Sensitive Method Research Articles

Giant Cell Arteritis: Can Simple Ultrasound Examination Prevent Complex Consequences?

Giant cell arteritis (GCA) is a rare disease of the arteries, occurring mainly in the elderly. Although the involvement of temporal arteries can be mostly symptomatic, the occlusion of ophthalmic arteries has severe consequences. GCA affecting temporal arteries is an emergency requiring quick commencement of treatment with glucocorticoids due to the serious consequences of neglect—blindness. According to the new guidelines released by EULAR, ultrasound is the tool of choice in support of the clinical diagnosis of giant cell arteritis, replacing temporal artery biopsy (TAB), as it is a sensitive and non-invasive method that is widely available. The main limitation is that the reliability of this imaging is based on the technical expertise of ultrasonographers. However, performing imaging should not delay commencing the treatment. In this work, we present ultrasound images from a case report of a 74-year-old female patient where difficulties in establishing a diagnosis led to vision loss in both eyes. In this example, we describe the ultrasound findings in giant cell arteritis, emphasizing its usefulness in supporting a diagnosis of GCA.

A Novel Ferrocene-Linked Thionine as a Dual Redox Mediator for the Electrochemical Detection of Dopamine and Hydrogen Peroxide

We introduce a novel dual redox mediator synthesized by covalently linking ferrocene dicarboxylic acid (FcDA) and thionine (TH) onto a pre-treated glassy carbon electrode. This unique structure significantly enhances the electro-oxidation of dopamine (DA) and the reduction of hydrogen peroxide (H2O2), offering a sensitive detection method for both analytes. The electrode exhibits exceptional sensitivity, selectivity, and stability, demonstrating potential for practical applications in biosensing. It facilitates rapid electron transfer between the analyte and the electrode surface, detecting H2O2 concentrations ranging from 1.5 to 60 µM with a limit of detection (LoD) of 0.49 µM and DA concentrations from 0.3 to 230 µM with an LoD of 0.07 µM. The electrode’s performance was validated through real-sample analyses, yielding satisfactory results.

Graphene-Based Virus Enrichment Protocol Increases the Detection Sensitivity of Human Norovirus in Strawberry and Oyster Samples

Human noroviruses (HuNoVs), the most prevalent viral contaminant in food, account for a substantial proportion of nonbacterial gastroenteritis cases. Extensive work has been focused on the diagnosis of HuNoVs in clinical samples, whereas the availability of sensitive detection methods for their detection in food is lacking. Here, we developed a virus enrichment approach utilizing graphene-based nanocomposites (CTAB-rGO-Fe3O4) that does not rely on large instruments and is suitable for on-site food pretreatment. The recovery efficiency of the developed virus enrichment procedure for serially diluted GII.4 norovirus ranged from 10.06 to 72.67% in strawberries and from 2.66 to 79.65% in oysters. Furthermore, we developed a real-time recombinase polymerase amplification (real-time RPA) assay, which can detect as low as 1.22 genome copies µL−1 of recombinant plasmid standard and has no cross-reactivity with genomes of astrovirus, rotavirus, adenovirus, and MS2 bacteriophage. Notably, the combined virus enrichment and real-time RPA detection assay enhanced the detection limits to 2.84 and 37.5 genome copies g−1 in strawberries and oysters, respectively, compared to those of qPCR. Our strategy, the graphene-based virus enrichment method combined with real-time RPA, presents a promising tool for sensitively detecting HuNoVs in food samples.

A novel cystatin in Psoroptes ovis var. cuniculi: molecular characterization, serodiagnostic potential, and its anti-inflammatory property on rabbit peripheral blood mononuclear cells

