Amr M. Mahmoud Mohamed Bekhet

Abstract In this work, simple, reliable, eco-friendly, and quantitative electrochemical sensors were developed to detect chlorhexidine Digluconate (CHX) in a variety of dosage forms, including mouthwashes and intimate douches, as well as chlorhexidine in spiked human saliva. Without any sample pre-treatment or extraction processes, CHX was measured in colored aqueous formulations. Based on carbon screen-printed electrodes, two potentiometric sensors (sensors I and II), utilizing graphene nanocomposites (Gr-NC), were designed (SPEs). An ionophore, 2-hydroxypropyl-?-cyclodextrin, was doped into the Poly Vinyl Chloride (PVC) polymeric membrane to improve sensor selectivity.

One of the most promising new avenues in eco-friendly analytical techniques is the creation of ion-selective electrodes (ISEs). Today, several techniques are being developed to create electrodes that are at once easy to use, very sensitive, long lasting, and remarkably stable. Here, an eco-friendly solid-state microfabricated copper electrode was constructed using readily available, low-cost copper printed circuit boards, and then a Prussian blue analogue (PBA) thin coating was applied for potentiometric determination of safinamide mesylate (SAF). The surface morphology, crystallinity and chemical structure of the produced PBA thin film were investigated using scanning electron microscopy (SEM), X-ray diffractometer (XRD) and X-ray photoelectron spectroscopy (XPS) respectively. The proposed sensor was effectively implemented for the quantification of SAF in pure and pharmaceutical dosage form. There was no statistically significant difference between the results obtained using the suggested technique and the results obtained using the reported one. Also, green-metrics (including GAPI and the AGREE algorithm, which relies on 12 principles of green analytical chemistry) were used to provide a full evaluation of the investigated method greenness. Finally, the newly announced RGB12 model was used to compare the whiteness degree of the proposed and reported methods.

INTRODUCTION: Clomipramine is a tricyclic antidepressant acting as a serotonin reuptake inhibitor. Its maximum plasma concentration (C) is 13-310 ng/mL, the therapeutic range is 220-500 ng/mL and its toxic effect appears in doses above 900 ng/mL.

OBJECTIVES: The fabrication of eco-friendly solid-contact ion-selective electrodes to evaluate the concentration of Clomipramine in different matrices based on disposable screen-printed carbon electrode.

METHODS: Disposable screen-printed carbon electrode was utilized as a substrate to fabricate the proposed sensors. The sensors were optimized to determine Clomipramine using calix[4]arene as an ionophore into PVC polymeric membrane to enhance selectivity towards the target analyte. The solid-contact sensor potential stability was improved by the incorporation of graphene nanoparticles transducer layer.

RESULTS: The sensors were assessed as per the IUPAC recommendations. The linearity range was 1 × 10 to 1 × 10 M. The sensors were successfully applied to determine CLM in the pharmaceutical formulation. Furthermore, the ion selective electrodes were applied for Clompiramine assay in spiked plasma for the purpose of Point-of-Care testing to be a diagnostic tool for therapeutic monitoring of the cited central nervous system agent. The findings were statistically compared to the reported method showing no statistically significant difference.

CONCLUSION: This work was concerned with developing a green analytical method for the determination of Clomipramine. The proposed SC-ISE was mixed with graphene nanocomposite transducer interlayer. The graphene layer succeeded in preventing the formation of an aqueous layer so resulted in a stable, reproducible standard potential besides the rapid response time.

Environmental appearance of antimicrobials due to frequent use of personal care products as recommended by WHO can cause serious flare-up of antimicrobial resistance. In this work, three eco-friendly microfabricated copper solid-state sensors were developed for measuring triclosan in water. Multi-walled carbon nanotubes were incorporated in sensor 2 and 3 as hydrophobic conductive inner layer. Meanwhile, β-cyclodextrin was incorporated in sensor 3 as an ionophore for selective binding of TCS in presence of interfering compounds. The obtained linear responses of sensors 1, 2 and 3 were (1 × 10-1 × 10 M), (1 × 10-1 × 10 M) and (1 × 10- 1 × 10 M), respectively. Limit of detection was 9.87 × 10 M, 9.62 × 10 M, and 9.94 × 10 M, respectively. The miniaturized sensors were utilized for monitoring of triclosan in water samples.

