Journal of Pharmaceutical Analysis

Recent applications and chiral separation development based on stationary phases in open tubular capillary electrochromatography (2019-2022)

Xinyu Li, Qianjie Ma, Xiangtai Zheng, Qin Chen, Xiaodong Sun

2023, 13(4): 323-339. doi: 10.1016/j.jpha.2023.01.003

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Abstract:
Capillary electrochromatography (CEC) plays a significant role in chiral separation via the double separation principle, partition coefficient difference between the two phases, and electroosmotic flow-driven separation. Given the distinct properties of the inner wall stationary phase (SP), the separation ability of each SP differs from one another. Particularly, it provides large room for promising applications of open tubular capillary electrochromatography (OT-CEC). We divided the OT-CEC SPs developed over the past four years into six types: ionic liquids, nanoparticle materials, microporous materials, biomaterials, non-nanopolymers, and others, to mainly introduce their characteristics in chiral drug separation. There also added a few classic SPs that occurred within ten years as supplements to enrich the features of each SP. Additionally, we discuss their applications in metabolomics, food, cosmetics, environment, and biology as analytes in addition to chiral drugs. OT-CEC plays an increasingly significant role in chiral separation and may promote the development of capillary electrophoresis (CE) combined with other instruments in recent years, such as CE with mass spectrometry (CE/MS) and CE with ultraviolet light detector (CE/UV).

Recent progress in aptamer-based microfluidics for the detection of circulating tumor cells and extracellular vesicles

Duanping Sun, Ying Ma, Maoqiang Wu, Zuanguang Chen, Luyong Zhang, Jing Lu

2023, 13(4): 340-354. doi: 10.1016/j.jpha.2023.03.001

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Abstract:
Liquid biopsy is a technology that exhibits potential to detect cancer early, monitor therapies, and predict cancer prognosis due to its unique characteristics, including noninvasive sampling and real-time analysis. Circulating tumor cells (CTCs) and extracellular vesicles (EVs) are two important components of circulating targets, carrying substantial disease-related molecular information and playing a key role in liquid biopsy. Aptamers are single-stranded oligonucleotides with superior affinity and specificity, and they can bind to targets by folding into unique tertiary structures. Aptamer-based microfluidic platforms offer new ways to enhance the purity and capture efficiency of CTCs and EVs by combining the advantages of microfluidic chips as isolation platforms and aptamers as recognition tools. In this review, we first briefly introduce some new strategies for aptamer discovery based on traditional and aptamer-based microfluidic approaches. Then, we subsequently summarize the progress of aptamer-based microfluidics for CTC and EV detection. Finally, we offer an outlook on the future directional challenges of aptamer-based microfluidics for circulating targets in clinical applications.

Neutrophil elastase: From mechanisms to therapeutic potential

Weilin Zeng, Yingqiu Song, Runze Wang, Rong He, Tianlu Wang

2023, 13(4): 355-366. doi: 10.1016/j.jpha.2022.12.003

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Abstract:
Neutrophil elastase (NE), a major protease in the primary granules of neutrophils, is involved in microbicidal activity. NE is an important factor promoting inflammation, has bactericidal effects, and shortens the inflammatory process. NE also regulates tumor growth by promoting metastasis and tumor microenvironment remodeling. However, NE plays a role in killing tumors under certain conditions and promotes other diseases such as pulmonary ventilation dysfunction. Additionally, it plays a complex role in various physiological processes and mediates several diseases. Sivelestat, a specific NE inhibitor, has strong potential for clinical application, particularly in the treatment of coronavirus disease 2019 (COVID-19). This review discusses the pathophysiological processes associated with NE and the potential clinical applications of sivelestat.

Development of a CLDN18.2-targeting immuno-PET probe for non-invasive imaging in gastrointestinal tumors

Yan Chen, Xingguo Hou, Dapeng Li, Jin Ding, Jiayue Liu, Zilei Wang, Fei Teng, Hongjun Li, Fan Zhang, Yi Gu, Steven Yu, Xueming Qian, Zhi Yang, Hua Zhu

