查看更多>>摘要：ALKBH3 is an important marker for early diagnosis and histopathological grading of prostate cancer. However, the lack of a rapid and sensitive method to quantify the enzyme's activity in the current time necessitates the development of a new quantitative assay. Herein, we first tried to quantitative assay for ALKBH3 activity using an electrochemical method based on the degradation of the signal probe due to alkyl group of the m1A removal by ALKBH3. A strong electrochemical signal can be obtained when the ferrocene (Fc) labeled dsDNAs with 1methyladenine are immobilized on the electrode. In the presence of ALKBH3, the 3 ' blunt of DNA can be formed because of the removal of alkyl group of the Fc-DNA probe, which can be recognized and degraded by Exonuclease III (Exo III). As a result, the electrochemical signal produced by Fc greatly decreases, and the activity of ALKBH3 can be easily detected via changes in electrochemical signal. Quantitative analysis of ALKBH3 activity showed a wide detection range (0.1 and 20 ng/mL) and low detection limit (0.04 ng/mL). Furthermore, the method can be applied to detect 1-methyladenine through ALKBH3 in cell lysates and tissue samples, providing a new method for clinical detection of prostate cancer.
查看更多>>摘要：Under controlled dispersion conditions, sample injection towards a detector opened essential fields for the Analytical Chemistry fast development methods. Flow injection analysis (FIA) and batch injection analysis (BIA) systems are crucial for injecting the sample in these analytical methods. The BIA system eliminated the flow manifold, with samples injected directly onto the detector inside the batch injection cell. Paper was slightly evaluated coupled to FIA, and no reports were found associated with BIA. Still, it can potentially reduce the BIA manifold by removing the batch injection cell based on the capillarity properties to disperse the injected solution over the detection system. Hence, this article reported the first work coupling batch-injection analysis and microfluidic paper-based analytical device (BIA-mu PAD) with pencil-drawn electrodes directly attached to the paper using a CO2 laser pre-treated chromatographic paper. The laser pretreatment of the paper (optimized conditions: 6.5% laser power, 12 mm s-1 scan rate, and 12 mm output distance) was essential to enhance the electrochemical response for ferri/ferrocyanide redox couple and paracetamol (PAR), as shown by spectroscopic and electrochemical techniques. The proposed BIA-mu PAD was evaluated using pharmaceutical paracetamol samples as proof-of-concept (optimized conditions: 15 mu L injected volume and 6.4 mu L s-1 dispensing rate), obtaining good linearity (R = 0.9961) and recovery values ranging from 95 to 103%. Repeatability (n = 16) and reproducibility (n = 9) tests with 1 mmol L-1 PAR also presented well relative standard deviation (RSD) results of 5.1% and 6.6%, respectively. A sampling frequency of 76 h-1 was obtained, which is a similar value compared with conventional BIA apparatus. Limits of detection and quantification were estimated in 0.046 and 0.154 mmol L-1, respectively. Additionally, an improvement in the current response and the sample throughput was observed when comparing FIA and BIA-mu PADs, attesting the applicability of the proposed device and opening for new possibilities related to paper-based devices coupled with flow techniques.
查看更多>>摘要：The overuse or abuse of organophosphorus pesticides (OPs) can bring about severe contamination problems in foodstuff and the environment, which will seriously threaten human health and the ecosystem's cycle. Hence, it is in high demand to establish sensitive, portable, specific, and cost-effective methods for monitoring OPs to control food safety, protect the ecosystem, and prevent disease. The optical biosensor with enzyme as biorecognition elements has been an effective alternative for OPs detection. Herein, we firstly introduce various enzymes, sensing mechanisms, advantages and disadvantages used as bio-recognition elements in optical sensing for OPs detection. Then, we review various optical biosensing strategies based on enzymes as recognition elements that were ingeniously designed and successfully utilized for OPs detection, with a particular emphasis on photoluminescence (PL), chemiluminescence (CL), electrochemiluminescence (ECL), and colorimetric (CM) biosensing strategies. We not only highlight the state-of-art developments and the construction strategies of the enzyme-based optical biosensing method but also summarize the existing deficiencies, current challenges, and the future perspectives of OPs detection.
Negrete, Missael Antonio ArroyoWrobel, KazimierzBarrientos, Eunice YanezEscobosa, Alma Rosa Corrales...
