A colorimetric sensor for rapid discrimination of tea polyphenols and tea authentication based on Rh-decorated Pd nanocubes with high peroxidase-like activity.

The rapid development of nanozymes has offered substantial opportunities for the fields of biomedicine, chemical sensing, and food safety. Among these applications, multichannel sensors, with the capability of simultaneously detecting multiple target analytes, hold promise for the practical application of nanozymes in chemical sensing with high detection efficiency. In this study, Rh-decorated Pd nanocubes (Pd-Rh nanocubes) with significantly enhanced peroxidase-like activity are synthesized through the mediation of underpotential deposition (UPD) and subsequently employed to develop a multichannel colorimetric sensor for discriminating tea polyphenols (TPs) and tea authentication. Based on a single reactive unit of efficient catalytic oxidation of 3,3',5,5'-tetramethylbenzidine dihydrochloride (TMB), the nanozyme-based multichannel colorimetric sensor responds to each analyte in as short as 1 min. With the aid of principal component analysis (PCA) and hierarchical cluster analysis (HCA), various TPs and types of tea can be accurately identified. This work not only provides a new type of simply structured and highly active nanozymes but also develops a concise and rapid multichannel sensor for practical application in tea authentication and quality inspection.

Insight into the interaction and binding mechanism of a natural nonnutritive sweetener mogroside V with soybean protein isolates based on multi-spectroscopic techniques and computational simulations.

As a natural low-calorie sweetener, Mogroside V (Mog-V) has gradually become one of the alternatives to sucrose with superior health attributes. However, Mog-V will bring unpleasant aftertastes when exceeding a threshold concentration. To investigate the possibility of soy protein isolates (SPIs), namely ß-conglycinin (7S), and glycinin (11S) as flavor-improving agents of Mog-V, the binding mechanism between Mog-V and SPIs was explored through multi-spectroscopy, particle size, zeta potential, and computational simulation. The results of the multi-spectroscopic experiments indicated that Mog-V enhanced the fluorescence of 7S/11S protein in a static mode. The binding affinity of 7S-Mog-V was greater compared with 11S-Mog-V. Particle size and zeta potential analysis revealed that the interaction could promote aggregation of 7S/11S protein with different stability. Furthermore, computational simulations further confirmed that Mog-V could interact with the 7S/11S protein in different ways. This research provides a theoretical foundation for the development and application of SPI to improve the flavor of Mog-V, opening a new avenue for further expanding the market demand for Mog-V.

Single-step batch fabrication of microfluidic paper-based analytical devices with a 3D printer and their applications in nanoenzyme-enhanced visual detection of dopamine.

Wax printing is the most widely used method for fabricating microfluidic paper-based analytical devices (µPADs), but it still suffers from disadvantages like discontinuation of wax printers and need for additional equipment for heating treatment. To address these issues, this work initially describes a new class of wax printing approach for high-precision, batch fabrication of µPADs using a household 3D printer. It only involves a one patterning step of printing polyethylene wax into rice paper body. Under optimized parameters, a fabrication resolution, namely the minimum hydrophilic channel width, down to ~189 ± 30 µm could be achieved. In addition, the analytical applicability of such polyethylene wax-patterned µPADs was demonstrated well with enhanced colorimetric detection of dopamine as a model analyte by combining metal-organic framework (MOF) based nanoenzymes (ZIF-67) with a smartphone (for portable quantitative readout). The developed nanosensor could linearly detect dopamine over a concentration range from 10 to 1000 µM, with a detection limit of ca. 2.75 µM (3σ). The recovery results for analyzing several real samples (i.e., pig feed, chicken feed, pork and human serum) were between 91.82 and 102.79%, further validating its good detection accuracy for potential practical applications in food safety and medical diagnosis.

Simple, sensitive, colorimetric detection of pyrophosphate via the analyte-triggered decomposition of metal-organic frameworks regulating their adaptive multi-color Tyndall effect.

