• Title/Summary/Keyword: marker constituents

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Changes in Antioxidant Activity and Total Phenolic Content of Water Spinach (Ipomoea aquatic Forsk.) under In Vitro Biomimicking System

  • Lee, A-Young;Kim, Young-Suk;Shim, Soon-Mi
    • Food Engineering Progress
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    • v.14 no.4
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    • pp.342-345
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    • 2010
  • The purpose of current study was to examine bioaccessibility of antioxidant activity and total phenolic content in each part of water spinach (Ipomoea aquatic Forsk.). In vitro biomimicking system simulated human digestive fluid was employed in order to measure bioavailable anti-oxidative effect and phenolic content. Antioxidant activity and total phenolic content was measured by using the DPPH method and the Folin-Ciocalteu assay, respectively. Stem of water spinach had a higher DPPH free radical scavenging effect (5.43 mg/mL for $IC_{50}$) than leaf (5.95 mg/mL for $IC_{50}$), while leaf had a greater level of total phenolic content (287.45 ${\mu}g$ GAE/mL) than stem (216.45 ${\mu}g$ GAE/mL). Bioaccessible antioxidant capacity and digestive stability of total phenolic content showed a similar pattern to what found in raw materials. Our result also indicated that total phenolic content was not found to be a major marker for prediction of antioxidant activity. It is plausible that other constituents such as vitamin E and C in water spinach could be contributors for antioxidant activities.

Inhibition of Interleukin-4 and β-Hexosaminidase Release in RBL-2H3 Cells by Compounds Isolated from Lobelia chinensis

  • Kim, Tae Young;Jo, Beom-Geun;Park, No-Jun;Park, Young-Hun;Kim, Su-Nam;Yang, Min Hye
    • Natural Product Sciences
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    • v.27 no.4
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    • pp.251-256
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    • 2021
  • Lobelia chinensis Lour. has commonly been used in traditional Chinese medicine for the treatment of antidote, diuretic, diarrhea, and inflammation. This study aimed to identify the active compounds in an aqueous extract of L. chinensis responsible for its anti-atopic effect in vitro using RBL-2H3 cells. A chemical investigation of secondary metabolites in an aqueous extract of L. chinensis led to the isolation of nine chemical constituents, which included the four marker compounds, and these were evaluated for their inhibitory effects on IL-4 mRNA expression and the release of β-hexosaminidase in propidium iodide-induced RBL-2H3 cells. We found diosmetin and fraxidin inhibited cellular IL-4 mRNA expression, and that diosmetin and 6,8-dimethoxycoumarin inhibited DNP-specific IgE-induced degranulation in these cells. Our study suggests that diosmetin, fraxidin, and 6,8-dimethoxycoumarin are potential candidates for the treatment of atopic diseases.

Effects of Watercress Containing Rutin and Rutin Alone on the Proliferation and Osteogenic Differentiation of Human Osteoblast-like MG-63 Cells

  • Hyun, Hanbit;Park, Heajin;Jeong, Jaehoon;Kim, Jihye;Kim, Haesung;Oh, Hyun Il;Hwang, Hye Seong;Kim, Ha Hyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.4
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    • pp.347-352
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    • 2014
  • Most known osteoporosis medicines are effective for bone resorption, and so there is an increasing demand for medicines that stimulate bone formation. Watercress (N. officinale R. Br.) is widely used as a salad green and herbal remedy. This study analyzed a watercress extract using ultra-performance liquid chromatography/mass spectrometry, and identified a rutin as one of its major constituents. Osteogenic-related assays were used to compare the effects of watercress containing rutin (WCR) and rutin alone on the proliferation and differentiation of human osteoblast-like MG-63 cells. The reported data are expressed as percentages relative to the control value (medium alone; assigned as 100%). WCR increased cell proliferation to $125.0{\pm}4.0%$ ($mean{\pm}SD$), as assessed using a cell viability assay, and increased the activity of alkaline phosphatase, an early differentiation marker, to $222.3{\pm}33.8%$. In addition, WCR increased the expression of collagen type I, another early differentiation marker, to $149.2{\pm}2.8%$, and increased the degree of mineralization, a marker of the late process of differentiation, to $122.9{\pm}3.9%$. Rutin alone also increased the activity of ALP (to $154.4{\pm}12.2%$), the expression of collagen type I (to $126.6{\pm}6.2%$), and the degree of mineralization (to $112.3{\pm}5.0%$). Daidzein, which is reported to stimulate bone formation, was used as a positive control; the effects of WCR on proliferation and differentiation were significantly greater than those of daidzein. These results indicate that WCR and rutin can both induce bone formation via the differentiation of MG-63 cells. This is the first study demonstrating the effectiveness of either WCR or rutin as an osteoblast stimulant.