BackgroundThe ectoparasite Psoroptes ovis var. cuniculi causes substantial economic losses to the global rabbit industry. Currently, microscopy for identifying Psoroptes mite in skin scrapings, as the “diagnosis gold standard,” remains a challenge owing to its poor sensitivity in detecting low-level and/or early stage mite infestations. Additionally, Psoroptes infestations rapidly trigger cutaneous inflammation, thus the mites might produce some molecules to deal with the harmful effects of inflammation for their long-time survival on the host skin, but these molecules are still mostly unknown.MethodsTo seek a sensitive diagnostic method and illuminate the new antiinflammatory molecules, we characterized a novel cystatin of P. ovis var. cuniculi (PsoCys) using bioinformatics and molecular biology methods.ResultsThe results showed that PsoCys comprised the classical features of the type II cystatin superfamily including an N-terminal glycine residue, a central QXVXG motif, and a C-terminal LW motif. In mixed stages of mites, the transcription level of PsoCys was significantly higher in “fed” mites than in “starved” mites (P < 0.001), and among the different life-cycle stages of “fed” mites, the expression of PsoCys was higher in adult males than in larva, nymph, and adult females (P < 0.001). The established indirect ELISA based on recombinant PsoCys (rPsoCys-iELISA) presented 95.4% sensitivity and 95.7% specificity. The area under the receiver operating characteristic curve (AUC) for this method was 0.991, indicating its excellent diagnostic performance. Moreover, rPsoCys-iELISA had advantages over microscopy for detecting low-level and/or early stage mite infestations (90% versus 40% in artificial infestation cases at 3 weeks post-infestation; 61.9% versus 22.6% in clinical cases). In addition, rPsoCys could inhibit the activity of papain and cathepsin B in vitro, and significantly suppressed mRNA levels of toll-like receptors (TLR 1, 2, 4, and 6) and downstream molecules (NF-κB, p38, MyD88, IL-10, and IFN-γ) in LPS-stimulated rabbit PBMCs, indicating its anti-inflammatory property.ConclusionsOur findings indicated that PsoCys was a novel type II cystatin of Psoroptes mites, and it served as a potential serological diagnostic antigen for detecting low-level and/or early stage mite infestations, as well as a novel anti-inflammatory molecule of Psoroptes mites.Graphical abstract

Adapting lipidomic sample processing methods for boars housed in commercial settings

Abstract Multiple reaction monitoring (MRM) profiling is a sensitive method of lipid screening that has the capability to distinguish between different fertility phenotypes in gilts. However, MRM profiling has not yet been utilized to evaluate fertility phenotypes in boars. Markers indicative of fertility status in boars would be valuable as inclusion of subfertile boars in breeding programs results in a loss of efficiency and negative economic consequences. In addition, semen samples for lipidomic analysis are transported in liquid nitrogen or on dry ice to suspend metabolic activity within the sperm cells, however, these cryopreservation techniques are not commonly available at commercial boar studs. Therefore, the objective of this study was to develop a method of sample processing for MRM profiling that suspends metabolic activity within semen without freezing the sample. Five, sexually mature boars of similar genetics enrolled in a commercial breeding program were collected for the study. Following collection, ejaculates were aliquoted into methanol to suspend metabolic activity and shipped to Purdue University overnight for lipid extraction. Lipids were extracted using the Bligh &amp; Dyer method and MRM profiling was used for lipid screening. A total of 329 ion transitions (MRMs) related to lipids were detected with most lipids being characterized as plasma membrane lipids (74%) which were comprised of phosphatidylcholines (40%), ceramides (16%), phosphatidylethanolamines (11%), and phosphatidylserines (7%). acylcarnitines (AC) represented approximately 8% of the ejaculate lipidome. Hierarchical cluster and principal component analysis revealed that boars have a distinct ejaculate lipidome profile based on major plasma membrane lipid classes. In addition, we observed that one boar was unique in his abundance of AC which are related to progressive motility and sperm cell metabolism. These results indicate that this method of sample processing for MRM profiling is suitable to be used to evaluate the lipidome of ejaculates from commercial boars and has potential for broader applications across different livestock species in commercial environments

Phosphaturic mesenchymal tumor demonstrated by 68Ga-DOTATATE PET/CT in a patient: a case report

Tumor-induced osteomalacia (TIO) is a paraneoplastic syndrome caused by abnormally high levels of fibroblast growth factor 23 (FGF-23), most commonly produced and secreted by small phosphaturic mesenchymal tumors (PMT). These tumors can show various anatomic locations throughout soft tissue and bone. The presence of the tumor itself rarely causes symptoms. Nonspecific symptoms such as muscle weakness and musculoskeletal pain are related to the developing hypophosphatemia and osteomalacia as a secondary effect of the increased circulating levels of FGF-23. Therefore, as the initial presentation can mimic a wide range of metabolic or inflammatory diseases, proper diagnosis is often delayed. Localization of the tumor is crucial, as its complete surgical resection is the only curative treatment. Whole-body functional imaging targeting the overexpression of somatostatin receptors (SSTR) on the surface of the PMT cells, is a highly specific and sensitive imaging method to detect the primary tumor site. Here, we discuss a case of TIO in a patient initially presenting with symptoms of inflammatory spondyloarthritis. SSTR positron emission imaging using 68Ga-DOTATATE was central in diagnosing and localizing the primary tumor.

