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A ultrasensitive split-type photoelectrochemical (PEC) sensor was constructed for ampicillin (AMP) detection, utilizing a metal-organic framework (MOF)-confined In 2 S 3 /PCN-224 Z-scheme heterojunction as the photoactive material. The prepared In 2 S 3 /PCN-224 was demonstrated with high charge separation efficiency and a stable PEC signal response due to the unique electron flow direction of the Z-scheme configuration. To further enhance the detection sensitivity, target-mediated in-situ ion exchange via Cd 2+ ions was employed to modulate the photoactivity of In 2 S 3 /PCN-224. In the presence of AMP, the aptamers labeled with CdCO 3 were released from the DNA double-strand and then dissociated into Cd 2+ ions after acid treatment. Ion exchange reactions will occur upon introducing the solution into the In 2 S 3 /PCN-224 surface. Another photoactive material may be produced on the electrode surface to amplify the original PEC signal. The resulting split-type PEC sensor exhibited an impressive linear range (0.5-200 ng mL -1 ) with a low limit of detection (LOD, 0.09 pg mL -1 , S/N = 3). This work presents a promising strategy for the development of PEC biosensors, offering practical applications in the environmental analysis of antibiotics., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

C-ZIF-67@PAN nanofibers are prepared as an active material for ampicillin (AMP) detection. Metal organic frameworks (MOFs) can provide larger specific surface area and more binding sites. The PAN fiber in the further carbonization process can make ZIF-67 orderly arranged, enhance the conductivity of the material, and make the response more sensitive. Gold nanoparticles are incorporated into the material, and the aptamer is closely combined with the material by using the Au-S bond to further enhance the conductivity of the material. Under optimized conditions, the sensor has a wide detection range (0.001-100 μM) and a low detection limit (0.67 nM). In addition, the C-ZIF-67@PAN@Au@Apta sensor is successfully applied to AMP residues in milk samples with a recovery rate of 92.77-101.95%.

Antibiotic residue is a growing concern for human health, and exploring a method for antibiotic detection is imperative Herein, a trimetallic metal-organic framework (NH2-MIL-53(Al)@Mo/Zn-MOF) with an adsorption effect was prepared by growing Mo/Zn-MOF on the surface of NH 2 -MIL-53(Al) for simultaneous pre-concentration and detection of ampicillin (AMP). The NH 2 -MIL-53(Al)@Mo/Zn-MOF showed a large specific surface area and stable crystal structure, which is conducive to improving the adsorption efficiency and detection sensitivity. The adsorption process of NH 2 -MIL-53(Al)@Mo/Zn-MOF to AMP was simulated by a quasi-second-order kinetic model and Langmuir model. Moreover, a ratiometric fluorescent sensor was established based on fluorescence donors of NH 2 -MIL-53(Al)@Mo/Zn-MOF and CdTe QDs@SiO 2 and a quencher of mitoxantrone (MIT). With the increasing concentration of AMP, the fluorescence of CdTe QDs@SiO 2 was gradually quenched by MIT through the inner filter effect (IFE) and photoinduced electron transfer (PET) process. At the same time, that of NH 2 -MIL-53(Al)@Mo/Zn-MOF was maintained. With this strategy, the sensor achieved an ultra-sensitive detection of AMP with a low detection limit of 0.69 nM. Moreover, the constructed sensor exhibited satisfactory accuracy and reliability for AMP detection in food and environmental samples, which provides a new idea for developing integrated sensors for simultaneous adsorption and detection of antibiotics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

A novel copper and iron doped containing chitosan and heparin sodium carbon dots (CS-Cu,Fe/HS) nanozyme was formulated through a single-step microwave digestion method. CS-Cu,Fe/HS exhibits excellent peroxidase (POD)-like activity and positive charge characteristics, and it can oxidize the negatively charged 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) in the presence of H 2 O 2 to produce a green compound (ox-ABTS). Furthermore, CS-Cu,Fe/HS enhances electron transfer and provides additional active sites through the valence state transformations of Fe 2+ /Fe 3+ and Cu 2+ /Cu + . Interestingly, the POD-like activity of CS-Cu,Fe/HS is inhibited with the introduction of ampicillin (AMP), which may be because the Cu and Fe ions in CS-Cu,Fe/HS form complexes with AMP, leading to changes in the structure or surface properties of the nanozyme, thereby reducing the number of active sites on the nanozyme. Drawing from this, a straightforward and reliable colorimetric sensor was constructed for AMP detection, featuring a linear range of 0.033 to 110 μg/mL and a detection limit as low as 11.6 ng/mL. The proposed detection method for AMP performed well in real samples, with recoveries ranging from 94.8% to 110.2%., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

A biosensor is reported for electrochemical detection of ampicillin based on Au 5 Pt and DNA cycle dual-signal amplification strategy. Firstly, Au 5 Pt is prepared by reduction of chloroauric acid and chloroplatinic acid with serine-functionalized graphene quantum dot (SGQD). The resulting Au 5 Pt shows an excellent catalytic activity because of the synergy of Au and Pt. Then, Au 5 Pt is covalently combined with hairpin DNA and thionine molecule to form a redox probe. The probe was used for construction of the ampicillin biosensor coupling with DNA cycling. In the presence of ampicillin, the DNA cycle was triggered and leads to many redox probes being carried to the biosensor. This produces a significant signal amplification by oxidation and reduction of thionine molecules in these probes. The combination of Au 5 Pt catalysis with DNA cycle achieve ultrahigh sensitivity, selectivity, and stability for the electrochemical detection of ampicillin. Differential pulse voltammetry current linearly increases with the increase of ampicillin concentration in the range 1 × 10 -18 -1 × 10 -12  M with a detection limit of 3.2 × 10 -19  M (S/N = 3). The proposed analytical method has been satisfactorily used for electrochemical detection of ampicillin in milk., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Ampicillin (AMP) is a β-lactam antibiotic that can inhibit bacterial wall synthesis. The overuse and misuse of AMP makes it micropollutant that commonly found in food and various environmental media. In this work, a fluorescence polarization sensor was designed to sensitive detection of trace ampicillin based on click chemistry, using graphene oxide (GO) as a fluorescence polarization (FP) signal enhancement element. First, when ampicillin binds to its aptamer (apt), the adjacent alkyne and azide groups are separated, hindering the click-linking reaction. When Carboxyfluorescein (FAM) fluorophore-labeled probe (C-FAM) is added, its protruding 3-terminal FAM is recognized and cleaved by exonuclease I (EXO I), releasing fluorophores free that could not be adsorbed on GO, resulting in a lo0wer polarization signal. If there is no AMP in the system, aptamer probe is connected to its complementary chain ends by a click reaction. After C-FAM hybridizes with apt, the apt/P duplex is opened and the prominent single-stranded ends adsorb on the GO, leading a significantly enhanced FP signal. According to the relationship between the difference in FP values and the concentrations of AMP, the limit of detection of proposed method is as low as 80 pg/mL. This assay has a wide linear range plus excellent selectivity, and has been applied to detect AMP in milk and river water samples with satisfactory results, which demonstrates that the FP sensor has great potential for practical applications in food safety and environmental protection fields., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Water-soluble N-doped fluorescent (FL) carbon dots (ACDs) were successfully fabricated hydrothermally using ampicillin sodium as sole precursor. The produced ACDs exhibit satisfactory optical behavior, favorable photostability, and acceptable water solubility. With bright blue emission at 450 nm, the ACDs were utilized for multivariate sensing Fe 3+ and Cu 2+ based on the synergistic effect of the inner filter effect (IFE) and static quenching with detection limits of 0.31 μM and 0.26 μM, respectively. The practicality of ACDs has been verified by the successful determination  of Fe 3+ and Cu 2+ in real water and living cells. These findings confirm the feasibility of the proposed ACDs as FL sensors for efficient and selective detection of Fe 3+ and Cu 2+ , which present promising prospects for real-time monitoring these two metal ions  in environmental and biological systems., Competing Interests: Declarations. Ethics declarations: This research did not involve human or animal samples. Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

