• Title/Summary/Keyword: reverse-phase high performance liquid chromatography

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High Purity Extraction and Simultaneous High-performance Liquid Chromatography Analysis of Curcuminoids in Turmeric (강황에서 curcuminoids의 고순도 추출 및 고성능 액체 크로마토그래피 동시분석)

  • Lee, Kwang-Jin;Ma, Jin-Yeul;Kim, Young-Sik;Kim, Dong-Seon;Jin, Yinzhe
    • Journal of Applied Biological Chemistry
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    • v.55 no.1
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    • pp.61-65
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    • 2012
  • Three major curcuminoids in turmeric (Curcuma longa), curcumin, demethoxycurcumin, and bisdemethoxycurcumin, were efficiently extracted by optimizing extraction condition and simultaneously analyzed and identified using reverse phase high-performance liquid chromatography, thin-layer chromatography, and liquid chromatography-mass spectrometry method. The highest yield of extraction amount 0.279 g, 9.30% was obtained by dipping method with extraction time of 7 h.

Efficient Isolation of Dihydrophaseic acid 3'-O-β-D-Glucopyranoside from Nelumbo nucifera Seeds Using High-performance Countercurrent Chromatography and Reverse-phased High-performance Liquid Chromatography

  • Rho, Taewoong;Yoon, Kee Dong
    • Natural Product Sciences
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    • v.24 no.4
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    • pp.288-292
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    • 2018
  • High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase highperformance liquid chromatography (RP-HPLC) method was developed to isolate dihydrophaseic acid 3'-O-${\beta}$-D-glucopyranoside (DHPAG) from the extract of Nelumbo nucifera seeds. Enriched DHPAG sample (2.3 g) was separated by HPCCC using ethyl acetate/n-butanol/water system (6:4:10, v/v/v, normal-phase mode, flow rate: 4.0 mL/min) to give 23.1 mg of DHPAG with purity of 88.7%. Further preparative RP-HPLC experiment gave pure DHPAG (16.3 mg, purity > 98%). The current study demonstrates that utilization of CCC method maximizes the isolation efficiency compared with that of solid-based conventional column chromatography.

Determination of Isoflavonoid Glucosides in Rhizomes of Belamcanda chinensis by High Performance Liquid Chromatography

  • Cui, Jiong-Mo;Chung, Ha-Sook;Woo, Won-Sick
    • Korean Journal of Pharmacognosy
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    • v.24 no.4
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    • pp.309-312
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    • 1993
  • A new method for separation and quantitative determination of isoflavone glucosides in rhizomes of Belamcanda chinensis (Iridaceae) by high performance liquid chromatography was elaborated. A reverse-phase system with a Spheri-5 RP-18 column using MeOH:HOAc:$H_2O$(24 : 5 : 71) as a mobile phase was developed. The isoflavonoids were detected at 268 nm and the analysis was successfully carried out within 15 min.

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Reverse-Phase High Performance Liquid Chromatographic Determination of Amino Acids after Precolumn Derivatization with 1-Dimethyl-aminonaphthalene-5-sulfonyl chloride (Dansyl 유도체화와 역상 고성능액체크로마토그래피에 의한 아미노산의 정량)

  • 나혜경;전덕영;홍윤호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.815-822
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    • 1993
  • This experiment was carried out to determine the separation condition of 1-dimethylaminonaphthalene-5-sulfony(Dansyl) derivatives of amino acids by reverse-phase high performance liquid chromatography with Nova-Pak C18 column. Determined solvent system was solvent A(200mA phosphate buffer pH 6.8 15%, acetonitrile 11%, water 74%) and solvent B(acetonitrile 65%, methanol 28%, water 7%). Linear gradient of solvent B was applied from 12% to 80% for 50min. Complete separation of 20 amino acids including asparagine and glutamine which constitute protein was achieved within 50min. As the detection limit was the range of picomole, the resolution power was excellent. Reproducibility of the retention time was less than mean $\pm$0.05min. According to the above optimum chromatographic conditions, the amino acid composition of some food and human blood was examined. The most affluent amino acid was alanine in human blood, aspartic acid and glutamic acid in soy sauce, alanine and threonine in soy milk and proline in milk and yoghurt.

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Determination of Thiamine in Pharmaceutical Preparations by Reverse Phase Liquid Chromatography Without Use of Organic Solvent

  • Suh, Joon Hyuk;Kim, Junghyun;Jung, Juhee;Kim, Kyunghyun;Lee, Seul Gi;Cho, Hyun-Deok;Jung, Yura;Han, Sang Beom
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1745-1750
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    • 2013
  • A novel green aqueous mobile phase modified with room temperature ionic liquids (RTILs) was employed in the absence of volatile organic solvents or ion-pairing reagents to analyze thiamine, a very polar compound, by reverse phase high performance liquid chromatography (RP-HPLC). Due to its strongly hydrophilic nature, thiamine was eluted near the column dead time ($t_0$) using a mobile phase without adding RTILs or ion-pairing reagents, even if a 100% aqueous mobile phase, which has weak elution power under reverse phase conditions, was used. Thus, 1-ethyl-3-methyl-imidazolium hexafluorophosphate ([EMIM][$PF_6$]), which has the strongest chaotropic effect, was selected as a mobile phase additive to improve retention and avoid baseline disturbances at $t_0$. Various mobile phase parameters such as cation moiety, chaotropic anion moiety, pH and concentration of RTILs were optimized to determine thiamine at the proper retention time. Method validation was performed to assess linearity, intra- and inter-day accuracy and precision, recovery and repeatability; all results were found to be satisfactory. The developed method was also compared to the current official United States Pharmacopoeia (USP) and Korean Pharmacopoeia (KP) methods using an organic mobile phase containing an ionpairing reagent by means of evaluating various chromatographic parameters such as the capacity factor, theoretical plate number, peak asymmetry and tailing factor. The results indicated that the proposed method exhibited better efficiency of thiamine analysis than the official methods, and it was successfully applied to quantify thiamine in pharmaceutical preparations.

