• Title/Summary/Keyword: Reversed-Phase HPLC

Search Result 357, Processing Time 0.027 seconds

HPLC Study for Egg White Analysis (달걀 단백질 분석을 위한 HPLC 연구)

  • Jeon, Young-Ju;Lee, Eun;Kim, In-Ho
    • KSBB Journal
    • /
    • v.22 no.2
    • /
    • pp.119-122
    • /
    • 2007
  • Since egg white contains various protein, it is important to research the protein distribution of egg white. Specially, lysozyme and ovalbumin important proteins, are used in medicine and food industry. Reversed phase high performance liquid chromatography has been used for separation of egg white, and column of RP-HPLC is available in variety. We have used C4, C8 and C18 columns to obtain chromatograms by varying carbon chain length of stationary phase. Long carbon chain length of stationary phase has good separation of egg white. Also, we have changed the composition of mobile phase (acetonitrile, water, and trifluoroacetic acid) to find optimum chromatograms. Acetonitrile and water composition of 50 : 50 show many peaks from egg white. Isocartic and gradient elution in RP-HPLC were used to compare the chromatography of egg white.

Recent Developments in High-performance Liquid Chromatography of Lipids

  • Christie, William W.
    • Journal of the Korean Applied Science and Technology
    • /
    • v.10 no.1
    • /
    • pp.1-8
    • /
    • 1993
  • The possibilities for HPLC analysis of lipids have been revolutionised by the availability of evaporative light-scattering detectors, with which the response is independent of the nature of the mobile phase and does not depend On the presence of specific chromophores in the lipids. It was thus possible to develop an HPLC procedure, involving ternary gradient elution, for separating all the lipid classes in animal tissues in a single step. Although reversed-phase HPLC has been widely used for the analysis of molecular species of lipids, sliver ion chromatography can be a valuable alternative. For example, a stable silver ion column for HPLC was developed which permitted resolution of molecular species of triacylglycerols, even from such complex samples as fish oils, again With light-scattering detection and gradient elution. The capacity for HPLC resolution of diastereomeric diacyl-sn-glycerol derivatives, prepared from triacylglycerols. has lead to a new simple method for stereospecific analysis of the latter.

Separation of Egg White Using HPLC with Change of Mobile Phase and Temperature (HPLC에서 이동상 변화와 온도에 따른 난백의 분리)

  • Do, Jin-Sun;Song, Shin-Young;Cho, Ki-Jung;Kim, In-Ho
    • Korean Chemical Engineering Research
    • /
    • v.49 no.6
    • /
    • pp.829-834
    • /
    • 2011
  • Lysozyme in egg white functions as bacteriolysis agent and ovalbumin plays a role as antigen in immune system. Egg white analysis methods usually include electrophoresis, gel permeation chromatography and reversed-phase HPLC(RP-HPLC). Among them, RP-HPLC was selected for rapid analysis and C18 column(Agilent, USA) was used as HPLC column. Optimum conditions were searched by changing mobile phase and temperature. Capacity factor and resolution were calculated and compared for various elution conditions. In the isocratic elution, mobile phase volume ratio was changed from 30/70/0.1 to 60/40/0.1(Acetonitrile(ACN)/Distilled water(DW)/Trifluoroacetic acid(TFA)). ACN composition was increased by 10% and temperature was set as $20^{\circ}C$. In the gradient elution, ACN/DW ratio was changed from 10/90 to 60/40 during 20 minute and temperature was varied as 20, 30 and $40^{\circ}C$. In the isocratic elution, three peaks were separated at 50/50/0.1. Lysozyme and ovalbumin were confirmed as first and third peak in three peaks respectively. In the gradient elution, four peaks were separated at $30^{\circ}C$. Lysozyme and ovalbumin were confirmed as first peak and third peak in four peaks respectively.

