Novel dual-recognition electrochemical biosensor for the sensitive detection of AFM1 in milk.

Aflatoxin M1 (AFM1) is a highly carcinogenic and toxic compound that is commonly found in dairy foods such as milk. We developed an electrochemical MIP (molecularly imprinted polymer) and aptamer (Apt) based aptasensor (Apt-Au@PEIM/AFM1/MIP-Apt/AuNPs)/GCE) with dual-recognition signal amplification to detect AFM1 in milk. The MIP-Apt/AuNP/GCE sensor was prepared by synthesizing an MIP film through the electropolymerization of resorcinol on gold nanoparticles-modified electrodes. The dropwise addition of our cApt-Au@PEIM signal amplification probe could enhance the current signal response and improve the sensitivity of the sensor to detect AFM1. The AFM1 present in milk could prevent the probe from binding to the sensor, thereby drastically reducing the current signal. Under optimal experimental conditions, the sensor exhibits a linear range of 0.01-200 nM and a limit of detection of 0.07 nM (S/N = 3). The developed dual-recognition aptasensor would provide a novel and rapid method for specific detection of AFM1 in milk.

A novel differential ratiometric molecularly imprinted electrochemical sensor for determination of sulfadiazine in food samples.

Herein, we report a novel differential ratiometric molecularly imprinted polymer (MIP) electrochemical sensor for sulfadiazine (SDZ). An MIP membrane with double templates, SDZ and propyl gallate (PG), was fabricated on glassy carbon electrode (GCE) modified by CuInS2/ZnS nanocomposites. After adding PG in the samples as the reference, the current differences between MIP@CuInS2/ZnS/GCE and non-imprinted polymer@CuInS2/ZnS/GCE at the potentials of 0.18 V (ΔIPG) and 0.92 V (ΔISDZ) were measured. The ratio of ΔISDZ/ΔIPG was used for SDZ determination in the differential and ratiometric dual-mode. The influence of the variations in electrode modification and sample enrichment conditions on the determination of SDZ can be suppressed by 2.8 âˆ¼ 13.2-fold, enhancing the reproducibility and stability of the MIP sensor. The interference level was reduced by one order of magnitude compared with the normal MIP mode. The proposed sensors were used to determine SDZ in food samples, with the detection limit of 2.1 nM.

An electrochemical AChE-based biosensor for organophosphate pesticides using a modified CuNWs/rGO nanocomposite on a screen-printed carbon electrode.

An enzymatic electrochemical biosensor was built for the indirect detection of organophosphates (OPs), based on acetylcholinesterase inhibition. The biosensor was fabricated for enhanced performance on a screen-printed carbon electrode (SPCE), modified with copper nanowires (CuNWs) composited with reduced graphene oxide (rGO). The oxidation current was measured using the cyclic voltammogram (CV) method, as generated by the enzymatic interaction between acetylcholinesterase (AChE) and its substrate, acetylthiocholine (ATCh). The biosensing response is the reduction in signal caused by the inhibition of acetylcholinesterase in the presence of an organophosphate inhibitor. Benchmarking shows that the CuNWs/rGO nanocomposite enhanced the signal current considerably and decreased the oxidation potential for electrochemical detection of the OP chlorpyrifos, exhibiting a wide linear detection rangefrom 10 µg/L-200 µg/L, with a limit of detection of 3.1 µg/L and limit of quantification of 12.5 µg/L. This sensor is useful for the analysis of chlorpyrifosin drinking water and orange juice, with high recovery rates and no interference effects.

Enzymatic construction Au NPs-rGO based MIP electrochemical sensor for adulteration detection of bovine-derived allergen in camel milk.

In this work, a high-performance molecularly imprinted polymer (MIP) sensor for the determination of ß-lactoglobulin (ß-LG) was fabricated by using trypsin as a template removal reagent. Gold nanoparticles (Au NPs) and reduced graphene oxide (rGO) designed for electrode modification accelerate the heterogeneous electron transfer rate to enhance the sensitivity of the prepared sensor. With enzymatic hydrolysis, ß-LG templates were effectively digested into short peptides without damage to the MIP so that the imprinted cavities of the MIP were preserved with a complete spatial structure exhibiting high selectivity. Based on the optimization of the protein removal time and pH, the prepared MIP electrochemical sensor could recognize ß-LG in the range of 4-100 ng/mL with a low detection limit (3.58 ng/mL). The sensor also expressed excellent selectivity and was successfully applied to real sample detection. The results demonstrate that the proposed MIP electrochemical sensor may be a promising candidate for camel milk adulteration detection.

