• Title/Summary/Keyword: Gradient elution

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Separation of liquiritin, glycyrrhizic acid and glabridin from licorice by RP-HPLC (RP-HPLC를 이용한 감초에서 liquiritin, glycyrrhizic acid, glabridin의 분리)

  • Tian, Minglei;Yan, Hongyuan;Row, Kyung Ho
    • Analytical Science and Technology
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    • v.21 no.2
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    • pp.102-108
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    • 2008
  • Reversed-phase high performance liquid chromatography (RP-HPLC) was used for the simultaneous determination of liquiritin (LQ), glycyrrhizic acid (GA) and glabridin from licorice. An optimized run condition was selected with a binary gradient elution of methanol-water which ramped 35/65 to 80/20 (vol. %) in 0.0-8.0 min and a flow rate of 1.0 mL/min. A good linearity was obtained between 0.2 mg/mL and 1.0 mg/mL for LQ and GA, and 0.01 mg/mL-0.2 mg/mL for glabridin with the relative standard deviations less than 0.90% (n=5). The developed method was successfully applied to determination of the three components from licorice samples. The mean recoveries of three components are 80.79% for liquiritin, 89.71% for glycyrrhizic acid and 72.50% for glabridin.

Isolation and Identification of Inhibitory Compounds from Morus alba cv. Kuksang on α-amylase and α-glucosidase (국상(Kuksang) 뽕잎(Morus alba L.)으로부터 α-amylase와 α-glucosidase 저해 물질 분리 및 동정)

  • Choi, Moo-Young;Cho, Young-Je
    • Journal of Life Science
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    • v.25 no.8
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    • pp.870-879
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    • 2015
  • The objective of this research was to evaluate the inhibitory activities of phenolic compounds isolated from mulberry (Morus alba) leaves of 109 types against α-amylase and α-glucosidase. The inhibitory activity of the water extracts from Morus alba cv. Kuksang against α-amylase and α-glucosidase were determined as 93.8% and 48.7% respectively. The total phenolic content of extracts from Morus alba cv. Kuksang was 9.7±0.2 mg/g soluble in water and 14.3±0.2 mg/g soluble in ethanol. The inhibitory activity of the water extracts from Morus alba cv. Kuksang at 200 μg/ml phenolics concentration against α-amylase and α-glucosidase were determined as 100% and 82.6% respectively. The purification of inhibitory compounds was carried out by Sephadex LH-20 and MCI-gel CHP-20 column chromatography using a gradient elution procedure by nomal phase type (EtOH→distilled water) and reverse phase type (distilled water→MeOH). The quercetin was confirmed to be the chemical structure of the inhibitory compound against α-amylase and α-glucosidase by spectroscopic analysis of FAB-MS, NMR and IR spectrum.

Rapid Determination of Ginsenosides Rb1, Rf, and Rg1 in Korean Ginseng Using HPLC (HPLC를 이용한 고려인삼 중 진세노사이드 Rb1, Rf 및 Rg1의 신속분석 방법 개발)

  • Hong, Hee-Do;Choi, Sang-Yoon;Kim, Young-Chan;Lee, Young-Chul;Cho, Chang-Won
    • Journal of Ginseng Research
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    • v.33 no.1
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    • pp.8-12
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    • 2009
  • A simple gradient HPLC method for rapid determination of major ginsenosides ($Rg_1$ and $Rb_1$) and unique ginsenoside (Rf) of Korean ginseng (Panax ginseng C.A. Meyer) was developed. Within 50min, three ginsenosides have been separated and identified on $\mu$-Bondapak $C_{18}$ column ($3.9{\times}300\;mm$, $10{\mu}m$) with gradient elution using water and acetonitrile as a mobile phase. The method was validated in terms of linearity, accuracy, and precision. The correlation coefficients ($r^2$) for calibration curves of ginsenosides were over 0.9997. The developed HPLC method was successfully applied to the analysis of ginseng samples and the recoveries of ginsenosides were in the range of $101.1{\sim}115%$ with RSD<3.2%. The developed method could be used for rapid evaluation of the ginsenosides $Rg_1$, $Rb_1$, and Rf.