To report the testing signal of an immunochromatographic assay for on-site quantitative detection, a portable and user-friendly smartphone-based biosensing platform is developed in this study. This innovative system is composed of an ambient light sensor inherent smartphone reader and a 3D-printed handhold device, a quantitative tool capable of directly interpreting carbon nanoparticle (CNP)-conjugated immunochromatographic strips. To showcase the platform capability, the smartphone-based immunochromatography system (SPICS) reader and device were successfully used in CNP strips for rapid detection of the early pregnancy marker human chorionic gonadotropin in female urine (HCG; limit of detection [LOD]: 0.30 mIU mL -1 ), prostate-specific antigen in patient blood (PSA; LOD: 0.28 ng mL -1 ) and ampicillin residue in animal milk (AMP; LOD: 0.23 ng mL -1 ). The results were fully correlated with conventional commercial instruments ( R 2 = 0.99). The SPICS platform exhibits significant advantages, including portability, cost-effectiveness, easy operation, and rapid and quantitative detection, making it a valuable on-site diagnosis tool for use in home and community healthcare facilities.

Timely and rapid detection of antibiotic residues in the environment is conducive to safeguarding human health and promoting an ecological virtuous cycle. A foldable paper-based photoelectrochemical (PEC) sensor was successfully developed for the detection of ampicillin (AMP) based on glutathione/zirconium dioxide hollow nanorods/aptamer (GSH@ZrO 2 HS@apt) modified cellulose paper as a reactive zone with laser direct-writing lead sulfide/cadmium sulfide/graphene (PbS/CdS/LIG) as photoelectrode and cobalt hydroxide (CoOOH) as a photoresist material. Initially, AMP was introduced into the paper-based reaction zone as a biogate aptamer, which specifically recognized the target and then left the ZrO 2 HS surface, releasing glutathione (GSH) encapsulated inside. Subsequently, the introduction of GSH into the reaction region and etching of CoOOH nanosheets to expose the PbS/CdS/LIG photosensitive material increased photocurrent. Under optimal conditions, the paper-based PEC biosensor showed a linear response to AMP in the range of 5.0 - 2 × 10 4 pM with a detection limit of 1.36 pM (S/N = 3). In addition, the constructed PEC sensing platform has excellent selectivity, high stability and favorable reproducibility, and can be used to assess AMP residue levels in various real water samples (milk, tap water, river water), indicating its promising application in environmental antibiotic detection., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Monitoring methods for beta-lactam (β-lactam) antibiotics, especially for ampicillin (AMP), with simple operation and sensitivity for realtime applications are highly required. To address this need, antioxidant carbon dots (E-CDs) with excellent fluorescence properties were synthesized using citric acid and ethylenediamine as raw materials. With a quantum yield of 81.97%, E-CDs exhibited a specific and sensitive response to ˙OH. The quenched fluorescence of E-CDs by the formed ˙OH could be restored through a competition reaction with AMP. Leveraging the signal-quenching strategy of E-CDs, H 2 O 2 , and Fe 2+ , a fluorescence signal-on strategy was developed using AMP as the fluorescence recovery agent for the sensitive detection of AMP. The mechanism of the quenching of E-CDs by ˙OH was attributed to the damaging effect of ˙OH on E-CDs. Under optimal conditions, the detection limit of this method for AMP was determined to be 0.38 μg mL -1 . This method was successful in drug quality control and the spiked detection of AMP in lake water, milk, and sea cucumber, presenting a viable option for convenient and rapid antibiotic monitoring methods.

To develop a sensitive and simple ampicillin (AMP) sensor for trace antibiotic residue detection, the influencing factors of the modification effect of nanogold-functionalized nucleic acid sequences (Adenine: A, Thymine: T) were comprehensively analyzed in this study, including the modification method, base length and type. It was found that under the same base concentration, longer chains are more likely to reach saturation than shorter chains; and when the base concentration and length are both the same, A exhibits a higher saturation modification level compared to T. Based on these research findings, a highly sensitive fluorescence aptamer sensor for detecting ampicillin was constructed using the optimized functionalized sequence (ployA6-aptamer) and experimental conditions (6 hours binding time between nucleic acid aptamer and complementary strand, pH 7 working solution, 20 minutes detection time) based on the principle of fluorescence resonance energy transfer. The sensor has a detection range of 0.18 ng ml -1 to 3.11 ng ml -1 for ampicillin, with a detection limit of 0.04 ng ml -1 . It exhibits significant selectivity and achieves an average recovery rate of 98.71% in tap water and 91.83% in milk. This method can be used not only for residual ampicillin detection, but also for highly sensitive detection of various antibiotics and small biological molecules by replacing the aptamer type. It provides a research basis for the design of highly sensitive fluorescence aptamer sensors and further applications of nanogold@DNA composite structures.

The global threat of antibiotic resistance has increased the importance of the detection of antibiotics. Conventional methods to detect antibiotics are time-consuming and require expensive specialized equipment. Here, we present a simple and rapid biosensor for detecting ampicillin, a commonly used antibiotic. Our method is based on the fluorescent properties of chitosan-coated Mn-doped ZnS micromaterials combined with the β-lactamase enzyme. The biosensors exhibited the highest sensitivity in a linear working range of 13.1-72.2 pM with a limit of detection of 8.24 pM in deionized water. In addition, due to the biological specificity of β-lactamase, the proposed sensors have demonstrated high selectivity over penicillin, tetracycline, and glucose through the enhancing and quenching effects at wavelengths of 510 nm and 614 nm, respectively. These proposed sensors also showed promising results when tested in various matrices, including tap water, bottled water, and milk. Our work reports for the first time the cost-effective (Mn:ZnS)Chitosan micromaterial was used for ampicillin detection. The results will facilitate the monitoring of antibiotics in clinical and environmental contexts., (© 2024. The Author(s).)

