Mn2+-Mediated Modulation of PfAgo Activity for Biosensing.

Argonaute (Ago) as a powerful enzyme has provided new insights into biosensing due to its programmability, high sensitivity, and user-friendly operation. However, current strategies mainly rely on phosphorylated guide DNA to modulate the cleavage activity of Ago, which is limited in versatility and simplicity. Herein, the authors report the Mn2+-enhanced cleavage activity of Ago and employ Mn-ions with variable valence to regulate the activity of Pyrococcus furiosus Ago (PfAgo) for biosensing applications. The conversion of Mn ions with different valence states through MnO2 nanoflowers enables the sensitive detection of ascorbic acid, alkaline phosphatase, and arsenic with limits of detection of 2.5 nmol L-1, 0.009 U L-1, and 0.4 ng mL-1, respectively. A PfAgo-based immunoassay is further developed that allows for the detection of diverse targets, thus providing a promising toolbox to broaden PfAgo-based sensors into versatile bioanalytical and biomedical applications.

A Wash-Free Spheres-on-Sphere Strategy for On-Site and Multiplexed Biosensing.

Respiratory infections and food contaminants pose severe challenges to global health and the economy. A rapid on-site platform for the simultaneous detection of multiple pathogens is crucial for accurate diagnosis, appropriate treatment, and a reduced healthcare burden. Herein, we present a spheres-on-sphere (SOS) platform for multiplexed detection using a portable Coulter counter, which employs millimeter- and micron-sized spheres coupled with antibodies as multitarget probes. The assay allows for quantitative detection of multiple analytes within 20 min by simple mixing, enabling on-site detection. The platform shows high accuracy in identifying three respiratory viruses (SARS-CoV-2, influenza A virus, and parainfluenza virus) from throat swab samples, with LOD of 50.7, 32.4, and 49.1 pg/mL. It also demonstrates excellent performance in quantifying three mycotoxins (aflatoxin B1, deoxynivalenol, and ochratoxin A) from food samples. The SOS platform offers a rapid on-site approach with high sensitivity and specificity for applications in resource-limited settings.

"Four-In-One" Multifunctional Dandelion-Like Gold@platinum Nanoparticles-Driven Multimodal Lateral Flow Immunoassay.

The traditional lateral flow immunoassay (LFIA) with a single signal output mode may encounter challenges such as low sensitivity, poor detection range, and susceptibility to external interferences. These limitations hinder its ability to meet the growing demand for advanced LFIA. To address these issues, the rational development of multifunctional labels for multimodal LFIA emerges as a promising strategy. Herein, this study reports a multimodal LFIA using "four-in-one" multifunctional dandelion-like gold@platinum nanoparticles (MDGP). The inherent properties of MDGP, such as the broad absorption spectrum, porous dandelion-like nanostructure, and bimetallic composition with gold and platinum, endow them with capacities in dual spectral-overlapped fluorescence quenching, optical readout, catalytic activity, and photothermal effect. Benefiting from their multifunctional properties, the MDGP-LFIA enables multimodal outputs including fluorescent, colorimetric, and photothermal signals. This multimodal MDGP-LFIA allows for the detection of acetamiprid at a range of 0.01-50 ng mL-1 , with the lowest qualitative and quantitative detection results of 0.5 and 0.008 ng mL-1 , respectively, significantly better than the traditional gold nanoparticles-based LFIA. The diversity, complementarity, and synergistic effect of integrated output signals in this multimodal MDGP-LFIA improve the flexibility, practicability, and accuracy of detection, holding great promise as a point-of-care testing platform in versatile application scenarios.

Tuning the plasmonic and catalytic signals of Au@Pt nanoparticles for dual-mode biosensing.

Dual-mode biosensors have gained great attention due to their excellent detection accuracy provided by the mutual verification of output signals. However, current strategies for dual-mode sensing mainly rely on a signal probe exhibiting dual properties that may encounter unreliability. Herein, we report a dual-mode biosensing strategy by modulating the plasmonic and catalytic activities of nanoparticles through a surface growing approach. Ascorbic acid enables the growing of Au shell on Au@Pt NPs to tune both their peroxidase-like activity and plasmonic signal. Enzyme-catalyzed reactions can generate ascorbic acid to modulate the plasmonic and catalytic activities of nanoparticles. Combined with enzyme-linked immunosorbent assay, it enables dual-mode immunoassays of carbofuran with both a colorimetric readout by a spectrometer down to 0.1 ppb and a naked-eye readout of 5 ppb. This dual-mode biosensing technique advantages in tunable sensitivity and robustness, holding promise as an analytical platform for biomedical diagnosis and food safety.