This paper describes initially the application of the Tyndall effect (TE) of metal-organic framework (MOF) materials as a colorimetric signaling strategy for the sensitive detection of pyrophosphate ion (PPi). The used MOF NH2-MIL-101(Fe) was prepared with Fe3+ ions and fluorescent ligands of 2-amino terephthalic acid (NH2-BDC). The fluorescence of NH2-BDC in MOF is quenched due to the ligand-to-metal charge transfer effect, while the NH2-MIL-101(Fe) suspension shows a strong TE. In the presence of PPi analyte, the MOFs will undergo decomposition because of the competitive binding of Fe3+ by PPi over NH2-BDC, resulting in a significant decrease in the TE signal and fluorescence restoration from the released ligands. The results demonstrate that the new method only requires a laser pointer pen (for TE creation) and a smartphone (for portable quantitative readout) to detect PPi in a linear concentration range of 1.25-800 µM, with a detection limit of ~210 nM (3σ) which is ~38 times lower than that obtained from traditional fluorescence with a spectrophotometer (linear concentration range, 50-800 µM; detection limit, 8.15 µM). Moreover, the acceptable recovery of PPi in several real samples (i.e., pond water, black tea, and human serum and urine) ranges from 97.66 to 119.15%.

LC-QTOF/MS-based non-targeted metabolomics to explore the toxic effects of di(2-ethylhexyl) phthalate (DEHP) on Brassica chinensis L.

Di(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer known to pose health risks to humans upon exposure. Recognizing the toxic nature of DEHP, our study aimed to elucidate the response mechanisms in Brassica chinensis L. (Shanghai Qing) when subjected to varying concentrations of DEHP (2 mg kg-1, 20 mg kg-1, and 50 mg kg-1), particularly under tissue stress. The findings underscored the substantial impact of DEHP treatment on the growth of Brassica chinensis L., with increased DEHP concentration leading to a notable decrease in chlorophyll levels and alterations in the content of antioxidant enzyme activities, particularly superoxide dismutase (SOD) and peroxidase (POD). Moreover, elevated DEHP concentrations correlated with increased malondialdehyde (MDA) levels. Our analysis detected a total of 507 metabolites in Brassica chinensis L., with 331 in shoots and 176 in roots, following DEHP exposure. There was a significant difference in the number of metabolites in shoots and roots, with 79 and 64 identified, respectively (VIP > 1, p < 0.05). Metabolic pathway enrichment in Brassica chinensis L. shoots revealed significant perturbations in valine, leucine, and isoleucine biosynthesis and degradation, aminoacyl-tRNA, and glucosinolate biosynthesis. In the roots of Brassica chinensis L., varying DEHP levels exerted a substantial impact on the biosynthesis of zeatin, ubiquinone terpenoids, propane, piperidine, and pyridine alkaloids, as well as glutathione metabolic pathways. Notably, DEHP's influence was more pronounced in the roots than in the shoots, with higher DEHP concentrations affecting a greater number of metabolic pathways. This experimental study provides valuable insights into the molecular mechanisms underlying DEHP-induced stress in Brassica chinensis L., with potential implications for human health and food safety.

Varietal Authenticity Assessment of QTMJ Tea Using Non-Targeted Metabolomics and Multi-Elemental Analysis with Chemometrics.

In this paper, a combination of non-targeted metabolomics and multi-element analysis was used to investigate the impact of five different cultivars on the sensory quality of QTMJ tea and identify candidate markers for varietal authenticity assessment. With chemometric analysis, a total of 54 differential metabolites were screened, with the abundances significantly varied in the tea cultivars. By contrast, the QTMJ tea from the Yaoshan Xiulv (XL) monovariety presents a much better sensory quality as result of the relatively more abundant anthocyanin glycosides and the lower levels of 2'-o-methyladenosine, denudatine, kynurenic acid and L-pipecolic acid. In addition, multi-elemental analysis found 14 significantly differential elements among the cultivars (VIP > 1 and p < 0.05). The differences and correlations of metabolites and elemental signatures of QTMJ tea between five cultivars were discussed using a Pearson correlation analysis. Element characteristics can be used as the best discriminant index for different cultivars of QTMJT, with a predictive accuracy of 100%.

Impact of storage time on non-volatile metabolites and fungal communities in Liupao tea using LC-MS based non-targeted metabolomics and high-throughput sequencing.