Early transition metal carbides (MXene) hybridized by precious metals open a door for innovative electrochemical biosensing device design. Herein, we present a facile one-pot synthesis of gold nanoparticles (AuNPs)-doped two-dimensional (2D) titanium carbide MXene nanoflakes (Ti 3 C 2 T x /Au). Ti 3 C 2 T x MXene exhibits high electrical conductivity and yields synergistic signal amplification in conjunction with AuNPs leading to excellent electrochemical performance. Thus Ti 3 C 2 T x /Au hybrid nanostructure can be used as an electrode platform for the electrochemical analysis of various targets. We used screen-printed electrodes modified with the Ti 3 C 2 T x /Au electrode and functionalized with different biorecognition elements to detect and quantify an antibiotic, ampicillin (AMP), and a mycotoxin, fumonisin B1 (FB1). The ultralow limits of detection of 2.284 pM and 1.617 pg.mL -1 , which we achieved respectively for AMP and FB1 are far lower than their corresponding maximum residue limits of 2.8 nM in milk and 2 to 4 mg kg -1 in corn products for human consumption set by the United States Food and Drug Administration. Additionally, the linear range of detection and quantification of AMP and FB1 were, respectively, 10 pM to 500 nM and 10 pg mL -1 to 1 µg mL -1 . The unique structure and excellent electrochemical performance of Ti 3 C 2 T x /Au nanocomposite suggest that it is highly suitable for anchoring biorecognition entities such as antibodies and oligonucleotides for monitoring various deleterious contaminants in agri-food products., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Olive trees are one of the most widely cultivated fruit trees in the world. The chemical compositions and biological activities of olive tree fruit and leaves have been extensively researched for their nutritional and health-promoting properties. In contrast, limited data have been reported on olive flowers. The present study aimed to analyse bioactive compounds in olive flower extracts and the effect of fermentation-assisted extraction on phenolic content and antioxidant activity. High-performance thin-layer chromatography (HPTLC) hyphenated with the bioassay-guided detection and spectroscopic identification of bioactive compounds was used for the analysis. Enzymatic and bacterial in situ bioassays were used to detect COX-1 enzyme inhibition and antibacterial activity. Multiple zones of antibacterial activity and one zone of COX-1 inhibition were detected in both, non-fermented and fermented, extracts. A newly developed HPTLC-based experimental protocol was used to measure the high-maximal inhibitory concentrations (IC 50 ) for the assessment of the relative potency of the extracts in inhibiting COX-1 enzyme and antibacterial activity. Strong antibacterial activities detected in zones 4 and 7 were significantly higher in comparison to ampicillin, as confirmed by low IC 50 values (IC 50 = 57-58 µg in zone 4 and IC 50 = 157-167 µg in zone 7) compared to the ampicillin IC 50 value (IC 50 = 495 µg). The COX-1 inhibition by the extract (IC 50 = 76-98 µg) was also strong compared to that of salicylic acid (IC 50 = 557 µg). By comparing the locations of the bands to coeluted standards, compounds from detected bioactive bands were tentatively identified. The eluates from bioactive HPTLC zones were further analysed by FTIR NMR, and LC-MS spectroscopy. Multiple zones of antibacterial activity were associated with the presence of triterpenoid acids, while COX-1 inhibition was related to the presence of long-chain fatty acids., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Phyllanthus emblica L. (syn. Emblica officinalis), popularly known as amla, Indian gooseberry, or the King of Rasyana, is a member of Phyllanthaceae family and is traditionally used in Ayurveda as an immunity booster. The present study aimed to investigate the synergistic interaction of Phyllanthus emblica (FPE) fruits and its selected phytocompounds with ampicillin against selected bacteria. Further, an in silico technique was used to find if major phytocompounds of FPE could bind to proteins responsible for antibiotic resistance in bacterial pathogens and enhance the bioactivity of ampicillin. FPE and all the selected phytocompounds were found to have synergistic antibacterial activity with ampicillin against tested bacteria in different combinations. However, ellagic acid and quercetin interactions with ampicillin resulted in maximum bioactivity enhancement of 32-128 folds and 16-277 folds, respectively. In silico analysis revealed strong ellagic acid, quercetin, and rutin binding with penicillin-binding protein (PBP-) 3, further supported by MD simulations. Ellagic acid and quercetin also fulfill Lipinski's rule, showing similar toxicity characteristics to ampicillin. FPE showed synergistic interaction with ampicillin, possibly due to the presence of phytocompounds such as gallic acid, ellagic acid, quercetin, and rutin. Molecular docking and MD simulations showed the strong interaction of ellagic acid and quercetin with PBP-3 protein. Therefore, these compounds can be explored as potential non-toxic drug candidates to combat bacterial antimicrobial resistance., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Three penicillin-based β-lactam antibiotics (benzylpenicillin, amoxicillin, and ampicillin) were extracted by electromembrane extraction (EME) and determined in the resulting extracts by capillary zone electrophoresis (CZE) with UV-Vis detection. The EME was optimized for the simultaneous clean-up of complex samples and preconcentration of the three antibiotics and employed 1-octanol as the organic phase interface (impregnated in the pores of a hollow fiber), acidified donor solution (pH 3), and phosphate buffer (pH 5.6) as the acceptor solution. The EMEs were carried out for 20 min at 300 V and constant stirring (750 rpm) of the donor solution. At the optimized EME-CZE conditions, the sensitivity of the analytical method was sufficient for the determination of the three β-lactam antibiotics in undiluted cow's milk at concentrations below the EU maximum residue limits (4 μg/L) in foodstuffs. The method was simple, rapid, and convenient and offered extraction recoveries of 13.5 - 87.3 %, enrichment factors of 23.6 - 152.8, repeatability (RSD values) better than 7.6 %, linear analytical response in the 1 - 100 μg/L (3 - 100 μg/L for benzylpenicillin) concentration range with correlation coefficients ≥ 0.9997, and limits of detection from 0.2 to 1.2 μg/L. The proposed analytical concept was used for the rapid control of milk quality (i.e. assessment of excessive use of antibiotics in dairy animals), moreover, it was further extended to the trace determination of β-lactam antibiotics in other complex samples, such as in wastewater., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Objective: To synthesise the gold nanoparticles (AuNPs) using Acacia catechu through biogenic synthesis and evaluate their antimicrobial efficacy against S. mutans and E. coli in vitro., Methods: Green synthesised AuNPs were characterised using the ultraviolet-visible (UV-Vis) spectroscopy, and the size and shape of the synthesised nanoparticles were evaluated using the transmission electron microscopy (TEM). The antimicrobial efficacy of AuNPs (30/60/100 μl) against S. mutans/E. coli was evaluated on the Mueller-Hinton agar by measuring the zone of inhibition (ZOI) with ampicillin (15 μl) as a positive control., Results: The synthesised AuNPs were confirmed using the UV-Vis spectroscopy with peaks at 540 nm, and the size of the particle estimated using the TEM was between 5 and 15 nm. The antimicrobial efficacy of AuNPs was comparable to that of ampicillin against S. mutans/E. coli, but the difference was not significant. The antimicrobial effects increased in a dose-dependent fashion but were comparable across all concentrations and ampicillin., Conclusion: Green synthesised AuNPs exhibited significant antibacterial activity against S. mutans and E. coli at par with commercial ampicillin and demonstrated the potential towards anticariogenic agent for future use in dentistry., Competing Interests: None

The rupture of the Córrego do Feijão dam in Brumadinho (January 25, 2019) caused serious damage to the Paraopeba River and compromised the quality of its waters for human consumption. However, the possible effects of the dam collapse on the river microbiome and its antibiotic resistance profiles are unknown. The present study aims to analyse the possible shifts in microbial diversity and enhancement of antibiotic resistance in the Paraopeba River. To this end, two sampling campaigns (February and May 2019) were performed to obtain water across the entire Paraopeba River (eight sampling locations: Moeda, Brumadinho, Igarapé, Juatuba, Varginha, Angueretá, Retiro Baixo and Três Marias; ~464 km). This sampling scheme enabled determining the effects of the disaster on the river microbiome. Total DNA and microbial isolation were performed with these water samples. The 16S rRNA-based microbiome analyses (n = 24; 2.05 million 16S rRNA reads) showed changes in microbial diversity immediately after the disaster with the presence of metal-indicating bacteria (Acinetobacter, Bacillus, Novosphingobium, and Sediminibacterium). Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) identification of bacterial isolates (n = 170) also disclosed possible indicators of faecal contamination across the Paraopeba (Cloacibacterium, Bacteroides, Feaecalibacterium, Bifidobacterium, Citrobacter, Enterobacter, Enterococcus and Escherichia). Antibiotic resistance increased significantly to ampicillin, ampicillin/sulbactam, amoxicillin/clavulanate, ceftriaxone, and cefalotin among isolates obtained in May after the disaster. The effects of toxic mud on microbiomes were felt at all points sampled up to Anguereta. The ore mud may have exacerbated the growth of different antibiotic-resistant, metal-resistant, and faecal-indicating bacteria in the Paraopeba River., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Abstract: An integrated semi-continuous rector, with simultaneous crystallization of the antibiotic, is the most promising alternative for the enzymatic synthesis of semi-synthetic penicillins. Closed-loop optimization of the reactor operation requires the measurement of its state variables. An efficient method for on-line monitoring of these variables, i.e., the species concentrations, which is consistent with the dynamics of the process, is presented here. Multivariate calibration has been used to reduce times of analysis in multicomponent systems, and this is the approach of this work. Three multivariate calibration methods are compared for prediction of the concentrations of the components present in the enzymatic synthesis of ampicillin. UV spectra and their first-order derivatives were used as input data for the models. Results showed that all methods (SPA, PCR and PLS) had similar performances for this system. The use of first-order derivatives of the spectra did not improve substantially the performance of the methods. Bench assays of synthesis of ampicillin using immobilized penicillin G acylase (PGA) as catalyst were carried out to validate the methodology. Its results were compared with HPLC analyses. The technique showed to be very accurate for monitoring the concentrations of ampicillin (desired product) and 6-aminopenicillanic acid (6-APA, most expensive substrate), with precision similar to those obtained by HPLC. Prior information of the initial reactor state may be used to improve the estimation of less sensitive components (phenylglycine methyl ester, PGME and phenylglycine, PG). Modifications on the biocatalyst must be made in order to use the proposed automatic sampler when crystallization of products occurs. [Copyright &y& Elsevier]

