• Title/Summary/Keyword: mobile phase

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Mobile Phase Compositions for Ceramide III by Normal Phase High Performance Liquid Chromatography

  • Hong, Seung-Pyo;Lee, Chong-Ho;Kim, Se-Kyung;Yun, Hyun-Shik;Lee, Jung-Heon;Row, Kyung-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.1
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    • pp.47-51
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    • 2004
  • Ceramide III was prepared by the cultivation of Saccharomyces cerevisiae. Ceramide III was partitioned from the cell extracts by solvent extraction and analyzed by Normal Phase High Performance Liquid Chromatography (NP-HPLC) using Evaporative Light Scattering Detector (ELSD). We experimentally determined the mobile phase composition to separate ceramide III with NP-HPLC. Three binary mobile phases of n-hexane/ethanol, n-hexane/lsoprophyl Alcohol(IPA) and n-hexane/n-butanol and one ternary mobile phase of n-hexane/IPA/methanol were demonstrated. For the binary mobile phase of n-hexane/ethanol, the first mobile phase composition, 95/5(v/v), was step-increased to 72/23(v/v) at 3 min. In the binary mobile phase, the retention time of ceramide III was 7.87min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition of n-hexane/IPA/methanol, 85/7/8(v/v/v), was step-increased to 75/10/15(v/v/v) at 3 min. However, in the ternary mobile phase, the more peak area of ceramide III was observed.

Study of Retention of Mono-Substituted Phenols in Reversed-Phase Liquid Chromatography Based on the Linear Solvation Energy Relationships Using the Solvatochromic Parameters for Mobile Phases, ${\pi}_m^{\ast}, {\alpha}_m$ and ${\beta}_m$

  • Park, Jung-Hag;Jang, Myung-Duk;Kim, Sang-Tae
    • Bulletin of the Korean Chemical Society
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    • v.11 no.4
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    • pp.297-302
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    • 1990
  • Retention of mono-substituted phenols in reversed-phase liquid chromatography has been studied based on the linear solvation energy relationships using the solvatochromic mobile phase parameters, ${\pi}_m^{\ast}, {\alpha}_m$ and ${\beta}_m$. It has been observed that retention behavior of phenols in RPLC were well represented by regression equations vs. solvatochromic mobile phase parameters even though the equations may be incomplete due to lack of an explicit cavity term. Dependence of retention of monosubstituted phenols on the mobile phase properties were varied depending on the type of the organic cosolvent in the mobile phase, e.g., ${\beta}_m$ and {\alpha}_m$ in methanol-water system, but ${\pi}_m^{\ast} and ${\beta}_m$ in THF-water system. It has been suggested that retention of phenols in methanol-water system is controlled by the solvophobicity of the mobile phase.

The Influence of Temperature, Ultrasonication and Chiral Mobile Phase Additives on Chiral Separation: Predominant Influence of β-Cyclodextrin Chiral Mobile Phase Additive Under Ultrasonic Irradiation

  • Lee, Jae Hwan;Ryoo, Jae Jeong
    • Bulletin of the Korean Chemical Society
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    • v.33 no.12
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    • pp.4141-4144
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    • 2012
  • This paper introduces a technique for resolving amino acids that combines the advantages of the conventional CSP (chiral stationary phase) method with the CMPA (chiral mobile phase additive) method. A commercially available chiral crown ether column, CROWNPAK CR(+), was used as the CSP and three cyclodextrins (${\beta}$-CD, ${\gamma}$-CD, HP-${\beta}$-CD) were used as the mobile phase additives. Chromatographic resolution was performed at $25^{\circ}C$ and $50^{\circ}C$ with or without sonication. A comparison of the chromatographic results under ultrasonic conditions with those under non-ultrasonic conditions showed that ultrasound decreased the elution time and enantioselectivity at all temperatures. In the case of the ${\beta}$-CD mobile phase additive, the elution time and enantioselectivity under ultrasonic condition were significantly higher than under non-sonic condition at all temperatures. Commercially available Chiralpak AD, Whelk-O2 and Pirkle 1-J columns were used as CSPs to examine more meticulously the effects of ultrasonication and temperature on the optical resolution. The optical resolution of some chiral samples analyzed at $25^{\circ}C$ and $50^{\circ}C$ with or without sonication was compared. As in the previous case, the enantioselectivity was lower at $25^{\circ}C$ but similar enantioselectivity was observed at $50^{\circ}C$.

