Home About us Contact
|
Home About us Contact | ||
|
|
||
Elution Mode (elution + mode)
Kinds of Elution Mode Selected AbstractsCEC separation of heterocyclic amines using methacrylate monolithic columnsELECTROPHORESIS, Issue 11 2007Elena Barceló-Barrachina Abstract Two methacrylate-based monolithic columns, one with a negatively charged group (sulfonic group) and another with a new monomer N,N -dimethylamino ethyl acrylate (DMAEA), were prepared and tested for the separation of basic compounds by CEC. This new monolithic stationary phase was prepared by the in situ polymerization of DMAEA with butyl methacrylate and ethylene dimethacrylate, using a ternary porogenic solvent consisting of water, 1-propanol and 1,4-butanediol. The performance of this column was evaluated by means of the analysis of a family of heterocyclic amines. Separation conditions such as pH, amount of organic modifier, ionic strength and elution mode (normal or counterdirectional flow) were studied. At the optimal running electrolyte composition, and using the counterdirectional mode, symmetrical electrochromatographic peaks were obtained, with the number of theoretical plates up to 30,000 and a good resolution between closely related peaks. The 2-acrylamido-2-methyl-1-propane-sulfonic acid column was used for CEC-MS, taking advantage of the compatibility of its elution mode (normal flow) with the MS coupling. [source] Simultaneous determination of melamine and related compounds by hydrophilic interaction liquid chromatography,electrospray mass spectrometryJOURNAL OF SEPARATION SCIENCE, JSS, Issue 17-18 2010Jingen Xia Abstract A hydrophilic interaction liquid chromatography coupled to ESI-MS (HILIC/ESI-MS) method for the simultaneous determination of melamine and related compounds, i.e. ammeline, ammelide and cyanuric acid in foods was developed and validated. The separation was accomplished on a Venusil HILIC column with a mobile phase of acetonitrile + 10,mM ammonium formate buffer solution at pH 3.5 (88:12, v/v) under isocratic elution mode. For the detection of the targets, the ESI probe worked in the positive and negative switching mode. For each compound, three ions were selected as qualitative ions to obtain high specificity, and the most abundant ion of each compound was selected for quantification to obtain high sensitivity. The target compounds were quantified using SIM with 15N3-melamine and 13C3-cyanuric acid being used as an internal standards in the positive and negative modes, respectively. Compared with RP separation mode, HILIC has merits such as high separation and anti-interference efficiency. The method validation including linearity, LOD, LOQ, precision and recovery proved that the method has merits such high sensitivity, specificity and simplicity versus the other methods reported in the literature. [source] Mixed-mode ion-exchangers and their comparative chromatographic characterization in reversed-phase and hydrophilic interaction chromatography elution modesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2008Michael Lämmerhofer Abstract A set of particulate silica-supported mixed-mode RP/weak anion-exchangers (RP/WAX) (obtained by bonding of N -undecenoylated 3-aminoquinuclidine, 3-aminotropane and 2-dimethylaminoethylamine as well as of N -butenoyl-(2S,4S,5R)-2-aminomethyl-5-[(2-octylthio)ethyl]-quinuclidine to thiol-modified silica) were chromatographically characterized in comparison to selected commercially available columns using two distinct isocratic elution modes, viz. an aqueous-rich RP-type elution mode (with 40% ACN and 60% buffer) as well as an organic solvent-rich hydrophilic interaction chromatography (HILIC)-type elution mode (95 and 90% ACN). The mixed-mode RP/WAX phases showed multimodal applicability, unlike a polar embedded RP material (Synergi Fusion RP), amino phases (Luna NH2, BioBasic AX) or typical HILIC packings (ZIC-HILIC, TSKGel Amide-80). Principal component analysis (PCA) of the RP test data confirmed that the in-house developed RP/WAX columns as well as the Acclaim Mixed-Mode WAX-1 phase resemble each other in their chromatographic characteristics having slightly lower hydrophobic selectivity (,CH2 of 1.5) than the tested Synergi Fusion RP (,CH2 ,1.8). In contrast, a decrease in mixed-mode character due to lowered ion-exchange capacity and concomitantly increased RP-like behavior could be identified for other mixed-mode phases in the order of Obelisc R > Primesep B2 > Uptisphere MM3. PCA on HILIC data revealed that the RP/WAX phases behave dissimilar to TSKGel Amide-80, ZIC-HILIC and polysulfoethyl A under the chosen elution conditions. Hence, they may be regarded as complementary to these commercial stationary phases with applicability profiles for hydrophilic but also hydrophobic solutes. [source] An initial assessment of the use of gradient elution in microemulsion and micellar liquid chromatographyJOURNAL OF SEPARATION SCIENCE, JSS, Issue 17-18 2004Simon M. Bryant Abstract Novel microemulsion and micellar HPLC separations have been achieved using gradient elution and columns packed with reverse phase