The presence of antibiotics and their metabolites in milk and dairy products is a serious concern because of their harmful effects on human health. In the current study, a novel synergistic bimetallic nanocluster with gold and silver as an emission fluorescence probe was investigated for the simultaneous determination of tetracycline (TC), ampicillin (AMP), and sulfacetamide (SAC) antibiotics in the milk samples using excitation-emission matrix fluorescence (EEMF) spectroscopy. The multivariate curve resolution-alternating least squares (MCR-ALS) method was implemented to analyze augmented EEMF data sets to quantify the multicomponent systems in the presence of interferences with considerable spectral overlap. A pseudo-univariate calibration curve of the resolved emission spectra intensity against the concentration of the mentioned antibiotics was linear in the range of 5-5000 ng mL -1 for AMP and 50-5000 ng mL -1 for TC and SAC. The calculated values of the limit of detection ranged between 1.4 and 14.6 ng mL -1 with a relative standard deviation (RSD) of less than 4.9%. The obtained results show that the EEMF/MCR-ALS methodology using an emission fluorescence probe is a powerful tool for the simultaneous quantification of TC, AMP, and SAC in complex matrices with highly overlapped spectra.

The novelty of this work is the simultaneous analysis of sulbactam (SUL), ampicillin (AMP), and paracetamol (PARA) in human urine samples, using the environmentally benign RP-HPLC method. A C18 column was used in chromatographic separation using potassium dihydrogen phosphate (10 mmol L -1 , pH 5)/ethanol (90 %, V / V ) as the mobile phase; flow rate was 1.00 mL min -1 . UV detection at 220 nm was used for quantification. The proposed method showed good linearity in the concentration ranges of 2.20-250.00 μg mL -1 for SUL, 2.50-250.00 μg mL -1 for PARA, and 14.50-250.00 μg mL -1 for AMP. Direct injection of urine samples with no prior extraction was performed. This method was found successful in moving towards greener studies of drugs' urinary excretion, by decreasing hazardous solvent consumption and waste. Moreover, the method was applied to investigate the urinary excretion of the drugs and possible interaction between ampicillin and paracetamol., (© 2022 Basma M. Selim., published by Sciendo.)

The presence of antibiotics in the aquatic environment is becoming one of the main research focus of scientists and policy makers. Proof of that is the inclusion of four antibiotics, amongst which is amoxicillin, in the EU Watch List (WL) (Decision 2020/1161/EU)) of substances for water monitoring. The accurate quantification of amoxicillin in water at the sub-ppb levels required by the WL is troublesome due to its physicochemical properties. In this work, the analytical challenges related to the determination of amoxicillin, and six related penicillins (ampicillin, cloxacillin, dicloxacillin, penicillin G, penicillin V and oxacillin), have been carefully addressed, including sample treatment, sample stability, chromatographic analysis and mass spectrometric detection by triple quadrupole. Given the low recoveries obtained using different solid-phase extraction cartridges, we applied the direct injection of water samples using a reversed-phase chromatographic column that allowed working with 100% aqueous mobile phase. Matrix effects were evaluated and corrected using the isotopically labelled internal standard or correction factors based on signal suppression observed in the analysis of spiked samples. The methodology developed was satisfactorily validated at 50 and 500 ng L  -   1 for the seven penicillins studied, and it was applied to different types of water matrices, revealing the presence of ampicillin in one surface water sample and cloxacillin in three effluent wastewater samples., Competing Interests: Declaration of Competing Interest The authors declare that they do not have competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Herein, we propose an element probe based CRISPR/Cas14 detection platform and apply it to the detection of non-nucleic-acid targets. Combining metal isotope detection and CRISPR/Cas14 biosensing, the sensitive detection of non-nucleic-acid targets could be realized. We designed and optimized the element probe, which proved that Cas14 has a preference for longer lengths in element probe cleavage. Using this method, the quantitative detection of trace aqueous ampicillin can be achieved within 45 minutes at room temperature (25 °C). A detection limit as low as 2.06 nM is obtained with excellent performance in anti-interference tests and complex matrix detection.

We report the design of an electrochemical aptasensor for ampicillin detection, which is an antibiotic widely used in agriculture and considered to be a water contaminant. We studied the transducing potential of nanostructure composed of MoS2 nanosheets and conductive polypyrrole nanoparticles (PPyNPs) cast on a screen-printed electrode. Fine chemistry is developed to build the biosensors entirely based on robust covalent immobilizations of naphthoquinone as a redox marker and the aptamer. The structural and morphological properties of the nanocomposite were studied by SEM, AFM, and FT-IR. High-resolution XPS measurements demonstrated the formation of a binding between the two nanomaterials and energy transfer affording the formation of heterostructure. Cyclic voltammetry and electrochemical impedance spectroscopy were used to analyze their electrocatalytic properties. We demonstrated that the nanocomposite formed with PPyNPs and MoS2 nanosheets has electro-catalytic properties and conductivity leading to a synergetic effect on the electrochemical redox process of the redox marker. Thus, a highly sensitive redox process was obtained that could follow the recognition process between the apatamer and the target. An amperometric variation of the naphthoquinone response was obtained regarding the ampicillin concentration with a limit of detection (LOD) of 10 pg/L (0.28 pM). A high selectivity towards other contaminants was demonstrated with this biosensor and the analysis of real river water samples without any treatment showed good recovery results thanks to the antifouling properties. This biosensor can be considered a promising device for the detection of antibiotics in the environment as a point-of-use system.

In this work, a simple and rapid method based on the lateral flow assay (LFA) has been developed for the detection of dual antibiotics. To achieve the quantitative assay and to reduce the non-specific adsorption, an internal system has been developed. A non-specific DNA was exploited as an internal standard and could be recognized by the DNA marker that was coated at the internal line. Two different kinds of aptamers were applied to recognize ampicillin (AMP) and kanamycin (KAM), and the distance between the detection line and conjugate pad was then optimized. Under the optimum conditions, the quantitative assays of AMP (R 2 = 0.984) and KAM (R 2 = 0.990) were achieved with dynamic ranges of 0.50 to 500.0 ng/L, and of 0.50 to 1000.0 ng/L, respectively. The LOQs of AMP and KAM were 0.06 ng/L and 0.015 ng/L, respectively. Finally, the proposed method has been successfully applied to analyze aquaculture water, tap water, and lake water, and hospital wastewater, indicating the established method could be used to monitor the environment.

