Effect Of Column Temperature Research Articles

Development and Validation of Ultra-Performance Liquid Chromatography (UPLC) Method for Simultaneous Quantification of Hydrochlorothiazide, Amlodipine Besylate, and Valsartan in Marketed Fixed-Dose Combination Tablet

Fixed-dose combination therapy is considered a practical approach in the treatment of various diseases, as it can simultaneously target different mechanisms of action that achieve the required therapeutic efficacy through a synergistic effect. A combination of hydrochlorothiazide (HTZ), amlodipine (AMD), and valsartan (VLS) has been created for the treatment of hypertension. Therefore, the aim of this study was to develop an optimized UPLC method for the simultaneous quantification of this combination. A DoE at a level of 32 was used to investigate the effects of column temperature (20, 30, and 40 °C) and formic acid concentration (0.05, 0.15, and 0.25%) on the retention time of each active pharmaceutical ingredient (API), the peak area, and the peak symmetry, as well as the resolution between HTZ-AMD and AMD-VLS peaks. The optimized analytical method was validated and used to extract the three APIs from the marketed product. The optimized analytical condition with a column temperature of 27.86 °C and a formic acid concentration of 0.172% showed good separation of the three APIs in 1.62 ± 0.006, 3.59 ± 0.002, and 3.94 ± 0.002 min for HTZ, AMD, and VST, respectively. The developed method was linear with the LOQ for a HTC, AMD, and VST of 0.028, 0.038, and 0.101 ppm, respectively. Moreover, the developed assay was sustainable and robust, with an RSD % of less than 2%. The application of this method in the extraction of HTZ, AMD, and VST from the Exforge® marketed product showed good separation with a measurable drug content of 23.5 ± 0.7, 9.68 ± 0.1, and 165.2 ± 5.2 mg compared to the label claims of 25/10/160 for HTZ, AMD, and VST, respectively.

Toward an Ultimate Solution for Peptide Retention Time Prediction: The Effect of Column Temperature on Separation Selectivity.

We studied the effect of the column temperature on the selectivity of reversed-phase peptide separation in bottom-up proteomics. The number of peptide identifications from 2 h liquid chromatography with tandem mass spectrometry (LC-MS/MS) acquisitions reaches a plateau at 45-55 °C, driven simultaneously by improved separation efficiency, a gradual decrease in peptide retention, and possible on-column degradation of peptides at elevated temperatures. Performing 2D LC-MS/MS acquisitions at 25, 35, 45, and 55 °C resulted in the identification of ∼100,000 and ∼120,000 unique peptides for nonmodified and tandem mass tags (TMT)-labeled samples, respectively. These peptide collections were used to investigate the temperature-driven retention features. The latter is governed by the specific temperature response of individual residues, peptide hydrophobicity and length, and amphipathic helicity. On average, peptide retention decreased by 0.56 and 0.5% acetonitrile for each 10 °C increase for label-free and TMT-labeled peptides, respectively. This generally linear response of retention shifts allowed the extrapolation of predictive models beyond the studied temperature range. Thus, (trap) column cooling from room temperature to 0 °C will allow the retention of an additional 3% of detectable tryptic peptides. Meanwhile, the application of 90 °C would result in the loss of ∼20% of tryptic peptides that were amenable to MS/MS-based identification.

Chiral metal–organic framework [Zn2(D-Cam)2(4,4′-bpy)]n@SiO2 core–shell composite for HPLC separation