Long-term storage of Liupao tea (LPT) is usually believed to enhance its quality and commercial value. The non-volatile metabolites variations and the fungal succession play a key role for organoleptic qualities during the storage procedure. To gain in-depth understanding the impact of storage time on the quality of LPT, two different brands of LPT with different storage time, including Maosheng LPTs (MS) with 0, 5, 10 and 15 years and Tianyu LPTs (TY) with 0, 3, 5, 8 and 10 years, were resorted to investigate the changes of non-volatile metabolites and fungi as well as their correlation by multi-omics. A total of 154 and 119 differential metabolites were identified in these two different brands of MS and TY, respectively, with the aid of high-performance liquid chromatography with quadrupole-time-of-flight mass spectrometry. In both categories of LPTs, the transformation of differential metabolites in the various stages referred to the formation of alkaloids, increase of organic acids, biosynthesis of terpenoids as well as glycosylation and methylation of flavonoids. Thereinto, glycosylation and methylation of flavonoids were the critical stages for distinguishing MS and TY, which were discovered in MS and TY stored for about 10 and 8 years, respectively. Moreover, the results of high-throughput sequencing showed that the key fungal genera in the storage of LPTs consisted of Eurotium, Aspergillus, Blastobotrys, Talaromyces, Thermomyces and Trichomonascus. It was confirmed on the basis of multivariate analysis that the specific fungal genera promoted the transformation of metabolites, affecting the tea quality to some extent. Therefore, this study provided a theoretical basis for the process optimization of LPT storage.

A comprehensive review of the current trends and recent advancements on the authenticity of honey.

The authenticity of honey currently poses challenges to food quality control, thus requiring continuous modernization and improvement of related analytical methodologies. This review provides a comprehensively overview of honey authenticity challenges and related analytical methods. Firstly, direct and indirect methods of honey adulteration were described in detail, commenting the existing challenges in current detection methods and market supervision approaches. As an important part, the integrated metabolomic workflow involving sample processing procedures, instrumental analysis techniques, and chemometric tools in honey authenticity studies were discussed, with a focus on their advantages, disadvantages, and scopes. Among them, various improved microscale extraction methods, combined with hyphenated instrumental analysis techniques and chemometric data processing tools, have broad application potential in honey authenticity research. The future of honey authenticity determination will involve the use of simplified and portable methods, which will enable on-site rapid detection and transfer detection technologies from the laboratory to the industry.

Quantitative colorimetric sensing of heavy metal ions via analyte-promoted growth of Au nanoparticles with timer or smartphone readout.

This work describes two new colorimetric nanosensors for label-free, equipment-free quantitative detection of nanomolar copper (II) (Cu2+) and mercury (II) (Hg2+) ions. Both are based on the analyte-promoted growth of Au nanoparticles (AuNPs) from the reduction of chloroauric acid by 4-morpholineethanesulfonic acid. For the Cu2+ nanosensor, the analyte can accelerate such a redox system to rapidly form a red solution containing dispersed, uniform, spherical AuNPs that is related to these particles' surface plasmon resonance property. For the Hg2+ nanosensor, on the other hand, a blue mixture consisting of aggregated, ill-defined AuNPs with various sizes can be created, showing a significantly enhanced Tyndall effect (TE) signal (in comparison with that produced in the red solution of AuNPs). By using a timer and a smartphone to quantitatively measure the time of producing the red solution and the TE intensity (i.e., the average gray value of the corresponding image) of the blue mixture, respectively, the developed nanosensors are well demonstrated to achieve linear ranges of 6.4 nM to 100 µM and 6.1 nM to 1.56 µM for Cu2+ and Hg2+, respectively, with detection limits down to 3.5 and 0.1 nM, respectively. The acceptable recovery results obtained from the analysis of the two analytes in the complex real water samples including drinking water, tap water, and pond water ranged from 90.43 to 111.56%.

A bimetallic (Ni/Co) metal-organic framework with excellent oxidase-like activity for colorimetric sensing of ascorbic acid.

A novel nanozyme of bimetallic (Ni/Co) metal-organic framework (Ni/Co-MOF) was synthesized using a simultaneous precipitation and acid etching method with a zeolitic imidazolate framework ZIF-67 as the template. The as-synthesized Ni/Co-MOF catalyst presented a three-dimensional hollow nanocage structure and exhibited excellent intrinsic oxidase-like activity. It was demonstrated that Ni/Co-MOF could directly catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to produce a blue product (oxidized TMB, oxTMB) in the absence of H2O2. The mechanisms and kinetics of this nanozyme activity were investigated, and it was determined that the catalytic activity of Ni/Co-MOF was closely related to temperature and solution pH. Owing to its strong reducibility, ascorbic acid (AA) could reduce oxTMB, and the blue color of the reaction mixture faded over time. Therefore, a novel colorimetric platform was constructed to detect AA based on the oxidase-like activity of Ni/Co-MOF. Under optimal conditions, the absorbance of ox-TMB at 652 nm decreased linearly over the 0.015-50 µM AA range with a detection limit of 0.004 µM.

