• Title/Summary/Keyword: High-performance liquid chromatography/Photodiode array detector

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Radical Scavenging Activity and Content of Cynarin (1,3-dicaffeoylquinic acid) in Artichoke (Cynara scolymus L.)

  • Jun, Neung-Jae;Jang, Ki-Chang;Kim, Seong-Cheol;Moon, Doo-Young;Seong, Ki-Cheol;Kang, Kyung-Hee;Tandang, Leoncia;Kim, Phil-Hoon;Cho, So-Mi K.;Park, Ki-Hun
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.244-248
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    • 2007
  • The contents of total phenol and total flavonoid of artichoke (Cynara scolymus L.) were measured. The antioxidant activity of the artichoke was evaluated based on its potential as a scavenging the ABTS radical. These results showed the antioxidant activity of artichoke has a close relationship with the total flavonoid content. The compound showing antioxidant activity was isolated from the artichoke by repeated column chromatography and recrystallization. Based on the spectrometric studies, the compound was identified as 1,3-dicaffeoylquinic acid, known as cynarin. The content of cynarin from heads and leafs of the artichoke determined by $C_{18}$ reversed phase HPLC (high-performance liquid chromatography) coupled with photodiode array detector was 10.15 and 0.67 mg/g, respectively. This compound showed potent antioxidant activities against DPPH and ABTS radicals ($EC_{50}$ = 14.09 and 28.85 ${\mu}M$, respectively).

Composition and Quantitative Analysis of Stilbenoids in Mulberry (Morus alba L.) Leaves and Fruits with DAD/UV HPLC (DAD/UV HPLC를 이용한 뽕잎과 오디(Morus alba L.)에 함유된 Stilbenoids 조성 및 함량 분석)

  • Kim, Ji-Sun;Ha, Tae-Youl;Ahn, Ji-Yun;Kim, Hyun-Ku;Kim, Sun-A
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.1
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    • pp.124-128
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    • 2008
  • This study was aimed to analyze the composition and to quantify the contents of stilbenoids in the leaves and fruits of Morus alba L. using high performance liquid chromatography with phodtodiode array detector and UV detector. Optimal wavelength for the detection of various stilbenoids such as resveratrol, piceatannol, rhapontigenin, astringin, pterostilbene, piceid, rhaponticin and vitisin A was screened by DAD detector and set to 308 nm. Seven kinds of stilbenoids except vitisin A were identified in fruits, while 5 kinds of stilbenoids in leaves. Total stilbenoids contents were $609.15{\pm}7.24$ mg/100 g d.w. in fruits and $188.57{\pm}1.70$ mg/100 g d.w in leaves. Stilbenoids contents in fruits were 3 times higher than those in leaves. Rhaponticin was the most profound stilbene, analyzed to $389.26{\pm}5.22$ mg/100 g d.w. (63.8% of total stilbenoids) in fruits and $99.17{\pm}2.79$ mg/100 g d.w. (52.5% of total stilbenoids) in leaves. Astringin and piceatannol were only detected in fruits and vitisin A was not detected. Contents of piceid and rhaponticin were higher than those of aglycone forms, rhapontigenin and resveratrol.

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.

The influences of extraction time and pressure on the chemical characteristics of Gyejibokryeong-hwan decoctions

  • Kim, Jung-Hoon;Lee, Nari;Shin, Hyeun-Kyoo;Seo, Chang-Seob
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.1-6
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    • 2014
  • Objectives : This study was aimed to compare Gyejibokryeong-hwan (GBH) decoctions produced using different pressure levels for various extraction times to find the optimal extraction conditions through extraction yield, total soluble solids content (TSSC), hydrogen ion concentration (pH), and the contents of chemical compounds. Methods : Decoctions of GBH were prepared under the pressure levels of 0 or $1kgf/cm^2$ for 30-180 min using water as extraction solvent. The extraction yield, TSSC, and pH were measured, and the amounts of the chemical compounds were determined using high performance liquid chromatography-photodiode array detector. Results : The higher pressure and longer extraction time increased the values of TSSC and extraction yield, while decreased the pH value. The decoctions produced in 180 min by pressurized method and produced in 150 min by non-pressurized method showed maximum values of extraction yield and TSSC with minimum value of pH. The amounts of chemical compounds showed variations in pressurized and non-pressurized decoction during overall extraction times. The influences of pressure and extraction time on extraction yield, TSSC, pH, and the contents of chemical compounds were confirmed by regression analysis, which showed that all extraction values were significantly affected by at least one of two extraction factors, pressure and extraction time. Conclusions : This study suggests that the pressure and extraction time can significantly affect the extraction efficiency of components from GBH decoctions. However, optimal extraction conditions could not be chosen due to the variation of the amounts of chemical compounds.