Simultaneous Determination of Vitamin A and E in Infant Formula by Rapid Extraction and HPLC with Photodiode Array Detection (신속추출법 및 PDA-HPLC에 의한 조제분유 중 비타민 A, E의 동시분석)

  • Kwak, Byung-Man;Lee, Ki-Woong;Ahn, Jang-Hyuk;Kong, Un-Young
    • Korean Journal of Food Science and Technology
    • /
    • v.36 no.2
    • /
    • pp.189-195
    • /
    • 2004
  • Rapid and simple method was developed for simultaneous determination of vitamins A and E contents in infant formula. Vitamins A and E were extracted by PDA-HPLC with reversed phase column using organic solvent, and their contents in Certified Reference Material (CRM) and infant formula were determined and compared with results of Food Standards Codex and AOAC method for evaluation of developed method, Vitamins A and E contents in CRM determined by developed method were within certified range of standard values. Developed method has great advantages of simple and rapid sample preparation and simultaneous determination of vitamins A and E by PDA-HPLC using reversed phase column.

Simultaneous Determination of the Water Soluble Vitamins in Multi-Nutrient Tablets by Reversed-Phase High-Performance Liquid Chromatography

  • Kim, Hyung-Soo;Jang, Duck-Kyu;Woo, Dong-Kyun;Woo, Kang-Lyung
    • Preventive Nutrition and Food Science
    • /
    • v.7 no.1
    • /
    • pp.12-17
    • /
    • 2002
  • Simultaneous determination of nine water-soluble vitamins contained in multi-nutrient tablets was carried out by reversed phase high-performance liquid chromatography (RP-HPLC) equipped with analytical $C_{18}$ column and UV (270 nm) detector. Those standard vitamins were successfully separated within 23 minutes by gradient elution with solvent A (0.5 M potassium phosphate monobasic) and solvent B (0.25 M potassium phosphate monobasic-methanol, 1:1). Calibration curves showed good linealities with correlation coefficients (> 0.92) in tested ranged respectively. The detection limits were considered to be 2.1 ng for ascorbic acids 60 ng for Vit B$_{6}$ 3 ng for p-aminobenzoic acid, 9 ng for niacinamide, 9 ng for thiamin, 5.0 ng for folic acid and 1.5 ng for riboflavin at 0.05 a.u.f.s. Solid phase extraction through Sep-Pak (C$_{18}$ ) cartridge was successfully applied for purification of water soluble vitamins in commercial multi-nutrient tablets.ts.

Mutation Analysis in β2-Adrenergic Receptor Gene by Denaturing High Performance Liquid Chromatography (DHPLC) (DHPLC를 이용한 β2-교감신경수용체 유전자에서의 돌연변이 분석)

  • Park, Sang-Bum;Oh, Chung-Hun;Kim, Jong-Wan;Jang, Won-Cheoul
    • Analytical Science and Technology
    • /
    • v.15 no.3
    • /
    • pp.190-195
    • /
    • 2002
  • We analysed mutation of ${\beta}_2$-adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed-phase high performance liquid chromatography (IP-RP-HPLC). We extracted genomic DNA from 50 asthma patients, amplified DNA using PCR, and analysed PCR product by DHPLC. As a result, we obtained that mutation frequency was 15 (30%) among 50 cases. Consequently DHPLC mutation detection was confirmed that the result of direct sequencing was coincide exactly.

Quantitation and Validation of Atorvastatin using HPLC-UV

  • Heine, Daniel;Yong, Chul-Soon;Kim, Jung-Sun
    • Journal of Pharmaceutical Investigation
    • /
    • v.37 no.3
    • /
    • pp.187-192
    • /
    • 2007
  • A reversed phase HPLC analysis of atorvastatin (AS) standard solution was performed using diclofenac (DF) as internal standard. Column oven temperature, flow rate and the composition of the mobile phase were varied in order to determine a practical system setup using a C18 column and UV detector. Two C18 columns of different length were compared regarding their influence on the AS peak shape. Based on these preliminary experiments a validation study was performed utilizing a C18 column at $62^{\circ}C$ with a mobile phase consisting of sodium phosphate buffer (0.05 M, pH 4.0), methanol and acetonitrile (40:50:10, v/v/v). The detection limit for AS was $0.1{\mu}g/ml$ and inter- and intra-day calibration curves were linear over a concentration range of $0.2-50{\mu}g/ml$. Accuracy and precision were satisfactory in the AS concentration range of $0.5-50{\mu}g/ml$.