Designing tannic acid-polyethyleneimine-modified electrode and novel affinity peptide for ß-lactoglobulin detection in milk.

Harmful substances that cause food allergies can pose a significant threat to consumers along with food safety. According to the World Health Organization (WHO), approximately 10 % of the global population is currently affected by food allergies. Therefore, there is an urgent need for the development of more accurate and precise biosensors capable of detecting these hazardous substances including beta-lactoglobulin. Although numerous detection and analysis methods have been developed, they still suffer from various limitations. In this study, a tannic acid-polyethyleneimine (TA-PEI) network modified screen-printed electrodes (SPE) are newly developed and the binding sequence of peptide against ß-LG was successfully screened using random peptide library. A novel affinity peptide with the desired sequence of S-L-S-P-S-L-W-Q-V-S-M-L-G-G-G-G-E-P-L-Q-L-K-M against ß-lactoglobulin (ß-LG) is designed and synthesized. The synthesized affinity peptide was immobilized on TA-PEI modified SPE to develop peptide-based sensor against ß-LG for the first time. Under successful optimization, the developed sensor exhibited a linear relationship between 50 and 750 ng, with a Kd of 213.9 ng. In addition, the sensor was able to detect ß-LG in cow and goat milk, with average recoveries of 88.5 % and 92.2 %, respectively.

Highly stable electrochemical sensing platform for the selective determination of pefloxacin in food samples based on a molecularly imprinted-polymer-coated gold nanoparticle/black phosphorus nanocomposite.

The overuse of pefloxacin (PEF) leaves residues in foods. Therefore, the development of robust analytical techniques for the selective detection of PEF is of great importance. In this study, a highly stable electrochemical sensing platform has been constructed, using molecularly imprinted polymer (MIP)-coated gold nanoparticle/black phosphorus nanocomposites (BPNS-AuNPs), for the selective detection of PEF. BPNS-AuNPs significantly enhance the black phosphorus (BP) stability and electrochemical activity and offer a larger surface area to accommodate more imprinted sites for selective PEF binding. MIP/BPNS-AuNPs exhibit a broad linear detection range (0.005-10 µM), low detection limit (0.80 nM), and high sensitivity (3.199 µA µM-1). The MIP/BPNS-AuNPs show a high binding affinity for PEF, even in the presence of structural analogs, and maintain stable voltammetric signals for at least 35 d. The MIP sensor exhibits consistent high sensitivity in the detection of PEF in real milk and orange juice samples.

Optimization of voltammetric parameters for sensitive and simultaneous determination of ferulic acid and vanillin using a glassy carbon electrode based on 2-aminonicotinic acid in the presence of surfactant media.

The electrochemical sensor for simultaneous determination of ferulic acid (FA) and vanillin (VA) was prepared by electrochemical deposition of 2-aminonicotinic acid (2-ANA) on the glassy carbon (GC) electrode. The voltammetric determination of FA and VA was performed in the BR buffer solution in the presence of sodium dodecyl sulfate as a surfactant with SWV. The parameters of the SWV technique were optimized by response surface methodology experimental design. Under optimized conditions, the 2-ANA/GC modified electrode presented a linear working range of 2.8 × 10-8 M to 7.50 × 10-6 M and 7.50 × 10-6 M to 2.45 × 10-5 M for FA, 3.06 × 10-8 M to 1.27 × 10-5M for VA. The detection limit (LOD) values for FA and VA were 6.87 nM and 9.23 nM, respectively. Using the 2-ANA/GC sensor, concentrations of FA and VA in instant coffee and milk samples were determined with %recovery values between 103.40 and 97.07 and %RSD values between 0.76 and 4.40.

Hierarchical N-doped porous carbon scaffold Cu/Co-oxide with enhanced electrochemical sensing properties for the detection of glucose in beverages and ascorbic acid in vitamin C tablets.