Super Antibacterial Capacity and Cell Envelope-Disruptive Mechanism of Ultrasonically Grafted N-Halamine PBAT/PBF Films against Escherichia coli.

Antibacterial materials are urgently needed to combat bacterial contamination, growth, or attachment on contact surfaces, as bacterial infections remain a public health crisis worldwide. Here, a novel ultrasound-assisted method is utilized for the first time to fabricate oxidative chlorine-loaded AH@PBAT/PBF-Cl films with more superior grafting efficiency and rechargeable antibacterial effect than those from conventional techniques. The films remarkably inactivate 99.9999% Escherichia coli and Staphylococcus aureus cells, inducing noticeable cell deformations and mechanical instability. The specific antibacterial mechanism against E. coli used as a model organism is unveiled using several cell envelope structural and functional analyses combined with proteomics, peptidoglycomics, and fluorescence probe techniques. Film treatment partially neutralizes the bacterial surface charge, induces oxidative stress and cytoskeleton deformity, alters membrane properties, and disrupts the expression of key proteins involved in the synthesis and transport of the lipopolysaccharide and peptidoglycan, indicating the cell envelope as the primary target. The films specifically target lipopolysaccharides, resulting in structural impairment of the polysaccharide and lipid A components, and inhibit peptidoglycan precursor synthesis. Together, these lead to metabolic disorders, membrane dysfunction, structural collapse, and eventual death. Given the films' antibacterial effects via the disruption of key cell envelope components, they can potentially combat a wide range of bacteria. These findings lay a theoretical basis for developing efficient antibacterial materials for food safety or biomedical applications.

Computer Vision-Based Artificial Intelligence-Mediated Encoding-Decoding for Multiplexed Microfluidic Digital Immunoassay.

Digital immunoassays with multiplexed capacity, ultrahigh sensitivity, and broad affordability are urgently required in clinical diagnosis, food safety, and environmental monitoring. In this work, a multidimensional digital immunoassay has been developed through microparticle-based encoding and artificial intelligence-based decoding, enabling multiplexed detection with high sensitivity and convenient operation. The information encoded in the features of microspheres, including their size, number, and color, allows for the simultaneous identification and accurate quantification of multiple targets. Computer vision-based artificial intelligence can analyze the microscopy images for information decoding and output identification results visually. Moreover, the optical microscopy imaging can be well integrated with the microfluidic platform, allowing for encoding-decoding through the computer vision-based artificial intelligence. This microfluidic digital immunoassay can simultaneously analyze multiple inflammatory markers and antibiotics within 30 min with high sensitivity and a broad detection range from pg/mL to µg/mL, which holds great promise as an intelligent bioassay for next-generation multiplexed biosensing.

Tailoring the Surface and Composition of Nanozymes for Enhanced Bacterial Binding and Antibacterial Activity.

With the advantages of diverse structures, tunable enzymatic activity, and high stability, nanozymes are widely used in medicine, chemistry, food, environment, and other fields. As an alternative to traditional antibiotics, nanozymes attract more and more attention from the scientific researchers in recent years. Developing nanozymes-based antibacterial materials opens up a new avenue for the bacterial disinfection and sterilization. In this review, the classification of nanozymes and their antibacterial mechanisms are discussed. The surface and composition of nanozymes are critical for the antibacterial efficacy, which can be tailored to enhance both the bacterial binding and the antibacterial activity. On the one hand, the surface modification of nanozymes enables binding and targeting of bacteria that improves the antibacterial performance of nanozymes including the biochemical recognition, the surface charge, and the surface topography. On the other hand, the composition of nanozymes can be modulated to achieve enhanced antibacterial performance including the single nanozyme-mediated synergistic and multiple nanozymes-mediated cascade catalytic antibacterial applications. In addition, the current challenges and future prospects of tailoring nanozymes for antibacterial applications are discussed. This review can provide insights into the design of future nanozymes-based materials for the antibacterial treatments.

