• Title/Summary/Keyword: HPLC Profiling

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Studies on Quality Control of Domestic Agastachis Herba (국내산 곽향(藿香)의 품질관리 연구)

  • Han, Min Woo;Park, Chae Kyu;Park, Chul Su;Ahn, Hee Jun;Seo, Jeong Bum;Lee, Young Jong
    • The Korea Journal of Herbology
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    • v.34 no.2
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    • pp.67-74
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    • 2019
  • Objectives : Agastachis herba is a aerial part of Agastache rugosa (Fischer et Meyer) O. Kuntze which has been used as traditional medicines. Methods : The samples were collected using a domestic cultivation of Agastachis Herba. We experiment on essential oil content, the content of ethanol soluble extracts and HPLC profiling. Results : Loss on drying of Agastachis Herba contents of harvest in June ~ July was 4.93~8.26% and contents of harvest in August ~ September was 3.88~5.38%. Ash of Agastachis Herba contents of harvest in June ~ July was 5.07~7.97% and contents of harvest in August ~ September was 4.55~7.61%. Acid-insoluble ash of Agastachis Herba contents of harvest in June ~ July was 0.43~1.62% and contents of harvest in August ~ September was 0.46~1.44%. Essential oil contents of Agastachis Herba contents of harvest in June ~ July was $0.2{\sim}0.4m{\ell}$ and contents of harvest in August ~ September was $0.2{\sim}0.6m{\ell}$. Ethanol soluble extracts of Agastachis Herba contents of harvest in June ~ July was 9.72~20.44% and contents of harvest in August ~ September was 11.00~18.70%. HPLC profiling was detected of tilianin(12.93 min), acacetin(15.84 min) and agastachoside(17.95 min). Conclusion : Agastachis Herba should be harvested from August ~ September, the time of flowering. This is because of the high content of active compounds.

Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling

  • Rohit Kumar;Jai Kumar Kaushik;Ashok Kumar Mohanty;Sudarshan Kumar
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1130-1142
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    • 2023
  • Objective: Cow urine possesses several bioactive properties but the responsible components behind these bioactivities are still far from identified. In our study, we tried to identify the possible components behind the antimicrobial activity of cow urine by exploring the peptidome and metabolome. Methods: We extracted peptides from the urine of Sahiwal cows belonging to three different physiological states viz heifer, lactation, and pregnant, each group consisting of 10 different animals. The peptides were extracted using the solid phase extraction technique followed by further extraction using ethyl acetate. The antimicrobial activity of the aqueous extract was evaluated against different pathogenic strains like Staphylococcus aureus, Escherichia coli, and Streptococcus agalactiae. The safety of urinary aqueous extract was evaluated by hemolysis and cytotoxicity assay on the BuMEC cell line. The urinary peptides were further fractionated using high-performance liquid chromatography (HPLC) to identify the fraction(s) containing the antimicrobial activity. The HPLC fractions and ethyl acetate extract were analyzed using nLC-MS/MS for the identification of the peptides and metabolites. Results: A total of three fractions were identified with antimicrobial activity, and nLC-MS/MS analysis of fractions resulted in the identification of 511 sequences. While 46 compounds were identified in the metabolite profiling of organic extract. The urinary aqueous extract showed significant activity against E. coli as compared to S. aureus and S. agalactiae and was relatively safe against mammalian cells. Conclusion: The antimicrobial activity of cow urine is a consequence of the feeding habit. The metabolites of plant origin with several bioactivities are eliminated through urine and are responsible for their antimicrobial nature. Secondly, the plethora of peptides generated from the activity of endogenous proteases on protein shed from different parts of tissues also find their way to urine. Some of these sequences possess antimicrobial activity due to their amino acid composition.