Simultaneous HPLC Determination of Marker Compounds for the Standardization of Hedyotis diffusa (백운풀의 지표성분 설정 및 품질표준화를 위한 정량 분석법)

  • Bang, Han-Yeol;Yang, Eun-Ju;Kim, Jeong-Ah;Song, Kyung-Sik
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1025-1031
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    • 2013
  • From a 95% ethanolic extract of H. diffusa, four marker compounds (HD1~HD4) were isolated, which were relatively unique and exist in comparably high contents. The structures of marker compounds were identified as digitolutein (1), 2-hydroxy-3-methylanthraquinone (2), (E/Z)-6-O-p-coumaroyl scandoside methyl ester (4:1 mixture) (3), and (E/Z)-6-O-p-methoxycinnamoyl scandoside methyl ester (4:1 mixture) (4), respectively, on the basis of $^{13}C$ and $^1H$-NMR analyses. The calibration curves of marker compounds showed high linearity, as their correlation coefficient ($R^2$) were in the range of 0.9991~0.9999. In addition, the limit of detection (LOD) and the limit of quantification (LOQ) were $0.03{\sim}0.07{\mu}g/ml$ and $0.099{\sim}0.231{\mu}g/ml$, respectively. The intra-day/inter-day precision and accuracy were 0.23~2.00%/0.25~1.16% and 94.60~108.44%/94.73-110.23%, respectively. The optimal HPLC conditions for the simultaneous quantification of HD1~HD4 were as follows: stationary phase; Merck Chromolith RP-18e ($100{\times}4.6mm$, $5{\mu}m$), column temp.; room temperature, UV detection at 280 nm, flow rate; 2.0 ml/min, injection volume; $10{\mu}l$, mobile phase; start with the mixture of 80% solvent A ($H_2O$ containing 0.5% acetic acid) and 20% solvent B (methanol containing 0.5% acetic acid) and gradually decrease solvent A to 40% in 9 min., then retain this condition to 18 min. Under the HPLC condition, the four marker compounds 1~4 were successfully separated without any interference of other constituents. The results obtained in this study are expected to be helpful for the development of nutraceutics and natural medicines and for the quality control of this plant.

Simple Sequence Repeat Markers Linked to Quantitative Trait Loci Controlling Seed Weight, Protein and Oil Contents in Soybean (콩에서 종실의 무게와 oil 및 단백질 함량을 조절하는 양적 형질 유전자좌와 연관된 simple sequence repeat marker)

  • Kim, Hyeun-Kyeung;Kang, Sung-Taeg;Choung, Myoung-Gun;Jung, Chan-Sik;Oh, Ki-Won;Baek, In-Youl;Son, Beung-Gu
    • Journal of Life Science
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    • v.16 no.6
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    • pp.949-954
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    • 2006
  • Soybean [Glycine max (L.) Merr.] is an important crop, accounting for 48% of the world market in oil crops. Improvement of the quality and quantity of soybean seed constituents is one of the most important objectives in soybean breeding. Protein content and seed size are important properties to determine the quality of tofu and soy sprouts respectively. The objective of this study was to identify quantitative trait loci (QTLs) that control seed weight, protein and oil content in soybean. The 117 $F_{2:10}$ recombinant inbred lines (RlL) developed from a cross of 'Keunolkong' and 'Shinpaldalkong' were used. Narrow-sense heritability estimates based on a plot mean on seed weight, protein and oil content were 0.8, 0.78 and 0.71, respectively. Four independent QTLs for seed weight were identified from linkage group (LG) F, I and K. Five QTL for protein content were located on LG D1b, E, H, I and L. Oil content was related with six QTLs located on LG D1b, E, G, I, J and N. Protein and oil content have three common QTLs on LG D1b, E and I. Thus, we identified major loci improving soybean seed quality.

