A review on metal nanozyme-based sensing of heavy metal ions: Challenges and future perspectives.

Large scale mining, manufacturing industries, exploitation of underground water, depletion of groundwater level, and uncontrolled discharge of industrial wastes have caused severe heavy metal ion pollution to the environment throughout the world. Therefore, the rapid detection of such toxic metal ions is inevitable. However, conventional methods require sophisticated instruments and skilled manpower and are difficult to operate in on-field conditions. Recently, metal nanozyme-based assays have been found to have the potential as an alternative to conventional methods due to their portability, simplicity, and high sensitivity to detect metal ion concentration to as low as parts per trillion (ppt). Metal nanozyme-based systems for heavy metal ions enable rapid and cheap screening on the spot with a very simple instrument such as a UV-vis absorption spectrophotometer and therefore, are convenient for use in field operations, especially in remote parts of the world. The sensing mechanism of a nanozyme-based sensor is highly dependent on its surface properties and specific interactions with particular metal ion species. Such method often encounters selectivity issues, unlike natural enzyme-based assays. Therefore, in this review, we mainly focus our discussion on different types of target recognition and inhibition/enhancement mechanisms, and their responses toward the catalytic activity in the sensing of target metal ions, design strategies, challenges, and future perspectives.

Platinum ions mediate the interactions between DNA and carbon quantum dots: diagnosis of MRSA infections.

In this study, we have developed a rapid and cost-effective method employing platinum ion (Pt4+)-capped fluorescent carbon quantum dots (CQDs) coupled with loop-mediated isothermal amplification (LAMP) to detect dual MRSA genes. We synthesized nitrogen- and chlorine-co-doped fluorescent CQDs (CQDSPDs) from spermidine trihydrochloride via a simple one-step pyrolysis. The CQDSPDs capped with Pt4+ ions through the cooperative coordination of the amine and chlorine groups on the surface of CQDs facilitated the double-stranded DNA (dsDNA)-induced fluorescence quenching of CQDs, and enabled the construction of the CQDSPDs/Pt4+ probe for the detection of as few as 10 copies of the MRSA gene (mecA and femA). The sensitivity and specificity of the CQDSPDs/Pt4+ probe for MRSA detection in clinical specimens (n = 24) were 94% and 86%, respectively. Our results reveal that the CQDSPDs/Pt4+ probe has great potential for the diagnosis of antibiotic-resistant superbugs with high sensitivity, specificity, and agreement.

DNA engineered copper oxide-based nanocomposites with multiple enzyme-like activities for specific detection of mercury species in environmental and biological samples.

In this paper, we report the synthesis and application of enzyme-like DNA-copper oxide/platinum nanoparticles for the separate quantification of inorganic and organomercury species in various real samples. We synthesized a series of poly(thymine) (T60)-copper oxide/metal nanocomposites (T60-CuxO/M NCs; M = Au, Ag or Pt) that exhibited enzyme-like activities [oxidase (OX), peroxidase (POX), and catalase (CAT)]. The enzyme-like activities are tunable due to the incorporation of various metals into the NCs. Among a series of synthesized CuxO/M NCs, T60-copper oxide-platinum nanocomposites (T60-CuxO/Pt NCs) exhibited the highest OX-like activity via the O2-mediated oxidation of substrates, such as Amplex Red (AR), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), o-phenylenediamine (OPD), and 3,3',5,5'-tetramethylbenzidine (TMB), to form fluorescent or colored products. Interestingly, inorganic mercury ions (Hg2+) and organomercury species, such as methylmercury (MeHg+), ethylmercury (EtHg+), and phenylmercury (PhHg+), significantly inhibited the OX-like activity of T60-CuxO/Pt NCs. For the selective detection of mercury species, we used ABTS in the T60-CuxO/Pt NCs system, and the ABTS/T60-CuxO/Pt NCs-based assay allowed for the detection of mercury ions at nanomolar concentrations based on the decrease in the catalytic activity caused by the mercury ions. To separately quantify the inorganic and organomercury species in a sample, we employed selenium nanoparticles (Se NPs) as a masking agent, as they preferentially bind with inorganic mercury species. The ABTS/T60-CuxO/Pt NCs-based assay with the masking agent of Se NPs further provided specificity for the detection of organomercury species in environmental water samples (tap water, river water, and seawater) and fish muscle samples (dogfish muscle DORM-II).

Detection of urinary spermine by using silver-gold/silver chloride nanozymes.

