• 제목/요약/키워드: Gradient array

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황련해독탕 중 3종 생리활성 물질의 HPLC-DAD 동시 정량분석법 확립 (Simultaneous Analysis of three Marker Components in Hwangryunhaedok-tang by HPLC-DAD)

  • 양혜진;원진배;마진열;마충제
    • 약학회지
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    • 제55권1호
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    • pp.64-68
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    • 2011
  • In this study, a high performance liquid chromatography-diode array detector method was established, for simultaneous determination of three compounds, berberine, palmatine and geniposide in Hwangryunhaedok-tang, To develop and validate method, $C_{18}$ column (5 ${\mu}M$, 4.6 mm${\times}$250 mm) was used with gradient mobile phase, water containing 0.1% trifluoroacetic acid (TFA) and MeOH at the column temperature of $30^{\circ}C$. UV wavelength was set at 230 and 280 nm. Validation of the chromatography method was evaluated by linearity, precision and accuracy test. Calibration curve of standard components showed good linearity ($R^2$ > 0.9999). The limits of detection (LOD) and limits of quantification (LOQ) varied from 0.05 to 0.17 ${\mu}g/ml$ and 0.15 to 0.53 ${\mu}g/ml$, respectively. The relative standard deviations (RSDs) data of intra-day and inter-day test were in less than 2.99% and 1.90%, respectively. The results of the accuracy test were in the range of 98.36 to 102.52% with RSDs values 0.32 to 1.98%. The results of validation indicated that this method was a very accurate and sensitive assay.

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

  • 이지혜;엄영란;박화용;이재훈;마진열
    • 한국한의학연구원논문집
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    • 제15권2호
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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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HPLC-DAD-MS/MS를 이용한 금은화 생리활성 물질의 동시분석 (Simultaneous Analysis of Bioactive Metabolites from Lonicera japonica Flower Buds by HPLC-DAD-MS/MS)

  • 유성광;전주은;강경원;강삼식;신종헌
    • 약학회지
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    • 제52권6호
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    • pp.446-451
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    • 2008
  • A high-performance liquid chromatography (HPLC) with diode array detector (DAD) and electrospray ionization mass spectrometry (ESI-MS) was established for the simultaneous determination of chlorogenic acid (1), sweroside (2), luteolin-7-O-glucoside (3), (E)-aldosecologanin (4) and 3,5-dicaffeoylquinic acid (5) from Lonicera joponica flower buds. The optimal chromatographic conditions were obtained on an ODS column (5 ${\mu}m$, 4.6${\times}$150 mm) with the column temperature $25^{\circ}C$. The mobile phase was composed of (A) water with 0.1% formic acid and (B) acetonitrile with 0.1% formic acid using a gradient elution, the flow rate was 0.3 ml/min. Detection wavelength was set at 250 nm. All calibration curves showed good linear regression ($r^2$>0.994) within test ranges. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.05${\sim}$1.95% and 0.15${\sim}$2.26%, respectively, and the overall recoveries of 97.71${\sim}$103.65% for the five compounds analyzed. The verified method was successfully applied to quantitative determination of the three types (phenolic compounds, iridoids and flavonoids) of bioactive compounds in 21 commercial L. japonica flower buds samples from different markets in Korea and China. The analytical results demonstrated that the contents of the five analytes vary significantly with sources.

MOLECULAR GAS AND RADIO JET INTERACTION: A CASE STUDY OF THE SEYFERT 2 AGN M51

  • MATSUSHITA, SATOKI;TRUNG, DINH-V;BOONE, FRDERIC;KRIPS, MELANIE;LIM, JEREMY;MULLER, SEBASTIEN
    • 천문학논총
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    • 제30권2호
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    • pp.439-442
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    • 2015
  • We observed multiple CO transition lines and the HCN(1-0) line at ~ 1" (~ 34 pc) or higher resolution toward the Seyfert 2 nucleus of M51 using the IRAM Plateau de Bure Interferometer (PdBI) and the Submillimeter Array (SMA). All the images show very similar overall molecular gas distribution; there are two discrete clouds at the eastern and western sides of the nucleus, and the western cloud exhibits an elongated distribution and velocity gradient along the radio jet. In addition, high HCN(1-0)/CO(1-0) brightness temperature ratios of about unity have been observed, especially along the radio jet, similar to those observed in shocked molecular gas in our Galaxy. This strongly indicates that the molecular gas along the jet is shocked, that the radio jet and the molecular gas are interacting, and the jet is entraining both diffuse (CO) and dense (HCN) molecular gas outwards from the circumnuclear region. This is the first clear imaging of the outflowing molecular gas entrained by the AGN jet, and showing the detailed physical status of outflowing molecular gas. Since a relatively high HCN(1-0)/CO(1-0) ratio has been observed in the high velocity wing of ultraluminous infrared galaxies, it can also be explained by a similar mechanism to those we describe here.

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

  • 엄영란;이지혜;마진열
    • 한국한의학연구원논문집
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    • 제16권1호
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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.

