Leech is a commonly traditional Chinese medicine with antithrombotic effects. The activity of antithrombotic active ingredients in leech is an important parameter of the quality. Herein, a novel point-of-care-testing method based on antithrombin activity for detecting antithrombotic active ingredients was developed via personal glucose monitor. Fe3O4-PEI was synthesized via chemical coprecipitation, followed by the sequential conjugation of thrombin, thrombin aptamer, streptavidin, and invertase. Invertase catalyzed sucrose into glucose, which can be quantitatively detected using a personal glucose meter. Hirudin discovered in leeches, is the most effective thrombin inhibitor in the clinical treatment of thrombotic diseases and was used as a model drug in this work. In the presence of hirudin, it inhibited the activity of thrombin, resulting in a signal reduction in glucose. This point-of-care-testing method for hirudin activity detection exhibited a good linear range from 0.05~0.5 U mL−1 with a detection limit of 0.048 U mL−1, providing a simple and sensitive method for the activity evaluation of antithrombotic ingredients of Chinese medicine.
- Open Access
- Article
Point-of-Care-Testing Method for the Detection of Antithrombotic Active Ingredients in Leech by Personal Glucose Monitor
- Ruoyu Ba,
- Xiangyu Sun,
- Fen Wan,
- Li Zhang *,
- Fangfang Cheng *
Author Information
Received: 11 Apr 2026 | Revised: 08 Jun 2026 | Accepted: 15 Jun 2026 | Published: 07 Jul 2026
Abstract
Keywords
leech | Hirudin | Fe3O4 | Point-of-Care-Testing | PGM
References
- 1.
Wei, Y.; Yuan, X.; Zhang, X.; et al. Recent advances of organic small molecule-based fluorescent probes in bioimaging and photo-mediated therapy for thrombotic diseases. Coord. Chem. Rev. 2026, 550, 217352.
- 2.
Yi, X.; Wu, X.; Zang, E.; et al. Construction of safer hirudin-derived peptides with enhanced anticoagulant properties based on C-terminal active residue adaptation and modification. J. Adv. Res. 2026, 80, 1083–1098.
- 3.
Johnson, P.H. Hirudin: Clinical potential of a thrombin inhibitor. Annu. Rev. Med. 1994, 45, 165–177.
- 4.
Chen, J.; Xie, X.; Zhang, H.; et al. Pharmacological Activities and Mechanisms of Hirudin and Its Derivatives-A Review. Front. Pharmacol. 2021, 12, 660757.
- 5.
Markwardt, F. Hirudin as alternative anticoagulant—A historical review. Semin. Thromb. Hemost. 2002, 28, 405–414.
- 6.
De Vincenzo, F.; Acquasaliente, L.; Pontarollo, G.; et al. Noncoded aminoacids in protein engineering: Structure–activity relationship studies of hirudin–thrombin interaction. Biotechnol. Appl. Biochem. 2018, 65, 69–80.
- 7.
Troisi, R.; Balasco, N.; Autiero, I.; et al. Exosite binding in thrombin: A global structural/dynamic overview of complexes with aptamers and other ligands. Int. J. Mol. Sci. 2021, 22, 10803.
- 8.
Li, B.; Ou, Y.; Wu, Y.; et al. Progress in analytical methods of hirudin. Chin. J. Pharm. Anal. 2010, 30, 755–760.
- 9.
Bikker, R.; Meyer, K.; Domberg, P.; et al. Development and evaluation of point-of-care testing recertification with e-learning. Scand. J. Clin. Lab. Investig. 2020, 80, 133–138.
- 10.
Haggerty, L.; Tran, D. Cholesterol Point-of-Care Testing for community pharmacies: A review of the current literature. J. Pharm. Pract. 2017, 30, 451–458.
- 11.
Fu, D.L.; Zhang, B.; Zhang, S.; et al. An electrochemical point-of-care testing device for specific diagnosis of the albinism biomarker based on paradigm shift designs. Biosens. Bioelectron. 2024, 264, 116645.
- 12.
Wang, Y.; Wang, M.L.; Cui, J.R.; et al. Portable devices and machine learning-assisted lateral flow assay for food safety analysis: Developments and perspectives. Trends Food Sci. Tech. 2025, 163, 105180.