Testing milk for antibiotics before acceptance into dairies is required by the U.S. Pasteurized Milk Ordinance. Technological advances in tests have reduced screening times and improved detection accuracy. This work describes the validation of the Charm Rapid One Step Assay Beta-Lactam 30 Second Test according to the U.S. Food and Drug Administration Center for Veterinary Medicine protocol for raw commingled milk. Milk is added to the lateral flow test strip in an incubator/reader to deliver a 30 second result. Independent laboratory validation followed sensitivity, interference, and incurred residue protocols. Sensitivity, in parts per billion (ppb = µg/kg), using a probit curve determined 90% percent detection with 95% confidence, which met National Conference of Interstate Milk Shipments (NCIMS) specifications. Six U.S. approved beta-lactam drugs were detected below, but within 50% of, target/tolerance levels for penicillin G 2.9 ppb, ampicillin 5.9 ppb, amoxicillin 5.8 ppb, cephapirin 13 ppb, cloxacillin 8.1 ppb, and ceftiofur metabolites 73 ppb. No interferences were observed from 33 animal drugs at 100 ppb, somatic cells at 1.2 million/mL, or bacterial levels of >300 000 CFU/mL. Incurred residue detection levels were similar to levels determined with the spiked parent compound. The data support NCIMS that the BL30SEC method met U.S. criteria for testing milk for beta-lactams., (© AOAC INTERNATIONAL 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Self-supported Fe 3 N-Co 2 N nanoarray with high electric conductivity and large surface area was prepared for growth of MIPs and further constructing a sensitive and stable electrochemical sensor. For the evaluation of its performance, Fe 3 N-Co 2 N is used as sensing electrode material, and AMP is used as template molecule to construct the MIP electrochemical sensor. Under the optimized conditions, the developed MIPs electrochemical sensor detects AMP with a low detection limit of 3.65 × 10 -10  mol L -1 and shows outstanding reproducibility and stability. When the MIPs electrochemical sensor was applied to detect AMP in milk samples via standard addition method, the recovery within 97.06-102.43% with RSD of 1.05-2.11% was obtained. The fabrication of MIPs electrochemical sensor is highly promising for sensitive and selective electrochemical measurement and food safety testing. This work can provide theoretical guidance for truly challenging problems. Graphical abstract Principle diagram of MIP-EC sensor for detecting AMP Molecular imprinted polymers (MIPs) are widely performed for construction of electrochemical (EC) sensors especially for detecting small molecules in complex environment. However, the large-scale and robust preparation of MIPs in situ on sensor platform limits their practical applications. We fabricated a MIPs EC sensor based on Fe 3 N-Co 2 N in situ grown on carbon cloth (CC) as the substrate platform (Fe 3 N-Co 2 N/CC) combining with MIPs as the target recognition element for the label-free detection of AMP. Under the optimal conditions, the developed MIPs EC sensor can detect AMP with a low detection limit of 3.65 × 10 -10  mol L -1 . When the AMP in milk is detected by the proposed EC sensor, it shows ideal results. Therefore, the use of self-supported Fe 3 N-Co 2 N nanoarray as the platform for the fabrication of MIPs EC sensors is highly promising for sensitive and selective EC measurement and point-of-care testing.

A novel optical immunosensor for the screening of ampicillin (Amp) residues has been developed. The biosensor is based on fiber optic particle plasmon resonance detection and uses an enhancement method called as fiber optic nanogold-linked immunosorbent assay (FONLISA) for the sensitive detection of antibiotics. A commercial antibody which had a higher affinity for ampicillin than for other β-lactam antibiotics was chosen. A surface competitive binding assay was used in which a fixed concentration of antibiotic-conjugated gold nanoparticles (AuNPs) competes with free unlabeled antibiotic molecules to measure the amount of binding with antibody molecules immobilized on an optical fiber. The synthesis of the 11-mercaptoundecanoic acid (MUA)-ampicillin conjugate facilitates the attachment of the Amp molecules to AuNPs via MUA which acts as a linker between them. This AuNP-Amp conjugate was then used for the detection of β-lactam antibiotics. The practical limit of detection obtained for Amp was 0.74 ppb (7.4 × 10 -10  g/mL) which is lower than the recommended maximum residue limit (MRL) for β-lactams. The method also shows a wide linear range of 4 orders. Its applicability to the determination of ampicillin in spiked milk samples has been demonstrated with good recovery and reproducibility. Graphical abstract.

Rationale: Paper spray mass spectrometry (PS-MS) was used to analyze and quantify ampicillin, a hydrophilic compound and frequently utilized antibiotic. Hydrophilic molecules are difficult to analyze via PS-MS due to their strong binding affinity to paper substrates and low ionization efficiency, among other reasons., Methods: Solvent and paper parameters were optimized to increase the extraction of ampicillin from the paper substrate. After optimizing these key parameters, a Resolution IV 1/16 fractional factorial design with two center points was employed to screen eight different design parameters simultaneously., Results: Pore size, sample volume, and solvent volume were the most significant factors affecting average peak area under the curve (AUC) and the signal-to-blank (S/B) ratio for the 1 μg/mL ampicillin calibrant. After optimizing the key parameters, a linear calibration curve with a range of 0.2 μg/mL to 100 μg/mL was generated (R 2  = 0.98) and the limit of detection (LOD) and lower limit of quantification (LLOQ) were calculated to be 0.07 μg/mL and 0.25 μg/mL, respectively., Conclusions: The statistical optimization procedure undertaken here increased the mass spectral signal intensity by more than a factor of 40. This statistical method of screening followed by optimization experiments proved faster and more efficient, and produced more drastic improvements than typical one-factor-at-a-time experiments., (© 2020 John Wiley & Sons, Ltd.)

Antibiotics are one of the most widely used types of drugs in pharmaceutics. However, efficiency of these drugs has decreased recently owing to the threat of antibiotic resistance. One of the important factors causing antibiotic resistance is the excessive use of antibacterials in animals. Therefore, detection of antibiotics in foods of animal origin is crucial. The aim of this study was to develop a novel whole-cell based bioassay to be used for detection of some antibiotics. Green fluorescent protein (GFP)-expressing Escherichia coli cells were used as a recognition agent, and antibiotic detection was carried out by pursuing the inhibition rate of fluorescence intensity as a result of the inhibition of viable cells by the time of progress. The performance of bioassay was tested for different antibiotics, and the obtained results showed that the developed method can be used successfully for detection of ampicillin, benzylpenicillin, gentamicin, neomycin, and tetracycline with the limit of detection (LOD) values of 3.33, 0.29, 28.00, 618.36, and 33.17 µg/L, respectively. The assay was also tested with antibiotic spiked milk samples (skimmed UHT, full-fat UHT, and whole raw milk). According to obtained recovery values, developed method was successful for all samples. The precision and bias values of the method were found between the range of 1.30% to 7.54% and -8.00% to 0.64%, respectively. The developed method, which is inexpensive and simple with detection limits in line with the regulatory limits, is promising for use in milk quality monitoring. Method has potential to be used as a screening method after comprehensive validation. PRACTICAL APPLICATION: This method could be used in animal husbandry to check whether the antibiotic prescribed for the treatment of sick animals is still present in their milk as residual. For dairy industry, detection of residual antibiotics in milk is crucial because of their inhibition effects on the fermentation processes. Therefore, the proposed method can be used for routine analysis of raw milk reception in dairy industries. In addition, it is considered to have a wide range of applications for all foods., (© 2020 Institute of Food Technologists®.)

