Colorimetric aptasensor for Listeria monocytogenes detection using dual functional Fe3O4@MIL-100(Fe) with magnetic separation and oxidase-like activities in food samples.

A novel colorimetric aptasensor assay based on the excellent magnetic responsiveness and oxidase-like activity of Fe3O4@MIL-100(Fe) was developed. Fe3O4@MIL-100(Fe) absorbed with aptamer and blocked by BSA served as capture probe for selective isolation and enrichment of Listeria monocytogenes one of the most common and dangerous foodborne pathogenic bacteria. The aptamer absorbed on Fe3O4@MIL-100(Fe) was further used as signal probe that specifically binds with target bacteria conjugation of capture probe for colorimetric detection of Listeria monocytogenes, taking advantages of its oxidase-like activity. The linear range of the detection of Listeria monocytogenes was from 102 to 107 CFU mL-1, with the limit of detection as low as 14 CFU mL-1. The approach also showed good feasibility for detection of Listeria monocytogenes in milk and meat samples. The spiked recoveries were in the range 81-114% with relative standard deviations ranging from 1.28 to 5.19%. Thus, this work provides an efficient, convenient, and practical tool for selective isolation and colorimetric detection of Listeria monocytogenes in food.

Optimization of pre-enrichment strategies for mouse hematopoietic stem cell isolation and metabolomic analysis.

Blood cell production arises from the activity of hematopoietic stem cells (HSCs), defined by their self-renewal capacity and ability to give rise to all mature blood cell types. The mouse remains one of the most studied species in hematological research, and markers to define and isolate mouse HSCs are well-established. Given the very low frequency of HSCs in the bone marrow, stem cell pre-enrichment by red blood cell lysis and magnetic cell separation is often performed as part of the isolation process to reduce sorting times. Several pre-enrichment strategies are available, differing in their speed, degree of enrichment, final cell yield and cost. In the current study, we performed a side-by-side comparison and provide a decision tree to help researchers select a pre-enrichment strategy for mouse HSC isolation depending on their downstream application. We then compared different pre-enrichment techniques in combination with metabolomics analysis of HSCs, where speed, yield and temperature during pre-enrichment are crucial factors, and found that the choice of pre-enrichment strategy significantly impacts the number of metabolites detected and levels of individual metabolites in HSCs.

An exonuclease III-driven dual-mode aptasensor based on Au-Pd@Fc nanozyme and magnetic separation pretreatment for aminoglycoside antibiotics detection.

A novel dual-mode aptasensor was constructed for aminoglycoside antibiotics (AAs) detection by using a broad-spectrum aptamer as a biorecognition element, and Au-Pd@Fc functionalized by signal DNA as nanoprobes. In electrochemical mode, the target-induced cyclic amplification reaction run under the action of exonuclease-III, which increased the number of nanoprobes on the electrode surface. AAs could be quantitatively detected with LOD of 0.0355 ± 0.00613 nM. In colorimetric mode, the Au-Pd@Fc nanozyme catalyzed the color reaction of 3,3',5,5'-tetramethylbenzidine. The blue-shifted absorbance will be observed with the change of AAs concentration, and the LOD was 0.0458 ± 0.00572 nM. Furthermore, a magnetic molecular-imprinted material capable of specific adsorption of AAs was prepared on milk sample pretreatment. The aptasensor was used to detect 10 kinds of AAs in milk and the recoveries were 97.19 ± 4.41% âˆ¼ 98.70 ± 4.45% and 96.38 ± 3.53%-97.54 ± 4.13% in electrochemical and colorimetric methods. This work provided a theoretical basis for the application of aptamers in simultaneous detection of antibiotics.

Multifunctional nanoenzyme lateral flow immunoassay strip for rapid and ultrasensitive detection of carbofuran in vegetables.