Simultaneous Determination of Baicalin and Glycyrrhizin in Eul-Ja-Tang by HPLC/DAD

  • Lee, Mi-Kyeong;Lee, Ki-Yong;Kim, Seung-Hyun;Park, Jung-Hyun;Cho, Jung-Hee;Oh, Mi-Hyun;Baek, Ju-Hyun;Kim, Hyo-JIn;Kim, Young-Choong;Sung, Sang-Hyun
    • Natural Product Sciences
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    • v.14 no.3
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    • pp.147-151
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    • 2008
  • A high performance liquid chromatographic (HPLC) method for the simultaneous determination of marker constituents, baicalin and glycyrrhizin was established for the quality control of traditional herbal medicinal preparation, Eul-Ja-Tang (EJT). Separation and quantification were successfully achieved with a Waters XTerra RP18 column ($5{\mu}m$, 4.6 mm I.D. ${\times}$ 150 mm) by gradient elution of a mixture of acetonitrile and water containing 0.03% phosphoric acid (pH 2.03) at a flow rate of 1.0 mL/min. The diode-array UV/VIS detector (DAD) was used for the detection and the wavelength for quantification was set at 250 nm. The presence of baicalin and glycyrrhizin in this decoction was ascertained by retention time, spiking with each authentic standard and UV spectrum. Both baicalin and glycyrrhizin showed good linearity ($r^2$ > 0.999) in a relatively wide concentration ranges. The R.S.D. for intra-day and inter-day precision was less than 5% and the limits of detection (LOD) were about 30 ng. The mean recovery of each compound was 99.5 - 101.2% with R.S.D. values less than 4.0%. This method was successfully applied to the determination of contents of baicalin and glycyrrhizin in three commercial products of EJT, which resulted in the difference in the contents of these compounds. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial EJT products.

Quantitative Analysis of the Fifteen Constituents in Hyangso-San by LC-MS/MS (LC-MS/MS를 이용한 향소산 중 15종 성분의 정량분석)

  • Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.47 no.4
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    • pp.381-388
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    • 2016
  • Hyangso-san is a traditional herbal medicine that consists of the seven herbal medicines, Cyperi Rhizoma, Perillae Folium, Atractylodis Rhizoma, Citri Unshius Pericarpium, Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma Crudus, and Allii Fistulosi Bulbus. Hyangso-san has long been clinically used to treat the influenza, including headache, ferver, chills, and pantalgia. In this study, we were performed the simultaneous analysis of the 15 marker compounds (liquiritin apioside, liquiritin, ferulic acid, naringin, hesperidin, rosmarinic acid, liquiritigenin, kaempferol, glycyrrhizin, nobiletin, 6-gingerol, elemicin, atractylenolide III, nootkatone, and atractylenolide I) in Hyangso-san using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS). Column for the separation of the 15 ingredients was used a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ by using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient condition. Identifications of all analytes were performed using a Waters ACQUITY TQD LC-MS/MS system. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}L$, respectively. Correlation coefficient of the calibration curve was ${\geq}0.9958$. The values of limits of detection and quantification of the 15 components were 0.002-4.29 and 0.01-12.88 ng/mL, respectively. The result of an analysis using the established LC-MS/MS method, kaempferol and atractylenolide I were not detected, while other 13 compounds were 0.08-56.87 mg/g in lyophilized Hyangso-san sample.

Development of an UPLC-DAD Method for Simultaneous Analysis of Eight Marker Compounds of Bulhwangeumjeonggi-san (UPLC-DAD를 이용한 불환금정기산의 다성분 동시분석법 개발)