Ampicillin is a widely used β-lactam antibiotic that has been detected in various effluents and can alter biological processes used in wastewater treatment such as nitrification. Physiological and kinetic behaviour of a nitrifying consortium in the presence of ampicillin (AMP) (10, 25, and 50 mg/L) was evaluated in batch cultures. Under the experimental conditions (320 ± 8 mg bacterial protein/L, C/N = 2.4, 24 h), the nitrifying behaviour was very similar among the controls without AMP and the assays with antibiotic, as there was no AMP effect on efficiency (E NH4+  = 99.7 ± 4.2%), yields (Y NO2 -  = 0, Y NO3 -  = 1.0 ± 0.1 mg N/mg NH 4 + -N consumed), neither specific rates of NH 4 + oxidation and NO 3 - formation. Therefore, nitrifying bacteria were insensitive to AMP presence. At all assayed concentrations, after 24 h, 70.5 ± 3.7% of AMP was removed from the cultures through abiotic (16.0-16.5%), biosorption (23.2-47.0%), and biotransformation (10.0-29.8%) processes. With the increase in the initial AMP concentration, a greater participation of the biotransformation process, associated to an increase in the specific AMP consumption rate was attained. The sludge was able to completely oxidize NH 4 + to NO 3 - by nitrification and eliminate AMP biologically, but without reaching its full mineralization.

Electromembrane extraction was used with high-performance liquid chromatography for preconcentration and determination of ampicillin residues in cow milk. Ampicillin is transferred from an aqueous solution through a thin layer containing octan-1-ol, silver nanoparticles, and reduced graphene oxide which serves as a supported liquid membrane. Inside the fiber impregnated with supported liquid membrane mixture was filled 10 µL of an acceptor phase. Experimental parameters were optimized for extraction efficiency of ampicillin. Under the optimized conditions, the proposed method provided acceptable linear range (2-100 µg/L), satisfactory repeatability (RSD% < 7.1), low limit of detection (0.6 µg/L), and a high enrichment factor (295) corresponding to extraction recovery of 37%. Consequently, the proposed method was successfully applied for the determination of ampicillin residues in different cow milks., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A simple and sensitive electrochemical method was developed for ampicillin detection based on the protective effect of aptamer-antibiotic conjugate towards endonuclease DpnII activity. Without ampicillin, DNA aptamer firstly hybridizes with the capture probe to form double strand DNA (dsDNA) structure. Then, dsDNA is cleaved by DpnII restriction endonuclease to form two dsDNA fragments. In which, one fragment is released from electrode surface and the other fragment is kept on electrode surface. Then, the dsDNA fragment kept on electrode surface is further digested by Exo III, which leads to the release of the dsDNA fragment from electrode surface. Thus, the electrochemical signal increases due to the decrease of the interface electron transfer resistance causing by the release of dsDNA from electrode surface. However, the formation of dsDNA is blocked when forming aptamer-ampicillin conjugate, which makes the obstruction of the digestion of DpnII and Exo III towards capture probe. Thus, a weak electrochemical signal is achieved due to the increase of the interface electron transfer resistance causing by the dsDNA on the electrode surface. Based on the relationship between ampicillin concentration and the decrease of the electrochemical signal, antibiotic is detected with low detection limit of 32 pM under optimal conditions, which is lower than the mandated maximum residue limit of European Union (9.93 nM). The developed method also presents good detection selectivity. Moreover, the applicability is confirmed by detecting antibiotic in milk and water samples with satisfactory results., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We designed and synthesized a novel covalent organic framework (COF) by condensation polymerization of 1,3,6,8-tetrakis(4-formylphenyl)pyrene and melamine through imine bonds (represented by Py-M-COF). The basic characterizations revealed that the Py-M-COF not only exhibited an extended π-conjugation framework, a large specific surface area (495.5 m 2 g -1 ), big pore cavities, and nanosheet-like structure but also possessed rich functional groups, such as C˭C, C˭N, C˭O, and NH 2 . These features endowed the Py-M-COF with high charge carrier mobility, further improving the strong immobilization of DNA aptamer strands via π-π stacking interaction and electrostatic interaction. As such, the Py-M-COF-based electrochemical aptasensors are ultrasensitive in detecting different antibiotics, including enrofloxacin (ENR) and ampicillin (AMP), yielding extremely low detection limits of 6.07 and 0.04 fg mL -1 (S/N = 3) toward ENR and AMP, respectively, along with other excellent sensing performances. This biosensing platform based on Py-M-COF has potential applications for the sensitive detection of antibiotics or other analytes by replacing the corresponding aptamers., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Surface plasmon resonance technique is highly sensitive to various processes taking place on a metal film and it has emerged as a powerful label-free method to study molecular binding processes taking place on a surface. Another important but less explored area of applications is the use of hybrid methods which combine electrochemistry with optical methods for better monitoring and understanding of biochemical processes. A detection method based on surface plasmon resonance was developed for ampicillin, applying electrochemical techniques for the elaboration and characterization of the aptasensing platform used in this study. Ampicillin is a broad-spectrum β-lactam antibiotic, used both in human and veterinary medicine for the treatment and prevention of primary respiratory, gastrointestinal, urogenital, and skin bacterial infections. It is widely used because of its broad spectrum and low cost. This widespread use can result in the presence of residues in the environment and in food leading to health problems for individuals who are hypersensitive to penicillins. The gold chip was functionalized through potential-assisted immobilization, using multipulse amperometry, first with a thiol-terminated aptamer, as a specific ligand and secondly, using the same procedure, with mercaptohexanol, used to cover the unoccupied binding sites on the gold surface in order to prevent the nonspecific adsorption of ampicillin molecules. After establishing the optimal conditions for the chip functionalization, different concentrations of ampicillin were detected in real time, in the range of 2.5-1000 μmol L -1 , with a limit of detection of 1 μmol L -1 , monitoring the surface plasmon resonance response. The selectivity of the aptasensor was proven in the presence of other antibiotics and drugs, and the method was successfully applied for the detection of ampicillin from river water. Graphical abstract ᅟ.

Ampicillin (AMP) is an antibiotic widely used in hospitals and veterinary clinics around the world for treating infections caused by bacteria. Therefore, it is common to find traces of this antibiotic in wastewater from these entities. In this work, we studied the mineralization of this antibiotic in solution as well as the elimination of its antimicrobial activity by comparing different electrochemical advanced oxidation processes (EAOPs), namely electro-oxidation with hydrogen peroxide (EO-H 2 O 2 ), electro-Fenton (EF), and photo electro-Fenton (PEF). With PEF process, a high degradation, mineralization, and complete elimination of antimicrobial activity were achieved in 120-min electrolysis with high efficiency. In the PEF process, fast mineralization rate is caused by hydroxyl radicals (·OH) that are generated in the bulk, on the anode surface, by UV radiation, and most importantly, by the direct photolysis of complexes formed between Fe 3+ and some organic intermediates. Moreover, some products and intermediates formed during the degradation of the antibiotic Ampicillin, such as inorganic ions, carboxylic acids, and aromatic compounds, were determined by photometric and chromatographic methods. An oxidation pathway is proposed for the complete conversion to CO 2 .