This research focuses on the development of a highly efficient electrocatalyst, CuxO/NPC@Co3O4/NPC-10-7, for detecting glucose and ascorbic acid. In a 0.1 M NaOH solution, the modified electrode exhibits a sensitivity of 3314.29 µA mM-1 cm-2 for glucose detection. The linear range for ascorbic acid sensing is 0.5 µM - 23.332 mM, with a detection limit as low as 0.24 µM. In a 0.1 M PBS solution, the linear range for ascorbic acid detection extends to 43.328 mM, which represents the best performance reported to date by chronoamperometry. Moreover, the electrode demonstrates high accuracy, with a recovery rate of 96.80 % - 103.60 % for glucose detection and a recovery rate of 95.25 % - 104.83 % for ascorbic acid detection. These results suggest that the CuxO/NPC@Co3O4/NPC-10-7 modified electrode shows significant potential for practical applications in food detection.

Block copolymer-derived recessed nanodisk-array electrodes for electrochemical detection of ß-lactam antibiotics.

Antimicrobial resistance (AMR) is one of the major socio-economic factors contributing to public health. ß-lactams are most commonly prescribed drugs for variety of bacterial infections. Frequent use of antibiotics leads to AMR in humans and animals. The present work is focused on developing an electro-immunosensor to control and regulate the excessive use of antibiotics in animal-based food products. An amphiphilic block co-polymer poly(ethylene oxide-block-methyl methacrylate)(PEO-b-PMMA) was used to fabricate recessed nano-disk array electrode (RNE) and immobilized with Pen-Ab and Cef-Ab antibodies. The Limit of detection (LOD) of RNE working electrode was found to be 14.8 pM for penicillin and 13.8 pM for cefalexin with good selectivity in presence of non-specific antibiotics. Fabricated RNE electrode could detect trace amounts of spiked antigen in real samples of milk, egg and meat extract. Further, mesoporous thin film and microarrays can eventually be used to develop point-of-care diagnosis of antibiotics in animal-based food products.

Ultrasensitive molecular imprinted electrochemical sensor for in vivo determination of glycine betaine in plants.

Glycine betaine (GB) is a bioactive molecule protecting plants from abiotic stress. This study fabricated an ultrasensitive molecular imprinted polymer (MIP) electrochemical sensor to perform in vivo measurements of GB. Polydopamine (PDA) was formed on the carboxylated multi-walled carbon nanotubes (COOH-MWCNTs) by spontaneous polymerisation of dopamine (DA). Then MIP-coated MWCNTs were fabricated on a Au nanoparticles (NP) and thionine (Thi) modified screen-printed electrode (SPE). The MIP-COOH-MWCNTs/pThi/AuNPs/SPE exhibited an ultrasensitive GB detection response between 1 fmol/L and 10 mmol/L (R2 = 0.996) with a low detection limit (0.707 fmol/L, S/N = 3). In vivo measurement of GB in cucumber seedling leaves under different salinity stress conditions confirmed the practical applicability of the MIP sensor. Thus, this study proposed a novel and promising fabrication method for an electrochemical MIP sensor that has broad application prospects in precision agriculture.

Pt nanoparticles/laser-engraved graphene-based integrated electrochemical platform for point-of-use determination of ponceau 4R, amaranth and tartrazine in food.

This study presents the first integrated electrochemical platform (IEP) with point-of-use and portable features for on-site determination of ponceau 4R (RP), amaranth (AM), and tartrazine (TZ) in food by integrating a mass-producible Pt nanoparticles/laser-engraved graphene (Pt NPs/LEG)-based sensing chip and a reusable electrochemical miniaturized workstation. The sensing chip utilizes Pt NPs/LEG with unique architectures as electrode material and exhibits desirable analytical performance towards RP, AM, or TZ with satisfied linear range (0.25-50 µM for RP, AM, and TZ), high sensitivity (7.29 µA µM-1 cm-2, 7.18 µA µM-1 cm-2, and 2.02 µA µM-1 cm-2 for RP, AM, and TZ, respectively) and low limit of detection (50 nM, 58 nM, and 204 nM for RP, AM, and TZ, respectively). The sensing chip shows excellent selectivity, high stability, outstanding reproducibility, and acceptable mechanical stability. IEP can be implemented to test RP, AM, or TZ in real samples with satisfactory accuracy and recoveries.

Detection of luteolin in food using a novel electrochemical sensor based on cobalt-doped microporous/mesoporous carbon encapsulated peanut-like FeOx composite.

