MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Danlin Zhang,Shanshan Zhong,Chunlan Peng,Liuyu Chen,Jun Luo,Xinchun Li
Digoxin is a crucial cardiotonic agent, which has been widely used for remedy of heart failure for decades in clinical practice. Unfortunately, digoxin behaves extremely narrow therapeutic index and huge individual difference, therefore alerting its concentration level is imperative regarding therapeutic efficacy and safety. In this work, we initially present a label-free ultrasensitive electrochemical sensor for digoxin by dual signal amplification strategy integrating catalytic hairpin assembly (CHA) and electrochemically mediated atom transfer radical polymerization (eATRP). A DNA hairpin structure (H1) bearing azide group was firstly immobilized on gold electrode via Au-S bond. Then, an aptamer-containing initiator triggered CHA reaction with the assistance of hairpin-structured helper (H2), thereby exposing the azide group to enable the introduction of functional element for eATRP by click chemistry. The eATRP was electrochemically initiated, producing abundant electroactive species tethering to the formed H1-H2 hybrid structure. Through DNA-mediating electron transfer mechanism, a strong electrochemical current signal can be generated. The peak current measured by square wave voltammetry (SWV) was proportional to digoxin concentration ranging from 1.0 pM to 40 pM, with the detection limit of 0.59 pM, outperforming currently widely used ELISA, FPIA and LC-MS/MS. The designed electrochemical sensor was used for detection of clinical blood samples and the analytical results were in line with traditional immunoassay. By taking the advantages including low-cost, high sensitivity, and reliable assays, the present sensor offers a means for digoxin therapeutic monitoring. More importantly, this work would enrich our design strategy of electrochemical sensors toward clinical biopharmaceutics.
研究领域:
生物医学工程
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临床药理学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2022
·
IF 1.193
Xiaoqian JIANG,Xinxin XU,Shanshan SONG,Hua KUANG,Liqiang LIU,Liguang XU,Chuanlai XU
Digitoxin is a cardiac glycoside used in the therapy of heart failure and arrhythmias, and has potential as an anti-cancer and anti-influenza treatment. However, digitoxin has the disadvantage of a narrow therapeutic window meaning that it is easy to cause toxicity. Consequently, there is an urgent need to accurately monitor the plasma concentration of patients. In this study, an anti-digitoxin monoclonal antibody (mAb) 5D2 was developed and purified, and found to have outstanding specificity, sensitivity, and affinity. The half inhibitory concentrations (IC 50 ) and the limit of detection (LOD) values of mAb 5D2 in human plasma determined by ic-ELISA were 0.205 ng/mL and 0.010 ng/mL, while the cross-reactivity with other analogs was negligible. Based on mAb 5D2, a colloidal gold-based immunochromatographic assay was developed for the purpose of quickly monitoring the concentration of digitoxin in human plasma. The detection results could be obtained within 10 min, and were in agreement with those of liquid chromatography-tandem mass spectrometry (LC-MS/MS). The strip could be evaluated semi-quantitatively by the naked eye, with a visual LOD (vLOD) of 0.5 ng/mL and a cut-off value of 10 ng/mL. Meanwhile, the linear range was between 0.319 ng/mL and 2.406 ng/mL, which was obtained quantitatively by the strip reader. Therefore, the immunochromatographic strip could offer a new tool for rapid, accurate, and portable detection of digitoxin.