The electropolymerized molecularly imprinted polymers (MIP) have enabled the utilization of various functional monomers with superior selective recognition of the target analyte template. Methyldopa is an attractive synthetic dopamine analogue which has phenolic, carboxylic, and aminic functional groups. In this research, methyldopa was exploited to fabricate selective MIPs, for the detection of sofosbuvir (SFB), by a simple electropolymerization step onto a disposable pencil graphite electrode (PGE) substrate. The interaction between methyldopa, as a functional monomer, and a template has been investigated experimentally by UV spectroscopy. A polymethyldopa (PMD) polymer was electrografted onto PGE in the presence of SFB as a template. X-ray photoelectron spectroscopy (XPS), electrochemical impedance spectroscopy (ESI), and cyclic voltammetry (CV) were used for the characterization of the fabricated sensor. Differential pulse voltammetry (DPV) of a ferrocyanide/ferricyanide redox probe was employed to indirectly detect the SFB binding to the MIP cavities. The sensor shows a reproducible and linear response over a dynamic linear range from 1.0 × 10 M to 1.0 × 10 M of SFB with a limit of detection of 3.1 × 10 M. The sensor showed high selectivity for the target drug over structurally similar and co-administered interfering drugs, and this enabled its application to detect SFB in its pharmaceutical dosage form and in spiked human plasma samples.

The objective of this study is to fabricate solid-contact ion selective electrodes (SC-ISEs) that have long term stable potential. Various conducting polymers such as polyaniline and its derivatives have been successfully employed to improve the potential stability in SC-ISEs. Recently, the role of hydrophobicity at the interface between the conducting polymer solid contact and the ion sensing membrane has been investigated and figured out that the hydrophobic interfaces preclude water layer formation that deteriorate the SC-ISEs potential stability and reproducibility. In this work, a hydrophobic polyaniline derivative was fabricated on the surface of a glassy carbon electrode by electropolymerization of perfluorinated aniline monomers in acidic solution. The electropolymerized hydrophobic polymer was characterized by electrochemical impedance spectroscopy and X-ray photoelectron spectroscopy. The fabricated electrode was employed for determination of midazolam-a model drug-in pharmaceutical formulation without prior extraction. The SC-ISEs performance was optimized, and the potential drift was compared to control SC-ISEs, the SC-ISE linear range was 1 × 10-1 × 10 M, LOD was estimated to be 9.0 × 10 M, and potential drift was reduced to 100 μV/h.

Fabrication of a novel ion selective electrode for determining alcaftadine was achieved. The glassy carbon electrode (GCE) was utilized as a substrate in fabrication of an electrochemical sensor containing polyaniline (PANI) as an ion-to-electron transducer layer. A PVC polymeric matrix and nitrophenyl-octyl-ether were employed in designing the ion-sensing membrane (ISM). Potential stability was improved and minimization of electrical signal drift was achieved for inhibition of water layer formation at the electrode interface. Potential stability was achieved by inclusion of PANI between the electronic substrate and the ion-sensing membrane. The sensor's performance was evaluated following IUPAC recommendations. The sensor dynamic linear range was from 1.0 × 10 to 1.0 × 10 mol L and it had a 6.3 × 10 mol L detection limit. The selectivity and capabilities of the formed alcaftadine sensor were tested in the presence of its pharmaceutical formulation excipients as well as its degradation products. Additionally, the sensor was capable of quantifying the studied drug in a rabbit aqueous humor. Method's greenness profile was evaluated by the means of Analytical Greenness (AGREE) metric assessment tool.

One of the most important reasons for an increased mortality rate of cancer is late diagnosis. Point-of-care (POC) diagnostic sensors can provide rapid and cost-effective diagnosis and monitoring of cancer biomarkers. Portable, disposable, and sensitive sarcosine solid-contact ion-selective potentiometric sensors (SC-ISEs) were fabricated as POC analyzers for the rapid determination of the prostate cancer biomarker sarcosine. Tungsten trioxide nanoparticles (WO NPs), polyaniline nanoparticles (PANI NPs), and PANI-WO nanocomposite were used as ion-to-electron transducers on screen-printed sensors. WO NPs and PANI-WO nanocomposite have not been investigated before as ion-to-electron transducer layers in potentiometric SC sensors. The designated sensors were characterized using SEM, XRD, FTIR, UV-VIS spectroscopy, and EIS. The inclusion of WO and PANI in SC sensors enhanced the transduction at the interface between the screen-printed SC and the ion-selective membrane, offering lower potential drift, a longer lifetime, shorter response time, and better sensitivity. The proposed sarcosine sensors exhibited Nernstian slopes over linear response ranges 10-10 M, 10-10 M, 10-10 M, and 10-10 M for control, WO NPs, PANI NPs, and PANI-WO nanocomposite-based sensors, respectively. From a comparative point of view between the four sensors, PANI-WO nanocomposite inclusion offered the lowest potential drift (0.5 mV h), the longest lifetime (4 months), and the best LOD (9.95 × 10 M). The proposed sensors were successfully applied to determine sarcosine as a potential prostate cancer biomarker in urine without prior sample treatment steps. The WHO ASSURED criteria for point-of-care diagnostics are met by the proposed sensors.