2023, 13(4): 367-375. doi: 10.1016/j.jpha.2023.02.011

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Abstract:
Claudin18.2 (CLDN18.2) is a tight junction protein that is overexpressed in a variety of solid tumors such as gastrointestinal cancer and oesophageal cancer. It has been identified as a promising target and a potential biomarker to diagnose tumor, evaluate efficacy, and determine patient prognosis. TST001 is a recombinant humanized CLDN18.2 antibody that selectively binds to the extracellular loop of human Claudin18.2. In this study, we constructed a solid target radionuclide zirconium-89 (⁸⁹Zr) labled-TST001 to detect the expression of in the human stomach cancer BGC823^CLDN18.2 cell lines. The [⁸⁹Zr]Zr-desferrioxamine (DFO)-TST001 showed high radiochemical purity (RCP, >99%) and specific activity (24.15±1.34 GBq/μmol), and was stable in 5% human serum albumin, and phosphate buffer saline (>85% RCP at 96h). The EC₅₀ values of TST001 and DFO-TST001 were as high as 0.413±0.055 and 0.361±0.058nM(P>0.05), respectively. The radiotracer had a significantly higher average standard uptake values in CLDN18.2-positive tumors than in CLDN18.2-negative tumors (1.11±0.02 vs. 0.49±0.03, P=0.0016) 2 days post injection (p.i.). BGC823^CLDN18.2 mice models showed high tumor/muscle ratios 96h p.i. with [⁸⁹Zr]Zr-DFO-TST001 was much higher than those of the other imaging groups. Immunohistochemistry results showed that BGC823^CLDN18.2 tumors were highly positive (+++) for CLDN18.2, while those in the BGC823 group did not express CLDN18.2 (-). The results of exvivo biodistribution studies showed that there was a higher distribution in the BGC823^CLDN18.2 tumor bearing mice (2.05±0.16 %ID/g) than BGC823 mice (0.69±0.02 %ID/g) and blocking group (0.72±0.02 %ID/g). A dosimetry estimation study showed that the effective dose of [⁸⁹Zr]Zr-DFO-TST001 was 0.0705 mSv/MBq, which is within the range of acceptable doses for nuclear medicine research. Taken together, these results suggest that Good Manufacturing Practices produced by this immuno-positron emission tomography probe can detect CLDN18.2-overexpressing tumors.

High-throughput transcriptional profiling of perturbations by Panax ginseng saponins and Panax notoginseng saponins using TCM-seq

Junyun Cheng, Jie Chen, Jie Liao, Tianhao Wang, Xin Shao, Jinbo Long, Penghui Yang, Anyao Li, Zheng Wang, Xiaoyan Lu, Xiaohui Fan

2023, 13(4): 376-387. doi: 10.1016/j.jpha.2023.02.009

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Abstract:
Panax ginseng (PG) and Panax notoginseng (PN) are highly valuable Chinese medicines (CM). Although both CMs have similar active constituents, their clinical applications are clearly different. Over the past decade, RNA sequencing (RNA-seq) analysis has been employed to investigate the molecular mechanisms of extracts or monomers. However, owing to the limited number of samples in standard RNA-seq, few studies have systematically compared the effects of PG and PN spanning multiple conditions at the transcriptomic level. Here, we developed an approach that simultaneously profiles transcriptome changes for multiplexed samples using RNA-seq (TCM-seq), a high-throughput, low-cost workflow to molecularly evaluate CM perturbations. A species-mixing experiment was conducted to illustrate the accuracy of sample multiplexing in TCM-seq. Transcriptomes from repeated samples were used to verify the robustness of TCM-seq. We then focused on the primary active components, Panax notoginseng saponins (PNS) and Panax ginseng saponins (PGS) extracted from PN and PG, respectively. We also characterized the transcriptome changes of 10cell lines, treated with four different doses of PNS and PGS, using TCM-seq to compare the differences in their perturbing effects on genes, functional pathways, gene modules, and molecular networks. The results of transcriptional data analysis showed that the transcriptional patterns of various cell lines were significantly distinct. PGS exhibited a stronger regulatory effect on genes involved in cardiovascular disease, whereas PNS resulted in a greater coagulation effect on vascular endothelial cells. This study proposes a paradigm to comprehensively explore the differences in mechanisms of action between CMs based on transcriptome readouts.

Quantitative characterization of cell physiological state based on dynamical cell mechanics for drug efficacy indication

Shuang Ma, Junfeng Wu, Zhihua Liu, Rong He, Yuechao Wang, Lianqing Liu, Tianlu Wang, Wenxue Wang

2023, 13(4): 388-402. doi: 10.1016/j.jpha.2023.03.002

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Abstract:
Cell mechanics is essential to cell development and function, and its dynamics evolution reflects the physiological state of cells. Here, we investigate the dynamical mechanical properties of single cells under various drug conditions, and present two mathematical approaches to quantitatively characterizing the cell physiological state. It is demonstrated that the cellular mechanical properties upon the drug action increase over time and tend to saturate, and can be mathematically characterized by a linear time-invariant dynamical model. It is shown that the transition matrices of dynamical cell systems significantly improve the classification accuracies of the cells under different drug actions. Furthermore, it is revealed that there exists a positive linear correlation between the cytoskeleton density and the cellular mechanical properties, and the physiological state of a cell in terms of its cytoskeleton density can be predicted from its mechanical properties by a linear regression model. This study builds a relationship between the cellular mechanical properties and the cellular physiological state, adding information for evaluating drug efficacy.