In this paper, we report a simple one-step synthesis method for silver-gold bimetallic nanoparticles deposition on silver chloride nanosheets to form Ag-Au/AgCl nanohybrid with oxidase-like and peroxidase-like catalytic activity. We used these nanohybrid in the detection of spermine. First, 13 nm-sized Au NPs were synthesized by citrate reduction of HAuCl4 solution, and then, Ag+ ions were added to the solution without any purification. The added Ag+ reacted with the Cl- ions in the dispersion, thus immediately forming AgCl nanosheets through a precipitation reaction, and the aurophilic interactions with the Au NPs resulted in the formation and in situ self-deposition of Ag-Au NPs on the AgCl nanosheets at room temperature. We investigated the enzyme-mimicking activity of the Ag-Au/AgCl nanohybrid in detail via the O2- or H2O2-Amplex Red (AR) redox system. The Ag-Au/AgCl nanohybrid exhibited at least 150-fold higher catalytic activity than that of Ag-Au NPs or AgCl nanosheets, due to synergistic effect. Spermine inhibited the enzyme-mimic activity of the Ag-Au/AgCl nanohybrid, thereby allowing for the construction of a probe for detecting nanomolar concentrations of spermine in urine samples. This cost-effective sensing system was used to easily and rapidly detect the concentrations of spermine in complex urine samples.

Graphene oxide membrane as an efficient extraction and ionization substrate for spray-mass spectrometric analysis of malachite green and its metabolite in fish samples.

A graphene oxide (GO) nanosheet-modified N+-nylon membrane (GOM) has been prepared and used as an extraction and spray-ionization substrate for robust mass spectrometric detection of malachite green (MG), a highly toxic disinfectant in liquid samples and fish meat. The GOM is prepared by self-deposition of GO thin film onto an N+-nylon membrane, which has been used for efficient extraction of MG in aquaculture water samples or homogenized fish meat samples. Having a dissociation constant of 2.17 × 10-9 M-1, the GOM allows extraction of approximately 98% of 100 nM MG. Coupling of the GOM-spray with an ion-trap mass spectrometer allows quantitation of MG in aquaculture freshwater and seawater samples down to nanomolar levels. Furthermore, the system possesses high selectivity and sensitivity for the quantitation of MG and its metabolite (leucomalachite green) in fish meat samples. With easy extraction and efficient spray ionization properties of GOM, this membrane spray-mass spectrometry technique is relatively simple and fast in comparison to the traditional LC-MS/MS methods for the quantitation of MG and its metabolite in aquaculture products.

Visual detection of cyanide ions by membrane-based nanozyme assay.

In this paper, we report a simple one-step synthesis of well-dispersed amorphous cobalt hydroxide/oxide-modified graphene oxide (CoOxH-GO) possessing peroxidase-like catalytic activity, and its application for the detection of H2O2, glucose, and CN- ions. CoOxH is formed and deposited in situ on the GO surface through the reaction between GO (size ~ 240nm) and Co2+ in basic solution at room temperature. We investigated the enzyme-mimicking activity of the CoOxH-GO nanohybrid in detail via the H2O2-mediated oxidation of Amplex Red (AR) to form fluorescent resorufin. The peroxidase-like activity of CoOxH-GO is utilized herein for the quantitation of H2O2 in a wide concentration range, from 100nM to 100µM. When coupled with glucose oxidase (GOD), the AR/CoOxH-GO system can determine glucose level in blood samples. Interestingly, cyanide ions (CN-) significantly inhibit the catalytic activity of the CoOxH-GO nanohybrid, which allows for the construction of a probe for the detection of CN- in water samples and laboratory wastes. We fabricated a membrane-based CoOxH-GO probe for the visual detection of CN- by preparing a thin film of CoOxH-GO on a positively charged and porous nylon membrane (N+M). The CoOxH-GO/N+M operates on the principle that CN- inhibits the catalytic activity of CoOxH-GO towards the H2O2-mediated oxidation of AR to form reddish resorufin on the membrane. The intensity of the red color of the membrane decreases with increasing CN- concentration, which can be easily observed with the naked eye at the nanomolar level. This cost-effective sensing system allows for the rapid and simple determination of the concentrations of CN- in complicated wastewater samples.

Pulse laser-induced fragmentation of carbon quantum dots: a structural analysis.