Antibiotics are widely used in human medicine and veterinary production. Residues of these compounds reach the water sources through waste or direct application (e.g. aquaculture). The constant input of the parent drugs and their transformation products into the environment leads these pharmaceuticals to be considered as emerging pollutants. For some molecules, the pathway of degradation and formation in products is less known. To assess the impact of these substances in the environment and in the human health, it is necessary to elucidate the transformation products and their kinetic of degradation to evaluate the possible risks. In the present report, the characterization and the degradation kinetic of two widely used β-lactams antibiotics - amoxicillin and ampicillin - was evaluated. Surface water samples containing these antibiotics were submitted to photolysis and analyzed by liquid chromatography coupled to mass spectrometry with Orbitrap detection in order to establish the profile of degradation and the formation of transformation products. Results showed that the degradation of amoxicillin and ampicillin is almost complete and reach their maximum at 48 h in river water. Moreover, a database containing >65 transformation products of amoxicillin and ampicillin was build and real samples of industrial wastewater were analyzed to investigate the occurrence of amoxicillin, ampicillin and their transformation products., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study examined functional bentonite-supported nanoscale Fe/Ni (B-Fe/Ni) for the simultaneous removal of β-lactam antibiotics such as amoxicillin (AMX), ampicillin (AMP) and penicillin (PEN). The results show only 94.6% of AMX, 80.6% of AMP and 53.7% of PEN were removed in the mixed antibiotic solution, while 97.5% of AMX, 85.1% of AMP and 74.5% of PEN were removed in individual antibiotic solution. The decreased removal in a mixed antibiotic solution was attributed to competition between antibiotics for: firstly, active sites of iron oxide for the adsorption; and secondly, accepted electrons for the degradation in passivation of the nZVI surface. These were confirmed by various characterization techniques. Kinetics studies of mixed antibiotics using B-Fe/Ni confirmed that adsorption and degradation occurred simultaneously as removing of antibiotics in the presence of particles. Furthermore, the stability and durability of B-Fe/Ni applied to remove β-lactam antibiotics was demonstrated. Finally, B-Fe/Ni was used to reduce the concentration of mixed antibiotics from pharmaceutical wastewater, which indicated B-Fe/Ni is a promising material for antibiotics wastewater treatment., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aptamer-based lateral flow assays (LFAs) are an emerging field of aptamer applications due to numerous potential applications. When compared to antibodies, potential advantages like cost effectiveness or lower batch to batch variations are evident. The development of LFAs for small molecules, however, is still challenging due to several reasons, primarily linked to target size and accessible interaction sites. In small molecule analysis, however, aptamers in many cases are preferable since immunogenicity is not required and they may exhibit even higher target selectivity. We report the first cross-recognition of a small molecule (ampicillin) and a protein (C-reactive protein), predicted by in-silico analysis, then experimentally confirmed - using two different aptamers. These features can be exploited for developing an aptamer-based LFA for label-free ampicillin detection, functioning also for analysis in milk extract. Most importantly, the principal setup denotes a novel, transferable and versatile general approach for detection of small molecules using competitive LFAs, unlikely to be generally realized by aptamer-DNA-binding otherwise.

Two accurate, precise and sensitive high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) methods were developed for assay of ampicillin (AMP) and dicloxacillin (DX) in the presence of their impurity, 6-aminopenicillanic acid (APA). Method (A) is HPTLC method; using silica gel HPTLC F254 plates as a stationary phase with methanol: chloroform: acetic acid (1:9: 0.2, by volume) as a developing system. All the bands were scanned at 220 nm. Method (B) is reversed phase- HPLC which depended on isocratic elution using C18 column and mobile phase consisting of acetonitrile: water (60:40, v/v), pH adjusted to 4 with orthophosphoric acid, at a flow rate of 1 mL min-1 and ultraviolet detection at 240 nm. The proposed methods were validated as per ICH guidelines and their linearity was evident in the ranges of 0.5-2 μg band-1, 0.4-2 μg band-1 and 0.2-1.2 μg band-1 for method (A) and 5-40 μg mL-1, 5-40 μg mL-1 and 2-16 μg mL-1 for method (B) for AMP, DX and APA, respectively. The proposed methods were successfully used for assay of AMP and DX in pure form and in pharmaceutical formulation where no interference from the excipients was detected.

The molecularly imprinted polymer (MIP) films were electropolymerized on the surface of Au electrodes with luminol and pyrrole (PY) as the two monomers and ampicillin (AM) as the template molecule. The electrochemiluminescence (ECL) intensity peak of polyluminol (PL) of the AM-free MIP films at 0.7V vs Ag/AgCl could be greatly enhanced by AM rebinding. In addition, the ECL signals of the MIP films could also be enhanced by the addition of glucose oxidase (GOD)/glucose and/or ferrocenedicarboxylic acid (Fc(COOH) 2 ) in the testing solution. Moreover, Fc(COOH) 2 exhibited cyclic voltammetric (CV) response at the AM-free MIP film electrodes. Based on these results, a binary 3-input/6-output biomolecular logic gate system was established with AM, GOD and Fc(COOH) 2 as inputs and the ECL responses at different levels and CV signal as outputs. Some functional non-Boolean logic devices such as an encoder, a decoder and a demultiplexer were also constructed on the same platform. Particularly, on the basis of the same system, a ternary AND logic gate was established. The present work combined MIP film electrodes, the solid-state ECL, and the enzymatic reaction together, and various types of biomolecular logic circuits and devices were developed, which opened a novel avenue to construct more complicated bio-logic gate systems., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The objective of this study was to determine minimum inhibitory concentrations (MIC) of frequently used antimicrobials for Escherichia coli and Trueperella pyogenes isolated from postpartum bovine uteri of cows with acute puerperal metritis (APM, n = 67), cows suspected to have APM (n = 37), and healthy cows (n = 37) and to evaluate possible differences in MIC according to clinical signs. Cows with APM had reddish-brown, fetid vaginal discharge and rectal temperature (RT) ≥39.5°C within 21 d in milk; cows suspected to have APM had either reddish-brown, fetid vaginal discharge or RT ≥39.5°C within 21 d in milk; and healthy cows had neither fetid discharge nor RT ≥39.5°C. Samples were collected from cows on commercial dairy herds (n = 7) using the cytobrush technique. A total of 37 T. pyogenes isolates and 85 E. coli isolates were tested. Ceftiofur, a third-generation cephalosporin that is often used to treat APM, was the focus of analysis. Trueperella pyogenes and E. coli were isolated more often from samples of cows with APM (46 and 90%, respectively) compared with samples from healthy cows (19 and 54%, respectively). Regarding cows suspected to have APM, T. pyogenes and E. coli were numerically more often isolated (30 and 70%, respectively) than in healthy cows (19 and 54%, respectively). Minimum inhibitory concentrations of ceftiofur were low. For T. pyogenes and E. coli, MIC 50 (concentration that inhibited growth of 50% of isolates) were 0.25 and 0.5 µg/mL and MIC 90 (concentration that inhibited growth of 90% of isolates) were 0.5 and 1 µg/mL, respectively. Although ceftiofur inhibited all T. pyogenes at the highest concentration tested (64 µg/mL), the growth of 5.9% of E. coli was not impaired. Recently, ampicillin has been suggested as an alternative treatment for APM. Although the T. pyogenes isolates exhibited low MIC in general (MIC 50 ≤0.015 µg/mL and MIC 90 = 0.06 µg/mL) and 81.1% of all T. pyogenes could be inhibited at the lowest ampicillin concentration tested, 11.8% of the E. coli isolates were not impaired at the highest concentration (64 µg/mL) tested in this study. The MIC 50 and MIC 90 of E. coli were 4 and ≥128 µg/mL, respectively. We detected no difference in the MIC distributions of ceftiofur or ampicillin among isolates from the 3 APM groups. In summary, E. coli with high MIC against ceftiofur as well as against ampicillin were found in this study., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The prevalence of two groups of antibiotics; namely penicillin and sulfonamides was studied in fresh milk available in Kathmandu Valley of Nepal. The milk samples (n = 140) were collected from three different sources; individual farmers, cottage dairies and organized dairies of Kathmandu valley. Qualitative and semi-quantitative analysis with rapid screening kits revealed that 23% samples were positive for antibiotic residues in the fresh milk for penicillin and sulfonamide groups (1-256 µg/kg). High performance liquid chromatography (HPLC) analyses detected 81% samples positive for amoxicillin (68-802 µg/kg), 41% for sulfadimethoxine (31-69 µg/kg), 27% for penicillin G (13-353 µg/kg), and 12% for ampicillin (0.5-92 µg/kg). Due to the precision and accuracy of liquid chromatography method, it detected more positive samples and consequently presented higher prevalence than the rapid screening kits. The antibiotic residues were found above the maximum residue limits that presented serious threat to consumer health and raised a serious concern regarding the implementation and monitoring of international regulations in developing countries.