研究领域:
医学
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药理学
·
免疫学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Jia-Yang He,Xin Shang,Chun-Li Yang,Si-Yu Zuo,Ruo Yuan,Wen-Ju Xu
Exploring the ratiometric fluorescence biosensing of DNA-templated biemissive silver nanoclusters (AgNCs) is significant in bioanalysis, yet the design of a stimuli-responsive DNA device is a challenge. Herein, using the anti-digoxin antibody (anti-Dig) with two identical binding sites as a model, a tweezer-like DNA architecture is assembled to populate fluorescent green- and red-AgNCs (g-AgNCs and r-AgNCs), aiming to produce a ratio signal via specific recognition of anti-Dig with two haptens (DigH). To this end, four DNA probes are programmed, including a reporter strand (RS) dually ended with a g-/r-AgNC template sequence, an enhancer strand (ES) tethering two same G-rich tails (G18), a capture strand (CS) labeled with DigH at two ends, and a help strand (HS). Initially, both g-AgNCs and r-AgNCs wrapped in the intact RS are nonfluorescent, whereas the base pairing between RS, ES, CS, and HS resulted in the construction of DNA mechanical tweezers with two symmetric arms hinged by a rigid “fulcrum”, in which g-AgNCs are lighted up due to G18 proximity (“green-on”), and r-AgNCs away from G18 are still dark (“red-off”). When two DigHs in proximity recognize and bind anti-Dig, the conformation switch of these tweezers resultantly occurs, taking g-AgNCs away from G18 for “green-off” and bringing r-AgNCs close to G18 for “red-on”. As such, the ratiometric fluorescence of r-AgNCs versus g-AgNCs is generated in response to anti-Dig, achieving reliable quantization with a limit of detection at the picomolar level. Based on the fast stimulated switch of unique DNA tweezers, our ratiometric strategy of dual-emitting AgNCs would provide a new avenue for a variety of bioassays.
研究领域:
生物分析
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纳米生物学
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分子诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Run-qin Wang,Jia Chen,Bei-bei Guo,Li Yang,Shuang Hu
A dispersive liquid–liquid microextraction utilizing alkanolamine-based deep eutectic solvent was firstly proposed and introduced to preconcentrate trace level of digoxin in human plasma prior to high performance liquid chromatography analysis with deslanoside as the internal standard. In the procedure, a deep eutectic solvent was prepared with N,N-dimethylethanolamine as a hydrogen bond acceptor and n -heptanol as a hydrogen bond donor. Chemical structures of the prepared deep eutectic solvent were characterized by infrared spectroscopy and proton nuclear magnetic resonance spectroscopy. The experimental parameters affecting the extraction performance, including deep eutectic solvent composition, volume of extraction solvent, pH of aqueous sample solution, salt concentration and extraction time, were investigated and optimized. Under the optimum conditions, the method showed good linearity in the range of 0.32–80 ng/mL, precision (relative standard deviations 1.0 %–15 %), accuracy (relative errors −6.4 %–8.5 %) and acceptable extraction recoveries (80.3 %–81.4 %). In addition, satisfactory lower limit of quantification (0.32 ng/mL) and stability were achieved, while matrix effect was not observed in human plasma for the proposed method. It showed that the developed method was efficient, fast and eco-friendly, which provides a potential alternative for the preconcentration and determination of digoxin in complex biological matrices.
研究领域:
分析化学
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药物分析
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环境科学
✓ 已验证
产品关联
DOI
高影响因子
ACS Omega
·
2020
·
IF 2.87
Fanqian Ling,Liqiang Liu,Hua Kuang,Gang Cui,Chuanlai Xu
Mouse–mouse hybridoma cell lines producing stable, highly specific monoclonal antibodies with good affinity for the cardiac glycoside digoxin (DIG) were established to construct an indirect enzyme-linked immunosorbent assay and lateral-flow immunochromatographic strip to detect DIG in human blood. The hapten DIG was coupled to bovine serum albumin or chicken ovalbumin by sodium periodate oxidation. The highest sensitivity and specificity antibody had a median inhibitory concentration (IC50) of 0.45 ng/mL, a linear range of detection of 0.293–0.7 ng/mL, and low cross-reactivity with several DIG analogues. The cut-off value of the lateral-flow immunochromatographic strip was 5 ng/mL when the strip was tested with human blood. The immunochromatographic lateral flow strip test provides a quick and convenient method for determining DIG in plasma which can be visually observed in only 5 min to promote rational drug use.