Misuse of illicit drugs is a serious problem that became the primary concern for many authorities worldwide. Point-of-care (POC) diagnostic tools can provide accurate and fast screening information that helps to detect illicit drugs in a short time. A portable, disposable and reproducible core-shell molecularly imprinted polymer (MIP) screen-printed sensor was synthesized as a POC analyzer for the assay of the date rape drug "ketamine hydrochloride" in different matrices. Firstly, the screen-printed electrode substrate was modified electrochemically with polyaniline (PANI) as an ion-to-electron transducer interlayer to improve the potential signal stability. Secondly, core-shell MIP was prepared, the core consisting of silica nanoparticles prepared by Stober's method, while the MIP shell was synthesized onto silica nanoparticles surface by copolymerizing methacrylic acid functional monomer and the crossing agent; ethylene glycol dimethacrylate in the presence of ketamine as a template molecule. Finally, the core-shell MIP was incorporated into the PVC membrane as an ionophore and drop-casted over PANI modified screen-printed carbon electrode. The imprinting process and the morphology of MIP were examined using scanning electron microscopy, Fourier-transform infrared and X-ray photoelectron spectroscopic methods. The sensor exhibited a short response time within 3-5 s in a pH range (2.0-5.0). The potential profile indicated a linear relationship in a dynamic concentration range of 1.0 × 10 M to 1.0 × 10 M with a slope of 54.7 mV/decade. The sensor was employed to determine ketamine in biological matrices and beverages.

This work is dedicated to the greenness estimation of three proposed spectrophotometric techniques [e.g., ratio difference (RD), mean centering of ratio spectra (MCR) and continuous wavelet transform of ratio spectra (CWT)] for the determination of a binary combination named Ofloxacin (OFL) and Ornidazole (ORN). Applying the green analytical chemistry methods to assess the proposed methods has widely attained the analytical community care. The greenness assessment was performed via three evaluation approaches; the "Analytical Eco-Scale", the "National Environmental Method Index" (NEMI) and "Green Analytical Procedure Index" (GAPI). Following the examination of the zero spectrum of OFL and ORN, it is observed that OFL and ORN spectra are overlapped, so they can be detected by the methods mentioned previously. The ratio difference method was carried out at wavelengths of 294.6 nm and 265.6 nm for OFL, 292 nm and 315 nm for ORN. The linear range was (2-15 µg/mL) for OFL and (3-30 µg/mL) for ORN. The MCR method based on the use of mean centered ratio spectra in dual steps and calculating the second ratio spectra mean centered values at 294.6 nm for OFL and 315 nm for ORN. The continuous wavelet transformation which carried out using MATLAB at wavelengths of 265 nm for OFL and 306 for ORN. These techniques were intended for the binary mixture analysis in bulk powder and pharmaceutical formulations with high recoveries. The developed methods were validated according to ICH guidelines. All techniques were statistically compared to either an official method for OFL or a reported method for ORN and the results indicate that there were not any significant differences.

Point of care (POC), also identified as on-site testing, has evolved as a rapid and accurate technique for drug of abuse screening and analysis. The aim of this work is to detect tropicamide (TPC) abuse in biological fluids; we selected rat plasma as example. We developed a disposal miniaturized, portable, green, and budget-friendly POC solid-state electrochemical sensor based on potentiometric transduction. To attain that, an innovative microfabricated electrode modified with conducting polymer poly(3-octylthiophene) (POT) has been placed on sensitized printed circuit board (PCB). A two-stage optimization process was implemented to develop the fabricated electrode. The first stage of the optimization process depends on screening various ionophores in order to enhance the sensor selectivity towards tropicamide. Copper nanoparticles exhibited the highest selectivity towards TPC. The second stage was utilizing a polymer as an ion-to-electron transducer layer between the copper nanoparticles impregnated ion sensing membrane and the microfabricated solid-contact ion-selective electrode. This polymer was added to boost the stability of the potential drift (1.2 mV/h) due to the hydrophobic behavior of the POT, which precludes the formation of an aqueous layer at the Cu electrode/polymeric membrane interface and improve the limit of detection (1.1 × 10 M). Nernstian potentiometric response was accomplished for TPC with a slope of 58.46 ± 0.43 mV/decade and E ∼ 189.39 ± 2.12 over the concentration range 1.0 × 10 to 1.0 × 10 M. The suggested sensor's intrinsic figure of merits include a quick response time (13 ± 2 s) and long life time (45 days). The proposed sensor has been successfully employed in the selective determination of TPC in pharmaceutical formulations, and biological fluids. When the results were compared to those of the official approach, there was no statistically significant difference. The Eco-Scale tool assessed and measured the greenness profile of the established method.