A lateral flow immunoassay strip (LFIAS) is one of the most frequently rapid test technologies for carbofuran (CAR). Nevertheless, the LFIAS has a poor quantitative capability and low sensitivity. And, it also requires often complex sample handling steps, making testing time longer. In this study, Fe3O4 nanoparticles were successively modified with MIL-100(Fe)-based metal-organic framework (MOF) and chloroplatinic acid hexahydrate to obtain a core-shell complex of Fe3O4-MOF-Pt. The complex had a peroxidase-mimicking activity catalytic function that enabled signal amplification and sensitivity enhancement. Upon coupling with carbofuran monoclonal antibody (CAR-mAb), the magnetic separation properties of the probe enabled target-specific enrichment. The LFIAS based on Fe3O4-MOF-Pt nanocomposites could detect CAR in the range of 0.25-50 ng mL-1 with a limit of detection (LOD) of 0.15 ng mL-1, enabling colorimetric and catalytic analysis. In addition, the method showed high specificity and stability for detecting CAR in various vegetables, and recovery rates of the spiked samples were 91.40%-102.40%. In conclusion, this study provided one-stop detection of "target enrichment-visual inspection". While lowering the LOD, it reduced the detection time and improved the detection efficiency. The multifunctional Fe3O4-MOF-Pt nanocomposite provides an idea for the construction of novel multifunctional probes to improve the detection performance of conventional LFIAS.

Effect of granulated biochar sphere on mineral nutrients in removing potentially toxic elements from contaminated agricultural soils.

Biochar (BC) granulation, yielding BC-based spheres, serves as an eco-friendly, cost-effective and efficient adsorbent for the removal of potential toxic elements (PTEs) from contaminated agricultural soils. The effect of BC-based spheres on mineral nutrients while effectively removing PTEs from contaminated soils is worth investigating. In this study, we utilized natural clay minerals, magnetic minerals and BC to produce water-hardened magnetic composite biochar sphere (WMBCS) that was capable of removing PTEs from composite contaminated agricultural soils. We explored the effect of WMBCS on minerals (Al, Ca, Fe, Mn, Na, Mg, Si, K, P, NH4+, and NO3-) in the removal of soil PTEs. WMBCS was a mineral nutrient-rich, recyclable, alkaline BC-based sphere that removes Cd (23.07-29.20 %), Pb (27.68-31.10 %), and As (26.17-37.48 %) from soils after three regeneration cycles. The effect of WMBCS on mineral nutrients varies depending on element type, BC and soil type. Compared to water-hardened magnetic composite phosphate modified biochar spheres (WMPBCS), water-hardened magnetic composite unmodified biochar spheres (WMUBCS) had more significant effect on Ca, Mg, Mn, Al and NH4+ in alkaline soils, but a greater effect on Ca, Mg, Mn, Fe and NO3- in acidic soils. Additionally, WMBCS displayed a more pronounced impact on mineral nutrients in alkaline soils than in acidic soils. The application of WMBCS reduced the accumulation of PTEs in wheat (18.40-84.70 %) and rice (27.96-88.66 %), but significantly inhibited seed germination and altered the uptake of mineral nutrients by seedlings due to its effects on soil physicochemical properties and mineral nutrient dynamics. Overall, WMBCS is suitable as a potential amendment for the remediation of soils co-contaminated with Cd, As, and Pb, but its effects on mineral nutrients cannot be overlooked, particularly in agricultural soils.

Dual magnetization and amination of cellulosic chains for the efficient adsorption of heavy metals.

Compounds functionalized with hydroxyl and amino groups were found to have good potential for the adsorption of different ions. In this work, a new system of cellulosic chains was amended with amine substitutions and bonded to a magnetic core of NiFe2O4@SiO2 to form NiFe2O4@SiO2-cellulose-NH2 system. The prepared sample showed suitable magnetic separation and was characterized via XRD, FT-IR, SEM, EDS, and TGA-DTA analyses. The adsorption potential of NiFe2O4@SiO2-cellulose-NH2 system has been investigated on the heavy metals (Cd, Ni, and Pb) removal from a synthetic wastewater environment. The results show that the magnetic property created by the magnetic core increased the recycling potential of the adsorbent and the magnetic core has a positive effect on the absorption potential of the polymer. The adsorption removal of Cd(II), Ni(II), and Pb(II) ions was studied using NiFe2O4@SiO2-cellulose-NH2 systems in different pH, temperatures, metal ion concentrations, and adsorbent dosages. The maximum adsorption capacities of single heavy metal ions were obtained as 406.44 mg/g (for Cd(II) ions), 411.63 mg/g (for Ni(II) ions), and 414.68 mg/g (for Pb(II) ions) under optimized conditions as pH = 6.5, ion concentration: 500 mg/L, adsorbent dosage: 1.2 g/L and room temperature.

Recent trends in magnetic spinel ferrites and their composites as heterogeneous Fenton-like catalysts: A review.