Point-of-Care Detection of Antioxidant in Agarose-Based Test Strip through Antietching of Au@Ag Nanostars.

Antioxidants are crucial for human health, and the detection of antioxidants can provide valuable information for disease diagnosis and health management. In this work, we report a plasmonic sensing approach for the determination of antioxidants based on their antietching capacity toward plasmonic nanoparticles. The Ag shell of core-shell Au@Ag nanostars can be etched by chloroauric acid (HAuCl4), whereas antioxidants can interact with HAuCl4, which prevents the surface etching of Au@Ag nanostars. We modulate the thickness of the Ag shell and morphology of the nanostructures, showing that the core-shell nanostars with the smallest thickness of Ag shell have the best etching sensitivity. Owing to the extraordinary surface plasmon resonance (SPR) property of Au@Ag nanostars, the antietching effect of antioxidants can induce a significant change in both the SPR spectrum and the color of solution, facilitating both the quantitative detection and naked-eye readout. This antietching strategy enables the determination of antioxidants such as cystine and gallic acid with a linear range of 0.1-10 µM. The core-shell Au@Ag nanostars are further immobilized in agarose gels to fabricate test strips, which can display different color changes in the presence of HAuCl4 from 0 to 1000 µM. The agarose-based test strip is also capable of detecting antioxidants in real samples, which allows naked-eye readout and quantitative detection by a smartphone.

Colorimetric Sensing Strategy through the Coordination Chemistry between Ascorbic Acid 2-Phosphate and Copper Ions.

The coordination chemistry between phosphorylated molecules and metal ions has been reported, while few studies focus on its sensing capability. Herein, we report a colorimetric sensing strategy through the coordination chemistry between ascorbic acid 2-phosphate (AAP) and copper ions. The phosphate group-containing AAP can coordinate with copper ions to induce a visible color change from blue to green in a rapid way, which can be easily read by the naked eye or a smartphone based on the blue-to-green (B/G) ratio. This coordination chemistry provides a facile and convenient strategy for designing colorimetric assays. Alkaline phosphatase can catalyze the hydrolysis of AAP to ascorbic acid (AA), thus modulating the AAP/AA transformation and the AAP-mediated coordination, offering a straightforward way for monitoring the enzymatic activity. This colorimetric sensing strategy shows good performances in stability, sensitivity, cost, and scale-up production, holding great promise as a point-of-care technique for diagnostic applications.

Identification of Antibiotic Resistance in ESKAPE Pathogens through Plasmonic Nanosensors and Machine Learning.

Antibiotic-resistant ESKAPE pathogens cause nosocomial infections that lead to huge morbidity and mortality worldwide. Rapid identification of antibiotic resistance is vital for the prevention and control of nosocomial infections. However, current techniques like genotype identification and antibiotic susceptibility testing are generally time-consuming and require large-scale equipment. Herein, we develop a rapid, facile, and sensitive technique to determine the antibiotic resistance phenotype among ESKAPE pathogens through plasmonic nanosensors and machine learning. Key to this technique is the plasmonic sensor array that contains gold nanoparticles functionalized with peptides differing in hydrophobicity and surface charge. The plasmonic nanosensors can interact with pathogens to generate bacterial fingerprints that alter the surface plasmon resonance (SPR) spectra of nanoparticles. In combination with machine learning, it enables the identification of antibiotic resistance among 12 ESKAPE pathogens in less than 20 min with an overall accuracy of 89.74%. This machine-learning-based approach allows for the identification of antibiotic-resistant pathogens from patients and holds great promise as a clinical tool for biomedical diagnosis.

Gold Nanomaterials-Implemented CRISPR-Cas Systems for Biosensing.