Quality Monitoring of Specification Standard of Gardeniae Fructus in the Korean Pharmacopoeia and Studies HPLC Standard Chromatogram (치자(梔子)의 규격 기준 모니터링 및 HPLC 표준크로마토그램 연구)

  • Kim, Kyoung Hee;Kim, Sun Mi;Shin, Seung Hoon;Lee, Young Jong;Baek, Wan Sook
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.97-105
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    • 2017
  • Objectives : Gardeniae Fructus is a ripe fruit of Gardenia jasminoides Ellis, which has been used as traditional medicines for anti-inflammatory, diuretic, antipyretic, and antibacterial activity. The aim of this study was to compare of Gardeniae Fructus in South Korea collected during three years according to the standards in monographs of the Korean Pharmacopoeia Eleventh edition (KP11). Methods : 30 items of Gardeniae Fructus from two cultivation regions were classified into dried(n=15) & steamed (n=15) and tested according to the standards in monographs of the KP11. Gardeniae Fructus was carried out identification(comparison of colors, thin layer chromatography), heavy metals, residual pesticides, total ash, and assay registered at KP11. Add to we tested loss on dry, contents of ethanol-soluble extracts, and HPLC profiling. Results : In TLC chromatogram of identification test, the spot of gardenoside and geniposide were observed at $R_f$ value of about 0.3 and 0.5. Heavy metals and residual pesticides met the requirements of the standards for all samples. The results of total ash of each samples are measured maximum 4.87 %. According to HPLC for assay, the samples contain 4.80~6.10 % of geniposide and 0.45~1.83 % of gardenoside. Conclusion : We have verified the current specification standard of Gardeniae Fructus and standard that is not set. By the results, it is proposed a new draft of loss on drying and confirmed the content of gardenoside revised. HPLC standard chromatogram of Gardeniae Fructus is proposed. We hope that it will help the standardization of Gardeniae Fructus.

Biological Activities and Metabolite Analysis of Various Extracts and Fractions from Red Ginseng Marc

  • Lee, Dong Gyu;Jang, Ik Soon;Kang, Young-Hwa
    • Korean Journal of Plant Resources
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    • v.33 no.6
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    • pp.597-603
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    • 2020
  • Red ginseng marc (RGM) has been used on primary industries using fertilizer or forage, and it mostly has been dumped. To improve utilization of RGM, the biological activities of RGM were examined. RGM was extracted and fractionated using various solvents and their biological activities were compared. The hexane fraction from the methanol extract of RGM (RGMMH) showed strong anti-cancer activity (58.56 ± 6.04% at 100 ㎍/mL) and anti-inflammatory effect (65.72 ± 1.33% at 100 ㎍/mL). But, oil extract of RGM extracted with hexane (RGMH) showed low activities (anti-cancer: 16.42 ± 3.33%, at 100 ㎍/mL, anti-inflammatory activity: 29.46 ± 2.10%, at 100 ㎍/mL). Their metabolites were analyzed using HPLC. Panaxydol known as anti-cancer compound of RGM was one of major compounds in RGMMH. Meanwhile, panaxydol was detected in trace amount in red ginseng marc oil (RGMH). In addition, RGMMH and RGMH showed big differences in HPLC profiling. This research suggests optimal extraction method of RGM oil.

Quality Monitoring of Specification of Crataegi Fructus in the Korean Pharmacopoeia and Studies HPLC Standard Chromatogram (산사(山楂)의 규격 기준 모니터링 및 HPLC 표준크로마토그램 연구)

  • Kim, Kyoung Hee;Kim, Sun Mi;Lee, Young Jong;Baek, Wan Sook
    • The Korea Journal of Herbology
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    • v.32 no.1
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    • pp.55-62
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    • 2017
  • Objectives : Crataegi Fructus has been used as traditional medicines for more digestion action, amenorrhea due to blood stasis, and hyperlipemia. The aim of this study was to compare of Crataegi Fructus in South Korea collected during three years according to the standards in monographs of the Korean Pharmacopoeia Eleventh edition (KP 11). Methods : Crataegi Fructus was carried out identification test (Qualitative reaction, Thin layer chromatography), heavy metal test, and total ash registered at KP. Add to we tested loss on dry, contents of ethanol-soluble extracts, and HPLC profiling. Results : Identification test (TLC) was on comparing with ursolic acid standard solution in $R_f$ value, all samples showed red purple spot ($R_f$ value 0.9). Ursolic acid spot in $R_f$ value 0.35 showed by changing mobile phase condition. Heavy metals showed contents for Pb, Cd, As, and Hg range of 0.0 ~ 0.5 ppm, 0.0 ~ 0.2 ppm, 0.0 ~ 0.3 ppm, and 0.0 ~ 0.1 ppm. Loss on drying was ranged from 5.5 to 11.9 %, total ash was between the range 2.7 ~ 4.0 %. Contents of ethanol-soluble extracts was ranged from 17.8 to 44.9 %. The content of chlorogenic acid was ranged from 0.0 to 0.1 % based on the chlorogenic acid standard curve. Conclusion : We have verified the current specification standard of Crataegi Fructus and standard that is not set. We hope that it will help the standardization of Crataegi Fructus.

