• 제목/요약/키워드: Reverse-phase high performance liquid chromatography(RP-HPLC)

검색결과 25건 처리시간 0.027초

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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    • 제24권4호
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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
    • 생약학회지
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    • 제24권4호
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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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RP-HPLC법에 의한 무우의 매운 성분인 4-methylthio-3-butenyl isothiocyanate의 분리 및 정제 (Purification of 4-Methylthio-3-butenyl isothiocyanate the Pungent Principle in Radish Roots by RP-HPLC)

  • 김미리;이혜수
    • 한국식품조리과학회지
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    • 제2권2호
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    • pp.16-20
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    • 1986
  • 무우 속의 매운 맛 성분을 용매추출법 ($CCl_4$,사용)으로 분리하여 RP-HPLC법에 의해 정제한 후에 관능검사에 의해 pungency test를 하고, UV spectrum분석, GC/MC 분석을 수행한 결과 4-methylthio-3-butenyl isothiocyanate 임을 확인하였다. RP-HPLC에서 mobile Phase로 70% acetonitrile을 사용하였는데, 정제된 4-methylthio-3-butenyl isothiocfanate는 단일 peak로 분리되어 나왔고 이때의 retention time은 5.2분이였다. 아울러 RP-TLC에서 Rf치는 0.9이었다.

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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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    • 제34권6호
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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.

Prevalence of Antibiotics in Nectar and Honey in South Tamilnadu, India

  • Solomon, RD. Jebakumar;Santhi, V. Satheeja;Jayaraj, Vimalan
    • Animal cells and systems
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    • 제10권3호
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    • pp.163-167
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    • 2006
  • Reverse-Phase High-performance Liquid Chromatography (RP-HPLC) based technique is one of the most sensitive techniques to detect the antibiotics present in honey. In the southern part of Tamilnadu, India, majority of the farmlands are occupied by plantations such as coconut, banana and rubber. A variety of antimicrobial compounds and antibiotics, which have been reported in pollen, nectar and other floral parts of the plant, gets accumulated in honey through honeybees (Apis mellifera). We have collected the nectar samples from banana (Musa paridasiaca) and rubber (Ficus elastica) flowers and the honey from honey hives of banana and rubber cultivated areas. The extracted nectar and honey samples are subjected to RP-HPLC analysis with authentic antibiotic standards. Nectar and honey samples showed 4-17, 11-29 ${\mu}g/kg$ of streptomycin, 2-29, 3-44 ${\mu}g/kg$ of ampicillin and 17-34, 26-48 ${\mu}g/kg$ of kanamycin respectively.

셀레늄이 강화된 이스트에서 셀레늄 단백질의 분리 및 확인 (Separation and identification of selenoproteins in selenium-enriched yeast)

  • 김경미;박용남
    • 분석과학
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    • 제26권6호
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    • pp.357-363
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    • 2013
  • Selenium-containing proteins were separated from selenium-enriched yeast (SEY) using Trizol$^{(R)}$ reagent followed by anion exchange (AE) chromatography. This method is simpler and less time consuming than electrophoresis. Five selenium containing proteins were identified by on-line AE HPLC-ICP/MS (high performance liquid chromatography-inductively coupled plasma/mass spectrometry). Each protein was enzymatically hydrolyzed to seleno-amino acids and separated with RP (reverse phase) HPLC for the identification of selenoproteins.

물리적 가공법을 이용한 강황으로부터 Curcuminoid의 고체상추출 (Solid-Phase Extraction of Curcuminoid from Turmeric Using Physical Process Method)

  • 이광진;양혜진;정상원;마진열
    • 생약학회지
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    • 제43권3호
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    • pp.250-256
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    • 2012
  • In order to extract the curcuminoid such as curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC) in turmeric (Curcuma longa), solvent extraction methods (dipping and ultrasonic extraction method) and solid-phase extraction (SPE) were used. RP-HPLC (reverse-phase high-performance liquid chromatography) and TLC (thin-layer chromatography) were used for identification and analysis the three curcuminoid. From the experimental results, it is evident that the percentage of curcuminoid extracted from turmeric by ultrasonic extraction method was higher than dipping method. The percentage of curcumin extracted from turmeric by pure methanol was higher than any aqueous methanolic composition. Moreover, the total peak area of three curcuminoid was above 92% in RP-HPLC using solid-phase extraction. These results will form a database for investigating the constituents of natural products and the resources of pharmaceutical, nutrition, and cosmetic products.

RP-HPLC법에 의한 무우의 4-methylthio-3-butenyl isothiocyanate의 정량 (Quantitative Determination of 4-methylthio-3-butenyl isothiocyanate in Radish Root by RP-HPLC)

  • 김미리;이혜수
    • 한국식품과학회지
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    • 제18권6호
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    • pp.475-478
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    • 1986
  • RP-HPLC방법에 의해 무우 속에 존재하는 MTBI의 함량을 정량적으로 측정 해 본 결과 10n moles/ml와 120 nmoles/ml사이에서 좋은 직선적인 상관관계(r=0.9997)를 나타내었으며, 편리하고 정확(오차 5%내외)하며 재현성(recovery 99%)이 높았다. 또, MTBI는 알카리용액 중에서는 서서히 분해되었으나 산성용액 중에서는 매우 빠르게 분해되었으며, 이 결과는 231nm에서 UV흡광도가 서서히 감소하는 결과와도 일치하였다. 한편, MTBI는 유기용매(65% acetenitrile)에서 비교적 안정하였다. 또한. 한국산 생무우 중에 함유된 MTBI는 최적추출조건을 PH8.5에서 1분내에 분석하였을때 $210\sim420{\mu}moles/100g$ 정도이었다.

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Determination of bioavailability of tolperisone HCI by HPLC

  • Yang, Sang-In;Choi, Sun-Hee;Lee, Seung-Jin;Jang, Choon-Gon;Lee, Seok-Yong
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.241.1-241.1
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    • 2002
  • Tolperisone hydrochloride is used as a muscle relaxant. Very few assay methods of tolperisone were reported. such as potentiometry. spectrophotometry and high performance thin layer chromatography. In addition. there is no report related to HPLC method to determine the tolperisone level in biological sample. In this study, A very sensitive reverse phase high performance liquid chromatographic (RP-HPLC) method for the determination of tolperisone HCI in plasma has been developed. (omitted)

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In vitro Folding of Recombinant Hepatitis B Virus X-Protein Produced in Escherichia coli: Formation of Folding Intermediates

  • Kim, Sun-Ok;Sohn, Mi-Jin;Jeong, Soon-Seog;Shin, Jeh-Hoon;Lee, Young-Ik
    • BMB Reports
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    • 제32권6호
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    • pp.521-528
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    • 1999
  • The folding of recombinant hepatitis B virus X-protein (rHBx) solubilized from Escherichia coli inclusion bodies was investigated. By sequential dialysis of urea, rHBx was folded into its native structure, which was demonstrated by the efficacy of its transcriptional activation of the adenovirus major late promoter (MLP), fluorescence spectroscopy, and circular dichroism (CD) analysis. The decrease in CD values at 220 nm and a corresponding blue shift of the intrinsic fluorescence emission confirmed the ability of rHBx to refold in lower concentrations of urea, yielding the active protein. Equilibrium and kinetic studies of the refolding of rHBx were carried out by tryptophan fluorescence measurements. From the biphasic nature of the fluorescence curves, the existence of stable intermediate states in the renaturation process was inferred. Reverse phase-high performance liquid chromatography (RP-HPLC) analysis further demonstrated the existence of these intermediates and their apparent compactness.

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