Arianne Soliven

Tocochromanols composition in kernels recovered from different apricot varieties: RP-HPLC/FLD and RP-UPLC-ESI/MSn study

Composition of tocochromanols in kernels recovered from 16 different apricot varieties (Prunus armeniaca L.) was studied. Three tocopherol (T) homologues, namely α, γ and δ, were quantified in all tested samples by an RP-HPLC/FLD method. The γ-T was the main tocopherol homologue identified in apricot kernels and constituted approximately 93% of total detected tocopherols. The RP-UPLC-ESI/MSn method detected trace amounts of two tocotrienol homologues α and γ in the apricot kernels. The concentration of individual tocopherol homologues in kernels of different apricots varieties, expressed in mg/100 g dwb, was in the following range: 1.38–4.41 (α-T), 42.48–73.27 (γ-T) and 0.77–2.09 (δ-T). Moreover, the ratio between individual tocopherol homologues α:γ:δ was nearly constant in all varieties and amounted to approximately 2:39:1.

Optimization of gradient reversed phase chromatographic peak capacity for low molecular weight solutes

A general protocol for optimizing peak capacity for the separation of low molecular weight molecules under gradient elution conditions has not yet been developed. By studying the effects of gradient time, flow rate, temperature, final eluent composition, and column length on peak capacity, a protocol has been developed for the optimization of a separation of small molecules such as those seen in metabolomic studies. The strategy developed employs the Linear-Solvent-Strength Theory (LSS Theory) to predict retention, building on an approach for the optimization of the peak capacity of large molecules (peptides) in fixed column format separations.

π-Selective stationary phases: (III) Influence of the propyl phenyl ligand density on the aromatic and methylene selectivity of aromatic compounds in reversed phase liquid chromatography

The retention characteristics of phenyl type stationary phases for reversed phase high performance liquid chromatography are still largely unknown. This paper explores the retention process of these types of stationary phases by examining the retention behaviour of linear PAHs and n-alkylbenzenes on a series of propyl phenyl stationary phases that have changes in their ligand density (1.23, 1.31, 1.97, 2.50 micromol m(-2)). The aromatic and methylene selectivities increased with increasing ligand density until a point where a plateau was observed, overall the propyl phenyl phases had a higher degree of aromatic selectivity than methylene selectivity indicating that these columns are suitable for separations involving aromatic compounds. Also, retention characteristics relating to the size of the solute molecule were observed to be influenced by the ligand density. It is likely that the changing retention characteristics are caused by the different topologies of the stationary phases at different ligand densities. At high ligand densities, the partition coefficient became constant.

Enhancing the separation performance of the first-generation silica monolith using active flow technology: parallel segmented flow mode of operation.

Active flow technology (AFT) columns are designed to minimise inefficient flow processes associated with the column wall and radial heterogeneity of the stationary phase bed. This study is the first to investigate AFT on an analytical scale 4.6mm internal diameter first-generation silica monolith. The performance was compared to a conventional first-generation silica monolith and it was observed that the AFT monolith had an increase in efficiency values that ranged from 15 to 111%; the trend demonstrating efficiency gains increasing as the volumetric flow to the detector was decreased, but with no loss in sensitivity.

Impact of Species and Variety on Concentrations of Minor Lipophilic Bioactive Compounds in Oils Recovered from Plum Kernels

The profile of bioactive compounds (carotenoids, tocopherols, tocotrienols, phytosterols, and squalene) in oils recovered from the kernels of 28 plum varieties of hexaploid species Prunus domestica L. and diploid plums Prunus cerasifera Ehrh. and their crossbreeds were studied. Oil yields in plum kernels of both P. cerasifera and P. domestica was in wide ranges of 22.6-53.1 and 24.2-46.9% (w/w) dw, respectively. The contents of total tocochromanols, carotenoids, phytosterols, and squalene was significantly affected by the variety and ranged between 70.7 and 208.7 mg/100 g of oil, between 0.41 and 3.07 mg/100 g of oil, between 297.2 and 1569.6 mg/100 g of oil, and between 25.7 and 80.4 mg/100 g of oil, respectively. Regardless of the cultivar, β-sitosterol and γ-tocopherol were the main minor lipophilic compounds in plum kernel oils and constituted between 208.5 and 1258.7 mg/100 g of oil and between 60.5 and 182.0 mg/100 g of oil, respectively. Between the studied plum species, significant differences were recorded for δ-tocopherol (p = 0.007), 24-methylenecycloartanol (p = 0.038), and citrostadienol (p = 0.003), but they were insufficient for discrimination by PCA.