Quantitative Determination of Lignans from Schizandra chinensis by HPLC (HPLC에 의한 오미자 리그난 성분의 정량분석)

  • Koo, Dong-Chil;Suh, Won-Se;Baek, So-Yoon;Shim, Sang-Hee
    • Korean Journal of Pharmacognosy
    • /
    • v.42 no.3
    • /
    • pp.233-239
    • /
    • 2011
  • A reversed phase HPLC method was established for quantitative determination of lignans including schizandrin (1), gomisin A (2), deoxyschizandrin (3), gomisin N (4), and wuweizisu C (5) from Schizandra chinensis using a binary gradient of acetonitrile and water as a mobile phase with UV detection at 254nm. In result, contents of schizandrin (1), gomisin A (2), deoxyschizandrin (3), gomisin N (4), and wuweizisu C (5) in Schizandra chinensis from Moonkyung were 8.41${\pm}$0.30, 3.01${\pm}$0.13, 1.06${\pm}$0.05, 7.69${\pm}$0.30, and 1.68${\pm}$0.06 mg/g, respectively.

Application of Reversed-Phase Solid Phase Extraction for the HPLC Analysis of Microcystins in Water (다량 시료중 마이크로시스틴의 농축 및 분석)

  • Kim, Myeong-Hee;Kim, Tae-Seung;Kim, Tae-Keun;Park, Sun Ku
    • Analytical Science and Technology
    • /
    • v.13 no.3
    • /
    • pp.399-402
    • /
    • 2000
  • To determine the concentrations of microcystins present in lake water or in tap water using high performance liquid chromatography, it is necessary to concentrate a large volume of water samples (about 20 L) into very small volume (0.1-0.3 mL). Concentration can be conveniently done when disc type solid phase extraction (SPE) apparatus is used. Using this apparatus we have investigated the recovery rates of three kinds of microcystins, RR, YR, LR. The recovery rates were relatively low and the reproducibilities were not good either. It is expected, however, that the appropriate selection of the disc conditioning and eluting solvents and reproducible reconcentration process after SPE will improve both the recovery rates and the reproducibilities.

  • PDF

Separation and Determination of Co(II) and Ni(II) Ion as their 4-(2-Pyridylazo) resorcinol Chelates by Reversed-Phase Capillary High-Performance Liquid Chromatography (역상 모세관-고성능 액체 크로마토그래피에 의한 코발트와 니켈 이온의 4-(2-피리딜아조)레조루신올 킬레이트로서의 분리 및 정량)

  • Chung, Yong-Soon;Chung, Won-Seog
    • Journal of the Korean Chemical Society
    • /
    • v.47 no.6
    • /
    • pp.547-552
    • /
    • 2003
  • Separation and determinations of Co(II) and Ni(II) ions as their 4-(2-pyridylazo)resorcinol(PAR) chelates by reversed-phase capillary high-performance liquid chromatography(RP-CpHPLC) were performed. Among many capillary columns, Vydac C4 column was selected and acetonitrile solution was used as mobile phase. The effect of pH and MeCN concentration(%) on the retention factor, k and peak intensity was examined and discussed. As a results, it was found that 22.5% MeCN and pH 5.60 was adequate as mobile phase for the separation of the two metal ions and determination of Co(II) ion, but the mobile phase condition for Ni(II) ion determination was 22.5% MeCN of pH 7.20. Detection limit(D.L., S/N=3) of Co(II) and Ni(II) ions were $2.0{\times}10{-7}$ M(14.9 ppb) and $1.0{\times}10{-6}$ M(59.2 ppb), respectively.