Rapid Preparation Method Research Articles

From non-affinity to high-affinity: Rapid preparation of nanobodies utilizing high-precision alphafold and structural-interaction analysis for detection of enrofloxacin in marine fish.

Traditional antibody preparation methods rely on animal immunization and experimental screening, often requiring 3-6 months, which cannot meet the urgent demands for antibodies in environmental emergencies or contamination events. Herein, a rapid antibody preparation method was proposed to transform anti-macromolecule nanobodies into high-affinity anti-small molecule (enrofloxacin) nanobodies, based on the high-precision predictive capabilities of AlphaFold, integrated with structural and interaction analysis. The high-precision prediction capability of AlphaFold ensured the accuracy of nanobody structural analysis and targeted modification. Binding structure design enabled the antigen and antibody to adopt an optimal binding conformation. Interaction analysis provided guidance for targeted modifications and served as a tool for evaluating the results. Molecular docking results revealed that the template nanobodies, which initially couldn't dock with enrofloxacin, formed 3-5 noncovalent bonds after modification. Bio-layer interferometry (BLI) assays demonstrated that the equilibrium dissociation constant (KD) of the modified nanobodies ranged from 5.56 ± 0.34 nM to 94.73 ± 4.35 nM, with half-maximal inhibitory concentration (IC50) between 231.9 and 3293.6 ng/mL. Based on the modified nanobodies and BLI platform, the detection of enrofloxacin in marine fish was achieved with a limit of detection (LOD) as low as 59.97 μg/kg. In summary, this study provided a novel perspective for the rapid nanobody preparation, showing significant potential in food safety and environmental monitoring.

Rapid sequencing and identification for 18-STRs long amplicon panel using portable devices and nanopore sequencer

STRs are the most commonly used forensic genetic markers for human identification. Nanopore sequencing has shown the advantages of high portability and large data throughput. Previous studies indicate it has great potential for profiling STRs based on the ligation library preparation method. However, this method, which requires more library preparation time and operations, is unsuitable for rapid STR profiling, particularly for field forensic applications. The transposase-based rapid library preparation method offers the possibility to perform human identification using portable instruments. However, the amplicons of conventional STR panels are too small and would be cut into scraps with rapid methods, making them impractical for genotyping. In this study, we developed an 18-STRs multiplex amplification panel with amplicons of ~1.4 Kbp. The PCR conditions were optimized to be finished within 2 h and 12 min, and the PCR products could undergo rapid methods that involved random fragmentation. We found that, on average, 29.16 % of reads from the long-amplicon panel and rapid library kit covered the whole STR region, sufficient for downstream STR profiling analysis. We conducted a small validation experiment on 24 samples using portable instruments powered by a 1.5 kW‧h portable power source. The entire process took 10.5 h and we obtained enough data from 24 samples to perform trustworthy pairwise identification analysis using the STR profiles. The overall accuracy of the analysis was 95.36 %. In sum, the study evaluated and demonstrated the viability and potential of nanopore sequencing for forensic application in the field.

Investigating the Use of Novel Blood Processing Methods to Boost the Identification of Biomarkers for Non-Small Cell Lung Cancer: A Proof of Concept.

Diagnosis of non-small cell lung cancer (NSCLC) currently relies on imaging; however, these methods are not effective for detecting early stage disease. Investigating blood-based protein biomarkers aims to simplify the diagnostic process and identify disease-associated changes before they can be seen by using imaging techniques. In this study, plasma and frozen whole blood cell pellets from NSCLC patients and healthy controls were processed using both classical and novel techniques to produce a unique set of four sample types from a single blood draw. These samples were analyzed using 12 immunoassays and liquid chromatography-mass spectrometry to collectively screen 3974 proteins. Analysis of all fractions produced a set of 522 differentially expressed proteins, with conventional blood analysis (proteomic analysis of plasma) accounting for only 7 of the total. Boosted regression tree analysis of the differentially expressed proteins produced a panel of 13 proteins that were able to discriminate between controls and NSCLC patients, with an area under the ROC curve (AUC) of 0.864 for the set. Our rapid and reproducible (<10% CV for technical replicates) blood preparation and analysis methods enabled the production of high-quality data from only 30 μL of complex samples that typically require significant fractionation prior to proteomic analysis. With our methods, almost 4000 proteins were identified from a single fraction over a 62.5 min gradient by LC-MS/MS.