Glycomacropeptide Hydrolysed from Bovine K-Casein ; II. Chromatographic Changes of k-Casein Macropetide as Related to Trichloroacetic Acid Concentration (우유의 k-Casein에서 분해된 Glycomacropeptide에 관한 연구; II. Trichloroacetic Acid의 농도에 따른 k-Casein Macropeptide 분별 특성의 변화)

  • Moon Yong-Il;Lee Wonjae;Oh Sejong
    • Food Science of Animal Resources
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    • v.25 no.4
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    • pp.478-482
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    • 2005
  • Bovine k-casein macropeptide was prepared by adding TCA (3, 6, and $12\%$) treatment after chymosin reaction. Each TCA soluble macropeptide was fractionated into five peak by ion exchange column chromatography. In proportiion to TCA concentrations, the ratio of peak area showed different the elution pattern. At the 6 and $12\%$ TCA concentration, area ratio of P-I which did not content carbohydrates was decreased to 19.9 and $17.0\%$ of total peak area respectively. The area of P-III was changed from $10.2\%\;to\;26.2\;and\;13.2\%$ when the TCA concentration was increased from 3 to 6 and $12\%$ Cholera toxin binding activity of k-casein macropeptide eluted at $0.17\~0.18M$ NaCl gradient was not inhibited by 6 and $12\%$ TCA treatments. The use of $6\%$ TCA as extraction buffer was feasible and led to an effective separation of the peak III.

Application of Isolated Tyrosinase Inhibitory Compounds from Persimmon Leaves (감나무 잎으로 부터 분리한 tyrosinase 억제물질의 응용)

  • Cho, Young-Je;An, Bong-Jeun;Kim, Jeung-Hoan
    • Journal of Life Science
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    • v.21 no.7
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    • pp.976-984
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    • 2011
  • Total phenolic content was the highest in 60% ethanol extracts at 21.91 mg/g, and inhibitory activity against tyrosinase of 60% ethanol extracts was higher than ethanol extracts of other concentration. The inhibitory compounds against tyrosinase from Persimmon leaves were purified using Sephadex LH-20, MCI-gel CHP-20 column chromatography with gradient elution. Two purified compounds were isolated as a result. The chemical structures of each compound were determined and identified using $^1H$-NMR and $^{13}C$-NMR, FAB-Mass. The compounds were confirmed as (+)-gallocatechin and prodelphinidin B-3. The tyrosinase inhibitory activities of purified (+)-gallocatechin and prodelphinidin B-3 were 29.5 and 40.2%, respectively. The inhibitory activities of (+)-gallocatechin and prodelphinidin B-3 against melanin biosynthesis in melanoma cell were 32.5 and 46.7%. The safety of essence with tyrosinase inhibitory compounds from persimmon leaves was also assessed by various safety profiles. First, changes in pH (4.90~4.95) and viscosity (23,000~26,000 cP) was not detected for 60 days. Essence also showed stability against temperature and light for 60 days. All these findings suggest that extracts from persimmon leaves have a great potential as a cosmetical ingredient with a potent whitening effect.

Development of a rapid HPLC method for the determination of penciclovir in human plasma using a monolithic column and its application to a bioequivalence study (모노리틱 칼럼을 이용한 혈장 중 펜시를로버의 HPLC 신속분석법 개발 및 이를 이용한 생물학적동등성시험)