Luteolin (Lu) is a dietary flavonoid that has attracted much attention due to its multiple health benefis effects. Herein, an ultrasensitive electrochemical sensor for Lu was constracted based on cobalt-doped microporous/mesoporous carbon (MMC) encapsulated peanut-like Fe2O3 composite. The FeOx-Co-MMC composite was obtained by pyrolyzing a precursor named Fe2O3-Co-microporous/mesoporous dopamine (Fe2O3-Co-MMPDA) which was synthesized by a soft template method. Under optimized conditions, the sensor exhibited good detection of Lu with a low limit of detection (LOD) of 0.031 nM and a wide linear range from 0.05 to 1000 nM in detecting Lu. It also demonstrated good reproducibility (RSD = 3.16%), stability (RSD = 2.34%), and anti-interference properties. The sensor successfully detected Lu in real food samples such as honeysuckle with recoveries ranging from 96.1% to 104.8% (RSD <3%, n = 3). The present study provides an alternative method for Lu detection in food.

A Review on Implantable Neuroelectrodes.

The efficacy of every neuromodulation modality depends upon the characteristics of the electrodes used to stimulate the chosen target. The geometrical, chemical, mechanical and physical configuration of electrodes used in neurostimulation affects several performance attributes like stimulation efficiency, selectivity, tissue response, etc. The efficiency of stimulation in relation to electrode impedance is influenced by the electrode material and/or its geometry. The nature of the electrode material determines the charge transfer across the electrode-tissue interface, which also relates to neuronal tissue damage. Electrode morphology or configuration pattern can facilitate the modulation of extracellular electric field (field shaping). This enables selective activation of neurons and minimizes side effects. Biocompatibility and biostability of the electrode materials or electrode coating have a role in glial formation and tissue damage. Mechanical and electrochemical stability (corrosion resistance) determines the long-term efficacy of any neuromodulation technique. Here, a review of electrodes typically used for implantable neuromodulation is discussed. Factors affecting the performance of electrodes like stimulation efficiency, selectivity and tissue responses to the electrode-tissue interface are discussed. Technological advancements to improve electrode characteristics are also included.

Voltammetric sensor for an anti-cancer drug cisplatin based on bismuth nanoparticles/graphene modified glassy carbon electrode.

A selective and sensitive voltammetric sensor for the measurement of the chemotherapy drug cisplatin was based on bismuth nanoparticles decorated on a graphene-modified glassy carbon electrode (BiNPs/Gr/GCE). The surface morphologies of electrode modifications were characterized using scanning electron microscopy (SEM). Electrochemical characteristics were evaluated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Differential pulse voltammetry (DPV) was employed to detect cisplatin oxidation. Under the optimal conditions, oxidation peak currents increased linearly with cisplatin concentration from 6.0 to 180 µM. The detection limit was calculated to be 4.4 µM and the limit of quantification was 14.7 µM. Analytical performance studies demonstrated the good electrode reproducibility and the selectivity of the BiNPs/Gr/GCE towards cisplatin detection. The developed cisplatin sensor was successfully utilized to measure cisplatin in samples of human serum. Recoveries ranged from 89 to 102% and the results from the developed sensor were concordant (P > 0.05) with the results of analysis by high-performance liquid chromatography coupled with ultraviolet detection.

Disposable electrochemical immunoplatform to shed light on the role of the multifunctional glycoprotein TIM-1 in cancer cells invasion.

Detecting overexpression of cancer biomarkers is an excellent tool for diagnostic/prognostic and follow-up of patients with cancer or their response to treatment. This work illustrates the relevance of interrogating the levels of T-cell immunoglobulin and mucin domain 1 (TIM-1) protein as a diagnostic/prognostic biomarker of high-prevalence breast and lung cancers by using an amperometric disposable magnetic microparticles-assisted immunoplatform. The developed method integrates the inherent advantages of carboxylic acid-functionalized magnetic beads (HOOC-MBs) as pre-concentrator support and the amperometric transduction at screen-printed carbon electrodes (SPCEs). The immunoplatform involves a sandwich-type immunoassay assembled on HOOC-MBs through the specific capture/labeling of TIM-1 using capture antibodies and horseradish peroxidase (HRP)-conjugated biotinylated detection antibodies as biorecognition elements. The magnetic immunoconjugates were confined onto the working electrode (WE) surface of the SPCEs for amperometric detection using the hydroquinone/hydrogen peroxide/HRP (HQ/H2O2/HRP) redox system. The method allows the selective detection of TIM-1 protein over the 87-7500 pg mL-1 concentration range in only 45 min, with a limit of detection of 26 pg mL-1. The developed bioplatform was successfully applied to the analysis of breast and lung cancer cell extracts, providing the first quantitative results of the target glycoprotein in these types of samples.