We report the design and fabrication of a "signal-on" electrochemical aptamer-based (E-AB) sensor for detection of ampicillin. The signaling of the sensor is based on target binding-induced changes in the conformation and flexibility of the methylene blue-modified aptamer probe. The sensor's response is fast; signal saturation can be reached in ~ 200s. Since all the sensor components are surface-immobilized, it is regenerable and can be reused for at least three times. It has demonstrated good specificity and is capable of differentiating between ampicillin and structurally similar antibiotics such as amoxicillin. More importantly, it is selective enough to be employed directly in complex samples, including serum, saliva, and milk. Although both alternating current voltammetry (ACV) and square wave voltammetry (SWV) are suitable sensor characterization techniques, our results show that ACV is better suited for target analysis. Even under the optimal experimental conditions, the limit of detection of the sensor obtained in ACV (1µM) is significantly lower than that obtained in SWV (30µM)., (Copyright © 2017 Elsevier B.V. All rights reserved.)

An aptamer based method is described for the electrochemical determination of ampicillin. It is based on the use of DNA aptamer, DNA functionalized gold nanoparticles (DNA-AuNPs), and single-stranded DNA binding protein (ssDNA-BP). When the aptamer hybridizes with the target DNA on the AuNPs, the ssDNA-BP is captured on the electrode surface via its specific interaction with ss-DNA. This results in a decreased electrochemical signal of the redox probe Fe(CN) 6 3- which is measured best at a voltage of 0.188 mV (vs. reference electrode). In the presence of ampicillin, the formation of aptamer-ampicillin conjugate blocks the further immobilization of DNA-AuNPs and ssDNA-BP, and this leads to an increased response. The method has a linear reposne that convers the 1 pM to 5 nM ampicillin concentration range, with a 0.38 pM detection limit (at an S/N ratio of 3). The assay is selective, stable and reproducible. It was applied to the determination of ampicillin in spiked milk samples where it gave recoveries ranging from 95.5 to 105.5%. Graphical abstract Schematic of a simple and sensitive electrochemical apta-biosensor for ampicillin detection. It is based on the use of gold nanoparticles (AuNPs), DNA aptamer, DNA functionalized AuNPs (DNA-AuNPs), and single-strand DNA binding protein (SSBP).

In this work the adsorption and the release of ampicillin - a β-lactam penicillin-like antibiotic - from MCM-41, SBA-15, and (amino functionalized) SBA-15-NH 2 ordered mesoporous silica (OMS) materials were investigated. The silica matrices differ for their pore size (SBA-15 vs. MCM-41) mainly, and also for surface charge (SBA-15 and MCM-41, vs. SBA-15-NH 2 ). OMS samples were characterized through small-angle X-rays scattering (SAXS), transmission electron microscopy (TEM), N 2 adsorption-desorption isotherms, Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and potentiometric titrations. The quantification of immobilized and released ampicillin was monitored by mean of UV-Vis spectroscopy. Experimental adsorption isotherms evidenced that ampicillin's loading is not related to the pore size (d BJH ) of the adsorbent. Indeed the maximal loadings were 237mg/g for SBA-15 (d BJH =6.5nm), 278mg/g for MCM-41 (d BJH =2.2nm), and 333mg/g for SBA-15-NH 2 (d BJH =5.6nm). Loading seems, instead, to be related to the surface charge density (σ) of the sorbent surface. Indeed, at pH 7.4 ampicillin drug is negatively charged and likely prefers to interact with SBA-15-NH 2 (σ SBA-15-NH2 =+0.223Cm -2 ) rather than the slightly negatively charged silicas (σ SBA-15 =-0.044Cm -2 and σ MCM-41 =-0.033Cm -2 ). Similarly, ampicillin release is affected by interfacial interactions. Indeed, we found a burst release from pure silica samples (SBA-15 and MCM-41), whereas a sustained one from SBA-15-NH 2 sample. We explain this behavior as a result of an attractive interaction between the protonated amino group of SBA-15-NH 2 and the negatively charged carboxylate group of ampicillin. In summary, in order to obtain a sustained drug release, the chemical nature of the matrix's surface plays a role which is more important than its textural features. SBA-15-NH 2 matrix is hence a suitable candidate for local sustained release of antibiotic drugs., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The aim of this study was to prepare molecularly imprinted polymers (MIPs) with ampicillin (AMP) and to evaluate the feasibility of these materials for being used as solid phase extraction sorbent for the selective preconcentration and determination of AMP in cow milk samples. MIPs were synthesized by bulk polymerization using methacrylic acid or methyl methacrylate as monomer and ethylene glycol dimethacrylate as cross-linker at different ratios. Characterization of the MIPs were carried out by Fourier-transform infrared spectrometry. The variables affecting the molecularly imprinted solid-phase extraction (MISPE) procedure were optimized. AMP recoveries were higher than 98%, and RSD less than 7%. A preconcentration factor of 20 was reached, which was sufficient to determine AMP at levels allowed by the EU (4μgkg -1 ) in cow milk. The selectivity of the AMP-MIP was evaluated in presence of other structurally related β-lactam antibiotics (amoxicillin, oxacillin, penicillin G)., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The inherent difficulty of discovering new and effective antibacterials and the rapid development of resistance particularly in Gram-negative bacteria, illustrates the urgent need for new methods that enable rational drug design. Here we report the development of 3D imaging cluster Time-of-Flight secondary ion mass spectrometry (ToF-SIMS) as a label-free approach to chemically map small molecules in aggregated and single Escherichia coli cells, with ∼300 nm spatial resolution and high chemical sensitivity. The feasibility of quantitative analysis was explored, and a nonlinear relationship between treatment dose and signal for tetracycline and ampicillin, two clinically used antibacterials, was observed. The methodology was further validated by the observation of reduction in tetracycline accumulation in an E. coli strain expressing the tetracycline-specific efflux pump (TetA) compared to the isogenic control. This study serves as a proof-of-concept for a new strategy for chemical imaging at the nanoscale and has the potential to aid discovery of new antibacterials.

The aim of this study was detections of antibiotic resistance and resistance mechanism in bacteria isolated from mosquitos (Culex pipiens) living near humans. Therefore, antibiotic resistance in bacteria isolated from Culex pipiens was investigated by disk diffusion test and MIC E-test in this study. MALDI-TOF mass spectrometry was used for detection of resistant mechanism. In this study, hydrolytic breakdown products after a few hours of incubation of the bacteria isolated from Culex pipiens were detected. Results show that enzymatic destruction of ampicillin by beta-lactamases is able to be detected by MALDI-TOF mass spectrometry from wild strains of potential pathogens. The MALDI-TOF mass spectrometry is useful method for routine detection of beta-lactamases resistant mechanism, but overnight incubation of pure culture is necessary. The results are important for proper and fast intervention to limit the spread of beta-lactamase-producing wild bacteria and provide information for appropriate initial therapy of the infections caused by these microbes.

A fast, inexpensive and sensitive approach for the simultaneous determination of chlortetracycline, ampicilline and sarafloxacin in milk was developed using capillary electrophoresis coupled with an electrochemiluminescence detector. Under the optimal detection conditions, the linear ranges for chlortetracyline, ampicilline and sarafloxacin were 0.030-5.0, 0.050-5.0 and 0.0040-2.0 μg ml -1 , respectively. The correlation coefficients of chlortetracycline, ampicilline and sarafloxacin were determined as 0.9997, 0.9952 and 0.9978, respectively. Detection limits (S/N = 3) of chlortetracycline, ampicilline and sarafloxacin were found as 0.017, 0.018 and 0.0013 μg ml -1 , respectively. This method was successfully applied for the determination of chlortetracycline, ampicilline and sarafloxacin in milk. The recoveries were between 95.3% and 100%. The relative standard deviations of the detection limit and recovery were less than 2.6% and 3.2%, respectively.