Rapid microwave preparation of nitrogen doped carbon dots and their applications as highly sensitive nanoprobe for hematin

In this work, nitrogen-doped carbon dots (NCDs) were obtained by microwave rapid preparation method with lipase and guanidine hydrochloride as precursors in only 6 min. In order to achieve high sensitivity detection of hematin, based on obtained NCDs as a nanoprobe, a new detection method based on photoluminescence (PL) change was developed. Further studies have shown that the sensing strategy relied on the inner filter effect to selectively quench the PL emission of NCDs at 379 nm by hematin under 320 nm excitation. The nanoprobe had a wide linear range (0.2–13 μmol/L) and a detection limit of 0.07 μmol/L. The method was effectively applied to the identification of hematin in human blood samples. The proposed nanoprobe has the characteristics of simple and efficient design. It can achieve rapid and accurate detection, which is very suitable for real-time analysis of blood samples to facilitate diagnosis.

Rapid Analysis of Sterols in Blood-Derived Samples.

Dysregulations of cholesterol biosynthesis are known to be associated with several pathologies. Due to the rapid growth of clinical investigations in this research area, a specific, fast, and valid method for analyzing cholesterol, its precursors, and metabolites is required. Here, we describe a rapid method for sample preparation, separation, and quantification of sterols in blood-derived samples using polymeric solid phase extraction followed by gas chromatography-mass spectrometry. The validated method demonstrates a reliable quantification of cholesterol, its precursors, and metabolites.

A low-cost dpMIG-seq method for elucidating complex inheritance in polysomic crops: a case study in tetraploid blueberry.

Next-generation sequencing (NGS) library construction often requires high-quality DNA extraction, precise adjustment of DNA concentration, and restriction enzyme digestion to reduce genome complexity, which results in increased time and cost in sample preparation and processing. To address these challenges, a PCR-based method for rapid NGS library preparation, named dpMIG-seq, has been developed and proven effective for high-throughput genotyping. However, the application of dpMIG-seq has been limited to diploid and polyploid species with disomic inheritance. In this study, we obtained genome-wide single nucleotide polymorphism (SNP) markers for tetraploid blueberry to evaluate genotyping and downstream analysis outcomes. Comparison of genotyping qualities inferred across samples with different DNA concentrations and multiple bioinformatics approaches revealed high accuracy and reproducibility of dpMIG-seq-based genotyping, with Pearson's correlation coefficients between replicates in the range of 0.91 to 0.98. Furthermore, we demonstrated that dpMIG-seq enables accurate genotyping of samples with low DNA concentrations. Subsequently, we applied dpMIG-seq to a tetraploid F1 population to examine the inheritance probability of parental alleles. Pairing configuration analysis supported the random meiotic pairing of homologous chromosomes on a genome-wide level. On the other hand, preferential pairing was observed on chr-11, suggesting that there may be an exception to the random pairing. Genotypic data suggested quadrivalent formation within the population, although the frequency of quadrivalent formation varied by chromosome and cultivar. Collectively, the results confirmed applicability of dpMIG-seq for allele dosage genotyping and are expected to catalyze the adoption of this cost-effective and rapid genotyping technology in polyploid studies.

One-step laser preparation of unidirectional liquid spontaneous transport structures

The Tilted Wedge Microcavity Array (TWMA) structure of the Nepenthes alata peristome allows for unidirectional fluid transport without any surface energy gradient. However, current methods of creating bionic Nepenthes alata peristome structures have significant improvement room in terms of structural shape, machining efficiency, and cost-effectiveness. To address these issues, in this paper, bionic Nepenthes alata peristome TWMA structures on 7075 aluminium alloy was built via a nanosecond laser with highly efficiency, low cost and flexible processing pattern. A unilateral tilted microcavity structure was fabricated by designing the laser processing path, and at the same time, the samples were tilted to change the angle at which the laser beam hit the working surface. This ensures that the microcavity is machined without destroying the microcavity structure underneath. The variation law of aluminum alloy ablation threshold under the influence of laser incidence angle was studied. The dimensional parameters of the TWMA structure were analysed in relation to laser incidence angle θ, laser processing power P, number of laser scans n, and laser scanning speed V. Additionally, liquid spreading tests were carried out on different surfaces of bionic structures, and the influence of surface wettability on the spreading results was investigated. The results indicate that the ablation threshold decreases initially and then increases with an increase in the laser incidence angle. This reflects the trend of the aluminium alloy absorption rate, which initially increases and then decreases with an increase in the incidence angle. For the prepared TWMA structure, the optimum speed of achieving the unidirectional spreading effect was found to be 2.41 mm s−1 with a surface contact angle (CA) of 35°. This study presents a low-cost and rapid preparation method for creating liquid unidirectional transport structures without any energy input.