Chiral metal–organic frameworks, an emerging type of chiral porous materials, possess many outstanding characteristics such as high surface area, tunable pore sizes, and good chemical stability. Herein, we prepared a chiral [Zn2(D-Cam)2(4,4′-bpy)]n@SiO2 core–shell composite on the SiO2-NH2 microspheres by an in-situ growth method. The prepared [Zn2(D-Cam)2(4,4′-bpy)]n@SiO2-packed column exhibited high-resolution separation of racemic compounds and positional isomers at a low column backpressure (9–12 bar). Twelve pairs of enantiomers including alcohols, amines, epoxides, ketones, and esters were well resolved with sharp peak shapes on the [Zn2(D-Cam)2(4,4́-bpy)]n@SiO2 chromatographic column. Compared the separation performance of [Zn2(D-Cam)2(4,4́-bpy)]n@SiO2-packed column and two chromatographic columns (commercial Chiralpak AD-H column and as-prepared SiO2-NH2 column), the [Zn2(D-Cam)2(4,4′-bpy)]n-packed column and the commercial Chiralpak AD-H column have a good complementarity in chiral separation, and it also offered better separation performance compared to the SiO2-NH2 column for separating positional isomers. The effects of column temperature and analyte mass on the HPLC separation were investigated. Additionally, the [Zn2(D-Cam)2(4,4́-bpy)]n@SiO2 packed column has good repeatability and stability for HPLC separation of enantiomers and positional isomers. The relatvive standard deviations (RSDs) for repeated separations of 2,2,2-trifluoro-1-(9-anthryl)ethanol and o, m, p-bromoaniline were less than 0.8 % and 1.2 % for the retention time and peak area, respectively. After continuous use for one month, the RSDs for the retention time and peak area of them were in the range of 1.1–1.3 % and 1.5–1.8 % (inter-day and intra-day, n = 5), respectively. This work also indicates that the chiral MOFs@SiO2 composites are promising for HPLC separation.

Performance evaluation of 2-undecylimidazole/propyl methacrylate bifunctional silica gel for mixed-mode reversed-phase/anion-exchange chromatography

In this work, 2-undecylimidazole (UI) and propyl methacrylate (PMA) dual-functionalized silica gel was prepared and applied as a mixed-mode chromatographic stationary phase. Successful preparation of this stationary phase (SiO2-UI-PMA) was proved through a range of characterization techniques. The chromatographic performance of the obtained SiO2-UI-PMA was investigated for the separation of environmental endocrine disruptors (phthalates, steroid hormones, bisphenols and alkylphenols), sulfonamides and food-borne stimulants in reversed-phase chromatography, as well as for the separation of inorganic anions in anion-exchange chromatography. The effect of column temperature, acetonitrile content, salt concentration and mobile phase pH on retention was studied to explore the separation mechanism, and the results indicated that this stationary phase provided multiple interactions with analytes. Compared with the commercial C18 column, the SiO2-UI-PMA demonstrated more preferable separation ability under the mobile phase conditions with a low content of acetonitrile, thereby acting as a good alternative candidate for the conventional reversed-phase stationary phase in chromatographic analysis. Furthermore, the SiO2-UI-PMA stationary phase exhibited high column efficiency (45 860 plates m−1 for progesterone) even though it was prepared from the non-serviceable silica based chromatographic packing materials.

Simultaneous Determination of Escitalopram Impurities including the R-enantiomer on a Cellulose tris(3,5-Dimethylphenylcarbamate)-Based Chiral Column in Reversed-Phase Mode.

A high-performance liquid chromatographic method was developed for the simultaneous determination of the related substances-three potential synthesis-related chemical impurities and the distomer-of escitalopram. The separation capacity of seven different polysaccharide-type chiral columns, including three amylose-based (Lux Amylose-1, Lux i-Amylose-1, Lux Amylose-2) and four cellulose-based columns (Lux Cellulose-1, Lux Cellulose-2, Lux Cellulose-3, and Lux Cellulose-4) were screened in the polar organic and reversed-phase modes. Lux Cellulose-1, based on cellulose tris(3,5-dimethylphenylcarbamate) as the chiral selector with an acetonitrile-water mixture containing 0.1% diethylamine was identified as the most promising separation system. Using the "one factor at a time" optimization approach, the effect of column temperature, flow rate, and mobile phase constituents on separation performance was evaluated, and the critical resolution values were determined. A U-shaped retention pattern was obtained when plotting the retention factors of the citalopram enantiomers versus the water content of the binary mobile phases on the Lux Cellulose-1 column. A thermodynamic analysis revealed enthalpy-driven enantioseparation in both the polar organic and reversed-phase modes. For further method optimizations, an L9 orthogonal array table was employed. Using the optimized parameters (Lux Cellulose-1 column with 0.1% (v/v) diethylamine in water/acetonitrile 55/45 (v/v); 0.8 mL/min flow rate at 25 °C), baseline separations were achieved between all compounds. Our newly developed HPLC method was validated according to the ICH guidelines and its application was tested with a commercially available pharmaceutical formulation. The method proved to be suitable for routine quality control of related substances and the enantiomeric purity of escitalopram.