In recent years, there has been a growing interest in utilizing spinel ferrite and their nanocomposites as Fenton-like catalysts. The use of these materials offers numerous advantages, including ability to efficiently degrade pollutants and potential for long-term and repeated use facilitated by their magnetic properties that make them easily recoverable. The remarkable catalytic properties, stability, and reusability of these materials make them highly attractive for researchers. This paper encompasses a comprehensive review of various aspects related to the Fenton process and the utilization of spinel ferrite and their composites in catalytic applications. Firstly, it provides an overview of the background, principles, mechanisms, and key parameters governing the Fenton reaction, along with the role of physical field assistance in enhancing the process. Secondly, it delves into the advantages and mechanisms of H2O2 activation induced by different spinel ferrite and their composites for the removal of organic pollutants, shedding light on their efficacy in environmental remediation. Thirdly, the paper explores the application of these materials in various Fenton-like processes, including Fenon-like, photo-Fenton-like, sono-Fenton-like, and electro-Fenton-like, for the effective removal of different types of contaminants. Furthermore, it addresses important considerations such as the toxicity, recovery, and reuse of these materials. Finally, the paper presents the challenges associated with H2O2 activation by these materials, along with proposed directions for future improvements.

Aggregation-Augmented Magnetism of Lanthanide-Doped Nanoparticles and Enabling Magnetic Levitation-Based Exosome Sensing.

Due to the presence of unpaired electron orbitals in most lanthanide ions, lanthanide-doped nanoparticles (LnNPs) exhibit paramagnetism. However, as to biosensing applications, the magnetism of LnNPs is so weak that can hardly be employed in target separation. Herein, it is discovered that the magnetism of the LnNPs is highly associated with their concentration in a confined space, enabling aggregation-augmented magnetism to make them susceptive to a conventional magnet. Accordingly, a magnetic levitation (Maglev) sensing system is designed, in which the target exosomes can specifically introduce paramagnetic LnNPs to the microbeads' surface, allowing aggregation-augmented magnetism and further leverage the microbeads' levitation height in the Maglev device to indicate the target exosomes' content. It is demonstrated that this Maglev system can precisely distinguish healthy people's blood samples from those of breast cancer patients. This is the first work to report that LnNPs hold great promise in magnetic separation-based biological sample sorting, and the LnNP-permitted Maglev sensing system is proven to be promising for establishing a new generation of biosensing devices.

Enhanced microfluidic multi-target separation by positive and negative magnetophoresis.

We introduce magnetophoresis-based microfluidics for sorting biological targets using positive Magnetophoresis (pM) for magnetically labeled particles and negative Magnetophoresis (nM) for label-free particles. A single, externally magnetized ferromagnetic wire induces repulsive forces and is positioned across the focused sample flow near the main channel's closed end. We analyze magnetic attributes and separation performance under two transverse dual-mode magnetic configurations, examining magnetic fields, hydrodynamics, and forces on microparticles of varying sizes and properties. In pM, the dual-magnet arrangement (DMA) for sorting three distinct particles shows higher magnetic gradient generation and throughput than the single-magnet arrangement (SMA). In nM, the numerical results for SMA sorting of red blood cells (RBCs), white blood cells (WBCs), and prostate cancer cells (PC3-9) demonstrate superior magnetic properties and throughput compared to DMA. Magnetized wire linear movement is a key design parameter, allowing device customization. An automated device for handling more targets can be created by manipulating magnetophoretic repulsion forces. The transverse wire and magnet arrangement accommodate increased channel depth without sacrificing efficiency, yielding higher throughput than other devices. Experimental validation using soft lithography and 3D printing confirms successful sorting and separation, aligning well with numerical results. This demonstrates the successful sorting and separating of injected particles within a hydrodynamically focused sample in all systems. Both numerical and experimental findings indicate a separation accuracy of 100% across various Reynolds numbers. The primary channel dimensions measure 100 µm in height and 200 µm in width. N52 permanent magnets were employed in both numerical simulations and experiments. For numerical simulations, a remanent flux density of 1.48 T was utilized. In the experimental setup, magnets measuring 0.5 × 0.5 × 0.125 inches and 0.5 × 0.5 × 1 inch were employed. The experimental data confirm the device's capability to achieve 100% separation accuracy at a Reynolds number of 3. However, this study did not explore the potential impact of increased flow rates on separation accuracy.

Facile Preparation of Magnetically Separable Fe3O4/ZnO Nanocomposite with Enhanced Photocatalytic Activity for Degradation of Rhodamine B.

