for aflatoxins ($B_1$, $B_2$, $G_1$ and $G_2$) in edible oil using a high performance liquid chromatography with florescence detector (HPLC/FLD). Aflatoxins were extracted from edible oil samples by means of MSPD (Matrix solid phased dispersion), utilizing $C_{18}$ as dispersing material and purified by using immunoaffinity column. The gression line coefficients were above 0.999. The recoveries for aflatoxins ranged from 85.9 to 93.0%, and relative standard deviations were below 5.7%. The new developed method of aflatoxins effectively enhanced recoveries by using MSPD-Immunoaffinity column compared with liquid extraction. The analytical method for liquid extraction of aflatoxin was appropriate for infant-children food. Reviewing the current method, the recoveries of aflatoxins ($B_1$, $B_2$, $G_1$ and $G_2$) were 89.5~92.3%.

Monitoring of Aflatoxins on Commercial Herbal Medicines (유통생약의 아플라톡신 모니터링)

  • Park, Seung-Young;Moon, Hyun-Ju;Cho, Soo-Yeul;Lee, Jun-Gu;Lee, Hwa-Mi;Song, Ji-Young;Cho, Ok-Sun;Cho, Dae-Hyun
    • Journal of Food Hygiene and Safety
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    • v.26 no.4
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    • pp.315-321
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    • 2011
  • This study was performed to investigate contamination levels of aflatoxins, the secondary metabolites produced by fungi Aspergillus flavus and A. parasiticus, in herbal medicine. Herbs is susceptible to these fungi infections through its growth harvest, transport and storage. This study determine the aflatoxin $B_1$, $B_2$, $G_1$ and $G_2$ levels by HPLC-florescence detector coupled with photochemical enhancement in 558 samples herbal medicine distributed in Korea and China. Also, We checked a transfer ratio of aflatoxins from raw herbal medicines to herbal medicine extract. Hot water extraction of herbal medicines was prepared by air pressure and high pressure condition. The analytical method for aflatoxins was validated in this method. In results recoveries of the analytical method were ranged from 67.4% to 96.2% and, limits of detection and quantitation for aflatoxins were $0.015{\sim}0.138\;{\mu}g/kg$ and $0.046{\sim}0.418\;{\mu}g/kg$, respectively. According to the results of monitoring on aflatoxins in herbal medicine, aflatoxins 1.7 ug/kg $B_1$ and 0.9 ug/kg $G_1$ were detected in only one sample of Strychni Ignatii Semen, and 0.8 ug/kg $G_1$ in Strychni Semen. About 13.6~51.3% of aflatoxins were transferred to hot water extract. Although the detected levels are under the permitted levels for aflatoxins in herbal medicine, these amounts should be considered in regard to overall daily exposure to mycotoxins.