Excellent oxidation resistance of rapidly crystallizing Cr2AlC coatings at high temperatures: Effect of microstructure

Two kinds of Cr2AlC coatings with different microstructures were prepared by direct crystallization during oxidation at 1000 ℃ and conventional crystallization at 700 ℃ for 1 h for amorphous Cr-Al-C coatings prepared by magnetron sputtering, respectively. The effect of microstructure on the oxidation behavior of the coatings at 1000 ℃ was investigated. The grain boundary density and dislocation density of the Cr2AlC coatings crystallized directly during oxidation were much higher, which provided much more sites for heterogeneous nucleation of α-Al2O3 and fast diffusion paths for Al. Therefore, the coating formed complete pure α-Al2O3 scales earlier with better antioxidant properties. More importantly, the present work demonstrates feasibility and advantages of directly applying amorphous Cr-Al-C coatings at high temperatures without additional crystallization treatments, which simplifies preparation process of Cr2AlC coatings, facilitates their industrial applications, and provides a new method for rapid preparation of MAX-phase coatings.

Boosting Thermoelectric Performance via Weakening Carrier-Phonon Coupling in BiCuSeO-Graphene Composites.

BiCuSeO is a promising oxygen-containing thermoelectric material due to its intrinsically low lattice thermal conductivity and excellent service stability. However, the low electrical conductivity limits its thermoelectric performance. Aliovalent element doping can significantly improve their carrier concentration, but it may also impact carrier mobility and thermal transport properties. Considering the influence of graphene on carrier-phonon decoupling, Bi0.88Pb0.06Ca0.06CuSeO (BPCCSO)-graphene composites are designed. For further practical application, a rapid preparation method is employed, taking less than 1h, which combines self-propagating high-temperature synthesis with spark plasma sintering. The incorporation of graphene simultaneously optimizes the electrical properties and thermal conductivity, yielding a high ratio of weighted mobility to lattice thermal conductivity (144 at 300K and 95 at 923K). Ultimately, BPCCSO-graphene composites achieve exceptional thermoelectric performance with a ZT value of 1.6 at 923K, bringing a ≈40% improvement over BPCCSO without graphene. This work further promotes the practical application of BiCuSeO-based materials and this facile and effective strategy can also be extended to other thermoelectric systems.

Dielectric Liquid Microlens Array with Tunable Focal Length Based on Microdroplet Array Created via Dip-Coating Method.

A dielectric liquid microlens array (LMA) with a tunable focal length was fabricated by using a microdroplet array generated through the dip-coating method. The process began with treating the octadecyltrichlorosilane (OTS) layer with selective UV/O3 irradiation for 20 min to establish a hydrophilic-hydrophobic patterning surface. The substrate was subsequently immersed in glycerol and then withdrawn at a constant rate to create a microdroplet array. Upon filling the cell with matching oil (SL5267) and placing it within a square array of a 200 μm diameter glycerol microdroplet array, the LMA was produced. The focal length ranged from approximately -0.96 to -0.3 mm within a voltage range of 0 to 60 Vrms. The glycerol microdroplets, characterized by their shapes, sizes, curvatures, and filling factors, can be precisely controlled by designing an OTS patterning or adjusting the dip-coating speed. This approach offers a rapid and high-throughput method for preparation. Our approach to fabricating tunable LMA offers several advantages, including simplicity of fabrication, uniform structural properties, cost-effectiveness, polarization independence, and excellent optical performance. These focus-tunable LMAs hold significant potential for applications in image processing, 3D displays, medical endoscopy, and military technologies.