Magnetic separation of photocatalysts holds great promise for water treatment. A magnetic separation method has a positive effect on the recovery of catalysts after degradation. In this paper, an efficient and reusable catalytic system is developed based on coating magnetic Fe3O4 by depositing Fe2+ on the surface of ZnO. The Fe3O4/ZnO nanocomposite exhibits enhanced performance for organic pollutant degradation. The Fe3O4/ZnO system demonstrates a high photocatalytic activity of 100% degradation efficiency in Rhodamine B (RhB) degradation under UV light irradiation for 50 min. The excellent photocatalytic activity is primarily due to the separation of photogenerated electron-hole pairs being facilitated by the strong interaction between Fe3O4 and ZnO. The induction of the magnetic Fe3O4 endows the Fe3O4/ZnO composite with superior magnetic separation capability from water. Experiments with different radical scavengers revealed that the hydroxyl radical (·OH) is the key reactive radical for the effective degradation of RhB. This work innovatively affords a common interfacial dopant deposition strategy for catalytic application in the degradation of organic dye pollutants and catalyst separation from wastewater efficiently.

Synthesis and fabrication of magnetically separable phosphate-modified magnetic chitosan composites for lead(II) selective removal from wastewater.

To address the urgent need for efficient removal of lead-containing wastewater and reduce the risk of toxicity associated with heavy-metal wastewater contamination, materials with high removal rates and easy separation must be developed. Herein, a novel organic-inorganic hybrid material based on phosphorylated magnetic chitosan (MSCP) was synthesized and applied for the selective removal of lead (II) from wastewater. From the characterization and the experimental results can be obtained that the magnetic saturation strength of MSCP reaches 14.65 emu/g, which can be separated quickly and regenerated readily, and maintains high adsorption performance even after 5 cycles, indicating that the adsorbent possesses good magnetic separation performance and durability. Also, MSCP showed high selective adsorption performance for lead in the multiple metal ions coexistence solutions at pH 6.0 and room temperature, with an adsorption coefficient SPb-MSCP of 78.85%, which was much higher than that of MSC (the SPb-MSC was 11.59%). Additionally, in the single lead system, the sorption characteristics of Pb(II) on MSCP and MCP had obvious pH-responsiveness, and their adsorption capacity increased with the increase of solution pH, reaching the maximal values of 80.19 and 72.68 mg/g, respectively. It is noteworthy that the acid resistance of MSCP with an inert layer coated on the core is significantly improved, with almost no iron leaching from MSCP over the entire acidity range, while MCP has 7.63 mg/g of iron leaching at pH 1.0. Significantly, MSCP exhibited a maximum adsorption capacity of 102.04 mg/g, which matches the Langmuir model at pH 6.0 and 298.15 K, and points to the pseudo-second-order kinetics of the chemisorption process of Pb(II) on MSCP. These findings highlight the great potential of MSCP for Pb(II) removal from aqueous solution, making it a promising solution for Pb(II) contamination in wastewater.

The Waelz Slag from Electric Arc Furnace Dust Processing: Characterization and Magnetic Separation Studies.

The Waelz slag generated during electric arc furnace dust processing is an iron-rich product with significant amounts of iron, zinc and copper. About 600-800 kg of the Waelz slag is generated per ton of the dust processed. The Waelz slag samples from two different plants were thoroughly characterized using inductively coupled plasma optical emission spectroscopy (ICP-AES), X-ray diffraction analysis (XRD), chemical phase analysis, Mössbauer spectroscopy and other supporting methods. The phase distribution of iron, zinc and copper was determined in the Waelz slag samples. Low-intensity wet magnetic separation was tested for the iron recovery from the Waelz slag samples. It was found that the Waelz slag samples have complex chemical and mineralogical compositions, which can impede the selective recovery of valuable elements. The obtained results indicate that the chemical and mineralogical composition of the Waelz slag samples has a considerable effect on the magnetic separation indexes. The experiments showed that the iron concentrates with Fe contents of 73% and 46.8% with the metallization degrees of 87.2% and 57.5% and the iron recovery degree of 54.8% and 52.9% were obtained at optimal conditions for two different samples, respectively, without selective segregation of Cu and Zn in the magnetic or non-magnetic fraction.

Synthesis, modifications, and applications of iron-based nanoparticles.