Carbon quantum dots (CQDs) have attracted enormous interest in recent years owing to their low cytotoxicity, excellent biocompatibility and strong fluorescence. They have been successfully employed in sensor, bio-imaging, and drug carrier applications. A complete understanding of their core-surface structure is essential for tuning their physical and chemical properties for various applications. Conventional characterizations of CQDs are conducted with electron microscopy or spectroscopy, such as transmission electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy and Raman spectroscopy. However, these techniques cannot fully resolve the core-surface structure of CQDs. In this study, we attempt to analyze the structures of CQDs by laser desorption/ionization mass spectrometry (LDI-MS) using three model CQDs synthesized from citric acid (CA-CQDs), diammonium citrate (AC-CQDs) and spermidine trihydrochloride (Spd-CQDs). Both CA-CQDs and AC-CQDs produced anionic carbon cluster ions ([Cn]-, n = 4-9) during the laser desorption/ionization process. Additionally, AC-CQDs produced fragments containing C, N, and O that appeared at m/z values of 41.999, 91.015, and 107.008, which were identified by 15N isotopes as [CNO]-, [CH3N2O3]-, and [CH3N2O4]-, respectively. By contrast, subjecting Spd-CQDs to the same analysis did not yield carbon cluster ions ([Cn]-); instead, strong chlorine-associated ions with a unique isotopic pattern were observed, strongly implying that Spd-CQDs contain chlorine. The lack of carbon cluster ion formation in nitrogen- and chlorine-doped Spd-CQDs indicates that nitrogen and chlorine are abundantly and homogenously doped in the CQDs. We also found a shot-dependent fragmentation behavior for AC-CQDs that produces nitrogen- and oxygen-containing ions and carbon cluster ions ([Cn]-) during initial fragmentation of the surface, with a gradual destruction of the nanocrystalline carbon core after additional shots. These results suggest that LDI-MS can be used as a tool for analyzing the core-surface structure of CQDs, particularly when it contains a heteroatom doped carbon core with various surface functional groups containing nitrogen, oxygen and halogens.

Immobilization of aptamer-modified gold nanoparticles on BiOCl nanosheets: Tunable peroxidase-like activity by protein recognition.

A self-assembled nanocomposite is prepared from an aqueous mixture of aptamer-modified gold nanoparticles (Apt-Au NPs), bismuth ions and chloride ions. The Apt-Au NPs are immobilized on bismuth oxychloride (BiOCl) nanosheets in situ to form Apt-Au NPs/BiOCl nanocomposites. The as-prepared nanocomposites exhibit high peroxidase-like activity for the catalytic conversion of Amplex Red (AR) to fluorescent resorufin in the presence of H2O2. The catalytic activity of Apt-Au NPs/BiOCl nanocomposites is at least 90-fold higher than that of Apt-Au NPs or BiOCl nanosheets, revealing synergistic effects on their activity. The catalytic activity of Apt-Au NPs/BiOCl nanocomposites is suppressed by vascular endothelial growth factor-A165 (VEGF-A165) molecules that specifically interact with the aptamer units (Del-5-1 and v7t-1) on the nanocomposite surface. The AR/H2O2-Apt-Au NPs/BiOCl nanocomposites probe shows high selectivity (>1000-fold over other proteins) and sensitivity (detection limit ~0.5nM) for the detection of VEGF-A165. Furthermore, the probe is employed for the detection of VEGF isoforms and for the study of interactions between VEGF and VEGF receptors. The practicality of this simple, rapid, cost-effective probe is validated by the analysis of VEGF-A165 in cell culture media, showing its great potential for the analysis of VEGF in biological samples.

Selective Colorimetric Detection of Hydrogen Sulfide Based on Primary Amine-Active Ester Cross-Linking of Gold Nanoparticles.

Hydrogen sulfide (H2S) is a highly toxic environmental pollutant and also an important gaseous transmitter. Therefore, selective detection of H2S is very important, and visual detection of it with the naked eye is preferred in practical applications. In this study, thiolated azido derivates and active esters functionalized gold nanoparticles (AE-AuNPs)-based nanosensors have been successfully prepared for H2S perception. The sensing principle consists of two steps: first, H2S reduces the azide group to a primary amine; second, a cross-linking reaction between the primary amine and active ester induces the aggregation of AuNPs. The AE-AuNPs-based nanosensors show high selectivity toward H2S over other anions and thiols due to the specific azide-H2S chemistry. Under optimal conditions, 0.2 µM H2S is detectable using a UV-vis spectrophotometer, and 4 µM H2S can be easily detected by the naked eye. In addition, the practical application of the designed nanosensors was evaluated with lake water samples.

Logic control of enzyme-like gold nanoparticles for selective detection of lead and mercury ions.