Analyte-triggered in situ "off-on" of Tyndall effect for smartphone-based quantitative nanosensing of Ag+ ions.

This work describes two new colorimetric methods for smartphone-based point-of-care nanosensing of toxic Ag+ ions. They were based on the analyte-triggered in situ "off-on" of Tyndall effect (TE) of non-plasmonic colloid or plasmonic metal nanoprobes. The first TE-inspired assay (TEA) focused on the initial analytical application of precipitation reactions where a non-plasmonic AgCl colloid could be formed once mixing the analyte with a NaCl solution. Such AgCl colloid displayed strong visual TE signals after their irradiation by a laser pointer pen, which unexpectedly achieved a detection limit of ~ 400 nM. The second TEA was further designed to reduce the limit down to ~ 78 nM using the analyte's oxidizability towards 3,3',5,5'-tetramethylbenzidine molecules. The redox reaction could create positively charged products that could make negatively charged plasmonic gold nanoparticles aggregate through electrostatic interactions to remarkably amplify their TE responses. Both limits were lower than the minimum allowable Ag+ level (~ 460 nM) in drinking water issued by the World Health Organization. The satisfactory recovery results for detecting Ag+ ions in river, pond, tap, and drinking water additionally demonstrated good selectivity, accuracy and practicality of the proposed methods for potential point-of-need uses in environmental analysis, public health, water safety, etc.

Investigation of the binding behavior of bioactive 7-methoxyflavone to human serum albumin by coupling multi-spectroscopic with computational approaches.

The natural flavonoids with bioactivity as secondary plant metabolites are mostly found in fruits, vegetables, tea and herbs, the distribution and bioavailability of which in vivo depends on the interaction and successive binding with carrier proteins in the systemic circulation. In this paper, the binding behavior of bioactive 7-methoxyflavone (7-MF) with human serum albumin (HSA) was studied with the aid of the combination of multi-spectroscopic methods, molecular docking and molecular dynamic simulation. The results of multi-spectroscopic experiments revealed that 7-MF interacted with HSA predominantly via fluorescence static quenching and the microenvironment around the fluorophore Trp residues in HSA became more hydrophilicity with the binding of 7-MF. Thermodynamic analysis demonstrated that hydrogen bonds and van der Waals forces played a dominant role in stabilizing the HSA-7-MF complex. Moreover, the docking experiment and molecular dynamic simulation further confirmed that 7-MF could enter the active cavity of HSA and caused more stable conformation and change of secondary structure of HSA through forming hydrogen bond. The exploration of the mechanism of 7-MF binding to HSA lights a new avenue to understand the stability, transport and distribution of 7-MF and 7-MF may hold great potential to be extended as a promising alternative of dietary supplements or pharmaceutical agents.

Metabolomic Analysis Reveals the Effect of Insecticide Chlorpyrifos on Rice Plant Metabolism.

Pesticides as important agricultural inputs play a vital role in protecting crop plants from diseases and pests; however, the effect of pesticides on crop plant physiology and metabolism is still undefined. In this study, the effect of insecticide chlorpyrifos at three doses on rice plant physiology and metabolism was investigated. Our results revealed that chlorpyrifos cause oxidative stress in rice plants and even inhibit plant growth and the synthesis of protein and chlorophyll at high doses. The metabolomic results suggested that chlorpyrifos could affect the metabolic profiling of rice tissues and a total of 119 metabolites with significant changes were found, mainly including organic acids, amino acids, lipids, polyphenols, and flavonoids. Compared to the control, the content of glutamate family amino acids were significantly disturbed by chlorpyrifos, where defense-related proline and glutathione were significantly increased; however, glutamic acid, N-acetyl-glutamic acid and N-methyl-glutamic acid were significantly decreased. Many unsaturated fatty acids, such as linolenic acid and linoleic acid, and their derivatives lysophospholipids and phospholipids, were significantly accumulated in chlorpyrifos groups, which could act as osmolality substances to help rice cells relieve chlorpyrifos stress. Three organic acids, aminobenzoic acid, quinic acid, and phosphoenolpyruvic acid, involved in plant defenses, were significantly accumulated with the fold change ranging from 1.32 to 2.19. In addition, chlorpyrifos at middle- and high-doses caused the downregulation of most flavonoids. Our results not only revealed the effect of insecticide chlorpyrifos on rice metabolism, but also demonstrated the value of metabolomics in elucidating the mechanisms of plant responses to stresses.