Highly sensitive site-specific SUMOylation proteomics in Arabidopsis.

SUMOylation-the attachment of a small ubiquitin-like modifier (SUMO) to target proteins-plays roles in controlling plant growth, nutrient signalling and stress responses. SUMOylation studies in plants are scarce because identifying SUMOylated proteins and their sites is challenging. To date, only around 80 SUMOylation sites have been identified. Here we introduce lysine-null SUMO1 into the Arabidopsis sumo1 sumo2 mutant and establish a two-step lysine-null SUMO enrichment method. We identified a site-specific SUMOylome comprising over 2,200 SUMOylation sites from 1,300 putative acceptors that function in numerous nuclear processes. SUMOylation marks occur on several motifs, differing from the canonical ψKxE motif in distant eukaryotes. Quantitative comparisons demonstrate that SUMOylation predominantly enhances the stability of SUMO1 acceptors. Our study delivers a highly sensitive and efficient method for site-specific SUMOylome studies and provides a comprehensive catalogue of Arabidopsis SUMOylation, serving as a valuable resource with which to further explore how SUMOylation regulates protein function.

Doping Control Analysis of Perfluorocarbons in Equine Plasma by Headspace Gas Chromatography-Tandem Mass Spectrometry.

In order to control the potential misuse of perfluorocarbons as an oxygen carrier in equine sports, a simple and sensitive method for detecting perfluorocarbons in equine plasma by gas chromatography/tandem mass spectrometry using negative chemical ionization with methane as reagent gas has been developed and fully validated. The method covers seven perfluorocarbons, which are the active components in blood substitute products, and shows good sensitivity and robustness. Limits of detection as low as 0.01 ng/mL could be achieved by the method. To the best of our knowledge, this is the first report of a detection method for the screening of perfluorocarbons in equine biological samples.

Aqueous Afterglow Dispersion Enabling On‐Site Ratiometric Sensing of Mercury Ions

AbstractPollution caused by heavy metal ions has become a global issue owing to their severe threat to the ecological environment and human health. However, it remains a considerable challenge to detect heavy metal ions in an efficient, selective, and high signal‐to‐noise ratio way. Herein, a portable and sensitive method is presented to probe Hg2+ by using an ultralong afterglow dispersion. The in situ encapsulation of phosphorescent carbon dots (CDs) within rigid hydrogen‐bonded organic frameworks (HOFs) leads to ultralong room temperature phosphorescence (RTP) in water with a maximum lifetime of up to 974.86 ms. Moreover, the resultant CDs@HOFs material exhibits robust and long‐term RTP emission with enhanced performance under strongly acidic or alkaline conditions, which contributes to the practical detection of Hg2+ in water. As such, an efficient and sensitive afterglow probe is facilely fabricated by integrating CDs@HOFs with a Hg2+ probe Rhodamine B derivative (RhBTh), demonstrating selective sensing of Hg2+ with greatly improved signal‐to‐noise ratios based on the triplet‐singlet Förster resonance energy transfer system (TS‐FRET). This work not only provides a reliable and versatile method for realizing robust RTP emission in water, but also expands the applications of afterglow materials in the field of optical sensing of toxic analytes.

Paraquat Poisoning: Insights from Autopsy, Histology, and Liquid Chromatography with Tandem Mass Spectrometry in Multidisciplinary Forensic Toxicology Practice

The herbicide paraquat (PQ) is responsible for a significant number of fatalities resulting from self-poisoning. Nevertheless, only a limited number of comprehensive studies focusing on fatal PQ poisoning, which include examination of autopsy findings, histopathology, and quantitative analysis of post-mortem samples, have been published. This study aimed to evaluate autopsy findings, histopathology, and quantitative analysis of PQ in post-mortem human serum samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS), a simple, sensitive, and specific method. Autopsies were performed on all deaths due to PQ poisoning, and serum samples were sent to the toxicology laboratory for chemical analysis. The method was successfully applied to seven human serum samples, and the results indicate its reliability for detecting PQ. The study reports fatal serum PQ levels ranging from 0.5 to 372.0 µg/mL. The comprehensive data presented in this study can be useful for further research and practical applications.

4D flow MRI velocity uncertainty quantification.

