Enhancing the Sustainability of Eco-Friendly Potentiometric Ion-Selective Electrodes for Stability-Indicating Measurement of Ethamsylate: Application in Bulk and Pharmaceutical Formulations.

BACKGROUND: Through the use of sustainable and green chemistry concepts, scientists need to decrease waste, conserve energy, and develop safe substitutes for hazardous compounds, all for protecting and benefiting society and the environment. OBJECTIVE: Four novel eco-friendly ion selective electrodes (ISE) were generated to determine Ethamsylate (ETM) in bulk powder and different pharmaceutical formulations. The present electrodes were fabricated to clearly distinguish ETM from a variety of inorganic, organic ions, sugars, some common drug excipients and the degradation product, hydroquinone (HQ) of ETM, and thus used for stability-indicating methods. METHODS: The electrodes fabrication was based on 2-nitrophenyl octyl ether (NPOE) that was employed as a plasticizer in electrodes 1, 2, and 3 within a polymeric matrix of polyvinyl chloride (PVC) except for electrode 4, in which dibutyl sebacate was used as a plasticizer. Electrodes 1 and 2 were fabricated using tetradodecylammonium bromide as an anionic exchanger, adding 4-sulfocalix-8-arene as an ionophore only to electrode 2 and preparing electrode 1 without incorporation of an ionophore. The fabrication of electrodes 3 and 4 was based on ethamsylate-tetraphenylborate (ETM-TPB) as an ion-association complex in a PVC matrix. The environmental sustainability was assessed using the green analytical procedure index (GAPI), and analytical greenness metric for sample preparation (AGREEprep). RESULTS: Electrodes 1 and 2 had linear dynamic ranges of 10-1-10-5 mol/L and 10-1-10-4 mol/L, respectively, with a Nernstian slope of 49.6 and 53.2 mV/decade, respectively. Electrodes 3 and 4 had linear dynamic ranges of 10-1-10-4 mol/L, with a Nernstian slope of 43.9 and 40.2 mV/decade, respectively. CONCLUSION: The electrodes' selectivity coefficients showed good selectivity for ETM. The utility of 4-sulfocalix-8-arene as an ionophore had a significant influence on increasing the membrane sensitivity and selectivity of electrode 2 compared to other electrodes. HIGHLIGHTS: Four novel eco-friendly ISEs were used for determination of ETM in bulk powder and different pharmaceutical formulations. Different experimental parameters were performed to optimize the determination conditions such as solvent mediators, dynamic response time, effect of pH, and temperature. Stability-indicating measurement of ETM in the presence of its degradate HQ and co-formulated drug tranexamic acid. Using new ecological assessment tools to determine whiteness and greenness profiles.

Exploitation of expired cellulose biopolymers as hydrochars for capturing emerging contaminants from water.

Expired chemicals pose a potential environmental threat to humans and living organisms. Herein, we proposed a green approach whereby expired cellulose biopolymers were converted to hydrochar adsorbents and tested for removing the emerging pharmaceutical contaminants of fluoxetine hydrochloride and methylene blue from water. A thermally stable hydrochar was produced with an average particle size of 8.1 ± 1.94 nm and a mesoporous structure that exhibited a larger surface area than the expired cellulose by 6.1 times. The hydrochar was efficient in removing the two contaminants with efficiencies that reached above 90% under almost neutral pH conditions. Adsorption exhibited fast kinetics and regeneration of the adsorbent was successful. The adsorption mechanism was hypothesized in view of the Fourier Transform Infra-Red (FTIR) spectroscopy and pH effect measurements to be mainly electrostatic. A hydrochar/magnetite nanocomposite was also synthesized, and its adsorption behavior for both contaminants was tested and it revealed an enhanced percent removal relative to the bare hydrochar by 27.2% and 13.1% for FLX and MB, respectively. This work supports the strategies for zero waste management and the circular economy.

Impedimetric Sensors for Cyclocreatine Phosphate Determination in Plasma Based on Electropolymerized Poly(o-phenylenediamine) Molecularly Imprinted Polymers.