Cytochrome P-450 2A6 Inhibitor Based on the Indole Moiety

  • Lee, Soo;Lee, Hyang-Yeol
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.3
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    • pp.435-442
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    • 2012
  • The cytochrome P-450 enzymes (CYP 2A6) regulate many endogenous signaling molecules and drugs. Aryl alkynes such as 2-ethynylnaphthalene are important P450 inhibitors which have been extensively studied as medicines or as an effective chemical probes for profiling mouse liver microsomal P-450. Here we have synthesized indole-based novel P450 inhibitor, 5-ethynyl indole 3, and showed that it has successfully inhibited CYP 2A6 by chemical inhibition reaction. By using HPLC equipped with a photo diode array(PDA) detector, all of the peaks derived from the enzymatic reaction have been characterized.

Perspectives on the Quality Assurance of Ginseng Products in North America

  • Fitzloff, J.F.;Yat, P.;Lu, Z.Z.;Awng, D.V.C.;Arnason, J.T.;Breemen, R.B.van;Ha, T.ll;Blumenthal, M.;Fong, H.H.S.
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.138-145
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    • 1998
  • A validated and reproducible HPLC method was developed for the profiling and quantitative analysis of ginsenosides in commercial products available in North America. Analysis of 280 Panax ginseng and Panax quinquefolius products showed profiles indicative of the presence of ginsenosides in the majority of these products. However, the quantitative contents of the products vary greatly, not only in products of different formulations, but also of products within each of the fromulations examined.

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Sugar content analysis and expression profiling of sugar related genes in contrasting Strawberry (Fragaria × ananassa) cultivars

  • Shanmugam, Ashokraj;Hossain, Mohammad Rashed;Natarajan, Sathishkumar;Jung, Hee-Jeong;Song, Jae-Young;Kim, Hoy-Taek;Nou, Ill-Sup
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.178-190
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    • 2017
  • $Fragaria{\times}ananassa$, a strawberry evolved from hybridization between F. virginiana and F. chiloensis, is a globally cultivated and consumed fruit crop valued for its flavor and nutritional value. Flavor and quality of fruits are determined by factors such as sugars and organic acids present during fruit development. These characteristics are highly subjective in different genotypes and affected by various environmental factors. In this study, we analyzed contents of major sugar compounds including fructose, glucose and sucrose by HPLC analysis in four cultivars namely, Maehyang, Seolhyang, Festival and Sweet Charlie. We identified 55 genes related to fructose, glucose, sucrose and soluble sugar regulation whose expression were analyzed in four cultivars at three developmental stages of the fruit namely, green, white and ripened stages. Expression of these genes across these progressive fruit developmental stages varied among cultivars. Among the 55 genes, genes FaFru3, FaSuc11 and FaGlu8 revealed differential patterns of expression along developmental stages of the fruit in high and low sugar-containing genotypes, respectively and may be putative candidates for sugar content in strawberries. Expression of genes are discussed with regard to corresponding sugar content in these genotypes. Further analysis and application of these genes may be valuable in developing high sugar containing cultivars via marker-assisted breeding.