Octadecylamine and serine-derived carbon dots-modified silica gel for reversed phase/hydrophilic interaction liquid chromatography

In order to meet the demand for the separation of complex samples, the development of mixed-mode chromatographic stationary phases has become one of the research priorities in chromatographic separation. In this work, carbon dots, Ser-OCDs, with amino, hydroxyl, carboxyl and alkyl chains were synthesized for the first time and modified on silica gel for RPLC/HILIC mixed-mode chromatography. The stationary phase, Sil-Ser-OCDs, enables the effective separation of hydrophobic and hydrophilic compounds such as alkylbenzenes, polycyclic aromatic hydrocarbons, nucleosides, bases and sulfonamides with better separation selectivity and higher column efficiency compared to classical C18 column. Polycyclic aromatic hydrocarbons, nucleosides, bases and sulfonamides can all be separated in less than 10 min, Moreover, the column efficiency (theoretical plates, N) of butylbenzene was even as high as 124,024 plates m−1. By investigating the effects of column temperature and concentration of buffer salt solution on retention and constructing the van't Hoff equation, hydrogen bonding, π-π conjugation, electrostatic repulsion, partitioning and hydrophobic interactions were found to be the main retention interactions. Sil-Ser-OCDs column was eluted for a long time at pH 3 and 8, and the relative retention remained basically unchanged, which proved that the stability of the column was excellent. This work broadens the variety of chromatographic stationary phase materials, at the same time, indicates that carbon dots have promising applications in mixed-mode chromatography.

Determination of panthenol enantiomers in cosmetic preparations using an achiral-chiral-coupled column HPLC system.

A new high-performance liquid chromatography (HPLC) method for separation and determination of panthenol enantiomers in hair care products was developed. Two types of detectors, low-wavelength ultraviolet (UV) and polarimetric, were used. Optimized conditions consisted of coupled achiral, amino type, and chiral, amylose tris(3,5-dimethylphenylcarbamate), stationary phases, mixture of n-hexane/ethanol (60:40, v/v) as mobile phase under isocratic conditions and flow rate 0.8 cm3 min-1 . The effect of column temperature on retention and resolution of enantiomers was studied. The analysis runtime was 10 minutes, and the average retention times for d- and l-panthenol were 7.10 ±0.1 minutes and 8.21 ±0.2 minutes, respectively. The resolution of enantiomers on coupled achiral-chiral columns was Rs = 2.7. The solid-phase extraction method was employed for extraction and purification of analytes. The validated method was selective, accurate, and linear (R2 > .998) over the concentration range of 0.001 to 1.0 mg cm-3 for both enantiomeric forms. The limits of detection (LOD) and quantitation (LOQ) of each enantiomer were 0.3 and 1.0 μg cm-3 , respectively. The results demonstrated the occurrence of d-panthenol in hair care products.