Simultaneous Determination of Liquiritin, Nodakenin, Hesperidin and Glycyrrhizin in Bojungikgi-tang Using HPLC-PDA (HPLC-PDA를 이용한 보중익기탕 중 Liquiritin, Nodakenin, Hesperidin 및 Glycyrrhizin의 동시분석)

  • Seo, Chang-Seob;Kim, Jung-Hoon;Shin, Hyeun-Kyoo
    • YAKHAK HOEJI
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    • v.57 no.3
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    • pp.187-193
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    • 2013
  • Bojungikgi-tang has been widely used for enhancement of physical fitness in Korea. The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of four marker compounds, liquiritin, nodakenin, hesperidin, and glycyrrhizin in Bojungikgi-tang (Buzhongyiqi-tang in Chinese, Hochuekkito in Japanese), a traditional Korean herbal prescription. The column for optimizing HPLC separation was used a Gemini $C_{18}$ column at column oven temperature of $40^{\circ}C$ with 1.0% (v/v) aqueous acetic acid (A) and 1.0% (v/v) acetic acid in acetonitirle (B) by gradient flow. The flow rate was 1.0 ml/min and the detector was a photodiode array (PDA) set at 254 nm, 280 nm, and 335 nm. Calibration curves of four components were acquired with $r^2$ values ${\geq}0.9999$. The recoveries were found to range 92.11~105.68% with relative standard deviations (RSDs, %) value less than 2.50%. The RSD values of intraand inter-day precision were 0.07~2.50% and 0.16~1.99%, respectively. The contents of liquiritin, nodakenin, hesperidin and glycyrrhizin in Bojungikgi-tang were 3.85~3.92 mg/g, 2.27~2.32 mg/g, 4.14~4.19 mg/g, and 3.39~3.45 mg/g, respectively. The established simultaneous analysis method will be effective for quality control of Bojungikgi-tang.

Simultaneous Analysis of Six Constituents in Oyaksungi-san using HPLC-PDA (HPLC-PDA를 이용한 오약순기산 중 6종 성분의 동시분석)

  • Seo, Chang-Seob;Kim, Jung-Hoon;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.20 no.2
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    • pp.37-46
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    • 2012
  • Objectives : Oyaksungi-san(Wuyaoshunqisan) has been used for treatment of stroke and rheumatoid arthritis in Korea. In this study, a simple and accurate high-performance liquid chromatography(HPLC) method was established for simultaneous determination of six main components, liquiritin, ferulic acid, naringin, hesperidin, neohesperidin, and glycyrrhizin in Oyaksungi-san, a traditional Korean herbal prescription. Methods : The analytical column for separation of six constituents was used a Gemini $C_{18}$ column maintained at $40^{\circ}C$. The mobile phase consisted of two solvent systems, 1.0% (v/v) acetic acid in $H_2O$ (A) and 1.0% (v/v) acetic acid in acetonitrile (B) by gradient flow. The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 254 nm for glycyrrhizin, 280 nm for liquiritin, naringin, hesperidin, and neohesperidin, and 320 nm for ferulic acid. Results : Calibration curves were acquired with $r^2$ values ${\geq}0.9998$. The results of recovery test were 91.58%-105.90% with a relative standard deviations (RSDs, %) value less than 2.0%. The values of RSD for intra- and inter-day precision were 0.03%-1.72% and 0.03%-1.63%, respectively. The contents of the six compounds in Oyaksungi-san were 0.33-9.30 mg/g. Conclusions : The newly established HPLC method will be helpful to improve quality control of Oyaksungi-san.

Quantitative Analysis of Marker Substances of Paeonia lactiflora by Solid Fermentation (작약의 고체발효에 따른 지표성분의 함량분석)

  • Lee, Ji-Hye;Um, Young-Ran;Park, Hwa-Yong;Lee, Jae-Hoon;Ma, Jin-Yeul
    • Korean Journal of Oriental Medicine
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    • v.15 no.2
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    • pp.119-124
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    • 2009
  • The purpose of this study was investigation of quantitative analysis of marker substances in Paeonia lactiflora extracts by solid fermentation. High performance liquid chromatography (HPLC) for the determination of albiflorin and paeoniflorin in P. lactiflora extracts by solid fermentation, the separation method was performed on C18 column ($250\;mm\;{\times}\;4.6\;mm$, $5\;{\mu}m$, RS tech) using gradient solvent mixtures of water-acetonitrile with photodiode array detector (230nm). The flow rate was 1.0 ml/min. Retention time of albiflorin and paeoniflorin was about 28.88, 31.92 min and linearity of calibration was showed good result(r2 = 0.9998, 0.9996), respectively. Content of albiflorin was $0.090\;{\pm}\;0.03%$ in P. lactiflora extract(control), $0.102\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Paecilomyces japonica, $0.056\;{\pm}\;0.01%$ in P. lactiflora extract fermented with Ganoderma lucidum, $0.093\;{\pm}\;0.00%$ in P. lactiflora extract fermented with honey and $0.046\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk. Content of paeoniflorin was $4.506\;{\pm}\;0.13%$ in control, $2.599\;{\pm}\;0.04%$ in P. lactiflora extract fermented with Paecilomyces japonica, $1.222\;{\pm}\;0.03%$ in P. lactiflora extract fermented with Ganoderma lucidum, $2.750\;{\pm}\;0.05%$ in P. lactiflora extract fermented with honey and $0.847\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk, respectively. Content of the marker substances did not increase in all fermentation experiment group.