Enhanced Indoor Conversion Efficiency of Dye-Sensitized Solar Cells by Optimizing Ball-Milling Process of TiO2 Paste

The rapid spread of the Internet of Things (IoT) along with the development of innovative low-power electronic devices has also driven the development of indoor photovoltaics. In this paper, we propose a simple and economically feasible solution that can improve the efficiency of dye-sensitized solar cells (DSSCs) under indoor light conditions by ~112%, without requiring a complex TiO2 photoanode architecture or the design of new dyes. The ball milling process of the TiO2 paste was optimized for indoor light conditions for the first time, both in terms of efficiency and production costs, by developing a rapid preparation method that can be used industrially for the application of DSSCs. A simple use of 12 mm diameter balls caused beneficial structural modifications, decreasing the size of the crystallites, and leading to a high OH generation on the TiO2 surface responsible for the improvement of energy conversion efficiency.

Optimal design of micromixer for preparation of nanoliposomes

Micromixers are essential components for controlling reactions in microfluidic devices, significantly enhancing their efficiency. This paper introduces an unbalanced separation and recombination micromixer with a circulating chamber and optimizes its geometric structure. The study investigates how the width ratio of the main subchannel and secondary subchannel, as well as the position of the circulating chamber, affect the mixing index and pressure drop at the micromixer's inlet and outlet. The enhanced mixing results from the unbalanced impact generated by the micromixer's separation and recombination, combined with the synergistic effect of fluid self-circulation. Simulation results indicate that the optimized micromixer achieves a mixing index close to 100 % when the Reynolds number exceeds 30, with a pressure drop of only 30 KPa at a Reynolds number of 50 in the micromixer's inlet and outlet. A microfluidic-based micromixer was fabricated using soft lithography and molding techniques. Liposomes were synthesized in a single step using soybean phospholipids, cholesterol, ethanol, and deionized water as raw materials. The micro-mixer exhibited exceptional mixing performance, enabling faster liposome synthesis compared to conventional methods. Additionally, the resulting liposomes exhibited smaller particle sizes and narrower size distributions. By adjusting the flow rates of the organic and water phases entering the micromixer, liposomes with a small particle size of 39 nm and a concentrated particle size distribution (PDI=0.104) were successfully produced. These findings highlight the microfluidic approach as a convenient, rapid, and efficient method for liposome preparation. Furthermore, the unbalanced separation and recombination micromixer with a circulating chamber demonstrated a simple structure, high mixing efficiency, and minimal pressure drop loss. It is versatile and suitable for various microreactor applications.

Extraction, rapid preparation and neuroprotective effect of texasin, a main active constituent from Caragana jubata (Pall.) Poir.

Searching for new anti-ischemic stroke (anti-IS) drugs has always been a hot topic in the pharmaceutical industry. Natural products are an important source of discovering anti-IS drugs. The aim of the present study is to extract, rapidly prepare and explore the neuroprotective effect of texasin, a main active constituent from Caragana jubata (Pall.) Poir., which is a kind of Tibetan medicine with a clear anti-IS effect. The results showed that 95% ethanol was the optimal extraction solvent. A three-step rapid preparation method for texasin was successfully established, with a purity of 99.2%. Texasin at the concentration of 25–100 µM had no effect on the viability of normal cultured PC12 cells; 12.5 and 25 µM texasin could enhance the viability of PC12 cells damaged by oxygen and glucose deprivation/reoxygenation (OGD/R), and their effects are comparable to the positive drug edaravone at the concentration of 50 µM. Compared with the normal group, the expression of Bcl-2 protein in OGD/R-injured PC12 cells was downregulated (p < 0.01), and that of PERK, eIF2α, ATF4, CHOP, Bax and Cleaved caspase-3 proteins were upregulated (p < 0.01, p < 0.001). Compared with the OGD/R group, 25 µM texasin could upregulate the expression of Bcl-2 protein (p < 0.01), and downregulate that of PERK, eIF2α, ATF4, CHOP, Bax and Cleaved caspase-3 proteins (p < 0.01, p < 0.001). The 7-OH and 1-O of texasin formed H-bonds with residues Cys891 of the hinge β-strand of PERK, which is crucial for kinase inhibitors. The above results suggest that the method established in the present study achieved rapid preparation of high-purity texasin. Texasin might inhibit neuronal apoptosis via the regulation of endoplasmic reticulum stress PERK/eIF2α/ATF4/CHOP signalling pathway to exert a protective effect on OGD/R-injured PC12 cells. Aiding by molecular docking, texasin was assumed to be a potential PERK inhibitor.

