Journal of Pharmaceutical Analysis

Personal glucose meters coupled with signal amplification technologies for quantitative detection of non-glucose targets: Recent progress and challenges in food safety hazards analysis

Feng He, Haijie Wang, Pengfei Du, Tengfei Li, Weiting Wang, Tianyu Tan, Yaobo Liu, Yanli Ma, Yuanshang Wang, A.M. Abd El-Aty

2023, 13(3): 223-238. doi: 10.1016/j.jpha.2023.02.005

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Abstract:
Ensuring food safety is paramount worldwide. Developing effective detection methods to ensure food safety can be challenging owing to trace hazards, long detection time, and resource-poor sites, in addition to the matrix effects of food. Personal glucose meter (PGM), a classic point-of-care testing device, possesses unique application advantages, demonstrating promise in food safety. Currently, many studies have used PGM-based biosensors and signal amplification technologies to achieve sensitive and specific detection of food hazards. Signal amplification technologies have the potential to greatly improve the analytical performance and integration of PGMs with biosensors, which is crucial for solving the challenges associated with the use of PGMs for food safety analysis. This review introduces the basic detection principle of a PGM-based sensing strategy, which consists of three key factors: target recognition, signal transduction, and signal output. Representative studies of existing PGM-based sensing strategies combined with various signal amplification technologies (nanomaterial-loaded multienzyme labeling, nucleic acid reaction, DNAzyme catalysis, responsive nanomaterial encapsulation, and others) in the field of food safety detection are reviewed. Future perspectives and potential opportunities and challenges associated with PGMs in the field of food safety are discussed. Despite the need for complex sample preparation and the lack of standardization in the field, using PGMs in combination with signal amplification technology shows promise as a rapid and cost-effective method for food safety hazard analysis.

Multidisciplinary strategies to enhance therapeutic effects of flavonoids from Epimedii Folium: Integration of herbal medicine, enzyme engineering, and nanotechnology

Yi Lu, Qiulan Luo, Xiaobin Jia, James P. Tam, Huan Yang, Yuping Shen, Xin Li

2023, 13(3): 239-254. doi: 10.1016/j.jpha.2022.12.001

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Abstract:
Flavonoids such as baohuoside I and icaritin are the major active compounds in Epimedii Folium (EF) and possess excellent therapeutic effects on various diseases. Encouragingly, in 2022, icaritin soft capsules were approved to reach the market for the treatment of hepatocellular carcinoma (HCC) by National Medical Products Administration (NMPA) of China. Moreover, recent studies demonstrate that icaritin can serve as immune-modulating agent to exert anti-tumor effects. Nonetheless, both production efficiency and clinical applications of epimedium flavonoids have been restrained because of their low content, poor bioavailability, and unfavorable in vivo delivery efficiency. Recently, various strategies, including enzyme engineering and nanotechnology, have been developed to increase productivity and activity, improve delivery efficiency, and enhance therapeutic effects of epimedium flavonoids. In this review, the structure-activity relationship of epimedium flavonoids is described. Then, enzymatic engineering strategies for increasing the productivity of highly active baohuoside I and icaritin are discussed. The nanomedicines for overcoming in vivo delivery barriers and improving therapeutic effects of various diseases are summarized. Finally, the challenges and an outlook on clinical translation of epimedium flavonoids are proposed.

Advances and challenges in using nirmatrelvir and its derivatives against SARS-CoV-2 infection