Adsorptive stripping voltammetric determination of carbofuran in food using novel type of modified carbon-based electrode with grafted layers of nickel.

A voltammetric determination of carbofuran (CBF) was developed using a novel type of carbon-containing electrode (CCE) modified with carbon ink (CI) and a chromatographic sorbent (CS) based of chromaton (Ch), polyethylene glycol and nickel acetylacetonate with grafted layers of nickel (NiCS, stands for Ni modified CS) further denoted as CI/NiCS/CCE. The surface morphology of this modified electrode was investigated by scanning electron microscopy (SEM) and by electrochemical impedance spectroscopy (EIS). CBF which is not electrochemically oxidizable was first hydrolyzed in alkaline medium to give anodically active phenolic analogue CBFP. The electrochemical reactions of CBFP at CI/NiCS/CCE were studied in phosphate buffer (PB) by cyclic voltammetry (CV) and linear sweep adsorptive stripping voltammetry (LSAdSV) using linear scan voltammetry in the first derivative mode (LSVFD). Linear concentration dependences in the concentration ranges from 0.1 to 10 µM and from 10 to 100 µM were obtained by the LSAdSV with limit of detection (LOD) and limit of quantification (LOQ) 0.06 and 0.19 µM, respectively. The novel modified CI/NiCS/CCE showed good stability and selectivity and was successfully used to determine CBF in real samples of vegetables and fruits with LOD 0.01 mg kg-1.

Toward the early diagnosis of tuberculosis: A gold particle-decorated graphene-modified paper-based electrochemical biosensor for Hsp16.3 detection.

Tuberculosis (TB) currently remains a major life-threatening disease as it can be fatal if not treated properly or in a timely manner. Herein, we first describe a disposable and cost-effective paper-based electrochemical biosensor based on a gold particle-decorated carboxyl graphene (AuPs/GCOOH)-modified electrode for detecting heat shock protein (Hsp16.3), which is a specific biomarker indicating the onset of TB infection. The device pattern was first engineered to facilitate detection procedures and printed on low-cost filter paper to create hydrophobic and hydrophilic regions using a wax printing technique. Immunoassays proceeded in a half-sandwich format because it is a reagent-less approach and requires no labeling step. The fabrication of the immunosensor began with GCOOH drop casting, the electrochemical deposition of AuPs, and the establishment of a biorecognition layer against Hsp16.3 utilizing 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS)-sulfo standard chemistry. The appearance of Hsp16.3 resulted in a substantial decrease in the electrochemical signal response of the redox probe employed [Fe (CN)6]3-/4- due to the created immunocomplexes that possess insulation properties. GCOOH enables direct antibody immobilization, and AuPs enhance the electrochemical properties of the sensor. This proposed immunosensor, while requiring only a miniscule sample volume (5 µL), achieved superior performance in terms of the limit of detection, measuring at 0.01 ng/mL. Our platform was confirmed to be highly specific to Hsp16.3 and can rapidly detect TB-infected sera without necessitating any pre-enrichment (20 min), making it an alternative and particularly suitable for the early diagnosis of TB in resource-scarce countries.

Conductive PEDOT:PSS copolymer electrode coatings for selective detection of dopamine in ex vivo mouse brain slices.

A novel voltammetric sensor was developed to selectively determine dopamine (DA) concentration in the presence of ascorbic acid (AA) and 3,4-dihydroxyphenylacetic acid (DOPAC). This sensor utilizes a modified pencil graphite electrode (PGE) coated with a newly synthesized poly (3,4-ethylene dioxythiophene) (PEDOT):poly (styrene sulfonate-co-2-(3-(6-Methyl-4-oxo-1,4-dihydropyrimidin-2-yl) ureido) ethyl methacrylate) (P(SS-co-UPyMA)) composite. The PEDOT:P(SS-co-UPyMA) (PPU) composite was characterized using nuclear magnetic resonance, X-ray photoelectron, and Raman spectroscopies. The PPU-coated PGE was characterized using electrochemical techniques, including cyclic and differential pulse voltammetry. Compared to uncoated, PPU-coated PGE demonstrated improved sensitivity and selectivity for DA. The sensor exhibited a dynamic linear range of 0.1-300 µM for DA, with a detection limit of 44.4 nM (S/N = 3). Additionally, the PPU-coated PGE showed high reproducibility and storage stability for four weeks. To demonstrate its practical applicability, the PPU-coated PGE sensor was used for ex vivo brain slice samples from control and Parkinson's disease model mice.