Cyclocreatine and its water-soluble derivative, cyclocreatine phosphate (CCrP), are potent cardioprotective drugs. Based on recent animal studies, CCrP, FDA-awarded Orphan Drug Designation, has a promising role in increasing the success rate of patients undergoing heart transplantation surgery by preserving donor hearts during transportation and improving the recovery of transplanted hearts in recipient patients. In addition, CCrP is under investigation as a promising treatment for creatine transporter deficiency, an X-linked inborn error resulting in a poor quality of life for both the patients and the caregiver. A newly designed molecularly imprinted polymer (MIP) material was fabricated by the anodic electropolymerization of o-phenylenediamine on screen-printed carbon electrodes and was successfully applied as an impedimetric sensor for CCrP determination to dramatically reduce the analysis time during both the clinical trial phases and drug development process. To enhance the overall performance of the proposed sensor, studies were performed to optimize the electropolymerization conditions, incubation time, and pH of the background electrolyte. Scanning electron microscopy, electrochemical impedance spectroscopy, and cyclic voltammetry were used to characterize the behavior of the developed ultrathin MIP membrane. The CCrP-imprinted polymer has a high recognition affinity for the template molecule because of the formation of 3D complementary cavities within the polymer. The developed MIP impedimetric sensor had good linearity, repeatability, reproducibility, and stability within the linear concentration range of 1 × 10-9 to 1 × 10-7 mol/L, with a low limit of detection down to 2.47 × 10-10 mol/L. To verify the applicability of the proposed sensor, it was used to quantify CCrP in spiked plasma samples.

Analytical comparability study of anti-CD20 monoclonal antibodies rituximab and obinutuzumab using a stability-indicating orthogonal testing protocol: Effect of structural optimization and glycoengineering.

Glycoengineering and biosimilarity are the key factors for growing, promising and progressive approaches in monoclonal antibodies development. In this study, the physicochemical stability of anti-CD20 rituximab (RTX); originator and biosimilar was compared to its glycoengineered humanized version; obinutuzumab (OBZ). An orthogonal stability-indicating protocol using a set of validated bioanalytical techniques; size exclusion high performance liquid chromatography (SE-HPLC), reversed phase liquid chromatography (RP-HPLC), quantitative gel electrophoresis by TapeStation, receptor binding assay and dynamic light scattering (DLS) was used to investigate the effect of different stress factors on the pattern and kinetics of degradation. SE-HPLC results supported with spectral purity showed similar degradation extent with a different pattern of degradation between RTX and OBZ. A lower tendency to form degraded fragments and a relatively higher favorability for degradation through aggregate formation has been revealed in case of OBZ. Results were in agreement with those of DLS and receptor binding assay which showed specificity to the intact antibodies in the presence of their degradation products. Furthermore, results were additionally confirmed through denaturing quantitative gel electrophoresis which suggested reducible covalent bonds as the mechanism for aggregates formation. RP-HPLC results showed two oxidized forms via excessive oxidation of RTX and OBZ with nearly the same degradation percent. Comparability data of RTX and OBZ using the applied methodologies showed that although glycoengineering; carried out to enhance the therapeutic and biological activity of OBZ altered the pattern of degradation but did not significantly affect the overall stability. Results showed also consistent stability profile between the biosimilar and its originator RTX products.

Label-Free Potentiometric Ion Flux Immunosensor for Determination of Recombinant Human Myelin Basic Protein: Application to Downstream Purification from Transgenic Milk.

Recombinant human myelin basic protein (rhMBP) produced in the milk of transgenic cows was found exclusively associated with milk caseins. This hindered its direct determination without extensive sample pretreatment. Here, a label-free potentiometric immunosensor was developed and validated for the determination of rhMBP. An ion flux was generated under zero-current based on surface blocking of the polymeric membrane ion-selective electrode by anti-hMBP antibody and tetrabutylammonium bromide as a marker ion. The immunosensor was successfully employed in the quantitative determination of hMBP in the range of 0.10-20.00 µg/mL with a limit of detection of 50.00 ng/mL. The applicability of the passive ion flux immunosensor for determination of target analyte in complex matrices was investigated. Downstream purification of rhMBP from the milk of transgenic cows was achieved using cation exchange chromatography, immobilized metal affinity chromatography, and immunoaffinity chromatography. The specificity of the immunosensor along with matrix effect of milk proteins were demonstrated. Results obtained using the rhMBP immunosensor were further cross-validated using an orthogonal testing protocol assembled of RP-HPLC and SE-HPLC. It should be noted that the proposed ion flux immunosensor provided a feasible and specific tool for monitoring rhMBP concentration/purity, immunogenic activity, and stability. Such approach provides an attractive economic alternative to sophisticated biosensors required for in-process quality control of biopharmaceutical products.