  • Kim, Jin Hee;Park, Ah Yeon;Jung, Eun Ha;Lee, Cheol Woo;Lee, Tae Ho;Youm, Jeong-Rok
    • Analytical Science and Technology
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    • v.20 no.4
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    • pp.323-330
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    • 2007
  • A simple and rapid HPLC method with fluorescence detection(FLD) for quantitation of penciclovir in human plasma using a monolithic column was developed and validated. Penciclovir and ganciclovir(internal standard, I.S.) were separated on a Chromolith column RP-18e ($4.6{\times}100mm$) with a mobile phase consisting of a mixture of (A) methanol/50 mM sodium phosphate buffer containing 200 mg/L sodium dodecyl sulfate (3/97, pH 2.5) and (B) methanol/50 mM sodium phosphate buffer containing 200 mg/L sodium dodecyl sulfate (50/50, pH 2.5) at a flow gradient of $1.6{\sim}4.0mL/min$. The retention times of penciclovir and internal standard were less than 4.0 min. Calibration curve was linear ($R^2=0.9994$) over a concentration range of $0.1{\sim}5{\mu}g/mL$. Intra-day precision, accuracy and inter-day precision were 1.36~8.55 %, 92.8~100.0 % and 0.93~5.62 %, respectively with a limit of quantitation at $0.1{\mu}g/mL$. The present HPLC-FLD method is sensitive, precise and accurate. The method described herein has been successfully used for the bioequivalence study of a famciclovir formulation product after oral administration to healthy Korean volunteers.

Method Development of Ellagic Acid as Marker Compound for Standardization of Gochang Bokbunja (Rubus coreanus Miquel) as Functional Ingredient (고창 복분자의 기능성원료 표준화를 위한 지표성분으로서 Ellagic Acid의 분석법 개발)

  • Kim, Yunjeong;Han, Song-Hee;Jeon, Ji-Yeong;Hwang, Minho;Im, Yong-Jin;Chae, Soo-Wan;Kim, Min-Gul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.11
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    • pp.1554-1558
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    • 2012
  • Method development and validation of ellagic acid for the standardization of Gochang Bokbunja as a functional ingredient and health food were accomplished. A Symmetry$^{(R)}$ (C18, $4.6{\times}250mm$, $5.0{\mu}m$) column was used with a gradient elution system of 1% formic acid in water and acetonitrile. This method was validated according to specificity, linearity, accuracy, precision test, and recovery test. Specificity was confirmed with identical retention time, and calibration curves of ellagic acid showed good linear regression ($R^2$ >0.9996). Relative standard deviations (RSD) of data from the intra- and inter-day experiments were less than 2.28% and 2.84%, except in the low limit of quality control (LLOQ, $1{\mu}g/mL$) sample. The results of the recovery test were from 89.17% to 97.92% with RSD values from 0.05 to 0.14%. Therefore, we performed analysis of ellagic acid as a marker compound in Gochang Bokbunja extracts. The amount of ellagic acid in Gochang Bukbunja was about $1.918{\mu}g/mg$ (0.192%) in the three times analysis, and RSD was less than 2.36% by the validated method. These results suggest that the developed HPLC method is simple, efficient, and could contribute to the quality control of Gochang Bokbunja extract as a functional ingredient.

Analytical Validation of Rosmarinic Acid in Water Extract of Perilla frutescens Britton var. acuta Kudo as Functional Health Ingredient (건강기능식품 기능성 원료로써 장흥 차조기 열수 추출물의 지표성분인 로즈마린산 분석법 검증)

  • Park, Sung-Yong;Kim, Jung-Eun;Choi, Chul-Yung;Lee, Dong-Wook;Kim, Ki-Man;Yoon, Goo;Yoon, In-Su;Moon, Hong-Seop;Cho, Seung-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.1
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    • pp.85-88
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    • 2015
  • This study attempted to establish an HPLC analysis method for determination of marker compounds as a part of material standardization for the development of health functional food materials from Perilla frutescens Britton var. acuta Kudo. The quantitative determination method of rosmarinic acid as a marker compound of P. frutescens Britton var. acuta Kudo extract (PFE) was optimized by HPLC analysis using a C18 column ($4.6{\times}150mm$, $5{\mu}m$) with 0.1% acetic acid as the elution gradient and methanol as the mobile phase at a flow rate of 1 mL/min and detection wavelength of 280 nm. The HPLC/UV method was applied successfully to quantification of the marker compound in PFE after validation of the method with linearity, accuracy, and precision. The method showed high linearity in the calibration curve at a coefficient of correlation ($R^2$) of 0.9995, and the limit of detection and limit of quantitation were $0.36{\mu}g/mL$ and $1.2{\mu}g/mL$, respectively. Relative standard deviation (RSD) values of data from intra- and inter-day precision were less than 3.21% and 1.43%, respectively. Recovery rate test at rosmarinic acid concentrations of 12.5, 25 and $50{\mu}g/mL$ scored between 97.04~98.98% with RSD values from 0.25~1.97%. These results indicate that the established HPLC method is very useful for the determination of marker compound in PFE to develop a health functional material.