Solriamfetol impurities: Synthesis, characterization, and analytical method (UPLC-UV) validation

Nafisah Al-Rifai, Anas Alshishani, Fouad Darras, Ola Taha, Shereen Abu-Jalloud, Lena Shaghlil, Yousef Al-Ebini

2023, 13(4): 403-411. doi: 10.1016/j.jpha.2023.02.012

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Abstract:
Given that impurities may affect the quality and safety of drug products, impurity identification and profiling is an integral part of drug quality control and is particularly important for newly developed medications such as solriamfetol, which is used to treat excessive daytime sleepiness. Although the high-performance liquid chromatography analysis of commercial solriamfetol has revealed the presence of several impurities, their synthesis, structure elucidation, and chromatographic determination have not been reported yet. To bridge this gap, we herein identified, synthesized, and isolated eight process-related solriamfetol impurities, characterized them using spectroscopic and chromatographic techniques, and proposed plausible mechanisms of their formation. Moreover, we developed and validated a prompt impurity analysis method based on ultrahigh-performance liquid chromatography with UV detection, revealing that its selectivity, linearity, accuracy, precision, and quantitation limit meet the acceptance criteria of method validation stipulated by the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use. Thus, the developed method was concluded to be suitable for the routine analysis of solriamfetol substances.

Breath-by-breath measurement of exhaled ammonia by acetone-modifier positive photoionization ion mobility spectrometry via online dilution and purging sampling

Lu Wang, Dandan Jiang, Lei Hua, Chuang Chen, Dongming Li, Weiguo Wang, Yiqian Xu, Qimu Yang, Haiyang Li, Song Leng

2023, 13(4): 412-420. doi: 10.1016/j.jpha.2023.02.008

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Abstract:
Exhaled ammonia (NH₃) is an essential noninvasive biomarker for disease diagnosis. In this study, an acetone-modifier positive photoionization ion mobility spectrometry (AM-PIMS) method was developed for accurate qualitative and quantitative analysis of exhaled NH₃ with high selectivity and sensitivity. Acetone was introduced into the drift tube along with the drift gas as a modifier, and the characteristic NH₃ product ion peak of (C₃H₆O)₄NH₄⁺ (K₀=1.45cm²/V·s) was obtained through the ion-molecule reaction with acetone reactant ions (C₃H₆O)₂H⁺ (K₀=1.87cm²/V·s), which significantly increased the peak-to-peak resolution and improved the accuracy of exhaled NH₃ qualitative identification. Moreover, the interference of high humidity and the memory effect of NH₃ molecules were significantly reduced via online dilution and purging sampling, thus realizing breath-by-breath measurement. As a result, a wide quantitative range of 5.87-140.92μmol/L with a response time of 40ms was achieved, and the exhaled NH₃ profile could be synchronized with the concentration curve of exhaled CO₂. Finally, the analytical capacity of AM-PIMS was demonstrated by measuring the exhaled NH₃ of healthy subjects, demonstrating its great potential for clinical disease diagnosis.

A chiral metal-organic framework {(HQA)(ZnCl₂)(2.5H₂O)}_n for the enantioseparation of chiral amino acids and drugs

Xiangtai Zheng, Qi Zhang, Qianjie Ma, Xinyu Li, Liang Zhao, Xiaodong Sun

2023, 13(4): 421-429. doi: 10.1016/j.jpha.2023.03.003

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Abstract:
Chiral metal-organic frameworks (CMOFs) with enantiomeric subunits have been employed in chiral chemistry. In this study, a CMOF formed from 6-methoxyl-(8S,9R)-cinchonan-9-ol-3-carboxylic acid (HQA) and ZnCl₂, {(HQA)(ZnCl₂)(2.5H₂O)}_n, was constructed as a chiral stationary phase (CSP) via an in situ fabrication approach and used for chiral amino acid and drug analyses for the first time. The {(HQA)(ZnCl₂)(2.5H₂O)}_n nanocrystal and the corresponding chiral stationary phase were systematically characterised using a series of analytical techniques including scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, circular dichroism, X-ray photoelectron spectroscopy, thermogravimetric analysis, and Brunauer-Emmett-Teller surface area measurements. In open-tubular capillary electrochromatography (CEC), the novel chiral column exhibited strong and broad enantioselectivity toward a variety of chiral analytes, including 19 racemic dansyl amino acids and several model chiral drugs (both acidic and basic). The chiral CEC conditions were optimised, and the enantioseparation mechanisms are discussed. This study not only introduces a new high-efficiency member of the MOF-type CSP family but also demonstrates the potential of improving the enantioselectivities of traditional chiral recognition reagents by fully using the inherent characteristics of porous organic frameworks.

2023 Vol. 13, No. 4