Liquid chromatographic enantioseparation of thalidomide and its derivatives on cyclodextrin-bonded stationary phases

Abstract The chiral separation of three racemic immunomodulatory drugs, thalidomide, pomalidomide and lenalidomide was studied, using three cyclodextrin bonded stationary phases (β-, hydroxypropyl-β- and carboxymethyl-β-CD) in reversed-phase and polar organic mode. In polar organic mode, using acetonitrile and methanol, no chiral separation was observed. In reversed-phase mode pomalidomide showed chiral interactions with all selectors, while lenalidomide showed no chiral interactions with any of the cyclodextrins employed. Thalidomide showed chiral interactions with β-and carboxymethyl-β-CD, only. Based on these observations it can be concluded that the oxo group at position two is necessary for chiral recognition, while the aromatic primary amine group enhances it. Orthogonal experimental design was used to investigate the effect of the eluent composition, flow rate, and the column temperature on chiral separation. Concentration of the organic modifier was the most important factor among the investigated three variables showing high impact on the chiral separations. In the case of thalidomide optimized parameters (β-cyclodextrin-based stationary phase, 0.1% acetic acid/acetonitrile 95/5 (v/v), 5 °C column temperature, 0.6 ml/min flow rate) resulted in a resolution of 1.68 ± 0.02 between enantiomers. For pomalidomide, this value was 2.70 ± 0.02, under the circumstances as follows: β-cyclodextrin-based stationary phase, 0.1% acetic acid/acetonitrile 90/10 (v/v), 15 °C column temperature and 0.8 mL/min flow rate. Utilizing the experimental conditions employed on an LC-MS/MS system, concentrations as low as 2 ng/mL could be determined from mouse plasma for both substances. Elution sequences were determined with enantiopure standards and in both cases the R-enantiomers eluted first. The methods developed are suitable for the chiral separation of the abovementioned compounds and are sound starting points for bioanalytical method development.

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Effect of column temperature on the retention of peropyrene-type polycyclic aromatic hydrocarbons on various chemically bonded stationary phases in reversed-phase liquid chromatography

Journal of Chromatography A ◽

10.1016/s0021-9673(00)94558-8 ◽

1988 ◽

Vol 436 ◽

pp. 1-10 ◽

Cited By ~ 31

Author(s):

K. Jinno ◽

T. Nagoshi ◽

N. Tanaka ◽

M. Okamoto ◽

J.C. Fetzer ◽

...

Keyword(s):

Polycyclic Aromatic Hydrocarbons ◽

Liquid Chromatography ◽

Aromatic Hydrocarbons ◽

Stationary Phases ◽

Reversed Phase ◽

Reversed Phase Liquid Chromatography ◽

Column Temperature ◽

Bonded Stationary Phases ◽

Polycyclic Aromatic ◽

Phase Liquid

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Selective separation of xanthones and saponins from the rhizomes of Anemarrhena asphodeloides by modulating the density of surface charges in C18-bonded stationary phases

Analytical Methods ◽

10.1039/c7ay01667k ◽

2017 ◽

Vol 9 (38) ◽

pp. 5604-5610 ◽

Cited By ~ 1

Author(s):

Jianfeng Cai ◽

Huaxia Xin ◽

Lingping Cheng ◽

Zhimou Guo ◽

Jiatao Feng ◽

...

Keyword(s):

Stationary Phase ◽

Stationary Phases ◽

Reversed Phase ◽

Selective Separation ◽

Separation Method ◽

Surface Charges ◽

Bonded Stationary Phases ◽

Robust Separation ◽

Anemarrhena Asphodeloides ◽

Positively Charged

A rapid and robust separation method based on the positively charged reversed-phase (PGRP) stationary phase was developed for selective separation of saponins and xanthones from the rhizomes of Anemarrhena asphodeloides.

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Direct Enantiomeric Resolution of Seventeen Racemic 1,4-Dihydropyridine-Based Hexahydroquinoline Derivatives by HPLC

International Journal of Molecular Sciences ◽

10.3390/ijms20102513 ◽

2019 ◽

Vol 20 (10) ◽

pp. 2513 ◽

Cited By ~ 1

Author(s):

Jiayi Sun ◽

Miyase Gözde Gündüz ◽

Junyuan Zhang ◽

Jia Yu ◽

Xingjie Guo

Keyword(s):

Stationary Phase ◽

High Performance ◽

Chiral Stationary Phase ◽

Reversed Phase ◽

Chiral Separations ◽

Racemic Mixture ◽

Enantiomeric Resolution ◽

Column Temperature ◽

Limit Of Quantitation ◽

Pharmacologically Active

1,4-Dihydropyridine (DHP) scaffold holds an outstanding position with its versatile pharmacological properties among all heterocyclic compounds. Although most of the commercially available DHPs are marketed as a racemic mixture, the chiral center at C-4 can lead to even opposite pharmacological activities between the enantiomers. In the present study, enantioseparation of seventeen DHP structural analogues, consisting of either pharmacologically active or newly synthesized derivatives, (M2-4, MD5, HM2, HM10, CE5, N11, N10, N7, M11, MC6-8, MC13, MD23, and 42IIP) by high-performance liquid chromatography was investigated using immobilized polysaccharide-based chiral stationary phase, Chiralpak IC column. Due to the solvent versatility of the covalently immobilized chiral stationary phase in enantiomer separation, multiple elution modes including standard normal phase, nonstandard mobile phase, and reversed phase were used to expand the possibility to find the optimum enantioselective conditions for the tested analytes. Under appropriate separation conditions, complete enantiomeric separation was obtained for nearly all compounds except MC6-8 and MC13 which contained two chiral centers. Additionally, the effects of the polar modifier, the additive, and column temperature on the chiral recognition were evaluated. The thermodynamic parameters calculated according to the linear van’t Hoff equation indicated that the chiral separations in this study were enthalpy-driven or entropy-driven. Some parameters of method validation such as linearity, limit of quantitation, and repeatability were also measured for all studied compounds to prove the reliability of the method.

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Stationary phase type and temperature effect on HPLC separation of lactic acid enantiomers

Acta Chimica Slovaca ◽

10.2478/acs-2021-0008 ◽

2021 ◽

Vol 14 (1) ◽

pp. 60-65

Author(s):

Laura Jánovová ◽

Katarína Hroboňová

Keyword(s):

Lactic Acid ◽

High Performance ◽

Stationary Phases ◽

Chiral Stationary Phases ◽

Reversed Phase ◽

Optical Isomers ◽

Column Temperature ◽

Retention Factors ◽

Chromatographic Parameters ◽

Macrocyclic Antibiotics

Abstract Lactic acid is a biologically important organic acid existing in two enantiomeric forms which are differently metabolized in the human body. In this paper, direct chiral separation of lactic acid by high performance liquid chromatography is presented. Five chiral stationary phases based on macrocyclic antibiotics were used for enantioseparation and chromatographic parameters, such as retention factors, resolution and selectivity factors, were determined under different column temperatures ranging from 5 to 45 °C. Optical isomers of lactic acid were efficiently separated using chiral stationary phases based on teicoplanin (R S = 1.9 ) and ristocetin (R S = 1.7 ) in reversed-phase separation mode at the column temperature of 25 °C.

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