Determination of the phase ratio of a dehydroabietic-acid-bonded silica-gel chromatographic stationary phase and its effect on separation thermodynamics.

Rosin-based chromatographic columns are widely used for separation purposes, but, to date, their phase ratios (Φ) have been imprecisely measured. This affects the understanding of their separation mechanism and the calculation of related thermodynamic parameters. In this study, a stationary phase was synthesized by bonding dehydroabietic acid (DA) to silica gel (Si-DO) and applied for reversed-phase liquid chromatography. The distribution coefficient (Kdm) of methyl dehydroabietate (MD), which has the same structure as the bonded phase of Si-DO, was used as a surrogate for the determination of the equilibrium coefficient (K) of Si-DO, and the Kdm values of MD in different mobile phases were measured and compared with the K values of Si-DO. It was found that the phase ratio of Si-DO varied with mobile phase composition and temperature, as shown by the Φ values: 0.039-0.122 for the methanol/water system and 0.051-0.116 for the acetonitrile/water system; in addition, the a indices were 0.552-0.757 and 0.564-0.674, respectively. The Kdm of MD was closer to the K of Si-DO than those of other surrogate models, including the octanol-water and octane-mobile phase partition coefficients. In addition, the thermodynamic parameters (ΔG°, ΔH°, and ΔS°) of n-alkylbenzenes on Si-DO were negative, indicating a spontaneous and enthalpy-driven separation process. Overall, the phase ratio of rosin-based columns is crucial for accurate thermodynamic analysis and interpretation of the separation mechanism. Finally, the MD surrogate model allows the estimation of phase ratio of Si-DO and other similar columns, providing a novel method for measuring the phase ratio of rosin-based columns and providing a validated concept and methodology for determining the phase ratios of HPLC columns.

Preparation, characterization, and separation mechanism of a dehydroabietic-acid-based shape-selective chromatographic stationary phase1.

Chromatographic stationary phases with molecular-shape selectivity are advantageous for the separation and analysis of geometric isomers. Herein, dehydroabietic acid is bonded on the surface of silica microspheres via 3-glycidoxypropyltrimethoxysilane to form a monolayer dehydroabietic-acid stationary phase (Si-DOMM) with a racket-shaped structure. Various characterization techniques indicate that Si-DOMM is successfully prepared, and the separation performance of a Si-DOMM column is evaluated. The stationary phase has a low silanol activity and metal contamination and a high hydrophobicity and shape selectivity. The resolutions of lycopene, lutein, and capsaicin on the Si-DOMM column confirm that the stationary phase exhibits high shape selectivity. The elution order of n-alkyl benzene on the Si-DOMM column indicates its high hydrophobic selectivity and suggests that the separation is an enthalpy-driven process. Repeatability experiments reveal highly stable preparation processes of the stationary phase and column and indicate that the relative standard deviations of retention time, peak height, and peak area are less than 0.26%, 3.54%, and 3.48%, respectively. Density functional theory calculations using n-alkylbenzenes, polycyclic aromatic hydrocarbons, amines, and phenols as model solutes provide an intuitive and quantitative description of the multiple retention mechanisms. The Si-DOMM stationary phase exhibits superior retention and high selectivity for these compounds via multiple interactions. The bonding phase of the monolayer dehydroabietic acid stationary phase with a racket-shaped structure has a unique affinity for benzene, strong shape selectivity, and good separation performance for geometrical isomers with different molecular shapes.

A new oleanolic-type triterpene glycoside from Anchusa italica.

A new oleanolic-type triterpene glycoside, (3ß,21ß)-21-[(ß-d-glucopyranosyl-(1→2)-ß-d-glucopyranosyl)oxy]-3-hydroxyolean-12-en-28-oic acid (1), together with five analogues, oleanazuroside 1 (2), oleanazuroside 2 (3), 24-hydroxytormentic acid ester glucoside (4), 24-epi-pinfaensin (5), and oleanolic acid 3-O-α-l-arabinoside (6) were isolated from the n-butyl alcohol extract of Anchusa italica. Their structures were determined spectroscopically and compared with previously reported spectral data. Compounds 3-4 and 6 were evaluated for their cytotoxic activities against five human cancer cell lines, but only compound 6 showed moderate cytotoxicity.