8页
查看更多>>摘要：In this work, a principle of flow injection analysis (FIA) was applied for sample introduction to an electrospray ionization - ion trap mass spectrometer (ESI-ITMS) with the aim to quantify chromium(III) picolinate (CrPic(3)) in commercial supplements by multiple reaction monitoring, and using cobalt(II) picolinate as internal standard (IS). FIA system was operated with ammonium formate 10 mmol L-1 in methanol-water (1:1, v/v) as a carrier solution at a flow rate 200 mu L min(-1); 100 mu L injections were performed in 2-min intervals. Setting ion transitions m/z 419 -> 270 and 304 -> 260 for the analyte and IS, respectively, and 100 ms integration time, the method detection and quantification limits 12 ng g (-1) and 40 ng g(-1) of Cr (as CrPic(3)) in the air-dried powder. Acetonitrile extracts of the real-world samples presented varying from sample-to-sample chemical composition and IS efficiently compensated for ionization interferences. Mean results from triplicate analysis of four different supplements were obtained with relative standard deviation 0.1-4.0%, indicating acceptable precision. Trueness of the proposed FIA-ESI-ITMS/MS procedure was demonstrated by 95.8-108% percentage recoveries attained in the analysis of the CrPic(3)-spiked samples. For comparative purposes, total Cr was determined by ICP-MS. The quantitative results obtained indicate the necessity of analytical control of Cr(III) supplements commercially available and demonstrate that the proposed FIA-ESI-ITMS/MS procedure is well-suited for the determination of CrPic(3) in such products.
查看更多>>摘要：Carbon monoxide (CO) is one of the most significant signal molecules and plays an important role in regulating human physiological and pathological processes. In this study, a novel Pd-based complex (Pd-BNP-OH) was developed for endogenous CO detection. The structure and morphology of Pd-BNP-OH was characterized by SEM, oS, and NMR analyses. When Pd-BNP-OH was reacted with CO, a strong fluorescence enhancement at 510 nm was observed. In addition, Pd-BNP-OH exhibited high stability and selectivity toward CO in PBS buffer. In biological experiments, Pd-BNP-OH exhibited little cytotoxicity in cellular environment, and a bright fluorescence turn on was observed in the presence of exogenous CO and endogenous generated CO. The probe was then applied for CO detection in live zebrafish by both one-photon and two-photon excitation. Significantly, PdBNP-OH has excellent two-photon property, controllable structure and high biocompatibility. These features enable the probe to detect endogenously generated carbon monoxide in live organisms successfully.
查看更多>>摘要：To accurately determine ultra-trace Pu isotopes in small environmental samples, we explored ICP-MS/MS in NH3-He mode, and investigated mechanism of U-238 interference removal and measurement sensitivity improvement for plutonium isotopes. The interference of uranium and uranium hydrides was effectively eliminated using 0.4 mL/min NH3 as reaction gas by shifting them to U(NHm)(n)(+) and UH(NHm)(n)(+). The overall interference of uranium was reduced to <2.4 x 10 (-7), while remaining excellent Pu-239 sensitivity (13,900 Mcps/(mg/L)) mainly due to ion focusing effect of Pu by helium gas. On this basis, the purification of plutonium using a single AG1- x 4 column was proved to be sufficient for accurate determination of plutonium isotopes by the developed detection method, and the detection limits for the method were estimated to be 0.16 fg (0.4 mu Bq) for Pu-239, 0.046 fg (0.4 mu Bq) for Pu-240 and 0.039 fg (0.15 mBq) for Pu-241. The method was validated by analyzing plutonium isotopes in certificated reference materials and reported environmental samples of only 1-2 g. The analytical results of ultra-trace Pu isotopes in small amounts (similar to 1 g) of lake sediments obtained by the developed method were successfully applied to sediment dating.