material. Initial attempts at gradient microemulsion liquid chromatography proved impossible on use of a microemulsion successfully used in capillary electrophoresis. Optimisation of the microemulsion composition allowed the generation of stable microemulsions to achieve separations in HPLC. The novel use of organic-solvent micellar chromatography in gradient elution mode was shown to give efficient separations. A range of efficient separations of pharmaceuticals and related impurities were obtained. Acidic, basic, and neutral solutes were resolved covering a wide range of water solubilities and polarities. Elution times were in the order of 4,15 minutes. Separations were briefly compared to those accomplished with a micellar HPLC system. It is proposed that gradient elution in both microemulsion and micellar HPLC can be regarded as a highly successful means of achieving resolution of complex mixtures and should be considered for routine analysis and further investigation. [source] Preparative isolation and purification of four flavonoids from the petals of nelumbo nucifera by high-speed counter-current chromatographyPHYTOCHEMICAL ANALYSIS, Issue 3 2010Xingfeng Guo Abstract Introduction , Flavonoids, the primary constituents of the petals of Nelumbo nucifera, are known to have antioxidant properties and antibacterial bioactivities. However, efficient methods for the preparative isolation and purification of flavonoids from this plant are not currently available. Objective , To develop an efficient method for the preparative isolation and purification of flavonoids from the petals of N. nucifera by high-speed counter-current chromatography (HSCCC). Methodology , Following an initial clean-up step on a polyamide column, HSCCC was utilised to separate and purify flavonoids. Purities and identities of the isolated compounds were established by HPLC-PAD, ESI-MS, 1H-NMR and 13C-NMR. Results , The separation was performed using a two-phase solvent system composed of ethyl acetate,methanol,water,acetic acid (4,:,1,:,5,:,0.1, by volume), in which the upper phase was used as the stationary phase and the lower phase was used as the mobile phase at a flow-rate of 1.0,mL/min in the head-to-tail elution mode. Ultimately, 5.0,mg syringetin-3- O-, -d-glucoside, 6.5,mg quercetin-3- O-, -d-glucoside, 12.8,mg isorhamnetin-3- O-, -d-glucoside and 32.5,mg kaempferol-3- O-, -d-glucoside were obtained from 125,mg crude sample. Conclusion , The combination of HSCCC with a polyamide column is an efficient method for the preparative separation and purification of flavonoids from the petals of N. nucifera. Copyright © 2009 John Wiley & Sons, Ltd. [source] On-line 2D-LC-ESI/MS/MS determination of rifaximin in rat serumBIOMEDICAL CHROMATOGRAPHY, Issue 11 2009R. Nageswara Rao Abstract A highly sensitive and selective on-line two-dimensional reversed-phase liquid chromatography/electrospray ionization,tandem mass spectrometry (2D-LC-ESI/MS/MS) method was developed and validated to determine rifaximin in rat serum by direct injection. The 2D-LC-ESI/MS/MS system consisted of a restricted access media column for trapping proteins as the first dimension and a Waters C18 column as second dimension using 0.1% aqueous acetic acid:acetonitrile as mobile phase in a gradient elution mode. Rifampacin was used as an internal standard. The linear dynamic range was 0.5,10 ng/mL (r2 > 0.998). Acceptable precision and accuracy were obtained over the calibration range. The assay was successfully used in analysis of rat serum to support pharmacokinetic studies. Copyright © 2009 John Wiley & Sons, Ltd. [source] High-performance liquid chromatography and LC-ESI-MS method for identification and quantification of two isomeric polyisoprenylated benzophenones isoxanthochymol and camboginol in different extracts of Garcinia speciesBIOMEDICAL CHROMATOGRAPHY, Issue 8 2009Satyanshu Kumar Abstract A rapid, sensitive and simple reverse-phase high-performance liquid chromatography,electrospray ionization mass spectrometric method has been developed for the identification and quantification of two isomeric polyisoprenylated benzophenones, isoxanthochymol and camboginol, in the extracts of the stem bark, seeds and seed pericarps of Garcinia indica and in the fruit rinds of Garcinia cambogia. The separation of isoxanthochymol and camboginol was achieved on a Perkin Elmer RP8 column (10 × 2.1 mm with 5.0 µm particle size) using a solvent system consisting of a mixture of acetonitrile,water (80:20, v/v) and methanol,acetic acid (99.0:1.0, v/v) as a mobile phase in a gradient elution mode. The limits of detection and quantification were 5 and 10 µg/mL for isoxanthochymol and 50 and 100 µg/mL for camboginol, respectively. The intra- and inter-day precisions were 2.34 and 3.41% for isoxanthochymol and 3.35 and 3.66% for camboginol. The identity of the two isomeric compounds in the samples was determined on a triple quadrupole mass spectrometer with ESI interface operating in the negative ion mode. The method was used to identify and quantify isoxanthochymol and camboginol in the different extracts of two