Magnetic molecularly imprinted polymer nanoparticles for simultaneous extraction and determination of 6-mercaptopurine and its active metabolite thioguanine in human plasma.

Cytotoxic drugs used in cancer chemotherapy require the continuous monitoring of their plasma concentration levels for dose adjustment purposes. Such condition necessitates the presence of a sensitive technique for accurate extraction and determination of these drugs together with their active metabolites. In this study a novel solid phase extraction technique using magnetic molecularly imprinted nanoparticles (MMI-SPE) is combined with liquid chromatography tandem mass spectrometry (LC-MS/MS) to extract and determine the anti-leukemic agent; 6-mercaptopurine (6-MP) and its active metabolite thioguanine (TG) in human plasma. The magnetic molecularly imprinted nanoparticles (Fe3O4@MIP NPs) were synthesized via precipitation polymerization technique and were characterized using different characterization methods A computational approach was adopted to help in the choice of the monomer used in the fabrication process. The Fe3O4@MIPs NPs possessed a highly improved imprinting efficiency, fast adsorption kinetics following 2nd order kinetics and good adsorption capacity of 1.0 mg/g. The presented MMI-SPE provided the optimum approach in comparison to other reported ones to achieve good extraction recovery and matrix effect of trace levels of 6-MP and TG from plasma. Chromatographic separation was carried out using a validated LC-MS/MS assay and recovery, matrix effect and process efficiency were evaluated. Recovery of 6-MP and TG was in the range of 85.94-103.03%, while, matrix effect showed a mean percentage recovery of 85.94-97.62% and process efficiency of 85.54-96.18%. The proposed extraction technique is simple, effective and can be applicable to the extraction and analysis of other pharmaceutical compounds in complex matrices for therapeutic drug monitoring applications.

Assessment of pectin-coated magnetite nanoparticles in low-energy water desalination applications.

Novel magnetite nanoparticles (NPs) modified with pectin coating were fabricated, characterized, and evaluated as potential draw solute in a forward osmosis (FO) process for water desalination applications. The prepared NPs had a spherical shape with an average diameter of 200 nm and saturation magnetization of 23.13 emu/g. Thermogravimetric analysis (TGA) and FTIR spectra elucidated the successful pectin coating on magnetite surface. The potential use of the fabricated NPs in water desalination was conducted via a newly developed lab-scale FO system. Deionized water, saline water (0.2, 0.5, and 1 g% NaCl solution), and real well water (TDS = 0.9 g%) were used as feed solutions. In all experiments, the water flux gradually decreased along with the extension of experimental time and NaCl rejection rate by the FO membrane was measured to be higher than 95%. Moreover, it was found that the pectin-coated magnetite NPs demonstrated to be able to draw clean water across the FO membrane from well water with a remarkable salt rejection of 97%. Thus, it is believed that the proposed FO system using pectin-coated magnetite NPs as draw solute can be a promising technique for desalination of well waters in an environmental-friendly and energy-saving manner.

In vitro stabilization and in vivo improvement of ocular pharmacokinetics of the multi-therapeutic agent baicalin: Delineating the most suitable vesicular systems.

Baicalin is a multi-purpose flavonoid used in the treatment of different ocular diseases. Owing to its poor stability in basic pH and its poor solubility, a suitable carrier system is needed to enhance its ocular therapeutic potential. Therefore, the objective of this work was to prepare and contrast different baicalin vesicular systems; namely liposomes, penetration enhancer vesicles PEVs and transfersomes. Results revealed that baicalin vesicles exhibited suitable particle size and zeta potential, high entrapment efficiency and controlled release. Depending on the vesicular composition, selected formulations were able to resist physical changes of particle size, zeta potential, entrapment efficiency and in vitro release after storage for 3 months, while retarding the degradation of baicalin. Selected vesicular formulations displayed equivalent or superior antioxidant potential compared to baicalin solution, with absolute superiority over ascorbic acid reference, while demonstrating sterilization endurance and safety on ocular tissues. Pharmacokinetic studies revealed that transfersomes displayed the fastest onset of action, while liposomes displayed the highest extent of absorption as concluded from the Tmax, Cmax, and AUC0-∞ values with 4-5 folds increase in bioavailability compared to baicalin control solution. This delineates baicalin vesicular systems as a promising platform for treatment of ocular diseases such as inflammation, cataract and diabetic retinopathy.