Functional logic gates based on lead ions (Pb(2+)) and mercury ions (Hg(2+)) that induce peroxidase-like activities in gold nanoparticles (Au NPs) in the presence of platinum (Pt(4+)) and bismuth ions (Bi(3+)) are presented. The "AND" logic gate is constructed using Pt(4+)/Pb(2+) as the input and the peroxidase-like activity of the Au NPs as the output; this logic gate is denoted as "Pt(4+)/Pb(2+)(AND)-Au NPPOX". When Pt(4+) and Pb(2+) coexist, strong metallophilic interactions (between Pt and Pb atoms/ions) and aurophilic interactions (between Au and Pb/Pt atoms/ions) result in significant increases in the deposition of Pt and Pb atoms/ions onto the Au NPs, leading to enhanced peroxidase-like activity. The "INHIBIT" logic gate is fabricated by using Bi(3+) and Hg(2+) as the input and the peroxidase-like activity of the Au NPs as the output; this logic gate is denoted as "Bi(3+)/Hg(2+)(INHIBIT)-Au NPPOX". High peroxidase-like activity of Au NPs in the presence of Bi(3+) is a result of the various valence (oxidation) states of Bi(3+) and Au (Au(+)/Au(0)) atoms on the nanoparticle's surface. When Bi(3+) and Hg(2+) coexist, strong Hg-Au amalgamation results in a large decrease in the peroxidase-like activity of the Au NPs. These two probes (Pt(4+)/Pb(2+)(AND)-Au NPPOX and Bi(3+)/Hg(2+)(INHIBIT)-Au NPPOX) allow selective detection of Pb(2+) and Hg(2+) down to nanomolar quantities. The practicality of these two probes has been validated by analysis of Pb(2+) and Hg(2+) in environmental water samples (tap water, river water, and lake water). In addition, an integrated logic circuit based on the color change (formation of reddish resorufin product) and generation of O2 bubbles from these two probes has been constructed, allowing visual detection of Pb(2+) and Hg(2+) in aqueous solution.

Logical regulation of the enzyme-like activity of gold nanoparticles by using heavy metal ions.

In this study we employed self-deposition and competitive or synergistic interactions between metal ions and gold nanoparticles (Au NPs) to develop OR, AND, INHIBIT, and XOR logic gates through regulation of the enzyme-like activity of Au NPs. In the presence of various metal ions (Ag(+), Bi(3+), Pb(2+), Pt(4+), and Hg(2+)), we found that Au NPs (13 nm) exhibited peroxidase-, oxidase-, or catalase-like activity. After Ag(+), Bi(3+), or Pb(2+) ions had been deposited on the Au NPs, the particles displayed strong peroxidase-like activity; on the other hand, they exhibited strong oxidase- and catalase-like activities after reactions with Ag(+)/Hg(2+) and Hg(2+)/Bi(3+) ions, respectively. The catalytic activities of these Au NPs arose mainly from the various oxidation states of the surface metal atoms/ions. Taking advantage of this behavior, we constructed multiplex logic operations-OR, AND, INHIBIT, and XOR logic gates-through regulation of the enzyme-like activity after the introduction of metal ions into the Au NP solution. When we deposited Hg(2+) and/or Bi(3+) ions onto the Au NPs, the catalase-like activities of the Au NPs were strongly enhanced (>100-fold). Therefore, we could construct an OR logic gate by using Hg(2+)/Bi(3+) as inputs and the catalase-like activity of the Au NPs as the output. Likewise, we constructed an AND logic gate by using Pt(4+) and Hg(2+) as inputs and the oxidase-like activity of the Au NPs as the output; the co-deposition of Pt and Hg atoms/ions on the Au NPs was responsible for this oxidase-like activity. Competition between Pb(2+) and Hg(2+) ions for the Au NPs allowed us to develop an INHIBIT logic gate-using Pb(2+) and Hg(2+) as inputs and the peroxidase-like activity of the Au NPs as the output. Finally, regulation of the peroxidase-like activity of the Au NPs through the two inputs Ag(+) and Bi(3+) enabled us to construct an XOR logic gate.

Peroxidase-mimic bismuth-gold nanoparticles for determining the activity of thrombin and drug screening.

Fibrinogen-modified bismuth-gold nanoparticles (Fib-Bi-Au NPs) are prepared and used as enzyme mimics for the H(2)O(2)-mediated reaction with Amplex Red (AR), which is further employed for determining thrombin activity and drug screening.