Multi-dimensional liquid chromatography (MD-LC) continues to gain in popularity for applications where conventional one-dimensional liquid chromatography is insufficient to solve the analytical problem at hand. In this review we have focused on articles published in the years 2019 to early 2023 and look for trends using our previous review published in 2018 as a baseline. We have also explored usage patterns related to involvement of industrial laboratories in the published research. The two major areas of technical development have been continued work on modulation strategies that help mitigate problems associated with mobile phase mismatch when coupling complementary separation mechanisms, and development of computer-aided method development strategies. Progress in these areas is making 2D-LC easier to use, and it appears that this is translating to a shift toward more involvement by industrial laboratories. Indeed, over 34% of the more than 200 publications on 2D-LC in the last four years have had at least one-industry affiliated author. A recent inter-laboratory comparison study focused on the performance of a sophisticated multi-stage, multi-dimensional separation for therapeutic protein characterization is an exemplary indication of the increasing investment of industrial laboratories to MD-LC, and we expect this trend to continue for the foreseeable future. 

Recent trends in two-dimensional liquid chromatography

Risperidone is an important antipsychotic agent used for treatment of both acute and chronic schizophrenia The drug has been effective in resolving the disease’s signs while keeping extrapyramidal side effects as low as possible To achieve a sustained and gradual release of risperidone from PLGA microspheres, we utilized an emulsifying agent; PLGA-PEG-PLGA triblock (M-triblock), instead of PVA (M-PVA), applying ring-opening polymerization of the triblock via supercritical carbon dioxide (scCO2) PVA and PLGA-PEG-PLGA (as emulsifiers) were successfully employed to prepare risperidone-loaded PLGA microspheres The scCO2 and microwave irradiation methods were used to construct PLGA-PEG-PLGA copolymers The microspheres were subjected to various analyses to determine the particles’ sizes, morphological characteristics, and structure (via XRD) and in vitro drug release kinetics Drug loading capacity and encapsulation percentage were also determined The results showed a mean diameter of 4334 ± 230 µm for M-PVA, and that of the M-triblock was 5749 ± 121 µm Risperidone encapsulation percentages were 728 ± 209 for M-PVAs and 734 ± 241 for the M-triblock Particle size, risperidone distribution, and drug loading and encapsulation percentages were similar between M-PVAs and the M-triblock In scanning electron microscopy (SEM), the morphology of the triblock emulsifier was more uniform than PVAs The triblock emulsifier showed slower initial drug release than PVAs due to the precipitation of the triblock hydrogel around PLGA microspheres Overall, the triblock used here can be a good alternative to PVA

The PLGA Microspheres Synthesized by a Thermosensitive Hydrogel Emulsifier for Sustained Release of Risperidone

A novel and sensitive heart-cutting two-dimensional liquid chromatography with ultraviolet detection method (2D-LC-UV) was developed and validated for determination of amisulpride in human plasma. The 2D-LC system consists of a first dimensional (1 D) LC column and a middle transfer column as well as a second-dimensional (2 D) LC column. After simple protein precipitation, the sample was directly injected into the introduction valve of the 2D-LC system. The 1 D column, playing a role of primary separation and preconcentration for complex plasma matrices, transferred the targets to the intermediate column. Following capture of targets on the middle column online, the analytes were transferred to the 2 D separation column by a six-port valve. The 2 D column, avoiding interference from the plasma matrix, completed further separation and quantification. An assistant pump was optimized for primary enrichment as well as final elution in the heart-cutting mode. The analytical time of amisulpride was 7.401 min. The accuracy was between 0.48 and 8.49%, while the intra- and inter-day precisions ranged from 0.9 to 3.1% and from 1.7% to 3.3%, respectively. The linear range of amisulpride was 48.15-2,407.59 ng/ml, while the extraction recovery was 98.7-101.3%. The strategy established in the study, which was successfully applied to therapeutic drug monitoring of amisulpride for routine clinical detection, displays high sensitivity, good repeatability, convenience and low cost.

A novel and sensitive method for determination of amisulpride in human plasma by two-dimensional liquid chromatography.