An automatic method is presented for estimating 4D flow MRI velocity measurement uncertainty in each voxel. The velocity distance (VD) metric, a statistical distance between the measured velocity and local error distribution, is introduced as a novel measure of 4D flow MRI velocity measurement quality. The method uses mass conservation to assess the local velocity error variance and the standardized difference of means (SDM) velocity to estimate the velocity error correlations. VD is evaluated as the Mahalanobis distance between the local velocity measurement and the local error distribution. The uncertainty model is validated synthetically and tested in vitro under different flow resolutions and noise levels. The VD's application is demonstrated on two in vivo thoracic vasculature 4D flow datasets. Synthetic results show the proposed uncertainty quantification method is sensitive to aliased regions across various velocity-to-noise ratios and assesses velocity error correlations in four- and six-point acquisitions with correlation errors at or under 3.2%. In vitro results demonstrate the method's sensitivity to spatial resolution, venc settings, partial volume effects, and phase wrapping error sources. Applying VD to assess in vivo 4D flow MRI in the aorta demonstrates the expected increase in measured velocity quality with contrast administration and systolic flow. The proposed 4D flow MRI uncertainty quantification method assesses velocity measurement error owing to sources including noise, intravoxel phase dispersion, and velocity aliasing. This method enables rigorous comparison of 4D flow MRI datasets obtained in longitudinal studies, across patient populations, and with different MRI systems.

Моделирование чувствительности литийсодержащих кристаллических сцинтилляторов к нейтронному излучению

This paper presents the results of Monte Carlo modelling of the sensitivity of the lithium containing detectors CLYC, NaIL, CLLB to thermal neutrons. The difference between the sensitivity and the neutron freepath depth of the CLYC scintillator enriched and unenriched with the 6Li isotope is shown. The analysis of the distribution of alpha particles and neutrons in the scintillator volume is performed, showing the inefficiency of the unenriched scintillator due to the fact that not all neutrons react with the crystal material. Mathematical modelling by the Monte Carlo method of the scintillator sensitivity and its comparison with respect to the irradiation direction has been carried out. It is shown that for the scintillator in the face and side geometry with the size more than 38 mm the difference in sensitivity at change of the neutron flux direction is connected only with the scintillator cross section and 6Li concentration with the same value of the specific sensitivity. The obtained model results were compared with available experimental data for NaIL and CLLB scintillators, which showed the reliability of the developed models and calculation methods.

Rapid analysis of amatoxins in human urine by means of affinity column chromatography and liquid chromatography-high-resolution tandem mass spectrometry

Analysis of amatoxins is of great importance as these cyclic peptides contribute to a high number of fatalities each year. Development of analytical approaches needs to focus on rapid, sensitive, and reliable methods. By establishing an affinity column chromatography-based assay using the monoclonal amanitin antibody AMA9G3 and liquid chromatography (LC) coupled to high-resolution mass spectrometry (HRMS) for the trace detection of α-, β-, and γ-amanitin in human urine samples to confirm ingestion, we report the first approach that extents the current status of amatoxin analysis. The presented procedure allows detection of amatoxins in human urine down to 1 ng/mL. The method was successfully validated qualitatively for α- and γ-amanitin according to international recommendations. A proof of concept was performed by analyzing 37 urine samples after suspected amatoxin consumption submitted for regular clinical toxicological analysis. Using this antibody-based enrichment strategy, acute amatoxin intoxications can be determined within 90 min and due to the high sensitivity and selectivity, a comparable approach using target specific antibodies may also be used for other toxicological relevant peptides.

Isotope-Labeled Chemoselective Probes for Labeling, Separation, and Comprehensive Quantitative Analysis of Sub-Metabolome.

The significance of small molecule metabolites as biomarkers for disease diagnosis and prognosis is growing increasingly evident, necessitating the development of highly sensitive qualitative and quantitative methods. Herein, multi-chemoselective probes are synthesized and applied for profiling metabolites, including carboxyl, phosphate, hydroxyl, amino, thiol, and carbonyl compounds. This approach seamlessly integrates magnetic solid-phase materials, orthogonal cleavage sites, isotopic tags, and selective coupling sites, minimizes matrix interference, and enhances quantitative accuracy. Meanwhile, a homemade program, High-Resolution Isotope-Assisted Identification and Quantitative (HRIAIQuant) is developed to process the data, which adeptly filters through 33,874 ion pairs present in human serum, leading to the identification of 701 known metabolites and a remarkable 1,062 potential novel ones. This method is successfully applied to analyze metabolites in multiple brain regions of SAMP8 and SAMR1 models, offering a novel tool for Alzheimer's disease research.