Tyndall-effect-based colorimetric assay with colloidal silver nanoparticles for quantitative point-of-care detection of creatinine using a laser pointer pen and a smartphone.

Herein, this paper initially reports a new colorimetric Tyndall effect-inspired assay (TEA) for simple, low-cost, sensitive, specific, and point-of-care detection of creatinine (an important small biomolecule) by making use of silver nanoparticles (AgNPs) as model colloidal nanoprobes for visual light scattering signaling. The naked-eye TEA method adopts negatively-charged citrate-capped AgNPs (Cit-AgNPs) prepared by sodium citrate reduction. In the presence of alkaline conditions, the creatinine analyte can form carbanion/oxoanion amino tautomers which in turn crosslink with carboxylate groups on the Cit-AgNPs via a hydrogen bonding network to mediate the aggregation of such colloidal nanoprobes showing a significantly-enhanced TE signal that was created and quantified by a hand-held laser pointer pen and a smartphone, respectively. The results demonstrate that the resulting equipment-free method with the TE readout could enable the portable quantification of creatinine with a detection limit of ∼55 nM, which was ∼90-2334 times lower than that obtained from AgNP-based colorimetric approaches with the most common localized surface plasma resonance signaling. Moreover, it shows a larger analytical sensitivity up to ∼580.8227 signal per nM, offering ∼2.4-232-fold improvement in comparison with many of the recent instrumental creatinine nanosensors. The accuracy and practicality of the developed nanosensing system was additionally confirmed with satisfactory recovery results ranging from ca. 98.52 to 100.36% when analyzing a set of real complex human urine samples.

Effects of chlorpyrifos on the metabolic profiling of Bacillus megaterium strain RRB.

Many microorganisms have been reported to degrade organic pollutants in the environment and plants, however, the specific information about the effect of organic pollutants on the metabolism of microorganisms is poorly investigated. In the present study, the effect of the pesticide chlorpyrifos on the metabolic profiling of Bacillus megaterium strain RRB was investigated using metabolomics. Our data show that chlorpyrifos acting as an energy source was readily concentrated in the strain RRB from the culture medium. During early cultivation, the shift in energy sources from tryptic soy broth to chlorpyrifos may temporarily cause the strain RRB to enter the starvation stage, where some synthesis-related amino acids and intermediates in the pathways of TCA cycle and pyridoxine metabolism were decreased. The increase of nucleotides and lysine may help the strain RRB cope with the starvation stage. During later cultivation, many metabolites including organic acids, nucleosides and sugar phosphates were gradually accumulated, which indicates that chlorpyrifos could be utilized by the stain RRB to generate metabolites bacteria needed. In addition, arginine acting as a nitrogen-storage amino acid was gradually decreased with later cultivation, suggesting that chlorpyrifos could not provide enough nitrogen for bacteria.

Tyndall-Effect-inspired assay with gold nanoparticles for the colorimetric discrimination and quantification of mercury ions and glutathione.

This work initially reports a new nanosening method for simple, sensitive, specific, visual detection of mercury (II) (Hg2+) and glutathione (GSH) using the Tyndall Effect (TE) of the same colloidal gold nanoparticle (GNP) probes for efficient colorimetric signaling amplification. For the TE-inspired assay (TEA) method, arginine (Arg) molecules are pre-modified on the GNPs' surfaces (Arg-GNPs). Upon the Hg2+ introduction, it can be specifically coordinated with the terminal -NH2 and -COOH groups of the Arg molecules to make the Arg-GNPs aggregate, producing a significantly-enhanced TE signal in the reaction solution after its irradiation by a 635-nm red laser pointer pen. On the other hand, the introduction of the GSH results in the production of the original Arg-GNPs' weak TE response, as it is able to bind such metal ion via mercury-thiol reactions to inhibit the above aggregation. Under the optimal conditions, the utility of the new TEA method is well demonstrated to quantitatively detect the Hg2+ and GSH with the aid of a smartphone as a portable TE reader during the linear concentration ranges of 50-3000 and 10-3000 nM, respectively. The detection limits for the Hg2+ and GSH are estimated to be as low as ∼3.5 and ∼0.3 nM, respectively. The recovery results obtained from the detection of Hg2+ in the complex tap and pond water samples and the assay of GSH in real human serum and urine samples are also satisfactory.