Chromatographic Separation of Some Phenol Derivatives Using $\alpha$-Cyclodextrin in Mobile Phase ($\alpha$-씨클로덱스트린을 이동상으로 사용한 몇 가지 페놀 유도체들의 크로마토그래피적 분리)

  • 문영자;김봉희
    • Environmental Analysis Health and Toxicology
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    • v.12 no.3_4
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    • pp.75-84
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    • 1997
  • Chromatographic retention behavior and separation of various phenol derivatives on a Partisil 10 ODS 3 column-with mobile phase containing $\alpha$-cyclodextrin-were systematically studied. The decrease in k' values caused by the addition of cyclodextrins in the mobile phase was based on the formation of an inclusion complex, resulting in weakening of the hydrophobic interaction between solutes and the stationary phase. The content of the organic solvent in the mobile phase also influenced k' values of the solutes, and k' values increased with a decrease of the content of organic solvent in the mobile phase. A simple equation has been derived that reveals the hyperbolic dependence of the capacity factor on the total concentration of cyclodextrin. A plot of the reciprocal of the capacity factor against (CD)$_T$ gives a straight line and the dissociation constant, K$_D$, of the inclusion complex can be calculated from the slope. The capacity factor decreased with increasing temperature. The enthalpy was calculated from the slope of van't Hoff plots. Under optimum conditions, some mixtures of phenol derivatives were able to separated successfully.

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Effect of Mobile Phase Additive on Enantiomer Resolution for Chiral Amines on Polysaccharide-derived Chiral Stationary Phases by High Performance Liquid Chromatography (고성능 액체크로마토그래피의 다당유도체를 기초로 한 키랄 고정상에서 이동상 첨가제가 키랄 아민의 광학분리에 미치는 영향)

  • Paik, Man-Jeong;Yoon, Hye-Ran;Lee, Wonjae
    • KSBB Journal
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    • v.29 no.3
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    • pp.205-209
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    • 2014
  • Chromatographic enantiomer resolution of chiral amines was performed on several covalently immobilized and coated chiral stationary phases (CSPs) based on polysaccharide derivatives under the mobile phase conditions containing base or acid or acid/base additive. The chromatographic parameters including separation factors and capacity factors were greatly influenced by the nature of the mobile phase containing base or acid or salt additive as well as the used CSPs. When 0.05% triethylamine/0.05% trifluoroacetic acid as an additive in the mobile phase was used on all CSPs in this study, the greatest enantiomer resolution was observed except for Chiralpak AD. Also, it was shown that the change of base additive into acid or salt in the mobile phase may directly affect chiral recognition mechanisms between the chiral selectors and analytes occurring during enantiomer separation, resulting in the change of elution orders.

System Phase Noise for Mobile Satellite Communication Service (이동형 위성통신 서비스를 위한 시스템 위상 잡음)

  • Kim Young-wan
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.9 no.8
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    • pp.1780-1786
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    • 2005
  • The phase error generated in the transmission system affects the performance of digital transmission signal. The phase error are generated by random phase noise and tracking phase error due to denier phenomenon. In the mobile satellite communication system that generates the doppler frequency, which is a system with a movement, the proper system phase noise spectrum should be designed based on analyses for phase noise and static phase error effects. Based on the analyses of the doppler frequency and the phase error for bilateral satellite communication system providing an asynchronous service, the phase noise spectrums for the mobile satellite communication are designed in this paper. Also, the available transmission services under the less doppler effect are proposed and the proper signal source units for a required transmission system can be designed under the proposed system phase noise spectrum.