  • Lee, Kyung-hee;Lamichhane, Ramakanta;Kumar, Sharma Dipak;Raj, Pandeya Prakash;Kim, Se-Gun;Jung, Hyun-Ju
    • Korean Journal of Pharmacognosy
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    • v.47 no.4
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    • pp.366-373
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    • 2016
  • Bulhwangeumjeonggisan (BHGJGS) is a traditional herbal formulation generally used in the treatment of cold and gastritis. BHGJGS consists of eight herbal plants; Atractylodis Rhizoma, Magnoliae Cortex, Citri Pericarpium, Glycyrrhizae Radix, Agastachis Herba, Pinelliae Rhizoma, Zingiberis Rhizoma and Zizyphi Fructus. Complete standardization of this formulation has not been done yet. So, a simple and accurate method was developed and validated using Ultra Performance Liquid Chromatography (UPLC) with Diode Array Detector (DAD) for the standardization of BHGJGS. UPLC conditions were optimized using a c18 RP-Amide column with mobile phase; 0.1% phosphate buffer and acetonitrile, detection wavelength; 210 and 325 nm. The linearities of calibration curves were acceptable ($R^2$>0.9994), and the limit of detection and quantification were within the ranges of 0.011-0.091 and $0.034-0.277{\mu}g/ml$ respectively. The relative standard deviation (RSD) of intra- and inter-day precisions were under 3.61%. The RSD of repeatability was under 0.68 %. The results of recovery test were 94.4-107.9%, and the RSD were under 4.6%. The developed method was used to find the contents of standard constituents in BHGJGS mix extract powder, and two commercial formulation (A and B). The data show that the developed method was specific, sensitive, accurate, and precise for analysis of BHGJGS components.

Simultaneous Determination of Seven Compounds by HPLC-PDA and Cytotoxicity of Samchulkunbi-tang (삼출건비탕의 HPLC-PDA 동시 분석법 설정 및 세포독성)

  • Seo, Chang-Seob;Lee, Mee-Young;Kim, Jung-Hoon;Lee, Jin-Ah;Shin, Hyeun-Kyoo
    • The Korea Journal of Herbology
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    • v.25 no.3
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    • pp.65-71
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    • 2010
  • Objectives:To develop and validate HPLC-PDA methods for simultaneous determination of seven constituents in Samchulkunbi-tang (SKT). Additionally, we investigated the cytotoxicity against BEAS-2B cell line and splenocytes of SKT. Methods:Reverse-phase chromatography using a Gemini $C_{18}$ column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 230, 254 and 280 nm, were used for quantification of the three marker components of SKT. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. The cytotoxicity of SKT were measured by the CCK-8 assay method. Results:Calibration curves were acquired with $r^2$>0.9999, and the relative standard deviation (RSD) values (%) for intra- and inter-day precision were less than 6.0%. The recovery rate of each compound was in the range of 86.89-109.78%, with an RSD less than 4.0%. The contents of seven compounds in SKT were 1.39-6.84 mg/g. SKT had no cytotoxicity effect at 50-200 ${\mu}g$/mL concentrations. Conclusions:The established method will be helpful to improve quality control and in vitro efficacy study of SKT.

Simultaneous Determination of Paeoniflorin and Glycyrrhizin in Sayuk-san by HPLC/DAD

  • Lee, Mi-Kyeong;Lee, Ki-Yong;Kim, Seung-Hyun;Jeon, Min-Ji;Cho, Jung-Hee;Oh, Mi-Hyun;Baek, Ju-Hyun;Kim, Hyo-Jin;Sung, Sang-Hyun
    • Journal of Pharmaceutical Investigation
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    • v.39 no.1
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    • pp.23-27
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    • 2009
  • A high performance liquid chromatographic (HPLC) method for the simultaneous determination of marker constituents, paeoniflorin and glycyrrhizin was established for the quality control of traditional herbal medicinal preparation, Sayuk-san (SYS). Separation and quantification were successfully achieved with a Waters XTerra RP18 column ($5{\mu}m$, $4.6mm\;I.D.{\times}150mm$) by gradient elution of a mixture of acetonitrile and water containing 0.03% phosphoric acid (pH 2.03) at a flow rate of 1.0 mL/min. The diode-array UV/vis detector (DAD) was used for the detection and the wavelength for quantification was set at 230 nm. The presence of paeoniflorin and glycyrrhizin in this decoction was ascertained by retention time, spiking with each authentic standard and UV spectrum. All two compounds showed good linearity ($r^2$>0.996) in a relatively wide concentration ranges. The R.S.D. for intra-day and inter-day precision was less than 7.3% and the limits of detection (LOD) were less than 55.7 ng. The mean recovery of each compound was $102.3{\sim}111.1%$ with R.S.D. values less than 4.6%. This method was successfully applied to the determination of contents of paeoniflorin and glycyrrhizin in three commercial products of SYS. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial SYS products.