HPLC를 이용한 이중탕 중 3종 활성성분의 동시분석법 확립 (Simultaneous Determination of Three Compounds in Ejung Tang by HPLC-DAD and LC-ESI-MS)

  • 이보형;마진열;원진배;양혜진;윤보라;마충제
    • 생약학회지
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    • 제43권1호
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    • pp.10-15
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    • 2012
  • An accurate and sensitive analysis method was established for simultaneous determination of three bioactive compounds (glycyrrhizin, 6-gingerol and ginsenoside Rg3) in the Ejung Tang with high-performance liquid chromatography (HPLC)-photodiode array detection (DAD)-electrospray ionization (ESI)-Mass spectrometry (MS). The optimizing chromatographic separations a were acquired by an $C_{18}$ column ($5{\mu}m$, $4.6I.D{\times}250mm$, SHISHEDO) using gradient elution with water comprising 0.1% TFA(trifluoroacetic acid) and acetonitrile at a performing temperature of $35^{\circ}C$. Flow rate was 1.0 ml/min. A detection UV wavelength set at 205 nm and 250 nm. The three compounds were identified by electrospray ionization mass spectrometry. All calibration curves indicated great linear regression within test ranges ($R^2>0.9997$). The established method provided acceptable precision and accuracy. The relative standard deviations (RSDs) of intra-day and inter-day were less than 2.00% and 3.00%, respectively. The recoveries were found to range from 94.49 to 101.10% for the three compounds analyzed. These results showed that this method was effective and reliable for quality control of Eiung-Tang.

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

  • 서창섭;김정훈;신현규;황석연;김병수
    • 생약학회지
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    • 제45권3호
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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.

Simultaneous Analysis of Bioactive Metabolites from Rehmannia glutinosa by HPLC-DAD-MS/MS

  • Won, Tae-Hyung;Ryu, Sung-Kwang;Kang, Sam-Sik;Shin, Jong-Heon
    • Natural Product Sciences
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    • 제16권2호
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    • pp.116-122
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    • 2010
  • A high-performance liquid chromatography (HPLC) with diode array detector (DAD) and electrospray ionization mass spectrometry (ESI-MS) was established for the simultaneous determination of five representative metabolites of the iridoid and phenolic classes from Rehmannia glutinosa. The optimal chromatographic conditions were obtained on an ODS column (5 mm, $4.6{\times}250\;mm$) with the column temperature at $25^{\circ}C$. The mobile phase was composed of water and acetonitrile using a gradient elution with the flow rate 0.3 mL/min. Detection wavelength was set at 205 nm. All calibration curves showed good linear regression ($r^2$ > 0.997) within test ranges. Limits of detection (LOD) and quantitation (LOQ) values were lower than 0.123 and $0.373\;{\mu}g/mL$, respectively. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.09 - 0.76% and 0.16 - 1.41%, respectively, and the overall recoveries of 99.03 - 102.67% for all of the compounds analyzed. In addition, effectiveness of diverse extraction methods was compared to each other for the development of standard analytic method. The verified method was successfully applied to the quantitative determination of five representative metabolites in twenty-one commercial Rehmannia glutinosa samples from different markets in Korea and China. The analytical results showed that the contents of the five analytes vary significantly with sources.

Simultaneous determination of 30 ginsenosides in Panax ginseng preparations using ultra performance liquid chromatography

  • Park, Hee-Won;In, Gyo;Han, Sung-Tai;Lee, Myoung-Woo;Kim, So-Young;Kim, Kyung-Tack;Cho, Byung-Goo;Han, Gyeong-Ho;Chang, Il-Moo
    • Journal of Ginseng Research
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    • 제37권4호
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    • pp.457-467
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    • 2013
  • A quick and simple method for simultaneous determination of the 30 ginsenosides (ginsenoside Ro, Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, 20(S)-Rg2, 20(R)-Rg2, 20(S)-Rg3, 20(R)-Rg3, 20(S)-Rh1, 20(S)-Rh2, 20(R)-Rh2, F1, F2, F4, Ra1, Rg6, Rh4, Rk3, Rg5, Rk1, Rb3, Rk2, Rh3, compound Y, compound K, and notoginsenoside R1) in Panax ginseng preparations was developed and validated by an ultra performance liquid chromatography photo diode array detector. The separation of the 30 ginsenosides was efficiently undertaken on the Acquity BEH C-18 column with gradient elution with phosphoric acids. Especially the chromatogram of the ginsenoside Ro was dramatically enhanced by adding phosphoric acid. Under optimized conditions, the detection limits were 0.4 to 1.7 mg/L and the calibration curves of the peak areas for the 30 ginsenosides were linear over three orders of magnitude with a correlation coefficients greater than 0.999. The accuracy of the method was tested by a recovery measurement of the spiked samples which yielded good results of 89% to 118%. From these overall results, the proposed method may be helpful in the development and quality of P. ginseng preparations because of its wide range of applications due to the simultaneous analysis of many kinds of ginsenosides.

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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    • 제25권3호
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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.