Advancing Lung Cancer Diagnosis through NH2-MON-SPME-GC-MS/MS: Enhanced Sensitivity in Aldehyde Biomarker Detection from Exhaled Breath.

The sensitive detection of trace biomarkers in exhaled breath for lung cancer diagnosis represents a critical area of research in life analytical chemistry, with profound implications for early disease detection, therapeutic intervention, and prognosis monitoring. Despite its potential, the analytical process faces significant challenges due to the ultratrace levels of disease biomarkers present and the complex, high-humidity composition of exhaled breath. This study introduces a highly sensitive method for detecting aldehyde biomarkers in exhaled breath by integrating the use of amino-functionalized microporous organic networks (NH2-MON) as a solid-phase microextraction (SPME) fiber coating with gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) analysis. The method innovatively combines sample collection and extraction, achieving a dual-step enrichment process that significantly enhances both the enrichment efficiency and reproducibility of biomarker detection while effectively mitigating the interference caused by water vapor in exhaled breath. The NH2-MON, utilized as an SPME fiber coating, demonstrates exceptional enrichment capacity for five key aldehyde biomarkers, facilitating the development of a highly sensitive detection approach for these biomarkers in exhaled breath. Compared to previously reported methods, the proposed technique exhibits significantly lower limits of quantification, ranging from 0.77 to 11.89 pg mL-1, and achieves substantially higher enrichment factors, ranging from 9156- to 35723-fold. The practicality and feasibility of the method were validated through the analysis of exhaled breath samples from lung cancer patients, underscoring its potential application in the early diagnosis and monitoring of lung cancer.

In-Depth Host Cell Protein Analysis and Viral Protein Impurity Monitoring in Adeno-Associated Virus-Based Gene Therapy Products Using Optimized Wide Window Data-Dependent Acquisition Method.

Compared to other protein therapeutics, there is currently limited knowledge about the residual host cell proteins (HCPs) in adeno-associated virus (AAV)-based gene therapy products. This is primarily due to the lack of a robust and sensitive mass spectrometry-based method for HCP analysis in AAV samples. Existing liquid chromatography-mass spectrometry methods used for analyzing HCPs in therapeutic monoclonal antibodies (mAbs) often cannot be directly applied to AAVs, due to some unique characteristics of AAV samples encountered during their development such as limited sample availability/protein concentration and the presence of surfactants. In this study, we have developed a novel workflow for robust and in-depth HCP analysis of AAV samples by combining wide-window data-dependent acquisition for improved low-abundance HCP detection with single-pot, solid-phase-enhanced sample preparation (SP3) for low-input sample preparation. Using this newly developed method, we were able to detect more than 650 HCPs in a commercial AAV1 sample with a high quantitative reproducibility. This represents a greater than 5-fold increase in HCP protein identification compared to an in-solution digestion method followed by traditional data-dependent acquisition. Similar benefits can also be achieved for other AAV serotypes that were produced internally and purified through different processes. The detection limit of this method is as low as 0.06 ng/mL, enabling more comprehensive HCP coverage in AAV samples. Moreover, for the first time, we have identified several process-related viral proteins, such as Rep 78 and E4. These proteins need to be closely monitored during AAV process development as they may present a greater risk for immunogenicity compared to HCPs that are derived from human HEK293 cells.

A New Highly Effective Development and Validation of Trastuzumab and Hyaluronidase-Oysk in Bulk and Pharmaceutical Dosage form by Using RP-HPLC Method

A simple, rapid, precise, sensitive, and reproducible Reverse phase High-performance liquid chromatography (RP-HPLC) method has been developed for the quantitative analysis of Trastuzumab and Hyaluronidase-OYSK in the pharmaceutical dosage form. Chromatographic separation of Trastuzumab and Hyaluronidase-OYSK was achieved on Waters Alliance-e2695by using Agilent Eclipse XDB (250x 4.6mm, 5µ) column and the mobile phase containing Acetonitrile: Ammonium formate pH-3.0/OPA in the ratio of 30:70% v/v. The flow rate was 1.0 ml/min; detection was carried out by absorption at 228nm using a photodiode array detector at ambient temperature. The number of theoretical plates and tailing factor for Trastuzumab and Hyaluronidase-OYSK were NLT 2000 and should not be more than 2 respectively. % Relative standard deviation of peak areas of all measurements always less than 2.0. The proposed method was validated according to ICH guidelines. The method was found to be simple, economical, suitable, precise, accurate &amp; robust method for quantitative analysis of Trastuzumab and Hyaluronidase-OYSK study of its stability.