Direct Evaluation of Unsymmetrical Dimethylhydrazine with Wide Concentration Range in Wastewater by Ion Chromatography

Current methods for measurement of unsymmetrical dimethylhydrazine (UDMH) in wastewater suffer from a small detection range and complex pretreatment. In this study, ion chromatography (IC) with conductivity detection was used to directly analyze UDMH in wastewater, without any derivatization pretreatment, and the measurement range was as wide as 0.5–1000 μg·mL⁻¹. The effects of column temperature and sample pH on the detection were investigated. Furthermore, the chromatography conditions were optimized by combining the effects of different combinations of nitric acid, 2,6-pyridinedicarboxylic acid (PDCA), and methanesulfonic acid (MSA) on the analysis of mixed samples of UDMH and alkali metal ions. The linearity of the detected peak height (H) and peak area (A) versus UDMH concentration was studied at low, medium, and high concentrations. A good linear relationship was observed in the range of 0.5–1000 μg·mL⁻¹, especially for A, the relative standard deviation (RSD) and correlation coefficient (R²) of which were 0.077–1.653% and 0.9998–1.0000, respectively. In addition, the method showed good sensitivity with a limit of detection (LOD) of 0.05 μg·mL⁻¹ and a limit of quantitation (LOQ) of 0.16 μg mL⁻¹. The precision and accuracy (expressed by recovery) were verified, where the RSD were less than 1.49% and 3.17%. Interference between hydrazines and alkali metal ions was discussed. Finally, actual UDMH samples were determined, confirming the applicability of the developed method for analyzing UDMH in wastewater during the degradation process.

Standartization of P. ginseng and P. quinquefolius root extracts by liquid chromatography-mass spectrometry

An approach to detection of triterpene glycosides in ginseng extracts is developed using high performance liquid chromatography/mass spectrometry (HPLC-MS). Enhanced selectivity compared to commonly used HPLC-UV techniques provides simultaneous registration of chromatographic peaks and determination of 23 major and minor ginsenosides. For this purpose, in addition to the use of highly selective MS detection of adduct ions of sodium ginsenoside molecules and fragmentary sapogenin ions, special conditions of chromatographic separation on a sorbent modified with pentafluorophenyl (PFP) groups were specified. The effect of column temperature and mobile phase composition on the separation selectivity of glycosides was also studied. Though we failed to achieve complete chromatographic separation of the peaks for several compounds (F4 and Rg6, Rk3 and Rh4), their determination appeared possible in case of their joint attendance due to registration of the signals that differ in the value ofm/zratio. For all studied compounds, the linearity ranges and calibration equations along with the metrological characteristics such as the detection limit and reproducibility were determined. The developed approach was tested in standardization of the reference extracts of Asian (P. ginseng) and American (P. quinquefolius) ginseng roots. For some ginsenosides, the content declared by the manufacture did not match the actual content, but for others the concentrations were close to the declared values. Moreover, we managed to expand the range of ginsenosides under control which is rather important for their use in medicine.

Simultaneous determination of twenty amino acids and six carbohydrates in soy sauce by ion chromatography with integrated pulsed amperometric detection

A method of gradient elution ion chromatography with integrated pulsed amperometric detection for simultaneous determination of twenty amino acids and six carbohydrates in soy sauce was established. The effects of column temperature, pH value of solution, and standing time were studied, and then the suitable gradient elution conditions were found for the determination. In the conditions that the flow rate is 0.25 mL/min, pH value of solution is in the range of 5.2-6.7, temperature of column is 35℃, the linear relationships of the 26 components are good and the correlation coefficients are not less than 0.995. Except glutamine, leucine, isoleucine and methionine, the limits of detection (S/N=3) of the other 22 components are less than 0.03 mg/L. The recoveries of the 26 components in soy sauce are 84.2%-109% at the spiked levels of 0.20, 0.50, 2.00 mg/L, and the relative standard deviations (RSDs) are in the range of 2.7%-7.8%. The method is efficient, easy, sensitive, and accurate for simultaneous determination of amino acids and carbohydrates in soy sauce, and can provide an effective research technique for adulterated soy sauce.

Characterization of Purified Red Cabbage Anthocyanins: Improvement in HPLC Separation and Protective Effect against H₂O₂-Induced Oxidative Stress in HepG2 Cells.