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Quantitative Analysis of Marker Substances in Solid Fermented Angelicae Gigantis Radix by HPLC (HPLC를 이용한 고체발효 당귀의 지표성분 분석)

  • Um, Young-Ran;Lee, Ji-Hye;Ma, Jin-Yeul
    • Korean Journal of Oriental Medicine
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    • v.16 no.1
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    • pp.173-178
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    • 2010
  • The purpose of this study was investigation of quantitative analysis of marker substances in solid fermented Angelicae Gigantis Radix by High performance liquid chromatography(HPLC). HPLC was performed for determination of nodakenin and decursin in solid fermented Angelicae Gigantis Radix extract, the separation method was performed on C18 column ($250\;mm\;{\times}\;4.6\;mm$, $5\;{\mu}m$, RS tech) using gradient solvent mixtures of water-acetonitrile with photodiode array detector (330 nm). The flow rate was 1.0 ml/min. Retention time of nodakenin and decursin was about 11.47, 46.79 min and linearity of calibration was showed good result(r2=0.9999, 0.9999), respectively. Content of nodakenin was $0.76\;{\pm}\;0.02%$ in control, $0.31\;{\pm}\;0.00%$ in Angelicae Gigantis Radix extract fermented with Paecilomyces japonica(SDT)(p<0.01), $0.51\;{\pm}\;0.02%$ in Angelicae Gigantis Radix extract fermented with Ganoderma lucidum(SYT)(p<0.01), $0.82\;{\pm}\;0.03%$ in Angelicae Gigantis Radix extract fermented with honey(SST)(p<0.05) and $0.88\;{\pm}\;0.01%$ in Angelicae Gigantis Radix extract fermented with Nuruk(SNT)(p<0.01). Content of decursin was $4.50\;{\pm}\;0.08%$ in control, $2.90\;{\pm}\;0.05%$ in Angelicae Gigantis Radix extract fermented with Paecilomyces japonica(SDT)(p<0.01), $2.65\;{\pm}\;0.08%$ in Angelicae Gigantis Radix extract fermented with Ganoderma lucidum(SYT)(p<0.01), $4.46\;{\pm}\;0.11%$ in Angelicae Gigantis Radix extract fermented with honey(SST) and $4.73\;{\pm}\;0.04%$ in Angelicae Gigantis Radix extract fermented with Nuruk(SNT)(p<0.05), respectively.

Quantitative Analysis of Anthraquinones in Cassiae Semen by Processing Method (수치에 따른 결명자 주요 Anthraquinone의 함량분석)

  • Seo, Chang-Seob;Kim, Jung-Hoon;Shin, Hyeun-Kyoo;Hwang, Seock-Yeon;Kim, Byoung-Soo
    • Korean Journal of Pharmacognosy
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    • v.45 no.3
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    • pp.200-208
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    • 2014
  • In this study, we performed quantification determination of four major components including aurantio-obtusin, emodin, chrysophanol, and physcion in the 70% ethanol extracts of non-processed Cassiae Semen and processed Cassiae Semen using a high-performance liquid chromatography coupled with photodiode array detector. The analytical column for separation of the 4 constituents used a Gemini $C_{18}$ column kept at $40^{\circ}C$ by the gradient elution with 1.0% (v/v) acetic acid in water and 1.0% (v/v) acetic acid in acetonitrile as mobile phase. The flow rate was 1.0 mL/min and the injection volume was $10{\mu}L$. The amount of aurantio-obtusin, emodin, chrysophanol, and physcion in non-processed Cassiae Semen were 0.07%, 0.02%, 0.25%, and 0.10%, respectively. The amount of aurantio-obtusin, emodin, chrysophanol, and physcion in processed Cassiae Semen were 0.04-0.14%, 0.01-0.03%, 0.02-0.42%, and 0.01-0.24%, respectively. Consequently, the optimal processing condition of Cassiae Semen for the improvements of amounts of four anthraquinone compounds was obtained by roasting at $240^{\circ}C$ for 15 min.

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.