Wujun Chen, Bing Liang, Xiaolin Wu, Ling Li, Chao Wang, Dongming Xing

2023, 13(3): 255-261. doi: 10.1016/j.jpha.2022.10.005

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Abstract:
On December 22, 2021, the United States Food and Drug Administration approved the first main protease inhibitor, i.e., oral antiviral nirmatrelvir (PF-07321332)/ritonavir (Paxlovid), for the treatment of early severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nirmatrelvir inhibits SARS-CoV-2 infection, but high doses or long-term treatment may cause embryonic developmental toxicity and changes in host gene expression. The chiral structure of nirmatrelvir plays a key role in its antiviral activity. Ritonavir boosts the efficacy of nirmatrelvir by inactivating cytochrome P450 3A4 expression and occupying the plasma protein binding sites. Multidrug resistance protein 1 inhibitors may increase the efficacy of nirmatrelvir. However, Paxlovid has many contraindications. Some patients treated with Paxlovid experience a second round of coronavirus disease 2019 (COVID-19) symptoms soon after recovery. Interestingly, the antiviral activity of nirmatrelvir metabolites, such as compounds 12–18 , is similar to or higher than that of nirmatrelvir. Herein, we review the advances and challenges in using nirmatrelvir and its derivatives with the aim of providing knowledge for drug developers and physicians in the fight against COVID-19.

Overcoming chemoresistance in non-angiogenic colorectal cancer by metformin via inhibiting endothelial apoptosis and vascular immaturity

Guang-Yue Li, Shu-Jing Zhang, Dong Xue, Yue-Qi Feng, Yan Li, Xun Huang, Qiang Cui, Bo Wang, Jun Feng, Tao Bao, Pei-Jun Liu, Shao-Ying Lu, Ji-Chang Wang

2023, 13(3): 262-275. doi: 10.1016/j.jpha.2023.02.001

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Abstract:
The development of chemoresistance which results in a poor prognosis often renders current treatments for colorectal cancer (CRC). In this study, we identified reduced microvessel density (MVD) and vascular immaturity resulting from endothelial apoptosis as therapeutic targets for overcoming chemoresistance. We focused on the effect of metformin on MVD, vascular maturity, and endothelial apoptosis of CRCs with a non-angiogenic phenotype, and further investigated its effect in overcoming chemoresistance. In situ transplanted cancer models were established to compare MVD, endothelial apoptosis and vascular maturity, and function in tumors from metformin- and vehicle-treated mice. An in vitro co-culture system was used to observe the effects of metformin on tumor cell-induced endothelial apoptosis. Transcriptome sequencing was performed for genetic screening. Non-angiogenic CRC developed independently of angiogenesis and was characterized by vascular leakage, immaturity, reduced MVD, and non-hypoxia. This phenomenon had also been observed in human CRC. Furthermore, non-angiogenic CRCs showed a worse response to chemotherapeutic drugs in vivo than in vitro. By suppressing endothelial apoptosis, metformin sensitized non-angiogenic CRCs to chemo-drugs via elevation of MVD and improvement of vascular maturity. Further results showed that endothelial apoptosis was induced by tumor cells via activation of caspase signaling, which was abrogated by metformin administration. These findings provide pre-clinical evidence for the involvement of endothelial apoptosis and subsequent vascular immaturity in the chemoresistance of non-angiogenic CRC. By suppressing endothelial apoptosis, metformin restores vascular maturity and function and sensitizes CRC to chemotherapeutic drugs via a vascular mechanism.

3D Raman mapping as an analytical tool for investigating the coatings of coated drug particles

Georgia Koutentaki, Pavel Krýsa, Dan Trunov, Tomáš Pekárek, Markéta Pišlová, Miroslav Šoóš

2023, 13(3): 276-286. doi: 10.1016/j.jpha.2023.02.004

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Abstract:
The properties of dry-coated paracetamol particles (fast-dissolving model drug) with carnauba wax particles as the coating agent (dissolution retardant) were investigated. Raman mapping technique was used to non-destructively examine the thickness and homogeneity of coated particles. The results showed that the wax existed in two forms on the surface of the paracetamol particles, forming a porous coating layer: i) whole wax particles on the surface of paracetamol and glued together with other wax surface particles, and ii) deformed wax particles spread on the surface. Regardless of the final particle size fraction (between 100 and 800 μm), the coating thickness had high variability, with average thickness of 5.9 ± 4.2 μm. The ability of carnauba wax to decrease the dissolution rate of paracetamol was confirmed by dissolution of powder and tablet formulations. The dissolution was slower for larger coated particles. Tableting further reduced the dissolution rate, clearly indicating the impact of subsequent formulation processes on the final quality of the product.