In this study, the chemical profiles and antioxidant activities of red cabbage anthocyanin (RCA)-enriched extract are evaluated. The effects of column temperature on the HPLC resolution of the RCAs are studied. The HPLC resolutions became better as the column temperature increased from 20 °C–45 °C. An optimized HPLC condition was achieved at 45 °C and used for the quantification and qualification of the RCAs. The anthocyanins in the enriched powder are all derivatives of cyanidin (268 ± 2 μg/mg), mainly with 19% nonacylated, 51% monoacylated, and 31% diacylated structures with ferulic, sinapic, p-coumaric, and caffeic acids characterized by HPLC-MS. The RCA extracts markedly reduced intracellular oxidative stress production by H2O2 on HepG2 cells and consequently ameliorated cell apoptosis and improved viability. The analytical method and cellular antioxidant activity demonstration of the RCAs will greatly facilitate their functional applications.

Enantioselective separation of twelve pairs of enantiomers on polysaccharide-based chiral stationary phases and thermodynamic analysis of separation mechanism.

The enantioseparation of twelve pairs of structurally related 1-aryl-1-indanone derivatives was studied in the normal-phase mode using three different polysaccharide-type chiral stationary phases, namely Chiralpak IB, Chiralpak IC, and Chiralpak ID. n-Hexane/2-propanol and n-hexane/ethanol were employed as mobile phases. Among all the investigated chiral columns, Chiralpak IC exhibited the most universal and the best enantioseparation ability toward all the racemates, particularly with the mobile phase composed of n-hexane/2-propanol (90/10, v/v). Then the effects of column temperature on retention and enantioselectivity were examined in the range of 25-40°C. Satisfactory enantioseparation was obtained at ambient temperature. The natural logarithm of retention and separation factors (ln k and ln α) versus the reciprocal of absolute temperature (1/T) (Van't Hoff plots) were found to be linear for all racemates, indicating that the retention and separation mechanisms were independent of temperature in the range investigated. Then, the thermodynamic parameters (ΔΔH°, ΔΔS°, and ΔΔG°) were calculated from Van't Hoff plots. These values indicated that the solute transfer from the mobile to stationary phase was enthalpically favorable, and the process of enantioseparation was mainly enthalpy controlled. At last, the impact of small changes in molecular structures of the tested 1-indanone derivatives on enantioseparation was also discussed.

Aqueous size-exclusion chromatography of polyelectrolytes on reversed-phase and hydrophilic interaction chromatography columns

The size-exclusion separation of a water-soluble polyelectrolyte polymer, sodium polystyrene sulfonate (NaPSS), was demonstrated on common reversed-phase (C18, C4, phenyl, and cyano) and hydrophilic interaction chromatography (HILIC) columns. The effect of common solvents – acetonitrile (ACN), tetrahydrofuran (THF), and methanol (MeOH), used as mobile phase modifiers – on the elution of NaPSS and the effect of column temperature (within a relatively narrow range corresponding to typical chromatographic conditions, i.e., 10 °C–60 °C) on the partition coefficient, KSEC, were also investigated. Non-size-exclusion chromatography (non-SEC) effects can be minimized by the addition of an electrolyte and an organic modifier to the mobile phase, and by increasing the column temperature (e.g., to 50 °C or 60 °C). Strong solvents such as THF and ACN are more successful in the reduction of such effects than is the weaker solvent MeOH. The best performance is seen on medium polarity and polar stationary phases, such as cyanopropyl- and diol-modified silica (HILIC), where the elution of the NaPSS polyelectrolyte is by a near-ideal SEC mechanism. Hydrophobic stationary phases, such as C18, C4, and phenyl, require a higher concentration of a strong solvent modifier (THF) in the mobile phase to reduce non-SEC interactions of the solute with the stationary phase.