Computer-aided design of magnetic molecularly imprinted polymer nanoparticles for solid-phase extraction and determination of levetiracetam in human plasma.

Analytical methods should be accurate and specific to measure plasma drug concentration. Nevertheless, current sample preparation techniques suffer from limitations, including matrix interference and intensive sample preparation. In this study, a novel technique was proposed for the synthesis of a molecularly imprinted polymer (MIP) on magnetic Fe3O4 nanoparticles (NPs) with uniform core-shell structure. The Fe3O4@MIPs NPs were then applied to separate and enrich an antiepileptic drug, levetiracetam, from human plasma. A computational approach was developed to screen the functional monomers and polymerization solvents to provide a suitable design for the synthesized MIP. Different analysis techniques and re-binding experiments were performed to characterize the Fe3O4@MIP NPs, as well as to identify optimal conditions for the extraction process. Adsorption isotherms were best fitted to the Langmuir model and adsorption kinetics were modeled with pseudo-second-order kinetics. The Fe3O4@MIP NPs showed reasonable adsorption capacity and improved imprinting efficiency. A validated colorimetric assay was introduced as a comparable method to a validated HPLC assay for the quantitation of levetiracetam in plasma in the range of 10-80 µg mL-1 after extraction. The results from the HPLC and colorimetric assays showed good precision (between 1.08% and 9.87%) and recoveries (between 94% and 106%) using the Fe3O4@MIP NPs. The limit of detection and limit of quantification were estimated to be 2.58 µg mL-1 and 7.81 µg mL-1, respectively for HPLC assay and 2.32 µg mL-1 and 7.02 µg mL-1, respectively for colorimetric assay. It is believed that synthesized Fe3O4@MIP NPs as a sample clean-up technique combined with the proposed assays can be used for determination of levetiracetam in plasma.

Optimization of photocatalytic degradation of meloxicam using titanium dioxide nanoparticles: application to pharmaceutical wastewater analysis, treatment, and cleaning validation.

Meloxicam is a commonly prescribed nonsteroidal anti-inflammatory drug with analgesic and fever-reducing effects. In this study, photocatalytic degradation of meloxicam in the presence of TiO2 nanoparticles (TiO2NP) was optimized and applied for pharmaceutical wastewater treatment. A validated stability-indicating orthogonal testing protocol (reversed-phase (RP)-HPLC and capillary zone electrophoresis) was developed and validated for monitoring of meloxicam concentration in the presence of its photodegradation products. Fractional factorial design was employed in order to investigate the effects of pH, irradiation time, UV light intensity, TiO2NP loading, and initial meloxicam concentration on the efficiency of the process. The light intensity was found as the most significant parameter followed by irradiation time and concentration, respectively. The most influencing interactions were noted between irradiation time-concentration and irradiation time-light intensity. The kinetics of meloxicam degradation was investigated at the optimum set of experimental conditions. The protocol was successfully applied for treatment of incurred water samples collected during various cleaning validation cycles. A percentage degradation of 77.34 ± 0.02 % was achieved upon irradiation of samples containing 64.57 ± 0.09 µg/mL with UV light (1012 µW/cm(2), 8 h) in the presence of 0.4 mg/mL TiO2NP at pH 9.0 ± 0.05. Treatment of wastewaters collected during the cleaning validation of each product separately rather than the combined waste should result in a significant improvement in the economics of pharmaceutical wastewater treatment. This could be attributed to the relatively small waste volumes and the ability to tailor the experimental conditions to achieve maximum efficiency.

Comparative study of 2-hydroxy propyl beta cyclodextrin and calixarene as ionophores in potentiometric ion-selective electrodes for neostigmine bromide.