Magnetic nanoparticles (MNPs) are appealing materials as assistant to resolve environmental pollution issues and as recyclable catalysts for the oxidative degradation of resistant contaminants. Moreover, they can significantly influence the advancement of medical applications for imaging, diagnostics, medication administration, and biosensing. On the other hand, due to unique features, excellent biocompatibility, high curie temperatures and low cytotoxicity of the Iron-based nanoparticles, they have received increasing attention in recent years. Using an external magnetic field, in which the ferrite magnetic nanoparticles (FMNPs) in the reaction mixtures can be easily removed, make them more efficient approach than the conventional method for separating the catalyst particles by centrifugation or filtration. Ferrite magnetic nanoparticles (FMNPs) provide various advantages in food processing, environmental issues, pharmaceutical industry, sample preparation, wastewater management, water purification, illness therapy, identification of disease, tissue engineering, and biosensor creation for healthcare monitoring. Modification of FMNPs with the proper functional groups and surface modification techniques play a significant role in boosting their capability. Due to flexibility of FMNPs in functionalization and synthesis, it is possible to make customized FMNPs that can be utilized in variety of applications. This review focuses on synthesis, modifications, and applications of Iron-based nanoparticles.

Magnetic Ion-Imprinted Materials for Selective Adsorption of Cr(VI): Adsorption Behavior and Mechanism Study.

With the increase of hexavalent Cr(VI) wastewater discharged from industrial production, it seriously pollutes water bodies and poses a risk to human health. Adsorption is used as an effective means to treat Cr(VI), but its effectiveness is affected by pH, and the adsorption performance decreases when acidity is strong. Furthermore, research on the mechanism of Cr(VI) adsorption using DFT calculations needs to be developed. This study focuses on the development of magnetically responsive core-shell nano-ion imprinted materials (Fe3O4@GO@IIP) through magnetic separation and surface imprinting techniques. Characterization techniques including FT-IR, XRD, and EDS confirmed the core-shell nanostructure of Fe3O4@GO@IIP. Batch adsorption experiments and model simulations demonstrated the exceptional adsorption capacity of Fe3O4@GO@IIP for Cr(VI) in strongly acidic solutions (pH = 1), reaching a maximum of 89.18 mg/g. The adsorption mechanism was elucidated through XPS and DFT calculations, revealing that Fe3O4@GO@IIP operates through electrostatic interactions and chemical adsorption, with charge transfer dynamics quantified during the process. This research provides new insights for addressing Cr(VI) treatment in highly acidic environments.

Exploring the effective adsorption of polystyrene microplastics from aqueous solution with magnetically separable nickel/reduced graphene oxide (Ni/rGO) nanocomposite.

Microplastics (MPs) are a potential threat to both humans and aquatic environment as they serve as carriers of various contaminants necessitating the development of reliable, efficient, and ecofriendly techniques to remove MPs from water. In this study, reduced graphene oxide (rGO) magnetized using nickel nanoparticles was utilized as a potent adsorbent for the effective removal of microplastics from water. The synthesized nickel/reduced graphene oxide (Ni/rGO) nanocomposite was characterized by X-ray diffraction (XRD), Raman spectra, vibrating sample magnetometer (VSM), scanning electron microscopy-energy-dispersive X-ray analysis (SEM-EDX), thermogravimetric analysis, and Brunauer-Emmett Teller (BET) analysis. Magnetic Ni/rGO nanocomposite exhibited significant adsorption capability for polystyrene (PS) microspheres allowing the formation of PS-Ni/rGO complex which can be easily separated out using a magnet. The SEM images of PS-Ni/rGO complex confirmed the adsorption of PS microspheres onto the nano adsorbent due to hydrophobic interaction. The adsorbent demonstrated a maximum adsorption capacity of 1250 mg/g. The analysis of isotherm and kinetic models demonstrated that the adsorption mechanism conformed to the Langmuir isotherm and followed pseudo second order kinetics. This study paves a new pathway for the application of magnetically modified reduced graphene oxide for the expedient removal of microplastics from water with the ease of separation using a magnet. The adsorbent was recycled and reused for three times.

Magnetic CAR T cell purification using an anti-G4S linker antibody.