研究领域:
免疫学
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药物分析
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Huan Yao,Ting Huang,Zhifang Li,Huaxin Wu,Xueyao Li,Hua Tang,Fuhai Su,Xinliang Jin,Wei Zhang
Purity assessment is critical for the quality control of active pharmaceutical ingredients (APIs). However, there is a lack of rapid and accurate purity assessment methods that operate independently of target-specific certified reference materials (CRMs). Herein, we proposed a traceable quantification method, named SFC-ISC-qNMR, for purity assessment and quantitative determination of low-purity APIs, which was accomplished by tracing the test solution to the reference solution via supercritical fluid chromatography (SFC) collection with internal standard-correction (ISC), followed by an SI-traceable quantitative nuclear magnetic resonance (qNMR) analysis of the reference solution (SI = International System of Units). Digitoxin, a widely used cardiac glycoside in the treatment of heart failure and arrhythmias, was used as a model compound. By employing a mobile phase of supercritical CO2 modified with 24%–26% methanol, digitoxin with a purity exceeding 99.87% was isolated and collected. A national CRM of ethyl para-hydroxybenzoate was used as the IS for digitoxin. The SFC-ISC reference solution can be prepared in 15 min without the need for precise weighing, simply by reconstitution of the SFC collected high-purity analyte in deuterated methanol containing IS. The feasibility and reliability of the SFC-ISC-qNMR method were exhibited through the quantitative determination of digitoxin in chemical mixtures. The purity of a commercially available digitoxin reagent was determined to be 94.17% ± 0.15%, which was consistent with high-resolution qNMR analysis. The contents of spiked digitoxin in a digoxin analytical standard were measured at 1.06%, 1.37%, 2.39%, and 4.43%, respectively, showing high consistency with the gravimetric value with a bias less than 0.13%. In contrast, direct qNMR with deconvolution and HPLC analysis failed to provide accurate measurements. These results highlight the applicability of the SFC-ISC-qNMR method for drug evaluation and research, demonstrating its significant potential for the purity assessment of APIs.
研究领域:
分析化学
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药物分析
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色谱分析
·
核磁共振（NMR）
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Bioanalysis
·
2020
·Vol. 12
·期 9
·
IF 2.371
Peng Guo,Qiucheng Liang,Juntao Zheng,Jinmin Chen,Yanping Guan,Liang Ding,Fulin Jiang,Xuewen Chen,Min Huang,Ailan Chen,Guoping Zhong
Aim: Heart failure patients are frequently given comedication of digoxin and diuretics like spironolactone and tolvaptan. A UHPLC–MS/MS assay for determining canrenone (main active metabolite of spironolactone), digoxin and tolvaptan simultaneously should be developed so as to support related drug–drug interaction studies. Results: A UHPLC–MS/MS method for simultaneous determination of these three drugs in human plasma was established and fully verified as per CFDA guidelines. Chromatographic separation was achieved using a 4-min isocratic elution. Mass analyses were performed under positive electrospray ionization mode. The calibration curves were established over 1.0–400.0 ng/ml for canrenone and tolvaptan while over 0.1–40.0 ng/ml for digoxin. Conclusion: The developed method was feasible in detecting concentration and related drug–drug interaction studies.
研究领域:
药理学
·
分析化学
·
心脏病学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2017
·
IF 6.32
Shanglin Li,Yin Gu,Yi Lyu,Yan Jiang,Peng Liu
An integrated graphene oxide purification-lateral flow test strip (iGOP-LFTS) was developed for on-strip purifying and visually detecting polymerase chain reaction (PCR) products with an improved sensitivity as well as a more stringent specificity. PCR products amplified with a pair of biotin- and digoxin-labeled primers were directly pipetted onto GO pads, on which graphene oxide selectively adsorbed residual primers and primer-dimers with the aid of a running buffer containing MgCl2 and Tween 20. By stacking up three GO pads to increase the surface area for adsorption, 83.4% of double-stranded DNA with a length of 30 bp and 98.6% of 20-nt primers could be removed from a 10-μL DNA mixture. Since no primers interfered with detection, the increase of the sample loading volume from 5 to 20 μL could improve the signal-to-noise ratio of the test line 1.6 fold using the iGOP-LFTS while no changes were observed using the conventional LFTS. The limit of detection of the iGOP-LFTS was determined to be 30 copies of bacteriophage λ-DNA with naked eyes and this limit could be further decreased to 3 copies by loading 20 μL of the sample, which corresponded to a 1000-fold improvement compared to that of the LFTS detected by naked eyes. When the ImageJ analysis was employed, a 100-fold decrease of the detection limit can be obtained. In addition, due to the removal of the primer-dimers, the dim test line observed in the negative control of the LFTS was eliminated using the iGOP-LFTS. A mock clinical specimen spiked with defective HIV-1 (human immunodeficiency virus) viruses was successfully analyzed using a two-step reverse transcription-PCR with 30 amplification cycles followed by the iGOP-LFTS detection. These significant improvements were achieved without introducing any additional hands-on operations and instrumentations.