Due to their superiority in the simple design and precise targeting, clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have attracted significant interest for biosensing. On the one hand, CRISPR-Cas systems have the capacity to precisely recognize and cleave specific DNA and RNA sequences. On the other hand, CRISPR-Cas systems such as orthologs of Cas9, Cas12, and Cas13 exhibit cis-cleavage or trans-cleavage activities after recognizing the target sequence. Owing to the cleavage activities, CRISPR-Cas systems can be designed for biosensing by degrading tagged nucleic acids to produce detectable signals. To meet the requirements of point-of-care detection and versatile signal readouts, gold nanomaterials with excellent properties such as high extinction coefficients, easy surface functionalization, and biocompatibility are implemented in CRISPR-Cas-based biosensors. In combination with gold nanomaterials such as gold nanoparticles, gold nanorods, and gold nanostars, great efforts are devoted to fabricating CRISPR-Cas-based biosensors for the detection of diverse targets. This review focuses on the current advances in gold nanomaterials-implemented CRISPR-Cas-based biosensors, particularly the working mechanism and the performance of these biosensors. CRISPR-Cas systems, including CRISPR-Cas9, CRISPR-Cas12a, and CRISPR-Cas13a are discussed and highlighted. Meanwhile, prospects and challenges are also discussed in the design of biosensing strategies based on gold nanomaterials and CRISPR-Cas systems.

Nanotechnology-based diagnostic methods for coronavirus: From nucleic acid extraction to amplification.

The recent emergence of human coronaviruses (CoVs) causing severe acute respiratory syndrome (SARS) is posing a great threat to global public health. Therefore, the rapid and accurate identification of pathogenic viruses plays a vital role in selecting appropriate treatments, saving people's lives and preventing epidemics. Nucleic acids, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), are natural biopolymers composed of nucleotides that store, transmit, and express genetic information. Applications of nucleic acid detection range from genotyping and genetic prognostics, to expression profiling and detection of infectious disease. The nucleic acid detection for infectious diseases is widely used, as evidenced by the widespread use of COVID-19 tests for the containment of the pandemic. Nanotechnology influences all medical disciplines and has been considered as an essential tool for novel diagnostics, nanotherapeutics, vaccines, medical imaging, and the utilization of biomaterials for regenerative medicine. In this review, the recent advances in the development of nanotechnology-based diagnostic methods for coronavirus, and their applications in nucleic acid detection are discussed in detail. The techniques for the amplification of nucleic acids are summarized, as well as the use of magnetic nanoparticles for nucleic acid extraction. Besides, current challenges and future prospects are proposed, along with the great potential of nanotechnology for the effective diagnosis of coronavirus.

Cyclodextrin metal-organic framework by ultrasound-assisted rapid synthesis for caffeic acid loading and antibacterial application.

Cyclodextrin metal-organic framework by ultrasound-assisted rapid synthesis for caffeic acid (CA) loading and antibacterial application (U-CD-MOF) was successfully studied and this method shortened the preparation time to a few minutes. It was found that the ultrasonic power, reaction time and temperature would affect the morphology and size of the obtained crystal. Under the optimal conditions, U-CD-MOF had a cubic structure with uniform size of 8.60 ± 1.95 µm. U-CD-MOF was used to load the antibacterial natural product CA to form the composite (CA@U-CD-MOF) and the loading rate of CA@U-CD-MOF to CA could reach 19.63 ± 2.53%, which was more than twice that of γ-CD. Various techniques were applied to characterize the synthesized crystal, including Powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and N2 adsorption. In addition, antibacterial tests were performed on the obtained crystal. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CA@U-CD-MOF for Escherichia coli O157: H7 (E. coli O157: H7) were both 25 mg·mL-1, and the MIC for Staphylococcus aureus (S. aureus). was 25 mg·mL-1. The sustained release behavior of CA@U-CD-MOF to CA in ethanol fitted well to Higuchi model and the loading of CA was supported by molecular docking results. In general, U-CD-MOF was successfully achieved by ultrasound-assisted rapid synthesis and the obtained crystal was further evaluated for potential antibacterial application.

Nanobody and Nanozyme-Enabled Immunoassays with Enhanced Specificity and Sensitivity.

Immunoassay as a rapid and convenient method for detecting a variety of targets has attracted tremendous interest with its high specificity and sensitivity. Among the commonly used immunoassays, enzyme-linked immunosorbent assay has been widely used as a gold standard method in various fields that consists of two main components including a recognition element and an enzyme label. With the rapid advances in nanotechnology, nanobodies and nanozymes enable immunoassays with enhanced specificity and sensitivity compared with conventional antibodies and natural enzymes. This review is focused on the applications of nanobodies and nanozymes in immunoassays. Nanobodies advantage lies in their small size, high specificity, mass expression, and high stability. Nanozymes with peroxidase, phosphatase, and oxidase activities and their applications in immunoassays are highlighted and discussed in detail. In addition, the challenges and outlooks in terms of the use of nanobodies and the development of novel nanozymes in practical applications are discussed.