Separation of Madecassoside and Madecassic Acid Isomers by High Performance Liquid Chromatography Using β -Cyclodextrin as Mobile Phase Additive

  • Kai, Guiqing;Pan, Jian;Yuan, Chuanxun;Yuan, Yuan
    • Bulletin of the Korean Chemical Society
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    • v.29 no.3
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    • pp.551-554
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    • 2008
  • An improved HPLC method for the separation of madecassoside isomers (madecassoside and asiaticoside-B) has been developed. The isomers can be separated with high resolution from extracts of Centella asiatica by HPLC using $\beta$-cyclodextrin as a mobile phase added on a $C_{18}$ column. The result shows that the isomers can be separated with a mobile phase consisting of methanol:water (50:50, v/v) with 4 mmol/L $\beta$-CD. To elucidate the mechanism of the separation of madecassoside and asiaticoside-B, this paper studied the separation of their aglycon parts (madecassic acid and terminolic acid), another pair of isomers. The isomers can also be separated with high resolution with a mobile phase consisting of methanol:water (65:35, v/v) with 4 mmol/L $\beta$-CD and the pH of the mobile phase was adjusted to 4. The paper also studied the separation of the two isomers by HPLC using $\alpha$-CD and Glucosyl-$\beta$-CD as a mobile phase additive in order to elucidate the mechanism of the separation process.

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.

Problem-solving approach for salbutamol analysis by HPLC during pharmaceutical assay

  • Gholizadeh-Hashjin, Aiesheh;Hamishehkar, Hamed;Monajjemzadeh, Farnaz
    • Analytical Science and Technology
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    • v.35 no.5
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    • pp.189-196
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    • 2022
  • When cationic basic compounds are chromatographed using hydro-organic mobile phase, the presence of anionic free silanols in the silica-based stationary phases results in broad and asymmetrical peaks. The addition of an ionic reagent to the mobile phase prevents analytes from accessing free silanols, improving peak shape. In this study, the chromatographic behavior of salbutamol sulfate as a basic compound was investigated under various conditions, including the use of different columns, mobile phases, and ion-pair reagents such as triethanolamine (TEA) and sodium heptane sulfonate (SHS). The retention and peak shape of chromatograms were both evaluated. The results show that pre-conditioning the column with TEA and including it in the mobile phase can prevent cationic analytes from accessing anionic silanols, resulting in improved peak shape. Furthermore, buffering the mobile phase is an important factor in keeping the pH constant throughout the process. The chosen method was validated in part. This study could be helpful for researchers and analyst to solve such problems with cationic basic components.

Effect of acid or base additive in the mobile phase on enantiomer separation of amino alcohols using polysaccharide derived chiral columns (다당유도체를 기초로 한 키랄 컬럼에서 이동상에서의 산 또는 염기 첨가제가 아미노 알코올의 광학분리에 미치는 영향)

  • Huang, Hu;Jin, Jing Yu;Lee, Wonjae
    • Analytical Science and Technology
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    • v.22 no.4
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    • pp.313-318
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    • 2009
  • Liquid chromatographic enantiomer separation of amino alcohols was performed on several chiral columns based on polysaccharide derivatives under the mobile phase conditions containing acid or base additive. The chromatographic parameters were greatly influenced by the nature of the mobile phase containing acid or base additive as well as the used chiral columns. Compared to chromatographic results obtained in the mobile phase containing base additive (0.1% triethylamine), especially, Chiralcel OD showed dramatically enhanced separation factors and resolution factors with reduced capacity factors under the mobile phase condition containing acidic additive (0.1% trifluoroacetic acid). When the mobile phase containing 0.05% trifluoroacetic acid and 0.05% triethylamine was used on Chiralcel OD, the greatest separation factors and resolution factors among all other mobile phase conditions in this study were observed.