Alteration in Phenolic Compounds and Antioxidant Activities of Aronia melanocarpa Ethanol Extracts following Fermentation Using Different Strains of Leuconostoc mesenteroides to Develop Natural Antibiotic Alternative (항생제 대체 천연물질을 위한 아로니아 주정 추출물 개발에 있어 다양한 Leuconostoc mesenteroides 균주를 이용한 발효가 페놀계 화합물 및 항산화활성 변화에 미치는 영향)

  • Hwang, Joo Hwan;Kang, Ju Hui;Lee, Ki Hwan;Lee, Jae Hoon;Lee, Sang Moo;Kim, Nam Hyung;Kim, Joo Young;Kim, Eun Joong
    • Korean Journal of Organic Agriculture
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    • v.22 no.4
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    • pp.825-839
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    • 2014
  • Antioxidant activity is important for reducing oxidative stress that causes various metabolic disorders. Metabolic disorders are highly related to loss of productivity in livestock. Therefore, development of effective antioxidant compounds originating from plants is important for organic agriculture. Phenolic compounds in edible plants are regarded as major components relevant to antioxidant activity. The present study investigated the changes in antioxidant activity and phenolic compound profiles of Aronia (Aronia meloncarpa) by fermentation using different strains of Leuconostoc mesenteroides. A total of 5 strains of L. mesenteroides were used as starter cultures and their ${\beta}$-glucosidase activities were measured. A total of 6 experiment runs were prepared, one for control (uninoculated) and the others (inoculated) for treatments. For biological activity, antioxidant and antibacterial activities were measured. For phenolic compound profiling, TLC and HPLC analysis were performed. The strains of KACC12313 and KACC12315 showed greater enzyme activity than others. Treatment with KCCM35046 showed strong and broad antibacterial activity against to Listeria monocytogenes. Treatments with KCCM35046 and KACC12315 showed the highest total polyphenol content. The highest antioxidant activity was found in KACC12315 treatment. No remarkable alteration was found in thin layer chromatography (TLC) analysis. In phenolic compound profiling analysis, KCCM35046 showed notable alteration in compound area ratio compared to others and also showed the highest caffeic acid content. In chlorogenic acid, treatments with KCCM35046 and KACC12315 showed great content than others. Treatment with KACC12315 showed the greatest content of trans-ferulic acid. As a result of relative performance indexing analysis, L. mesenteroides KCCM35046 and KACC12315 were selected as the best strain for the fermentation of Aronia.

Cardioprotective effect of ginsenoside Rb1 via regulating metabolomics profiling and AMP-activated protein kinase-dependent mitophagy

  • Hu, Jingui;Zhang, Ling;Fu, Fei;Lai, Qiong;Zhang, Lu;Liu, Tao;Yu, Boyang;Kou, Junping;Li, Fang
    • Journal of Ginseng Research
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    • v.46 no.2
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    • pp.255-265
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    • 2022
  • Background: Ginsenoside Rb1, a bioactive component isolated from the Panax ginseng, acts as a remedy to prevent myocardial injury. However, it is obscure whether the cardioprotective functions of Rb1 are related to the regulation of endogenous metabolites, and its potential molecular mechanism still needs further clarification, especially from a comprehensive metabolomics profiling perspective. Methods: The mice model of acute myocardial ischemia (AMI) and oxygen glucose deprivation (OGD)-induced cardiomyocytes injury were applied to explore the protective effect and mechanism of Rb1. Meanwhile, the comprehensive metabolomics profiling was conducted by high-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (HPLC-Q/TOF-MS) and a tandem liquid chromatography and mass spectrometry (LC-MS). Results: Rb1 treatment profoundly reduced the infarct size and attenuated myocardial injury. The metabolic network map of 65 differential endogenous metabolites was constructed and provided a new inspiration for the treatment of AMI by Rb1, which was mainly associated with mitophagy. In vivo and in vitro experiments, Rb1 was found to improve mitochondrial morphology, mitochondrial function and promote mitophagy. Interestingly, the mitophagy inhibitor partly attenuated the cardioprotective effect of Rb1. Additionally, Rb1 markedly facilitated the phosphorylation of AMP-activated protein kinase α (AMPKα), and AMPK inhibition partially weakened the role of Rb1 in promoting mitophagy. Conclusions: Ginsenoside Rb1 protects acute myocardial ischemia injury through promoting mitophagy via AMPKα phosphorylation, which might lay the foundation for the further application of Rb1 in cardiovascular diseases.