Effect of parallel segmented flow chromatography on the height equivalent to a theoretical plate III – Influence of the column length, particle diameter, and the molecular weight of the analyte on the efficiency gain

The effects of column length on performance in segmented flow chromatography were tested. Column efficiencies were measured for 4.6mm I.D. 3, 5, 7.5 and 10 cm long columns packed with 3.0 μm Hypurity-C18 fully porous particles and of 4.6mm I.D. 5, 10, 15 and 25 cm long columns packed with 5 μm Hypersil GOLD C18 particles. For each column length and particle type, two different configurations were tested: (1) both the inlet and outlet column endfittings were standard and (2) the inlet endfitting was standard but the outlet endfitting allowed parallel segmentation of the exiting flow into a central and a peripheral coaxial region. The segmentation flow ratio was set at 45% (for 3 μm) and at 43% or 21% (for 5 μm). Four samples were used, naphthalene, toluene, butylbenzene, and insulin, which has a ten times smaller diffusion coefficient than the small molecules. The column performance for the low molecular weight compound is significantly improved at velocities above the optimum value when the outlet flow rate is segmented because longitudinal diffusion and mass transfer resistance of this compound in the stationary phase are negligible sources of band broadening at reduced linear velocities between 5 and 25. At high flow rate (4 mL/min), the long-range eddy dispersion terms are about 3.9, 3.2, 2.6, and 1.8h unit lower for the 3, 5, 7.5 and 10 cm long columns, respectively. The longer the column, the lower the efficiency improvement because the border effects are smaller. This result was not systematically observed for the columns packed with 5 μm particles because the transverse dispersion is larger. In contrast, the gain in column efficiency is marginal for insulin because the mass transfer mechanism of this compound is mostly controlled by the slow diffusivity of insulin across Hypurity-C18 particles.

Assessing the performance of curtain flow first generation silica monoliths

Analytical scale active flow technology first generation silica monolithic columns kitted out in curtain flow mode of operation were studied for the first time. A series of tests were undertaken assessing the column efficiency, peak asymmetry and detection sensitivity. Two curtain flow columns were tested, one with a fixed outlet ratio of 10% through the central exit port, the other with 30%. Tests were carried out using a wide range in inlet flow segmentation ratios. The performance of the curtain flow columns were compared to a conventional monolithic column. The gain in theoretical plates achieved in the curtain flow mode of operation was as much as 130%, with almost Gaussian bands being obtained. Detection sensitivity increased by as much as 250% under optimal detection conditions. The permeability advantage of the monolithic structure together with the active flow technology makes it a priceless tool for high throughput, sensitive, low detection volume analyses.

Comparison of core-shell particles and sub-2μm fully porous particles for use as ultrafast second dimension columns in two-dimensional liquid chtomatography

The peak capacity of small columns packed with 2.7μm core-shell particles and 1.8μm fully porous particles were compared at high temperatures using very steep (fast) gradient conditions and quite high linear velocities using the same instrument configuration as used to transfer first dimension effluent to the second dimension column as done in on-line comprehensive two-dimensional liquid chromatography. The experimental peak capacities of small columns (2.1mm×30mm) packed with both types of particles were measured with fast gradients (9s to 2min) at high temperature (95°C) using both the same flow rate (1.75mL/min) and then at different flow rates at the same pressure (400bar). Equal or slightly better peak capacities were achieved with the core-shell particle columns as compared to the fully porous particle columns at the same backpressure or the same flow rate. However, core-shell particles offer a real advantage over the smaller, fully porous particles because they can be operated at higher flow rates thus gradient mixer flush out and column reequilibration can be done in less time thereby allowing a greater fraction of the second dimension cycle time to be dedicated to the gradient time.

Lipophilic bioactive compounds in the oils recovered from cereal by‐products

BACKGROUND The by-products of seven different cereal grains were investigated as a source of extractable oil, rich in lipophilic bioactive compounds. RESULTS Oil yields (g kg(-1) DW) recovered from cereal by-products were as follows: 189 (rice bran) > 112 (wheat germ) > 74 (corn bran) > 58 (oat bran) > 41 (buckwheat bran) > 39 (spelt bran) > 33 (wheat bran) > 27 (rye bran). The main fatty acids identified in the studied oil samples were palmitic acid (11.39-17.23%), oleic acid (11.76-42.73%), linoleic acid (35.54-62.65%) and α-linolenic acid (1.05-9.46%). The range of total tocochromanols and phytosterols in the obtained oils was 0.369-3.763 and 1.19-35.24 g kg(-1) of oil, respectively. The oils recovered from buckwheat and corn bran, and wheat germ were dominated by tocopherols (99.9, 84.2 and 96.5%, respectively), whereas the oat, rice, rye, spelt, wheat bran oils were rich in tocotrienols (73.9, 79.6, 78.1, 90.6 and 73.8%, respectively). The campesterol and β-sitosterol constituted 10.1-32.5 and 30.4-63.7%, respectively, of total phytosterols contents identified in all of the studied samples. CONCLUSION The present study demonstrated that oils recovered from the cereal by-products are richer sources of bioactive compounds, compared with traditional oils.

Investigating retention characteristics of a mixed‐mode stationary phase and the enhancement of monolith selectivity for high‐performance liquid chromatography

The synthesis and chromatographic behavior of an analytical size mixed-mode bonded silica monolith was investigated. The monolith was functionalized by an in situ modification process of a bare silica rod with chloro(3-cyanopropyl)dimethyl silane and chlorodimethyl propyl phenyl silane solutions. These ligands were selected in order to combine both resonance and nonresonance π-type bonding within a single separation environment. Selectivity studies were undertaken using n-alkyl benzenes and polycyclic aromatic hydrocarbons in aqueous methanol and acetonitrile mobile phases to assess the methylene and aromatic selectivities of the column. The results fit with the linear solvent strength theory suggesting excellent selectivity of the column was achieved. Comparison studies were performed on monolithic columns that were functionalized separately with cyano and phenyl ligands, suggesting highly conjugated molecules were able to successfully exploit both of the π-type selectivities afforded by the two different ligands on the mixed-mode column.