Chimeric antigen receptor (CAR) redirected T cells are successfully employed in the combat against several hematological malignancies, however, are often compromised by low transduction rates making refinement of the CAR T cell products necessary. Here, we report a broadly applicable enrichment protocol relying on marking CAR T cells with an anti-glycine4-serine (G4S) linker antibody followed by magnetic activated cell sorting (MACS). The protocol is broadly applicable since the G4S peptide is an integral part of the vast majority of CARs as it links the VH and VL recognition domains. We demonstrate the feasibility by using the canonical second generation CARs specific for CEA and Her2, respectively, obtaining highly purified CAR T cell products in a one-step procedure without impairing cell viability. The protocol is also applicable to a dual specific CAR (tandem CAR). Except for CD39, T cell activation/exhaustion markers were not upregulated after separation. Purified CAR T cells retained their functionality with respect to antigen-specific cytokine secretion, cytotoxicity, and the capacity to proliferate and eliminate cognate tumor cells upon repetitive stimulation. Collectively, the one-step protocol for purifying CAR T cells extends the toolbox for preclinical research and specifically for clinical CAR T cell manufacturing.

Magnetically functionalized hydrogels for high-throughput genomic applications.

Single-cell genomics has revolutionized tissue analysis by revealing the genetic program of individual cells. The key aspect of the technology is the use of barcoded beads to unambiguously tag sequences originating from a single cell. The generation of unique barcodes on beads is mainly achieved by split-pooling methods, which are labor-intensive due to repeated washing steps. Towards the automation of the split-pooling method, we developed a simple method to magnetize hydrogel beads. We show that these hydrogel beads provide increased yields and washing efficiencies for purification procedures. They are also fully compatible with single-cell sequencing using the BAG-Seq workflow. Our work opens the automation of the split-pooling technique, which will improve single-cell genomic workflows.

Magnetic Nanoagent Coated with an Activated Macrophage Membrane for Colorimetric Detection of Bacteria.

The construction of cell mimics replicating the surface landscape and biological functions of the cell membrane offers promising prospects for biomedical research and applications. Inspired by the inherent recognition capability of immune cells toward pathogens, we have fabricated activated macrophage membrane-coated magnetic silicon nanoparticles (aM-MSNPs) in this work as an isolation and recognition tool for enhanced bacterial analysis. Specifically, the natural protein receptors on the activated macrophage membrane endow the MSNPs with a broad-spectrum binding capacity to different pathogen species. By further incorporation of a tyramide amplification strategy, direct naked-eye analysis of specific bacteria with a detection limit of 10 CFU/mL can be achieved. Moreover, application to the diagnosis of urinary tract infections has also been validated, and positive samples spiked with bacteria can be clearly distinguished with an accuracy of 100%. This work may enrich cell membrane-based architectures and provide an experimental paradigm for point-of-care testing (POCT) detection of bacteria.

Alkyl ammonium hydrogen sulfate immobilized on Fe3O4@SiO2 nanoparticles: a highly efficient catalyst for the multi-component preparation of novel tetrazolo[1,5-a]pyrimidine-6-carboxamide derivatives.

In this, a three-component reaction for the preparation of novel tetrazolo[1,5-a]pyrimidine-6-carboxamide derivatives from N,N'-(sulfonylbis(1,4-phenylene))bis(3-oxobutanamide), aldehydes and 1H-tetrazol-5-amine is reported. The application of Fe3O4@SiO2-(PP)(HSO4)2 (A) as a catalyst afforded the desired products (a1-a18) in high yields in DMF as solvent as well as under solvent-free conditions.

A magnetic calcium phosphate for selective capture of multi-phosphopeptides.

The specific enrichment of multi-phosphopeptides in the presence of non-phosphopeptides and mono-phosphopeptides was still a challenge for phosphoproteomics research. Most of these enrichment materials relied on Zn, Ti, Sn, and other rare precious metals as the bonding center to enrich multi-phosphopeptides while ignoring the use of common metal elements. The addition of rare metals increased the cost of the experiment, which was not conducive to their large-scale application in biomedical proteomics laboratories. In addition, multiple high-speed centrifugation steps also resulted in the loss of low-abundance multi-phosphopeptides in the treatment procedure of biological samples. This study proposed the use of calcium, a common element, as the central bonding agent for synthesizing magnetic calcium phosphate materials (designated as CaP-Fe3O4). These materials aim to capture multi-phosphopeptides and identifying phosphorylation sites. The current results demonstrate that CaP-Fe3O4 exhibited excellent selection specificity, high sensitivity, and stability in the enrichment of multi-phosphopeptides and the identification of phosphorylation sites. Additionally, the introduction of magnetic separation not only reduced the time required for multi-phosphopeptides enrichment but also prevented the loss of these peptides during high-speed centrifugation. These findings contribute to the widespread application and advancement of phosphoproteomics research.