Nuclear magnetic resonance-based solvent system selection for counter-current chromatography separation of compounds present in the same high-performance liquid chromatography peak: Flavonoids in barley seedlings as an example.

Partition coefficient is a key parameter for counter-current chromatography separation. High-performance liquid chromatography (HPLC) is the most commonly used tool for the screening of partition coefficients. However, HPLC technology is not applicable to the compounds present in the same chromatographic peak. Nuclear magnetic resonance (NMR) technology could easily distinguish compounds according to their characteristic absorption even if they exist in the same HPLC peak. In this study, two flavonoids present in the same HPLC peak were successfully purified by counter-current chromatography with a solvent system screened by NMR to show the great potential of NMR technology in the screening of the partition coefficient of co-efflux compounds. Through NMR screening, an optimized ethyl acetate/n-buthanol/water (7:3:10, v/v/v) system was applied in this study. As a result, two flavonoids, including 4.8 mg of 3'-methoxyl-6'''-O-feruloylsaponarin and 9.8 mg of 6'''-O-feruloylsaponarin were separated from 15 mg of the mixture. There is only one methoxy group difference between the two flavonoids. This study provides a new strategy for the screening of counter-current chromatography solvent systems and broadens the application scope of counter-current chromatography.

Systematic and practical solvent system selection strategy based on the nonrandom two-liquid segment activity coefficient model for real-life counter-current chromatography separation.

Solvent system selection is the first step toward a successful counter-current chromatography (CCC) separation. This paper introduces a systematic and practical solvent system selection strategy based on the nonrandom two-liquid segment activity coefficient (NRTL-SAC) model, which is efficient in predicting the solute partition coefficient. Firstly, the application of the NRTL-SAC method was extended to the ethyl acetate/n-butanol/water and chloroform/methanol/water solvent system families. Moreover, the versatility and predictive capability of the NRTL-SAC method were investigated. The results indicate that the solute molecular parameters identified from hexane/ethyl acetate/methanol/water solvent system family are capable of predicting a large number of partition coefficients in several other different solvent system families. The NRTL-SAC strategy was further validated by successfully separating five components from Salvia plebeian R.Br. We therefore propose that NRTL-SAC is a promising high throughput method for rapid solvent system selection and highly adaptable to screen suitable solvent system for real-life CCC separation.

Correlation and prediction of partition coefficient using nonrandom two-liquid segment activity coefficient model for solvent system selection in counter-current chromatography separation.

Selection of a suitable solvent system is the first and foremost step for a successful counter-current chromatography (CCC) separation. In this paper, a thermodynamic model, nonrandom two-liquid segment activity coefficient model (NRTL-SAC) which uses four types of conceptual segments to describe the effective surface interactions for each solvent and solute molecule, was employed to correlate and predict the partition coefficients (K) of a given compound in a specific solvent system. Then a suitable solvent system was selected according to the predicted partition coefficients. Three solvent system families, heptane/methanol/water, heptane/ethyl acetate/methanol/water (Arizona) and hexane/ethyl acetate/methanol/water, and several solutes were selected to investigate the effectiveness of the NRTL-SAC model for predicting the partition coefficients. Comparison between experimental results and predicted results showed that the NRTL-SAC model is of potential for estimating the K value of a given compound. Also a practical separation case on magnolol and honokiol suggests the NRTL-SAC model is effective, reliable and practical for the purpose of predicting partition coefficients and selecting a suitable solvent system for CCC separation.