The aim of the study was to evaluate the reference range of amisulpride for Chinese patients with schizophrenia and to assess its possible influencing factors based on therapeutic drug monitoring information. The relative adverse reactions of patients induced by amisulpride were also systematically investigated. A total of 425 patients with schizophrenia were assessed, including Positive and Negative Syndrome Scales, Treatment Emergent Symptom Scale, blood routine examination, hepatorenal function, lipids, hormones, as well as myocardial enzymes at baseline, and following treatment with amisulpride for 8 weeks. The steady-state plasma concentration of amisulpride was assayed using two-dimensional liquid chromatography. At the same dose, the amisulpride plasma concentration of patients combined with clozapine was higher than that without clozapine. The therapeutic reference range of amisulpride can be defined as 230.3–527.1 ng/ml for Chinese patients with schizophrenia. The potential side effects appear to be associated with significantly increased levels of LDH, CK, creatine kinase isoenzyme (CK-MB), TC and decreased level of E2, relative to the amisulpride plasma concentration. These findings could provide individualized medication and reduce the adverse effects of amisulpride for Chinese patients with schizophrenia. 

Amisulpride steady-state plasma concentration and adverse reactions in patients with schizophrenia: a study based on therapeutic drug monitoring data

The aim of the study was to evaluate the reference range of amisulpride for Chinese patients with schizophrenia and to assess its possible influencing factors based on therapeutic drug monitoring information. The relative adverse reactions of patients induced by amisulpride were also systematically investigated. A total of 425 patients with schizophrenia were assessed, including Positive and Negative Syndrome Scales, Treatment Emergent Symptom Scale, blood routine examination, hepatorenal function, lipids, hormones, as well as myocardial enzymes at baseline, and following treatment with amisulpride for 8 weeks. The steady-state plasma concentration of amisulpride was assayed using two-dimensional liquid chromatography. At the same dose, the amisulpride plasma concentration of patients combined with clozapine was higher than that without clozapine. The therapeutic reference range of amisulpride can be defined as 230.3–527.1 ng/ml for Chinese patients with schizophrenia. The potential side effects appear to be associated with significantly increased levels of LDH, CK, creatine kinase isoenzyme (CK-MB), TC and decreased level of E2, relative to the amisulpride plasma concentration. These findings could provide individualized medication and reduce the adverse effects of amisulpride for Chinese patients with schizophrenia.

A novel and robust two-dimensional liquid chromatography with ultraviolet detection method (2D-LC-UV) was developed and validated for high-throughput determination of the concentrations of valproic acid (VPA) in human plasma. This 2D-LC system was composed of a first-dimensional LC column, a second-dimensional LC column and an intermediate transfer column. The sample was directly injected into the 2D-LC system after an easy protein precipitation treatment. After online preconcentration and primary separation by the first-dimensional column, the target was captured by an intermediate column and then transferred to second-dimensional column for analysis. The system transferred the target through "central cutting" mode whereby the drug peak was not subject to interference from the matrix. The analysis cycle time was completed within 7.0 min. Compared with other methods that have been developed, the analysis time was reduced and the operation was much easier without any derivatization. The calibration curve was linear over the 5.90-188.94 μg/ml range for the VPA concentrations. The intra-day and inter-day precisions were <5.6%. The recoveries were in the range from 95.2 to 98.0%. This method appears to be sensitive, precise, rapid and low-cost for the quantification of VPA in serum sample.

A novel and nonderivatization method for the determination of valproic acid in human serum by two‐dimensional liquid chromatography

This study reports a novel heart-cutting two-dimensional high performance liquid chromatography (2D-LC) method with ultraviolet detection for the determination of risperidone and 9-hydroxyrisperido...

Determination of Risperidone and 9-Hydroxyrisperidone in Human Serum by Heart-Cutting Isocratic Two-Dimensional Liquid Chromatography

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A novel and nonderivatization method for the determination of valproic acid in human serum by two‐dimensional liquid chromatography

A novel and sensitive method for determination of amisulpride in human plasma by two-dimensional liquid chromatography.

Determination of Risperidone and 9-Hydroxyrisperidone in Human Serum by Heart-Cutting Isocratic Two-Dimensional Liquid Chromatography