A Study on Optimization for Separation of Phenols and Preconcentration-Separation of Trace Phenols in Reversed-Phase Liquid Chromatography (역상 액체 크로마토그래피에서 페놀류의 분리 최적화 및 미량 페놀류의 농축-분리에 관한 연구)

  • Lee Dai Woon;Lee Sung Won;So, Min Jeong;Cho Byung Yun
    • Journal of the Korean Chemical Society
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    • v.37 no.5
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    • pp.513-522
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    • 1993
  • The purpose of this study is to optimize the selectivity of mobile phase solvents for separation of 25 phenols in reversed phase liquid chromatography and to accomplish the simultaneous preconcentration and separation of trace phenols from water samples. Phenols used in this study were classified into three groups, chloro-, methyl-, and nitrophenols. Quaternary solvent mobile phases were employed to improve the selectivity. Overlapping resolution maps(ORM) as a statistical simplex techniques was used to predict the optimum solvent system. Additional criterion such as pH and temperature were also investigated. In order to improve the resolution and decrease the analysis time, isoselective multisolvent gradient elution system was employed with ORM-Prism method. The simultaneous preconcentration and separation of trace phenols from water samples were performed by using XAD-2/Dowex 1-X8 tandem column. When the extraction efficiency was evaluated by sampling up to 1 L of distilled water, recovery of the phenols, except phenol, was above 90% and the limit of detection of the phenols was 5 ppb. The XAD-2/Dowex 1-X8 method was superior to C18 cartridge in terms of recovery and selectivity.

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Simultaneous analysis of 12 kind maker components in Banhabaekchulchhonma-tang by RP-HPLC (RP-HPLC를 이용한 반하백출천마탕에서 12종 지표성분의 동시분석)

  • Lee, Kwang Jin;Yang, Hye Jin;Ma, Jin Yeul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.7
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    • pp.4682-4691
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    • 2014
  • Banhabaekchulchhonma-tang is a traditional Korean herbal prescription with a range of pharmacological activities. In this study, the simultaneous analysis of 12 kinds of marker components, Homogentisic acid, Hesperidin, Naringin, Alisol A, Atractylenolide II, Atractylenolide III, Ginsenoside Rg1, Formononetin, Gastrodin, Berberine, Palmatine and 6-gingerol, in Banhabaekchulchhonma-tang was performed using high-performance liquid chromatography (RP-HPLC). The standard sample of commercial $C_{18}$ reversed phase-column using water (0.1% TFA) and acetonitrile as the mobile phase with a step gradient elution mode. The flow rate (1.0mL/min), injection volume ($10{\mu}L$) and column oven temperature ($40^{\circ}C$) at a 200, 220, 280 and 340nm wavelength was conducted. All calibration curves of the standard components showed good linearity ($r^2$ >0.999). In addition, the limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.012 to $0.878{\mu}g/mL$ and 0.009 to $0.290{\mu}g/mL$. The precision intra-day and inter-day were ranged from 0.07 to 1.21% and 0.20 to 0.90%, respectively. The recoveries ranged from 97.17 to 108.40%. Each sample amount showed a very small change. These results highlight the efficient quality evaluation of Banhabaekchulchhonma-tang.