Rapid synthesis of MXenes and their potential risk to bacterial communities in the tomato rhizosphere

A rapid MXene preparation method has been developed, and the negligible impact of MXene on the soil environment has been confirmed through the evaluation of the sensitive rhizosphere bacterial community.

Photonic crystal gas sensors based on metal-organic frameworks and polymers.

A photonic crystal (PC) is an optical microstructure with an adjustable dielectric constant. The PC sensor was deemed a powerful tool for gas molecule detection due to its excellent sensitivity, stability, online use and tailorable optical performance. The detection signals are generated by monitoring the changes of the photonic band gap when the sensing behavior occurs. Recently, many efforts have been devoted to improving the PC sensor's detection performance and reducing technical costs by selecting and refining functional materials. In this case, metal-organic frameworks (MOFs) with a large specific surface, tunable structural properties and polymers with unique swelling properties have attracted increasingly attention. In this review, a systematic review of PC gas sensors based on MOFs and polymers was carried out for the first time. Firstly, the optical properties and gas sensing mechanism of PCs were briefly summarized. Secondly, a detailed discussion of the structural properties and rapid preparation methods of distributed Bragg reflectors (DBRs), opals and inverse opals (IOPCs) was presented. Thirdly, the recent advances in MOF, polymer and MOF/polymer-based PC sensors over the past few years were summarized. It should be noted that the sensitivity and selectivity enhancement strategy by appropriate material species selection, organic ligand functionalization, metal-ion doping, diverse functional material arrays, and multi-component compounding were analyzed in detail. Finally, prospects on PC gas sensors are given in terms of preparation methods, material functionalization and future applications.

Densification and grain growth in the flash sintering of high-entropy (La0.2Ce0.2Nd0.2Sm0.2Eu0.2)2Zr2O7 ceramics

Preparation of high-entropy ceramics (HECs) requires long duration at high temperatures, which limits their wide application. In this study, high-entropy (La0.2Ce0.2Nd0.2Sm0.2Eu0.2)2Zr2O7 ceramics was prepared at low temperatures of 762–885 °C in very short time of 60 s using the flash sintering. The effects of electric field strength and current limit on the densification and grain growth, as well as the phase transformation were systematically investigated. Both density and grain size were increased with the increase of the electric field strength and current limit. The phase transformation from defective fluorite to pyrochlore occurred at large current limit. The flash-sintered bulks exhibited good mechanical properties. This study demonstrates that flash sintering is an effective method for rapid preparation of HECs at low temperatures. Moreover, the microstructure of HECs can be controlled by adjusting the electric field strength and current limit.

Detection of Esca‐associated fungi in grapevine trunks using loop‐mediated isothermal amplification (LAMP) assays

AbstractEsca is a grapevine trunk disease (GTD) that is caused by filamentous fungi. It is responsible for considerable economic losses in viniculture on a global scale. Despite many unknown factors contributing to the development of symptoms in affected plants, Phaeoacremonium minimum (PMI), Phaeomoniella chlamydospora (PCH) and Fomitiporia mediterranea (FMED) are generally considered as the main causative fungal species. Early detection and specific identification of these pathogens therefore play an important role in disease control and evaluation of suitable countermeasures. In this study, loop‐mediated isothermal amplification (LAMP) assays were developed for each of the three pathogens. A genome‐based approach was applied for detection and selection of unique target DNA sequences. The designed primer sets showed overall good specificities, with some observed cross‐reactions towards closely related Phaeoacremonium species for the PMI primer set. The developed assays had detection limits of 100 pg (FMED, PMI) and 1 pg (PCH) per reaction (corresponding to 1460 [FMED]; 1950 [PMI]; 342 [PCH] genome copies per reaction). The application of the assays to field samples was demonstrated by testing individual infected grapevine trunks from two European viticultural regions using crude DNA obtained in a rapid sample preparation step. LAMP assay results matched those of PCR following a conventional DNA extraction protocol. The study showed that LAMP‐based rapid molecular detection of major Esca agents can serve as a useful tool for further research and surveillance of a highly devastating grapevine disease. The application of computer‐based whole genome comparison between target and non‐target species for the identification of unique target sequences as the basis for LAMP (or PCR) primer design was demonstrated to be a useful approach in species for which scarce sequence information is available. Moreover, the developed method for rapid DNA preparation from grapevine trunks may potentially be adapted to the DNA‐based detection also of other fungal species that cause grapevine trunk diseases.