研究领域:
分子生物学
·
分析化学
·
生物医学工程
·
病毒学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Gastroenterology Report
·
2020
·Vol. 8
·期 6
·465-475
·
IF 2.96
Hou Yong-Qiang,Wang Ying-Ying,Wang Xing-Can,Liu Yao,Zhang Chun-Ze,Chen Zhe-Sheng,Zhang Zhe,Wang Wei,Kong De-Xin
BackgroundColorectal cancer (CRC) is one of the leading causes of cancer death worldwide. Novel drugs for CRC therapy are urgently needed. Digoxin has been in clinical use for treatment of heart failure and atrial arrhythmias for many years. Fragmentary reports suggested that digoxin might have antitumor efficacy on CRC. Here, we aimed to investigate the antitumor effect of digoxin on human CRC cells and the underlying mechanism.MethodsCell viability was determined using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and plate colony formation assay. The effects of digoxin on cell-cycle distribution and apoptosis were analysed by flow cytometry. The anti-metastatic effect on tumor cells was determined by wound-healing assay and transwell assay. Anti-angiogenic effect was examined by determining the inhibition against proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs). Mechanism study was performed by Western blot, enzyme-linked immunosorbent assay (ELISA), and gelatin-zymography assay.ResultsDigoxin potently inhibited cell proliferation, induced G1-phase and G2/M-phase arrest in colorectal-cancer HCT8 and SW620 cells, respectively. No obvious apoptosis was observed in the treated cells. Anti-metastatic activities were shown on HCT8 cells by inhibiting the migration and invasion. Meanwhile, the expression of MMP2, MMP9, and phosphorylated Integrinβ1 were decreased. Digoxin inhibited the proliferation, migration, and tube formation of HUVECs and reduced HIF1α expression and vascular endothelial growth factor A (VEGF-A) secretion in HCT8 cells, suggesting anti-angiogenic activity. Furthermore, digoxin significantly reversed ABCB1-mediated multidrug resistance on SW620/Ad300 cells.ConclusionOur findings suggest that digoxin has the potential to be applied as an antitumor drug via inhibiting proliferation and metastasis as well as reversing the ABCB1-mediated multidrug resistance of colorectal cancer.
研究领域:
肿瘤学
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药理学
·
细胞生物学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
BIOSCIENCE REPORTS
·
2021
·Vol. 41
·期 10
·
IF 3.84
Wang Yingying,Hou Yongqiang,Hou Lanjiao,Wang Wei,Li Ke,Zhang Zhe,Du Bo,Kong Dexin
Lung cancer remains the leading cause of cancer mortality because of its metastatic potential and high malignancy. The discovery of new applications for old drugs is a shortcut for cancer therapy. We recently investigated the antitumor effect of digoxin, a well-established drug for treating heart failure, against nonsmall cell lung cancer A549 and H1299 cells. Digoxin inhibited the proliferation and colony-forming ability of the two cell lines and arrested the cell cycle at the G0/G1 phase in A549 cells and the G2/M phase in H1299 cells. Mitochondria-mediated apoptosis was induced in A549 cells but not in H1299 cells after treatment with digoxin. Moreover, digoxin inhibited the migration, invasion, adhesion and epithelial–mesenchymal transition of A549 and H1299 cells. Autophagy was induced in both cell lines after treatment with digoxin, with an increase in autophagosome foci. In addition, digoxin inhibited the phosphorylation of Akt, mTOR and p70S6K, signaling molecules of the PI3K/Akt pathway that are known to be involved in tumor cell survival, proliferation, metastasis and autophagy. Our findings suggest that digoxin has the potential to be used for therapy for human nonsmall cell lung cancer, but further evidence is required.