Ultrasensitive visual detection of Hg2+ ions via the Tyndall effect of gold nanoparticles.

This work reports a new methodology for naked-eye nanosensing of Hg2+ where the Tyndall effect of gold nanoparticles (GNPs) acts as a light scattering signalling readout. Its utility is demonstrated with ultrasensitive detection of the target with a limit down to 0.13 nM (∼5461-fold sensitivity improvement over conventional GNP-based methods with surface plasmon resonance signalling).

Enzymatic dephosphorylation-triggered self-assembly of DNA amphiphiles.

In this work, phosphorylated lipid-conjugated oligonucleotide (DNA-lipid-P) has been synthesized to develop an enzyme-responsive self-assembly of DNA amphiphiles based on dephosphorylation-induced increase of hydrophobicity. Since elevated ALP level is a critical index in some diseases, ALP-triggered self-assembly of DNA amphiphiles shows promise in disease diagnosis and cancer treatment.

A novel strategy for sensitive and rapid detection of ascorbic acid via the Tyndall effect of cobalt hydroxide nanoflakes.

Cobalt oxyhydroxide (CoOOH) nanoflakes, as nanoenzymes and fluorescence quenchers, have been widely used in colorimetric and fluorescent analysis. However, their promising light scattering property-the Tyndall effect (TE)-has never been applied in biosensors and biological analysis to date. Herein, we report for the first time a novel strategy for point-of-care detection of ascorbic acid (AA) with the TE of CoOOH nanoflakes providing colorimetric signaling. In this detection system, CoOOH nanoflakes exhibit a strong red TE signal under the illumination of a hand-held 635 nm laser pointer pen. However, the introduction of AA could induce a significant decrease of the TE because it could reduce CoOOH into Co2+ and results in the degradation of the CoOOH nanoflakes. The changes in the TE intensity could be read-out using a smartphone for the portable quantitative analysis of AA. The results showed that this CoOOH nanoflake-based TE-inspired assay (TEA) exhibited a good linear range from 0.25 µM to 40 µM for AA, with a detection limit of 12 nM. It also showed high selectivity toward AA over common potential interfering species. Importantly, this method possessed the advantages of simple operation, low consumption of time and equipment-free analysis and was successfully applied to the detection of AA in vitamin C tablets.

On-site, rapid and visual method for nanomolar Hg2+ detection based on the thymine-Hg2+-thymine triggered "double" aggregation of Au nanoparticles enhancing the Tyndall effect.

This work describes a new nanosensor for the simple, rapid, portable, colorimetric analysis of mercury(ii) (Hg2+) ions by combining the sensitive Tyndall effect (TE) of colloidal Au nanoparticles (AuNPs) with specific thymine-Hg2+-thymine (T-Hg2+-T) coordination chemistry for the first time. For the TE-inspired assay (TEA), in the presence of Hg2+ in a sample, the analyte can selectively mediate the hybridization of three types of flexible single-stranded DNAs (ssDNAs) to form stable rigid double-stranded DNAs (dsDNAs) via the T-Hg2+-T ligand interaction. Subsequent self-assembly of the dsDNAs with terminal thiol groups on the AuNPs' surfaces led to their "double" aggregation in addition to the lack of sufficient ssDNAs as the stabilizing molecules in a high-salt solution, resulting in a remarkably enhanced TE signal that positively relied on the Hg2+ level. The results demonstrated that such a TEA method enabled rapid naked-eye qualitative analysis of 625 nM Hg2+ within 10 min with an inexpensive laser pointer pen as an inexpensive handheld light source to generate the TE response. Making use of a smartphone for portable TE readout could further quantitatively detect the Hg2+ ions in a linear concentration range from 156 to 2500 nM with a limit of detection as low as 25 nM. Moreover, the developed equipment-free nanosensor was also used to analyze the Hg2+ ions in real samples including tap water, drinking water, and pond water, the obtained recoveries were within the range of 93.68 to 108.71%. To the best of our knowledge, this is the first report of using the AuNPs and functional nucleic acids to design a TE-based biosensor for the analysis of highly toxic heavy metal ions.