Polydimethylsiloxane Membranes Incorporating Metal-Organic Frameworks for the Sustained Release of Antibacterial Agents.

Cyclodextrin metal-organic frameworks (CD-MOFs) possess great potential in environmental applications due to their high specific surface area and good biocompatibility properties. However, the hydrophilicity of the CD-MOF hinders its ability to maintain a sustained release in water as a carrier. In this study, we prepared a CD-MOF that has codelivery ability for both phytochemicals [caffeic acid (CA)] and silver nanoparticles (Ag NPs) and further incorporated this material (CA@Ag@CD-MOF) into the polydimethylsiloxane (PDMS) matrix to construct a hybrid membrane. This hybrid membrane could effectively maintain the release capacity of the CD-MOF in water, while endowing PDMS with swelling ability in water. The hybrid membrane can achieve a sustained release for up to 48 h in water. In addition, the elastic modulus of the hybrid membrane increases by nearly 100%, and the swelling degree of the hybrid membrane in water increases by 42% compared with that of the pure PDMS membrane, indicating better mechanical properties. The hybrid membrane exhibits excellent antibacterial effects on Escherichia coli O157:H7 (E. coli O157:H7) and Staphylococcus aureus (S. aureus). We expect that this work will be beneficial to the delivery research of the CD-MOF in more environmental scenarios, especially in water treatment.

A New Strategy for Microbial Taxonomic Identification through Micro-Biosynthetic Gold Nanoparticles and Machine Learning.

Microorganisms can serve as biological factories for the synthesis of inorganic nanomaterials that can become useful as nanocatalysts, energy-harvesting-storage components, antibacterial agents, and biomedical materials. Herein, the development of biosynthesis of inorganic nanomaterials into a simple, stable, and accurate strategy for distinguishing microorganisms from multiple classification levels (i.e., kingdom, order, genus, and species) without gene amplification, biochemical testing, or target recognition is reported. Gold nanoparticles (AuNPs) biosynthesized by different microorganisms differ in color of the solution, and their features can be characterized, including the particle size, the surface plasmon resonance (SPR) spectrum, and the surface potential. The inter-relation between the features of micro-biosynthetic AuNPs and the classification of microorganisms are exploited at different levels through machine learning to establish a taxonomic model. This model agrees well with traditional classification methods that offers a new strategy for microbial taxonomic identification. The underlying mechanism of this strategy is related to the biomolecules produced by different microorganisms including glucose, glutathione, and nicotinamide adenine dinucleotide phosphate-dependent reductase that regulate the features of micro-biosynthetic AuNPs. This work broadens the application of biosynthesis of inorganic materials through micro-biosynthetic AuNPs and machine learning, which holds great promise as a tool for biomedical research.

Dietary exposure of copper and zinc oxides nanoparticles affect the fitness, enzyme activity, and microbial community of the model insect, silkworm Bombyx mori.

Copper and Zinc oxides nanoparticles (CuO and ZnO NPs, respectively) are among the most produced and commonly used engineered nanomaterials. They can be released into the environment, thereby causing health concerns and risks to biodiversity that indicate a need to evaluate their toxicological effects in a complex situation. Here, we used the insect model organism silkworm Bombyx mori to address the concerns about the biological effects associated with dietary exposure of CuO and ZnO NPs. ICP-MS analysis revealed significant accumulation of Cu and Zn (the latter being more accumulated) in silkworms' tissues (gut, fat body, silk gland, and malpighian tubule), and some elimination through feces in the respective NPs-exposed groups. NPs-exposures led to a decrease in larval body mass, survivorship, and cocoon production, where the effects of ZnO NPs were more pronounced. We also found that NPs-exposure induced gene expression changes (Attacin, lysozyme, SOD, and Dronc) and altered the activities of antioxidant enzymes (SOD, GST, and CAT), as well as impaired nutrient metabolism (alpha-amylase). Given their antibacterial property, CuO and ZnO NPs decreased species richness and diversity of the gut bacterial community and shifted their configuration to overt microbiome i.e., decreased abundance of probiotics (e.g., Acetobacter) and increased pathobionts (e.g., Pseudomonas, Bacillus, Escherichia, Enterococcus, Ralstonia, etc.) proportions. Overall, this integrated study revealed the unintended negative effects of CuO and ZnO NPs on silkworms and highlighted the potential to inevitably affect all living things due to intensive and possible mishandling of nanomaterials.