Organic solvent modifier and temperature effects in non-aqueous size-exclusion chromatography on reversed-phase columns

Common reversed-phase columns (C18, C4, phenyl, and cyano) offer inert surfaces suitable for the analysis of polymers by size-exclusion chromatography (SEC). The effect of tetrahydrofuran (THF) solvent and the mixtures of THF with a variety of common solvents used in high performance liquid chromatography (acetonitrile, methanol, dimethylformamide, 2-propanol, ethanol, acetone and chloroform) on reversed-phase stationary phase characteristics relevant to size exclusion were studied. The effect of solvent on the elution of polystyrene (PS) and poly(methyl methacrylate) (PMMA) and the effect of column temperature (within a relatively narrow range corresponding to typical chromatographic conditions, i.e., 10°C–60°C) on the SEC partition coefficients KSEC of PS and PMMA polymers, were also investigated. The bonded phases show remarkable differences in size separations when binary mixtures of THF with other solvents are used as the mobile phase. The solvent impact can be two-fold: (i) change of the polymeric coil size, and possible shape, and (ii) change of the stationary phase pore volume. If the effect of this impact is properly moderated, then the greatest benefit of optimized solute resolution can be achieved. Additionally, this work provides an insight on solvent-stationary phase interactions and their effects on column pore volume. The only effect of temperature observed in our studies was a decreased elution volume of the polymers with increasing temperature. SEC partition coefficients were temperature-independent in the range of 10°C–60°C and therefore, over this temperature range elution of PS and PMMA polymers is by near-ideal SEC on reversed-phase columns. Non-ideal SEC appears to occur for high molar mass PMMA polymers on a cyano column when alcohols are used as mobile phase modifiers.

Investigation of the effect of column temperature and back-pressure in achiral supercritical fluid chromatography within the context of drug impurity profiling

Twenty commercially available stationary phases were characterized in supercritical fluid chromatography (SFC) using a diverse set of pharmaceutical compounds. Six dissimilar phases were selected, and a benzodiazepine and a trimethoprim impurity mixture were screened on these phases. Two stationary phases were then selected for each mixture to study the effect of temperature and back-pressure on retention, separation and chromatographic efficiency using a response surface design approach. The maximal feasible domain for each phase was examined and column performance was monitored for stability during the duration of the study. Chromatographic responses of the individual mixture components, such as retention time, peak width and apparent plate count, were modelled as a function of temperature and back-pressure. The use of high temperatures led to improved separations and higher efficiencies while high back-pressures resulted in reduced retention. For the two mixtures, the response surface plots of the resolution of the worst-separated peak pair over the experimental domain allowed the identification of the temperature and back-pressure leading to the maximal resolution for the worst-separated peak pair. For the mixtures investigated, the use of high temperatures led to improved separations and high efficiencies, while high back-pressures resulted in reduced retention. These factors are fine-tuning parameters in SFC, and similarly to the modifier composition, they lead to local, rather than global selectivity differences.

Chromatographic profiles of tryptophan and kynurenine enantiomers derivatized with (S)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole using LC-MS/MS on a triazole-bonded column.

d- and l-Tryptophan (Trp) and d- and l-kynurenine (KYN) were derivatized with a chiral reagent, (S)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole (DBD-PyNCS), and were separated enantiomerically by high-performance liquid chromatography (HPLC) equipped with a triazole-bonded column (Cosmosil HILIC) using tandem mass spectrometric (MS/MS) detection. Effects of column temperature, salt (HCO2 NH4 ) concentration, and pH of the mobile phase in the enantiomeric separation, followed by MS detection of (S)-DBD-PyNCS-d,l-Trp and -d,l-KYN, were investigated. The mobile phase consisting of CH3 CN/10mM ammonium formate in H2 O (pH5.0) (90/10) with a column temperature of 50-60°C gave satisfactory resolution (Rs) and mass-spectrometric detection. The enantiomeric separation of d,l-Trp and d,l-KYN produced Rs values of 2.22 and 2.13, and separation factors (α) of 1.08 and 1.08, for the Trp and KYN enantiomers, respectively. The proposed LC-MS/MS method provided excellent detection sensitivity of both enantiomers of Trp and KYN (5.1-19nM).