A Simple and Rapid High-Performance Liquid Chromatography Method for Preparation and Content Detection of the Mainly Numbing Taste Substances of Zanthoxylum bungeanum Maxim.

As the characteristic numbing taste substances, hydroxyl-α-sanshool (HAS) and hydroxyl-β-sanshool (HBS) were considered vital indicators to evaluate the quality of Zanthoxylum bungeanum Maxim. However, it is very difficult to obtain their high-purity monomers individually, as the only difference between HAS and HBS is that C-6 cis-trans isomerism. In our study, a simple and rapid Ag +-HPLC method was developed to pure the standard chemicals of Z. bungeanum with numbing taste, and 1H NMR and 13C NMR were employed to determine the purity and structure. Moreover, an HPLC method was established to determine the content of numbing taste components of 16 varieties of Z. bungeanum from different regions. The analytical methods were validated for accuracy, precision, and linearity, respectively. The validated method was accurate (spiked recoveries 0.94-1.10), precise in terms of peak area (intra-day RSDs <1.25% and inter-day RSDs <1.61%), and linear (r2 ≥ 0.999). It was found that there were significant differences in the content of HAS and HBS among different types of Z. bungeanum, with HAS content ranging from 60.06 ± 1.14 to 164.13 ± 3.28mg/g and HBS ranging from 7.81 ± 0.36 to 21.11 ± 0.75mg/g. The RSDs of HAS range were 1.73-3.80% and that of HBS range 2.03-4.73% (RSDs ≤5%), which indicated that the measurements of HAS and HBS were reliable.

Patterned Liquid-Metal-Enabled Universal Soft Electronics (PLUS-E) for Deformation Sensing on 3D Curved Surfaces.

Liquid metals with metallic conductivity and infinitely deformable properties have tremendous potential in the field of conformal electronics. However, most processing methods of liquid metal electronics require sophisticated apparatus or custom masks, resulting in high processing costs and intricate preparation procedures. This study proposes a simple and rapid preparation method for patterned liquid-metal-enabled universal soft electronics (PLUS-E). The utilization of selective adhesion of the liquid metals on stretchable substrates and the adaptive toner mask enables rapid fabrication (<2 s/100 cm2), excellent stretchability (800% strain), and high forming accuracy (100 μm). Benefiting from the adaptive deformation of the substrate and toner mask, PLUS-E can be conformally applied to any shape of 3D surfaces. Besides, the stability of PLUS-E on 3D surfaces is improved by low-fluidity liquid metal composites. The finite element simulation is used to accurately forecast the deformation and resistance changes of the PLUS-E, and it provides guidance for device design and manufacturing. Finally, this method was utilized to develop various sensors for detecting human motion, catheter bending, and balloon expansion. All of them have obtained stable and reliable signal measurements, demonstrating the usefulness of PLUS-E in real-world applications.

An improved method for rapid and safe preparation and measurement of dimethylmercury using gas chromatography-atomic fluorescence spectrometry

Transformations between dimethylmercury (DMHg) and other mercury (Hg) species have been one of the critical knowledge gaps in the Hg global biogeochemical cycle due to the lack of detailed studies. The preparation and measurement of DMHg are challenging due to the high toxicity and volatility of DMHg. In this work, we invented a new DMHg generator for successfully preparing high-purity DMHg in a highly controllable and safe way. The DMHg could be spontaneously volatilized and diffused from the original preparation solution to the solution to be studied. The parameters for generating DMHg were optimized to be the pH value of 4.0 with a MeCo/Hg2+ molar ratio of 10 at 20 °C. The following measurement method of DMHg in the presence of various species of Hg was also investigated and optimized. Hg0 and DMHg could be separated effectively with the carrier gas flow rate of 15 mL min−1 and the gas chromatography column temperature of 30 °C. The interferences of Hg0, monomethylmercury and other species were excluded by systematic control experiments. A sensitive and reliable approach for quantifying trace DMHg in water was developed. Under the optimal conditions, the limits of detection for Hg0, MMHg and DMHg were 0.03, 0.002 and 0.024 ng L−1, respectively, with the relative standard deviation below 8.2%. The developed method was validated by the determination Hg species of different natural water samples. This work is expected to provide a new and safe strategy for DMHg preparation and a verified method for DMHg measurement.