Drug adulteration analysis based on complexation with cyclodextrin and metal ions using ion mobility spectrometry

Zhigang Liang, Huanhuan Wang, Fangling Wu, Longfei Wang, Chenwei Li, Chuan-Fan Ding

2023, 13(3): 287-295. doi: 10.1016/j.jpha.2022.11.002

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Abstract:
Drug adulteration and contamination are serious threats to human health therefore, their accurate monitoring is very important. Allopurinol (Alp) and theophylline (Thp) are commonly used drugs for the treatment of gout and bronchitis, while their isomers hypoxanthine (Hyt) and theobromine (Thm) have no effect and affect the efficacy of the drug. In this work, the drug isomers of Alp/Hyt and Thp/Thm are simply mixed with α-, β-, γ-cyclodextrin (CD) and metal ions and separated using trapped ion mobility spectrometry-mass spectrometry (TIMS-MS). TIMS-MS results showed that Alp/Hyt and Thp/Thm isomers could interact with CD and metal ions and form corresponding binary or ternary complexes to achieve their TIMS separation. Different metal ions and CDs showed different separation effect for the isomers, among which Alp and Hyt could be successfully distinguished from the complexes of [Alp/Hyt+γ-CD + Cu–H]⁺ with separation resolution (R_P–P) of 1.51; whereas Thp and Thm could be baseline separated by [Thp/Thm+γ-CD + Ca–H]⁺ with R_P–P of 1.96. Besides, chemical calculations revealed that the complexes were in the inclusion forms, and microscopic interactions were somewhat different, making their mobility separation. Moreover, relative and absolute quantification was investigated with an internal standard to determine the precise isomers content, and good linearity (R² > 0.99) was obtained. Finally, the method was applied for the adulteration detection where different drugs and urine were analyzed. In addition, due to the advantages of fast speed, simple operation, high sensitivity, and no chromatographic separation required, the proposed method provides an effective strategy for the drug adulteration detection of isomers.

Rapid authentication of different herbal medicines by heating online extraction electrospray ionization mass spectrometry

Zidong Qiu, Chaofa Wei, Xiang Li, Changjiangsheng Lai, Zhilai Zhan, Yan Jin, Li Zhou, Qingxiu Hao, Jian Yang, Shuanglong Wang, Liping Kang, Luqi Huang

2023, 13(3): 296-304. doi: 10.1016/j.jpha.2023.01.002

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Abstract:
The rapid and accurate authentication of traditional Chinese medicines (TCMs) has always been a key scientific and technical problem in the field of pharmaceutical analysis. Herein, a novel heating online extraction electrospray ionization mass spectrometry (H-oEESI-MS) was developed for the rapid and direct analysis of extremely complex substances without the requirement for any sample pretreatment or pre-separation steps. The overall molecular profile and fragment structure features of various herbal medicines could be completely captured within 10–15 s, with minimal sample (<0.5 mg) and solvent consumption (<20 μL for one sample). Furthermore, a rapid differentiation and authentication strategy for TCMs based on H-oEESI-MS was proposed, including metabolic profile characterization, characteristic marker screening and identification, and multivariate statistical analysis model validation. In an analysis of 52 batches of seven types of Aconitum medicinal materials, 20 and 21 key compounds were screened out as the characteristic markers of raw and processed Aconitum herbal medicines, respectively, and the possible structures of all the characteristic markers were comprehensively identified based on Compound Discoverer databases. Finally, multivariate statistical analysis showed that all the different types of herbal medicines were well differentiated and identified (R²X > 0.87, R²Y > 0.91, and Q² > 0.72), which further verified the feasibility and reliability of this comprehensive strategy for the rapid authentication of different TCMs based on H-oEESI-MS. In summary, this rapid authentication strategy realized the ultra-high-throughput, low-cost, and standardized detection of various complex TCMs for the first time, thereby demonstrating wide applicability and value for the development of quality standards for TCMs.