Spectrophotometric determination for green hydrophobic deep eutectic solvent-based microextraction of Brilliant Blue FCF (E133) from beverages

In this study, a simple, sensitive, and rapid method called green hydrophobic deep eutectic solvent-based liquid-liquid microextraction was developed to extract Brilliant Blue FCF dye from beverages. This method utilizes hydrophobic DES obtained by forming tetrabutylammonium bromide and 1-octanol in a 1:5 ratio as green extraction solvent. The transition of Brilliant Blue FCF to the DES phase occurred on its own, without the need for any reagents such as added salt or tetrahydrofuran. Several crucial factors were tried to get the best extraction efficiency, including species, DES volume and molar ratio, solution pH, ultrasonication, and centrifugation time. Under optimum conditions, extraction recoveries were achieved in the range of 95.1–101.3 % with the method developed for Brilliant Blue FCF. The detection and determination limits were observed to be 4.1 μg l-1 and 12.1 μg l-1, respectively. In addition, the relative standard deviation values for the method's accuracy were found to be 2.23 % and 3.48 % within and between days, respectively. It has been established that the developed method is highly environmentally friendly thanks to the application of the Analytical GREENness (AGREE) and Green Analytical Procedure Index (GAPI) tools. This study shows that DES applications can be carried out without the use of emulsifiers and dispersants by prioritizing the use of hydrophobic DES compounds as environmentally friendly and green extraction solvents in food samples.

Mn‐functionalized ZIF‐67 for Sensitive Colorimetric Detection of Artemisinin and Ascorbic Acid

AbstractArtemisinin (Art) is the first‐choice medicine for treating malaria. It also shows good therapeutic effects in antibacterial and antitumor treatment. Therefore, a simple and efficient method for detecting Art is of great significance. Mn‐functionalized ZIF‐67 (Mn@ZIF‐67) hybrid materials were synthesized using a post‐synthesis modification method. Due to the introduction of Mn2+, Mn@ZIF‐67 possesses higher peroxidase/oxidase like dual‐mimic enzymes activity than ZIF‐67, which can catalyze Art and generate peroxyl radicals. Based on this fact, Mn@ZIF‐67 was used to detect Art in the presence of TMB. A good linear relationship between absorption intensities at 652 nm and Art concentrations was obtained in the range of 0–22 μM with a detection limit of 200 nM. Ascorbic acid (AA), due to its reducibility, can inhibit peroxidase/oxidase mimetics activity of Mn@ZIF‐67 and the blue Mn@ZIF‐67‐TMB solution fades gradually with the addition of AA. Mn@ZIF‐67‐TMB acting as a colorimetric sensor, AA can be sensitively detected ranging from 0–30 μM, and the detection limit is 50 nM. Coexisting substances don't interfere with the detection of Art or AA. This sensing platform provides a highly sensitive and selective colorimetric method for monitoring AA and Art in serum, thereby demonstrating remarkable practicality.

Sensitive colorimetric detection of heparin using reverse modulation of peroxidase- and oxidase-mimetic activities in Fe3O4@PDA@MnO2 nanocomposites

Heparin, a widely studied glycosaminoglycan, plays crucial roles in the regulation of various physiological and pathological processes. Therefore, it's important to develop highly selective and sensitive methods for convenient monitoring of heparin levels in biological systems. We report the design and synthesis of Fe3O4@PDA@MnO2 nanoparticles (FPM-NPs), which exhibit dual enzymatic activities, enabling quantitative detection of heparin. The FPM-NPs feature a unique tri-layer spherical shell structure, possessing both peroxidase-like and oxidase-like activities, and catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence or absence of H2O2. Remarkably, upon co-incubated with heparin, the oxidase activity of FPM-NPs decreases, while the peroxidase activity increases. By leveraging these dual enzymatic properties of FPM-NPs, a highly sensitive and specific colorimetric detection of heparin is achieved, with a detection limit reaching 6.51 nM and a good linear response to quantify heparin ranging 10–800 nM. Additionally, the developed FPM-NPs are successfully applied to measure heparin in fetal bovine serum samples. We also extend this detection method to a paper-based chip, enabling portable detection of heparin through grayscale analysis of mobile phone photographs. The multi-nanozyme-based heparin detection approach provides a new perspective for future research on expanding the application of nanocomposite materials in biomedical detection and analysis.