The environmental emergence of unexpected contaminants has gained the attention of the scientific community. A broad spectrum antimicrobial compound named triclosan (TCS) was detected in the environment as an emerging contaminant. Owing to its inherent toxicity, we have proposed eco-friendly potentiometric liquid state sensors to be used for monitoring and quantifying TCS in environmental water samples. The proposed sensors have been optimized by modifying the inner filling solution using hydrophilic 2-hydroxypropyl β-cyclodextrin as a complexing agent to be capable of minimizing the trans-membrane ion flux and hence improving the selective and sensitive determination of TCS in environmental matrices with low LOD values. The obtained linear response of the optimized sensor was (1 × 10 to 1 × 10 M) compared to the control sensor (1 × 10 to 1 × 10 M). The obtained limit of detection (LOD) value was found to be 9.86 × 10 M compared to 9.78 × 10 M of the control sensor. The modification of the inner filling solution of the sensor with 2-hydroxypropyl β-cyclodextrin improves not only its sensitivity but also its response time to be only 5 seconds. The electrical performance of the proposed sensor was evaluated following IUPAC recommendations. Both the pH and temperature effects were studied and optimized. Two different greenness assessment tools, Analytical Eco-scale and Green Procedure Index, were adopted upon the evaluation of the proposed sensors' greenness.

Choline (Ch), is a vitamin-like essential water-soluble organic micronutrient. The US-FDA requires that infant formula not made from cow's milk must be supplemented with Ch. Direct determination of Ch in milk powders and infant formulas is a challenging task due to the lack of a detectable chromophore, its existence in free and complexed forms as well as the presence of multi-analytes in these complex matrices. Here, an enzyme-free potentiometric ion selective electrode (ISE) with high selectivity for Ch, a linear range from 0.03 μM up to 1 mM, a 0.061 μM detection limit (LOD) and a typical response time less than 5 and no greater than 60 s is developed for monitoring of Ch in infant formula and milk powders. To achieve these ISE parameters we relied on the ability of calixarenes and its derivatives to form host-guest complexes with the positively charged quaternary ammonium moiety of Ch. We employed a lipophilic (membrane-compatible) calixarene as an ionophore in the sensing membrane phase to provide a molecular receptor for Ch capable of selective binding; while utilizing, hydrophilic (water-soluble) p-sulfonated calixarene as a buffering agent to optimize the inner filling solution to reduce transmembrane Ch fluxes. All the calixarene structures and their complexes with Ch were optimized at the density functional theory (DFT) level and the Gibbs free energies for the inclusion of Ch into the calixarenes were calculated. The prepared sensor was shown to selectively respond only to Ch in the presence of all other interferents in the tested matrices with results that are not statistically significantly different for either accuracy or precision relative to the much more laborious official AOAC 1999 coupled enzymatic-spectrophotometric method. The proposed method is highly selective, non-enzymatic, requires no derivatization or incubation steps, offers a fast response time, and has the potential of portability for in situ analysis, while being relatively cost effective and non-laborious.

Thin layer chromatography (TLC) is a powerful and simple technique for screening and quantifying low quality and counterfeit pharmaceutical products. The detection methods used to detect and quantify separate analytes in TLC ranges from the densitometric method to mass spectrometric or Raman spectroscopic methods. This work describes the development and optimization of a simple and sensitive TLC method utilizing a smartphone CCD camera for verification of both identity and quantity of antibiotics in dosage form, namely ofloxacin and ornidazole. Mixtures of ofloxacin and ornidazole were chromatographed on a silica gel 60 F plate as a stationary phase. The optimized mobile phase is -butanol : methanol : ammonia (8 : 1 : 1.5 by volume). Iodine vapor has been used as a "universal stain" to visualize the spots on the TLC plates in order to obtain a visual image using the smartphone camera and a desk lamp as an illumination source, thus eliminating the need for a UV illumination source. The recorded images were processed to calculate the values ( values for ofloxacin and ornidazole were 0.12 and 0.76, respectively) which provide identity of the drugs while spot intensity was calculated using a commercially available smartphone app and employed for quantitative analysis of the antibiotics and "acetaminophen" as an example of a counterfeit substance. The smartphone TLC method yielded a linearity of ofloxacin and ornidazole in the range of 12.5-62.5 μg/band and 500-1000 μg/band, respectively. The limit of detection was found to be 1.6 μg/spot for ofloxacin and 97.8 μg/spot for ornidazole. The proposed method was compared with the bench top densitometric method for verification using a Camag TLC Scanner 3, the spot areas were scanned at 320 nm. The value of ofloxacin and ornidazole was calculated to be 0.12 and 0.76, respectively. The densitometric method yielded a linearity of ofloxacin and ornidazole in the range of 5-40 μg/band and 5-50 μg/band, respectively. The limit of detection was found to be 0.8 μg/spot for ofloxacin and 1.1 μg/spot for ornidazole. The proposed method has been successfully applied for the determination of ofloxacin and ornidazole present in more than one pharmaceutical dosage form and was comparable to the densitometric method.