Three novel neostigmine bromide (NEO) selective electrodes were investigated with 2-nitrophenyl octyl ether as a plasticiser in a polymeric matrix of polyvinyl chloride (PVC). Sensor 1 was fabricated using tetrakis(4-chlorophenyl)borate (TpClPB) as an anionic exchanger without incorporation of an ionophore. Sensor 2 used 2-hydroxy propyl ß-cyclodextrin as an ionophore while sensor 3 was constructed using 4-sulfocalix-8-arene as an ionophore. Linear responses of NEO within the concentration ranges of 10(-5) to 10(-2), 10(-6) to 10(-2) and 10(-7) to 10(-2) mol L(-1) were obtained using sensors 1, 2 and 3, respectively. Nernstian slopes of 51.6 ± 0.8, 52.9 ± 0.6 and 58.6 ± 0.4 mV/decade over the pH range of 4-9 were observed. The selectivity coefficients of the developed sensors indicated excellent selectivity for NEO. The utility of 2-hydroxy propyl ß-cyclodextrin and 4-sulfocalix[8]arene as ionophores had a significant influence on increasing the membrane sensitivity and selectivity of sensors 2 and 3 compared to sensor 1. The proposed sensors displayed useful analytical characteristics for the determination of NEO in bulk powder, different pharmaceutical formulations, and biological fluids (plasma and cerebrospinal fluid (CSF)) and in the presence of its degradation product (3-hydroxyphenyltrimethyl ammonium bromide) and thus could be used for stability-indicating methods.

Stability-indicating spectrophotometric and spectrodensitometric methods for the determination of diacerein in the presence of its degradation product.

Three sensitive, selective, and precise stability-indicating methods for the determination of the novel osteoarthritis drug, diacerein (DIA) in the presence of its alkaline degradation product (active metabolite, rhein) and in pharmaceutical formulation were developed and validated. The first method is a first derivative (D(1) ) spectrophotometric one, which allows the determination of DIA in the presence of its degradate at 322 nm (corresponding to zero crossing of the degradate) over a concentration range of 4-40 µg/mL with mean percentage recovery 100.21 ± 0.833. The second method is the first derivative of the ratio spectra (DD(1) ) by measuring the peak amplitude at 352 nm over the same concentration range as (D(1) ) spectrophotometric method, with mean percentage recovery 100.09 ± 0.912. The third method is a TLC-densitometric one, where DIA was separated from its degradate on silica gel plates using ethyl acetate:methanol:chloroform (8:1.5:0.5 v:v:v) as a developing system. This method depends on quantitative densitometric evaluation of thin layer chromatogram of DIA at 340 nm over a concentration range of 1-10 µg/spot, with mean percentage recovery 100.24 ± 1.412. The selectivity of the proposed methods was tested using laboratory-prepared mixtures. The proposed methods have been successfully applied to the analysis of DIA in pharmaceutical dosage forms without interference from other dosage form additives and the results were statistically compared with reference method.

Development and validation of PCR, PLS, and TLC densitometric methods for the simultaneous determination of vitamins B(1), B(6) AND B(12) in pharmaceutical formulations.

This work represents the simultaneous determination of thiamine hydrochloride (B(1)), pyridoxine hydrochloride (B(6)) and cyanocobalamine (B(12)) by two different methods namely spectrophotometry multivariate calibration and densitometry. The spectrophotometric numerical method depends on the use of spectrophotometric data coupled to PLS and PCR multivariate calibration methods for the simultaneous determination of (B(1)) and (B(6)) in the presence of (B(12)) in laboratory prepared mixtures and commercial tablets. A calibration set was prepared, where the three vitamins were modeled using a full factorial 23 with three center points experimental design. This calibration set was used to build the PLS and PCR models. The models were validated by testing their predictive ability on a validation set where low RMSEP, RSEP % were obtained for both models. Figures of merit were determined using the net analyte signal concept. The proposed models were applied successfully to simultaneous determination of B1 and B6 in presence of a low concentration of B12 in pharmaceutical dosage forms that contain simple excipients. The TLC densitometric method was based on the use of a developing system of chloroform: ethanol: water: acetic acid solution (2: 8: 2: 0.5 v/v) to separate the three vitamins. The separated spots were scanned at 242nm, 291nm and 360nm for B(1), B(6) and B(12) respectively. The proposed method was applied successfully to simultaneous determination of the three vitamins in their pure powder form in the range 0.1-1.5 (µg/spot), 0.5-3.5 (µg/spot), 0.1-1.5 (µg/spot) for B(1), B(6), and B(12) respectively and in their pharmaceutical formulations.