Covalent Organic Framework-Incorporated Nanofibrous Membrane as an Intelligent Platform for Wound Dressing.

Covalent organic frameworks (COFs) possess fascinating features that have sparked increasing interest as drug carriers in biomedical applications. However, the promising properties of COFs in wound healing have rarely been reported. Herein, a facile one-pot method is reported to prepare a curcumin-loaded COF (CUR@COF) by the condensation reaction and the Schiff base reaction and to further incorporate CUR@COF into polycaprolactone (PCL) nanofibrous membranes (CUR@COF/PCL NFMs) through electrospinning to develop a pH-triggered drug release platform for wound dressing. CUR@COF has a high CUR loading capacity of 27.68%, and CUR@COF/PCL NFMs exhibit increased thermal stability, improved mechanical properties, good biocompatibility, and enhanced antibacterial and antioxidant activities. More importantly, CUR@COF-based membranes show a pH-responsive CUR release profile by protonation under acidic conditions, suggesting the promotion of CUR release from membranes under an acidic extracellular microenvironment. The histopathological analysis and immunofluorescence staining of an in vivo skin defect model indicate that CUR@COF/PCL NFMs can accelerate wound healing and skin regeneration by reducing the expression of inflammatory factors (TNF-α) and enhancing the expression of angiogenesis (VEGF). This work provides a new strategy by employing COF-based drug-encapsulated nanocomposites for wound dressing applications.

A bio-inspired plasmonic nanosensor for angiotensin-converting enzyme through peptide-mediated assembly of gold nanoparticles.

Angiotensin-converting enzyme (ACE) can indicate blood pressure that relates to human health such as the cardiovascular disease. However, current methods are not competent to detect the ACE activity in a rapid and straightforward way. Plasmonic biosensors built on the modulation of metallic nanomaterials have emerged as novel tools for the detection of biomarkers. In this work, we report a bio-inspired strategy for the plasmonic detection of ACE in a rapid, sensitive, and selective way through peptide-mediated assembly of gold nanoparticles (AuNPs). In this biosensor, cysteine-angiotensin I-cysteine can assemble and aggregate AuNPs due to the Au-S bond. The presence of ACE can specifically catalyze the hydrolysis of angiotensin I, thus dissociating the cysteine-cysteine structure of the peptide that results in the disassembly and dispersion of AuNPs. This bio-inspired plasmonic nanosensor enables naked-eyed readout of ACE detection with great selectivity and high sensitivity with a LOD of 0.40 mU/mL. It also allows for the screening of ACE inhibitors and inhibitory peptides for the development of antihypertensive drugs or food. The biosensing technique developed in this work provides a new plasmonic approach that holds great promise as a point-of-care platform for biomedical diagnostics and the food industry.

Horseradish peroxidase-catalyzed formation of polydopamine for ultra-sensitive magnetic relaxation sensing of aflatoxin B1.

Aflatoxin B1 as one of the most toxic mycotoxins poses a major health risk to humans and animals. Highly sensitive detection methods of aflatoxin B1 are urgently required because of its low abundance in biological samples. In this work, we developed a magnetic relaxation sensing strategy using enzyme-catalyzed formation of polydopamine for signal amplification. Horseradish peroxidase can catalyze the reaction to generate polydopamine that assembles magnetic nanoparticles for magnetic relaxation sensing with a high signal-to-noise ratio. Combined with the specific antigen-antibody interaction, this magnetic sensor enables fast and ultra-sensitive detection of aflatoxin B1 by using transverse relaxation time (T2) as a readout. Under optimized conditions, the linear range of this magnetic sensor for detecting aflatoxin B1 is from 10 pg/mL to 10 ng/mL, and the limit of detection is 0.35 pg/mL. This sensor has been challenged for the quantitative analysis of aflatoxin B1 in animal feed samples that is promising for real-world applications.