An innovative and universal dual-signal ratiometric spectro-electrochemical imprinted sensor design for sandwich type detection of anticancer-drug, gemcitabine, in serum samples; cross validation via computational modeling.

An innovative and universal imprinted sensor design for sandwich type detection of gemcitabine (GMT) in human serum samples is described. GMT is widely used in the treatment of different tumors, such as lung, ovarian, pancreatic, and breast cancer. The serum albumin-drug interaction was translated to design a multifunctional, ratiometric and dual mode silver nanoparticle based probe (BSA-Ag nanoprobe), as a read out system. Subsequently, polypyrrol imprinted drug receptor sites was engineered to selectively capture the GMT on the transducer surface. The GMT was sandwiched between imprinted receptor surface and BSA-Ag nanoprobe to generate the spectro-electrochemical signals. The formation of nanoprobe was confirmed through various characterization techniques, including X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, micro-Raman spectroscopy, Dynamic light scattering (DLS), and UV-Visible (UV-Vis) analysis, while each step of sensor fabrication was characterized via field emission scanning electron microscope (FE-SEM), Static water Contact angle measurements, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). Different variable parameters were optimized to improve the analytical performance of the sensor design. Under optimal conditions, spectro-electrochemical sensor permitted linear ranges between 1 and 200 µmol L-1 and 0.5-200 µmol L-1, with limits of detection (LOD) of 0.4 µmol L-1 and 0.15 µmol L-1 respectively. Furthermore, the designed sensor successfully differentiated the serum samples of lung cancer patients and healthy volunteers. The obtained results were validated with standard Liquid chromatography-mass spectrometry (LC/MS) analysis of the patients and healthy volunteer's serum samples. Lastly, density functional theory (DFT) and molecular docking calculations revealed the enhanced GMT binding capability of molecularly imprinted polypyrrole and molecular level interaction between the GMT and BSA, to validate the sandwich sensor design.

Eco-friendly electrochemical sensor for determination of conscious sedating drug "midazolam'' based on Au-NPs@Silica modified carbon paste electrode.

Benzodiazepines (BZDs) are a group of drugs prescribed for their sedating effect. Their misuse and addictive properties stipulate different authorities for developing simple, fast and accurate analytical methods for instantaneous detection. Differential pulse voltammetric technique (DPV) was utilized for the selective assay of midazolam hydrochloride (MDZ) in the pure, parenteral dosage forms and plasma samples. A chemically modified carbon paste electrode (CPE) was implemented during the study. The method depended on the electroreduction of MDZ on the surface of the electrode over a potential range of 0.0 V to -1.6 V. The electrode was fabricated using silica nanoparticles (Si-NPs) which were incorporated into the composition of the CPE and used to enhance the electrode performance. Then, to enhance the sensitivity of the method, a chronoamperometric modification step was applied for depositing gold nanoparticles (Au-NPs) on the carbon paste electrode surface. Modification with Au-NPs showed a higher reduction current peak for MDZ with well-defined peaks. Various parameters such as pH of the media and measurements scan rate were investigated and optimized to enhance the sensor sensitivity. The sensor showed a dynamic linear response over a concentration range of 4.0 × 10-7 M to 2.9 × 10-4 M of MDZ with a LOD of 2.24 × 10-8 M using 0.1 M acetate buffer (pH 5.6). The sensor was validated in accordance with the ICH guidelines regarding accuracy, precision and specificity for the selective assay of MDZ in the presence of excipients. A greenness evaluation was performed using three different assessment tools, namely, the "Green Analytical Procedure Index" (GAPI), the "Analytical Greenness metric" (AGREE) and the "Whiteness Analytical Chemistry tool" (WAC) using the RGB12 model.