查看更多>>摘要：Cytokines are important factors in the early diagnosis of autoimmune diseases and require high sensitivity, high selectivity and quantitative detection. We proposed a miniaturized electrochemical magneto-immunosensor (EC-MIS) on portable interleukin-6 (IL-6) detection based on this requirement. Firstly, a micro-fabricated working electrode is electrochemically modified with a hybrid of reduced graphene oxide (rGO) and gold nanoparticles (AuNPs). Increased surface area and enhanced charge transfer rate improve the performance of this immuno-sensor on sensitivity. Secondly, magnetic beads attached with the capture antibody (cAb) are employed in sandwich immunoassay. This kind of immunoassay is immobilized on the working electrode surface by an external magnet to enrich the analyte IL-6. Thirdly, the last two features are combined and integrated on a microfluidic device in order to restrict the sample at certain areas and ease the operation of detection. With our prototypic EC-MIS operated in amperometric mode, we have achieved the detection of IL-6 with a linear range from 0.97 to 250 pg/mL and a limit of detection (LOD) of 0.42 pg/mL. Real serum samples were demonstrated and compared with benchtop equipment's results.
查看更多>>摘要：Nucleic acid testing (NAT) implemented on a portable, miniaturized, and integrated device with rapid and sensitive results readout is highly demanded for pathogen detection or genetic screening at resource-limited settings, especially after the outbreak of coronavirus disease 2019 (COVID-19). The integration of recombinase polymerase amplification (RPA) with emerging microfluidics, classified by paper-based microfluidics and chip-based microfluidics, shows great potential to perform laboratory independent NAT assays at point of care with minimal labor, time and energy consumption. This review summarizes the state-of-the-art of RPA integrated with paper-based microfluidics and chip-based microfluidics, and discusses their pros and cons. Finally, existing challenges and possible ways for optimization of microfluidics-based RPA are proposed.
查看更多>>摘要：Pseudomonas aeruginosa (P. aeruginosa), a ubiquitous opportunistic pathogen, can frequently cause chronic obstructive pulmonary disease, cystic fibrosis and chronic wounds, and potentially lead to severe morbidity and mortality. Timely and adequate treatment of nosocomial infection in clinic depends on rapid detection and accurate identification of P. aeruginosa and its early-stage antibiotic susceptibility test. Traditional methods like plating culture, polymerase chain reaction, and enzyme-linked immune sorbent assays are time-consuming and require expensive equipment, limiting the rapid diagnostic application. Advanced sensing strategy capable of fast, sensitive and simple detection with low cost has therefore become highly desired in point of care testing (POCT) of nosocomial pathogens. Within this review, advanced detection and sensing strategies for P. aeruginosa cells along with associated quorum sensing (QS) molecules over the last ten years are discussed and summarized. Firstly, the principles of four commonly used sensing strategies including localized surface plasmon resonance (LSPR), surface-enhanced Raman spectroscopy (SERS), electrochemistry, and fluorescence are briefly over viewed. Then, the advancement of the above sensing techniques for P. aeruginosa cells and its QS biomarkers detection are introduced, respectively. In addition, the integration with novel compatible platforms towards clinical application is highlighted in each section. Finally, the current achievements are summarized along with proposed challenges and prospects.
查看更多>>摘要：Considering the challenges of generating simple and efficient DNA (deoxyribonucleic acid) nanomachines for sensitive bioassays and the great potential of target-induced self-cycling catalytic systems, herein, a novel autocatalytic three-dimensional (3D) DNA nanomachine was constructed based on cross-catalytic hairpin assembly on gold nanoparticles (AuNPs) to generate self-powered efficient cyclic amplification. Typically, the DNA hairpins H1, H2, H3 and H4 were immobilized onto AuNPs first. In the presence of target microRNA-203a, the 3D DNA nanomachines were triggered to activate a series of CHA (catalytic hairpin assembly) reactions. Based on the rational design of the system, the products of the CHA 1 reaction were the trigger of the CHA 2 reaction, which could trigger the CHA 1 reaction in turn, generating an efficient self-powered CHA amplification strategy without adding fuel DNA strands or protein enzymes externally and producing high-efficiency fluorescence signal amplification. More importantly, the proposed autocatalytic 3D DNA nanomachines outperformed conventional 3D DNA nanomachines combined with the single-directional cyclic amplification strategy to maximize the amplification efficiency. This strategy not only achieves high-efficiency analysis of microRNAs (microribonucleic acids) in vitro and intracellularly but also provides a new pathway for highly processive DNA nanomachines, offering a new avenue for bioanalysis and early clinical diagnosis.
NSTL
Elsevier