研究领域:
肿瘤学
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药理学
·
细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
WORLD JOURNAL OF GASTROENTEROLOGY
·
2012
·
IF 2.471
Da-Peng Chen,Yong-Jian Xiong,Ze-Yao Tang,Qi-Ying Yao,Dong-Mei Ye,Sha-Sha Liu,Yuan Lin
AIM: To characterize the dual effects of deslanoside on the contractility of jejunal smooth muscle. METHODS: Eight pairs of different low and high contractile states of isolated jejunal smooth muscle fragment (JSMF) were established. Contractile amplitude of JSMF in different low and high contractile states was selected to determine the effects of deslanoside, and Western blotting analysis was performed to measure the effects of deslanoside on myosin phosphorylation of jejunal smooth muscle. RESULTS: Stimulatory effects on the contractility of JSMF were induced (45.3% ± 4.0% vs 87.0% ± 7.8%, P < 0.01) by deslanoside in 8 low contractile states, and inhibitory effects were induced (180.6% ± 17.8% vs 109.9% ± 10.8%, P < 0.01) on the contractility of JSMF in 8 high contractile states. The effect of deslanoside on the phosphorylation of myosin light chain of JSMF in low (78.1% ± 4.1% vs 96.0% ± 8.1%, P < 0.01) and high contractile state (139.2% ± 8.5% vs 105.5 ± 7.34, P < 0.01) was also bidirectional. Bidirectional regulation (BR) was abolished in the presence of tetrodotoxin. Deslanoside did not affect jejunal contractility pretreated with the Ca 2+ channel blocker verapamil or in a Ca 2+ -free assay condition. The stimulatory effect of deslanoside on JSMF in a low contractile state (low Ca 2+ induced) was abolished by atropine. The inhibitory effect of deslanoside on jejunal contractility in a high contractile state (high Ca 2+ induced) was blocked by phentolamine, propranolol and L-NG-nitro-arginine, respectively. CONCLUSION: Deslanoside-induced BR is Ca 2+ dependent and is related to cholinergic and adrenergic systems when JSMF is in low or high contractile states.
研究领域:
药理学
·
胃肠生理学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2024
·
IF 3.8
Yan Qiaofang,Huang Fei,Wang Chuyang,Zhan Min,Zhang Qiaoxuan,Huang Di,Yan Jun,Lin Haibiao,Huang Xianzhang,Han Liqiao
Digoxin, a cardiac glycoside, is widely used in the treatment of cardiovascular diseases. Due to its narrow therapeutic range, precise monitoring of its blood concentration is essential. A reference measurement procedure (RMP) is pivotal for ensuring result accuracy and comparability. The RMP for serum digoxin by ID-LC-MS/MS was optimized with sample pre-treatment and detection processes, and the bracketing calibration method was used, which facilitates more accurate measurement, especially for extreme concentrations. The performance of this optimized RMP was thoroughly evaluated. The limit of detection (LoD) was 0.05 ng/mL (0.06 nmol/L) and the lowest limit of quantification (LLoQ) was 0.10 ng/mL (0.13 nmol/L). The intra- and inter-assay imprecisions were 2.24%, 2.51%, 1.40% and 1.72%, 1.65%, 0.97% at 0.5, 2.0, 5.0 ng/mL, respectively. Recoveries were 99.63 to 101.42% and the linear response ranged from 0.1 to 10.0 ng/mL. The relative bias was 0.41% and 2.00% of our results compared with the median of all participating reference laboratories for IFCC-RELA (External Quality Assessment Scheme for Reference Laboratories in Laboratory Medicine) 2023A and 2023B. The uncertainty, calibration and measurement capability (CMC) of this method were also evaluated. The optimized RMP was applied in the Trueness Verification Plan of Southern China, which indicates significant differences among clinical systems, highlighting the need for standardization efforts. In addition, two commonly used clinical systems which employed immunoassay methods were compared with this optimized RMP, and 26 individual serum samples were analyzed. The good correlations indicate the feasibility of standardization for serum digoxin. The optimized RMP serves as an accurate reference baseline for routine methods, aiming to enhance the accuracy and precision of measurements in clinical laboratories. Graphical Abstract
研究领域:
临床化学
·
药理学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）