- 13.
Yang, D.; Geng, S.; Jing, R.; et al. Recent developments in personal glucose meters as point-of-care testing devices (2020–2024). Biosensors 2024, 14, 419.
- 14.
Huang, Q.; Chen, J.; Zhao, Y.; et al. Advancements in electrochemical glucose sensors. Talanta 2025, 281, 126897.
- 15.
Li, L.; Wang, T.; Zhong, Y.; et al. A review of nanomaterials for biosensing applications. J. Mater. Chem. B 2024, 12, 1168–1193.
- 16.
Girardet, T.; Bianchi, E.; Henrionnet, C.; et al. SPIONs magnetic nanoparticles for MRI applications: Microwave synthesis and physicochemical, magnetic and biological characterizations. Mater. Today Commun. 2023, 36, 106819.
- 17.
Ramimoghadam, D.; Bagheri, S.; Abd Hamid, S.B. Stable monodisperse nanomagnetic colloidal suspensions: An overview. Colloids Surf. B 2015, 133, 388–411.
- 18.
Zhang, Z.; Lv, M.; Hu, W.; et al. Peptide based covalent organic framework for the magnetic resonance imaging-mediated attenuation of amyloid-β fibrils. Adv. Funct. Mater. 2026, e29220.
- 19.
Wang, J.; Feng, K.; Yu, P.; et al. Iron-based nanozymes: From mechanisms to functional applications. Chem. Eng. J. 2026, 539, 177195.
- 20.
Zhang, R.; Li, Y.; Yan, H.; et al. Breaking the trade-off in MPI-guided magnetic hyperthermia by tailoring the dynamic magnetization of magnetic nanoparticles via site-selective trace doping. J. Am. Chem. Soc. 2026, 148, 18482–18496.
- 21.
Zhang, D.; Zhu, O.; Mou, F.; et al. Systemic-to-local nanorobot thrombolysis. Sci. Adv. 2026, 12, eaed9464.
- 22.
Chen, Z.; Lal, K.; Wang, A.; et al. Plasmonic magnetic nanoparticle-based photothermal sensor for sensitive detection of chlorantraniliprole. Food Chem. 2026, 508, 148487.
- 23.
Wang, T.; Hu, X.; Yang, Y.; et al. New insight into assembled Fe3O4@PEI@Ag structure as acceptable agent with enzymatic and photothermal properties. Int. J. Mol. Sci. 2022, 23, 10743.
- 24.
Zhang, R.; Yan, C.; Zong, Z.; et al. Taking glucose as intermediate bridge-signal-molecule for on-site and convenient detection of ochratoxin A in rice with portable glucose meter. Food Chem. 2023, 400, 134007.
- 25.
Bai, Y.; Li, Y.; Zhang, D.; et al. Enhancing the affinity of anti-human α-thrombin 15-mer DNA aptamer and anti-immunoglobulin E aptamer by polyT extension. Anal. Chem. 2017, 89, 9467–9473.
- 26.
Cao, X.; Luo, Y.; Liu, X.; et al. Aptamer-thrombin loaded magnetic microspheres for bio-specific extraction and precise detection of hirudin. Talanta 2024, 267, 125244.
- 27.
Ban, E.; Nam, H.S.; Yoo, Y.S. Competitive immunoassay for recombinant hirudin using capillary electrophoresis with laser-induced fluorescence detection. J. Chromatogr. A 2001, 924, 337–344.
- 28.
Jiang, S.; Jiao, J.; Zhang, T.; et al. Pharmacokinetics study of recombinant hirudin in the plasma of rats using chromogenic substrate, ELISA, and radioisotope assays. PLoS ONE 2013, 8, e64336.
- 29.
Dong, X.; Meng, Z.; Jin, J.; et al. Development, validation, and clinical pharmacokinetic application of ultra-performance liquid chromatography/tandem mass spectrometry method for simultaneously determining a novel recombinant hirudin derivative (Neorudin) and its active metabolite in human serum. J. Chromatogr. B 2017, 1063, 204–213.
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