Isolation and Purification of Bioactive Materials Using High-Performance Counter-Current Chromatography (HPCCC) (고속역류크로마토그래피 기술을 이용한 생리활성 물질의 분리 및 정제)

  • Jung, Dong-Su;Shin, Hyun-Jae
    • KSBB Journal
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    • v.25 no.3
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    • pp.205-214
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    • 2010
  • Many successive liquid-liquid extractions occur enabling purification of the crude material to occur. In high performance counter-current chromatography (HPCCC), crude material is partitioned between two immiscible layers of solvent phases. The stationary phase (SP) is retained by hydrodynamic force field effect and the mobile phase (MP) is pumped through the column. Purification occurs because of the different solubility of the components in the liquid mobile and stationary phases. There are many key benefits of liquid stationary phases such as high mass and volume injection loadings, total sample recovery, and easy scale-up. Many researchers showed that predictable scale-up from simple test is feasible with knowledge of the stationary phase retention for the planned process scale run. In this review we review the recent advances in HPCCC research and also describe the key applications such as natural products and synthetics (small or large molecules).

Determination of Ginseng Saponins by Reversed-Phase High Performance Liquid Chromatography (역상 고속액체크로마토그라피를 이용한 홍삼 사포닌의 정량)

  • Kim, Cheon-Suk;Kim, Se-Bong
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.1
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    • pp.21-25
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    • 2001
  • Major saponins in ginseng were analysed using reverse phase high performance liquid chromatography with binary mobile phase gradient control system instead of normal phase column. The optimum condition were as following : reverse phase column; ${\mu}{\beta}ondapak\;C_{18}$ column (Waters, $3.9mm{\times}300\;mm,\;5{\mu}m$), methyl cyanaide/water binary mobile phase gradient control system, solvent flow rate; 1.5 ml/min, and UV($203{\mu}m$ ) detector. The complete separation of ginsenoside $Rb_1,\;Rb_2,\;Rc,\;Rd,\;Re,\;Rf\;and\;Rg_1$ was achieved within 55 min. The Regression coefficients of the calibration curves for seven ginsenosides were 0.99.

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Development of High Performance Liquid Chromatography for Paclitaxel Purification from Plant Cell Cultures

  • Kim, Jin-Hyun;Choi, Hyung-Kyoon;Hong, Seung-Suh;Lee, Hyun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.204-210
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    • 2001
  • Paclitaxel can be produced in high yield and with a high degree of purify from plant cell cultures of Taxus chinensis. The complete purification method was systematically established and described. This method was an efficient procedure for the purification of paclitaxel from crude paclitaxel, consisting or reverse-phase chromatography, followed by a normal-phase chromatography. The two-stage HPLC purification scheme serves as an effective and economical approach for resolving paclitaxel from complex mixtures of taxoids, with high purify (>99%) and low impurities (<0.1%). The process is readily scalable to a pilot plant and eventually to a production environment where multikilogram quantities of material are expected to be produced. The process has been optimized to minimize solvent usage, complexity, and operating costs.

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Determination of Ginseng Saponins by Reversed-Phase High Performance Liquid Chromatography (역상 고속 액체크로마토그래피를 이용한 인삼 사포닌의 분석)

  • Jeong, Seung-Il;Kim, Choen-Suk;Lee, No-Woon;Choi, Kang-Ju;Lee, Yong-Gu;Kim, Il-Kwang
    • Analytical Science and Technology
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    • v.11 no.6
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    • pp.436-439
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    • 1998
  • Ginseng saponins were analysed using reversed-phase high performance liquid chromatography with several columns. The optimum conditions were as following : reverse phase column; Novapak $C_{18}$ ODS column ($3.9mm{\times}150mm$, $5{\mu}m$), acetonitrile/water binary mobile phase gradient controller system, solvent flow rate; 1.5 mL/min, and UV (203 nm) detector. The complete separation of ginsenoside $Rb_1$, $Rb_2$, Rc, Rd, Re, Rf and $Rg_1$ was achieved within 50 min. The regression coefficients of the calibration curves for seven ginsenosides were 0.98~0.99.

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Determination of Patulin in Commercial Apple Juice in Korea by High Performance Liquid Chromatography (High Performance Liquid Chromatography를 이용한 국내 시판 사과 주스의 Patulin 분석)

  • Cho, Wan-Il;Choi, Young-Boong;Moon, Tae-Wha
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.412-416
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    • 1997
  • Seven brands of commercial apple juice in Korea obtained from local stores were analyzed for patulin. Sep-Pak silica cartridge purification of the extract by ethyl acetate offered a good separation of patulin from other components. Patulin was determined by reverse phase liquid chromatography using a J'sphere $4\;{\mu}m$ ODS-H80 column with an ultraviolet detector set at 274 nm. Patulin concentrations in four samples ranged from 4 to $20\;{\mu}g/L$ and the other three samples from 30 to $45\;{\mu}g/L$. The limit of detection was $3\;{\mu}g/L$, and the recovery was $80{\sim}90%$ at the contamination level of $3.78{\sim}125.9\;{\mu}g/L$.

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