Fragmentation stability and retention time-shift obtained by LC-MS/MS to distinguish sialylated N-glycan linkage isomers in therapeutic glycoproteins

Chi Soo Park, Minju Kang, Ahyeon Kim, Chulmin Moon, Mirae Kim, Jieun Kim, Subin Yang, Leeseul Jang, Ji Yeon Jang, Ha Hyung Kim

2023, 13(3): 305-314. doi: 10.1016/j.jpha.2023.01.001

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Abstract:
Sialylated N-glycan isomers with α2-3 or α2-6 linkage(s) have distinctive roles in glycoproteins, but are difficult to distinguish. Wild-type (WT) and glycoengineered (mutant) therapeutic glycoproteins, cytotoxic T lymphocyte-associated antigen-4-immunoglobulin (CTLA4-Ig), were produced in Chinese hamster ovary cell lines; however, their linkage isomers have not been reported. In this study, N-glycans of CTLA4-Igs were released, labeled with procainamide, and analyzed by liquid chromatography-tandem mass spectrometry (MS/MS) to identify and quantify sialylated N-glycan linkage isomers. The linkage isomers were distinguished by comparison of 1) intensity of the N-acetylglucosamine ion to the sialic acid ion (Ln/Nn) using different fragmentation stability in MS/MS spectra and 2) retention time-shift for a selective m/z value in the extracted ion chromatogram. Each isomer was distinctively identified, and each quantity (>0.1%) was obtained relative to the total N-glycans (100%) for all observed ionization states. Twenty sialylated N-glycan isomers with only α2-3 linkage(s) in WT were identified, and each isomer's sum of quantities was 50.4%. Furthermore, 39 sialylated N-glycan isomers (58.8%) in mono- (3 N-glycans; 0.9%), bi- (18; 48.3%), tri- (14; 8.9%), and tetra- (4; 0.7%) antennary structures of mutant were obtained, which comprised mono- (15 N-glycans; 25.4%), di- (15; 28.4%), tri- (8; 4.8%), and tetra- (1; 0.2%) sialylation, respectively, with only α2-3 (10 N-glycans; 4.8%), both α2-3 and α2-6 (14; 18.4%), and only α2-6 (15; 35.6%) linkage(s). These results are consistent with those for α2-3 neuraminidase-treated N-glycans. This study generated a novel plot of Ln/Nn versus retention time to distinguish sialylated N-glycan linkage isomers in glycoprotein.

Ultrasensitive quantification of trace amines based on N-phosphorylation labeling chip 2D LC-QQQ/MS

Xiqing Bian, Yida Zhang, Na Li, Menglin Shi, Xiaolin Chen, Hui-Lu Zhang, Jie Liu, Jian-Lin Wu

2023, 13(3): 315-322. doi: 10.1016/j.jpha.2023.02.003

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Abstract:
Trace amines (TAs) are metabolically related to catecholamine and associated with cancer and neurological disorders. Comprehensive measurement of TAs is essential for understanding pathological processes and providing proper drug intervention. However, the trace amounts and chemical instability of TAs challenge quantification. Here, diisopropyl phosphite coupled with chip two-dimensional (2D) liquid chromatography tandem triple-quadrupole mass spectrometry (LC-QQQ/MS) was developed to simultaneously determine TAs and associated metabolites. The results showed that the sensitivities of TAs increased up to 5520 times compared with those using nonderivatized LC-QQQ/MS. This sensitive method was utilized to investigate their alterations in hepatoma cells after treatment with sorafenib. The significantly altered TAs and associated metabolites suggested that phenylalanine and tyrosine metabolic pathways were related to sorafenib treatment in Hep3B cells. This sensitive method has great potential to elucidate the mechanism and diagnose diseases considering that an increasing number of physiological functions of TAs have been discovered in recent decades.

2023 Vol. 13, No. 3