Garcinia species, Garcinia indica and Garcinia cambogia. Copyright © 2009 John Wiley & Sons, Ltd. [source] Identification and quantification of two antihepatotoxic coumarinolignoids cleomiscosin A and cleomiscosin B in the seeds of Cleome viscosa using liquid chromatography,tandem mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 4 2009Sunil K. Chattopadhyay Abstract A sensitive liquid chromatography/electrospray ionization tandem mass spectrometric (LC/ESI-MS/MS) method was developed for the identification and quantification of two antihepatotoxic coumarinolignoids cleomiscosin A and cleomiscosin B in different extracts of the seeds of Cleome viscosa. The separation of cleomiscosin A and cleomiscosin B was achieved on an RP18 column using a solvent system consisting of a mixture of acetonitrile,methanol (1:2, v/v) and acetonitrile,water,formic acid (5:95:0.3, v/v) as a mobile phase in a gradient elution mode. A multiple-reaction monitoring (MRM) method was developed for quantification of cleomiscosin A and cleomiscosin B in the seed extracts of Cleome viscosa. On the basis of signal-to-noise ratio of 3, the limit of detection in MRM mode for cleomiscosin A and cleomiscosin B were 1.0 and 4.0 ng/mL respectively. The method was validated in terms of linearity, accuracy and precision for 6 days. The method developed was found to be useful for identification and quantification of cleomiscosin A and cleomiscosin B in the different extracts of the seeds of Cleome viscosa. Copyright © 2008 John Wiley & Sons, Ltd. [source] High-performance liquid chromatography and LC-ESI-MS method for the identification and quantification of two biologically active isomeric coumarinolignoids cleomiscosin A and cleomiscosin B in different extracts of Cleome viscosaBIOMEDICAL CHROMATOGRAPHY, Issue 12 2008Sunil K. Chattopadhyay Abstract A rapid, sensitive and simple reverse-phase high-performance liquid chromatographic,electrospray ionization,mass spectrometry method for simultaneous determination of cleomiscosin A and cleomiscosin B has been developed and validated. The isomeric coumarinolignoids cleomiscosin A (1) and cleomiscosin B (2) were separated on a Waters symmetry C18 column with a solvent system composed of acetonitrile,methanol (1:2) and acetic acid,water (0.5 : 99.5) in a gradient elution mode. The absorption at 326 nm was chosen as the measuring wavelength in which resolution and baseline separation of compounds 1 and 2 could be obtained. The identity of the two isomeric compounds 1 and 2 in the samples were determined on a triple quadrupole mass spectrometer with ESI interface operating in the positive mode. Calibration curves were linear (r2 > 0.993) over the concentration range 20,200 µg/mL for cleomiscosin A and 10,200 µg/mL for cleomiscosin B with acceptable accuracy and precision, respectively. The intra-day and inter-day precision were 1.13 and 0.82% for cleomiscosin A and 1.78 and 1.28% for cleomiscosin B, respectively. The validated method was successfully applied for the analysis of the above two compounds in different extracts of Cleome viscosa. Copyright © 2008 John Wiley & Sons, Ltd. [source] Mixed-mode ion-exchangers and their comparative chromatographic characterization in reversed-phase and hydrophilic interaction chromatography elution modesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2008Michael Lämmerhofer Abstract A set of particulate silica-supported mixed-mode RP/weak anion-exchangers (RP/WAX) (obtained by bonding of N -undecenoylated 3-aminoquinuclidine, 3-aminotropane and 2-dimethylaminoethylamine as well as of N -butenoyl-(2S,4S,5R)-2-aminomethyl-5-[(2-octylthio)ethyl]-quinuclidine to thiol-modified silica) were chromatographically characterized in comparison to selected commercially available columns using two distinct isocratic elution modes, viz. an aqueous-rich RP-type elution mode (with 40% ACN and 60% buffer) as well as an organic solvent-rich hydrophilic interaction chromatography (HILIC)-type elution mode (95 and 90% ACN). The mixed-mode RP/WAX phases showed multimodal applicability, unlike a polar embedded RP material (Synergi Fusion RP), amino phases (Luna NH2, BioBasic AX) or typical HILIC packings (ZIC-HILIC, TSKGel Amide-80). Principal component analysis (PCA) of the RP test data confirmed that the in-house developed RP/WAX columns as well as the Acclaim Mixed-Mode WAX-1 phase resemble each other in their chromatographic characteristics having slightly lower hydrophobic selectivity (,CH2 of 1.5) than the tested Synergi Fusion RP (,CH2 ,1.8). In contrast, a decrease in mixed-mode character due to lowered ion-exchange capacity and concomitantly increased RP-like behavior could be identified for other mixed-mode phases in the order of Obelisc R > Primesep B2 > Uptisphere MM3. PCA on HILIC data revealed that the RP/WAX phases behave dissimilar to TSKGel Amide-80, ZIC-HILIC and polysulfoethyl A under the chosen elution conditions. Hence, they may be regarded as complementary to these commercial stationary phases with applicability profiles for hydrophilic but also hydrophobic solutes. [source] | ||||||||||