Molecularly imprinted polymer for glutathione by modified precipitation polymerization and its application to determination of glutathione in supplements

Molecularly imprinted polymers (MIP) particles for glutathione (GSH) with a narrow particle size distribution were prepared by modified precipitation polymerization using methacrylic acid as a functional monomer, divinylbenzene as a crosslinker and water as a co-solvent. The particle diameters of the MIP and non-imprinted polymer (NIP) prepared under the optimum conditions were 3.81±0.95 (average±standard deviation) and 3.39±1.22μm, respectively. The retention and molecular-recognition properties of the prepared MIP were evaluated using a mixture of acetonitrile and water as a mobile phase in hydrophilic interaction chromatography. With an increase of acetonitrile content, the retention factor of GSH was increased on the MIP. In addition to shape recognition, hydrophilic interactions seem to work for the recognition of GSH on the MIP. The MIP had a specific molecular-recognition ability for GSH, while glutathione disulfide, l-Glu, l-Cys, Gly-Gly and l-Cys-Gly could not be retained or recognized on the MIP. The effect of column temperature revealed that the separation of GSH on the MIP was entropically driven. Binding experiments and Scatchard analyses revealed that one binding sites were formed on both the MIP and NIP, while the MIP gave higher affinity and capacity for GSH than the NIP. Furthermore, the MIP was successfully applied for determination of GSH in the supplements.

High Performance Liquid Chromatography at −196 °C

Ultralow temperature high-performance liquid chromatography (HPLC) was developed using a liquefied gas as the mobile phase. HPLC separation of low molecular weight alkanes at -196 °C with liquid nitrogen mobile phase was successfully achieved, whereas their GC separation at -196 °C using helium gas mobile phase failed to elute the analytes due to strong adsorption. Prior to the further study of HPLC at -196 °C, the effect of column temperature on the chromatographic behavior was investigated, and it was found that the retention of analytes drastically increased when the column temperature was over the boiling point of the mobile phase. As the study of retention control in HPLC at -196 °C, the mobile phases of nitrogen and methane mixtures were investigated. The addition of methane to the nitrogen mobile phase suppressed the retention of the analytes (tetra-deuterated methane, ethane, and propane), that is, the retention on HPLC at ultralow temperature could be controlled by the mobile phase composition, akin to the typical retention in HPLC. The selectivity toward the n- and iso-alkane in HPLC at -196 °C was altered compared with that in GC separation at room temperature. A significant enhancement of retention of alkanes compared with alkanes were observed in HPLC at -196 °C.

Investigation of adsorption of phenols on detonation nanodiamonds using liquid column chromatography

Adsorption of phenols on microdisperse sintered detonation nanodiamond (MSDN) from water–acetonitrile solutions was investigated using high performance liquid chromatography (HPLC). Parabolic dependence between retention and organic solvent content in mobile phase was observed. The influence of physicochemical characteristic solutes including molecular volume (V), dipole moment (μ), hydrophobicity (logP), polarizability (α) and molecular refractivity (MR)) of phenols on their adsorption from water–acetonitrile solutions was investigated. Single- and multi-parameter correlations between retention factor and various physicochemical characteristics of sorbates were obtained. The effect of column temperature on adsorption of phenols on UDD was investigated. The changes in ΔH and ΔS for competitive adsorption of the test sorbates were determined.

Breakthrough Model for Adsorption of Paraffinic Mixtures in Mixed Phase Conditions

A model is developed that allows simulation of breakthrough adsorption experiments of n-alkanes dissolved in a short chain alkane solvent. The model is used to verify conclusions made in previous experimental work that studied competitive adsorption behavior of paraffinic mixtures in vapor and liquid phase conditions. In these types of breakthrough experiments, a pressure drop in the adsorption column results in a change in aggregation state and density changes inside the column, leading to a change in adsorption behavior. Simulations demonstrate the effect of column temperature, outlet pressure, and pellet size on competitive adsorption behavior, and the importance of the effect of density on adsorption behavior is also demonstrated.