Quality-by-design ecofriendly potentiometric sensor for rapid monitoring of hydroxychloroquine purity in the presence of toxic impurities.

Hydroxychloroquine (HCQ) is prescribed to treat malaria and certain autoimmune diseases. Recent studies questioned its efficiency in relieving COVID-19 symptoms and improving clinical outcomes. This work presents a quality-by-design approach to develop, optimize, and validate a potentiometric sensor for the selective analysis of HCQ in the presence of its toxic impurities (key starting materials), namely 4,7-Dichloroquinoline (DCQ) and hydroxynovaldiamine (HND). The study employed a custom experimental design of 16 sensors with different ion exchangers, plasticizers, and ionophores. We observed the Nernstian slopes, correlation coefficients, quantification limit, response time, and selectivity coefficient for DCQ and HND. The computer software constructed a prediction model for each response. The predicted responses strongly correlate to the experimental ones, indicating model fitness. The optimized sensor achieved 93.8% desirability. It proved a slope of 30.57 mV/decade, a correlation coefficient of 0.9931, a quantification limit of 1.07 × 10-6 M, a detection limit of 2.18 × 10-7 M, and a fast response of 6.5 s within the pH range of 2.5-8.5. The sensor was successfully used to determine HCQ purity in its raw material. The sensor represents a potential tool for rapid, sensitive, and selective monitoring of HCQ purity during industrial production from its starting materials.

A Box-Behnken design-based chemometric approach to optimize the sono-photodegradation of hydroxychloroquine in water media using the Fe(0)/S2O82-/UV system.

The huge utilization of hydroxychloroquine in autoimmune infections led to an abnormal increment in its concentration in wastewater, which can pose a real risk to the environment, necessitating the development of a pretreatment technique. To do this, we are interested in researching how hydroxychloroquine degrades in contaminated water. The main goal of this investigation is to optimize the operating conditions for the sono-photodegradation of hydroxychloroquine in water using an ultrasound-assisted Fe(0)/ S 2 O 8 2 - /UV system. To get adequate removal of HCQ, a chemometric method based on the Box-Behnken design was applied to optimize the influence of the empirical parameters selected, including Fe(0) dose, S 2 O 8 2 - concentration, pH, and initial HCQ concentration. The quadratic regression model representing the HCQ removal rate (η(%)) was evolved and validated by ANOVA. The optimal conditions as a result of the above-mentioned trade-off between the four input variables, with η(%) as the dependent output variable, were captured using RSM methodology and the composite desirability function approach. For HCQ full decomposition, the optimal values of the operating factors are as follows: S 2 O 8 2 - dose, 194.309 mg/L; Fe(0) quantity, 198.83 mg/L; pH = 2.017, and HCQ initial dose of 296.406 mg/L. Under these conditions, the HCQ removal rate, achieved after 60 min of reaction, attained 98.95%.

Análise das prescrições de pacientes com covid-19 em uso de hidroxicloroquina em uma unidade de terapia intensiva / Analysis of prescriptions of patients with COVID-19 using hydroxychloroquine in an Intensive Care Unit / Análisis de las prescripciones de pacientes con COVID-19 en uso hidroxicloroquina en una unidad de terapia intensiva

Mesmo em emergências sanitárias, quando terapias experimentais são empregadas, é importante prezar pela segurança e eficácia no uso de medicamentos, e a análise de prescrições médicas é uma das maneiras de monitorar aspectos de segurança. Objetivo: Quantificar e classificar as interações medicamentosas potenciais com hidroxicloroquina de acordo com o riscoem prescrições de pacientes com COVID-19 em pacientes com COVID-19 em uso de hidroxicloroquina admitidos em uma unidade de terapia intensiva de um Hospital de Ensino.Metodologia:Este estudo transversal baseou-se na análise de 162 prescrições de 38 pacientes admitidos em uma unidade de terapia intensiva de um Hospital de ensino entre abril e junho de 2020.O Micromedex® e o UpToDate® foram as bases de dados de apoio à conduta clínica utilizadas para estabelecer as interações medicamentosas potenciais. Resultados:A média de dias de internamento foi de 16,1 ± 14,0 e a média de dias em uso de hidroxicloroquina foi de 4,26 ± 1,74. 87,14% das prescrições apresentaram interações medicamentosas potenciais e a mais comum foi entre hidroxicloroquina e azitromicina. 76,4% das prescrições analisadas apresentaram interações medicamentosas potenciais com hidroxicloroquina. 73,5% das prescrições tiverampelo menos uma interação medicamentosa potencial entre medicamentos que prolongam o intervalo QT. Conclusões: Tendo em vista os riscos da exposição de pacientes críticos às interações medicamentosas, este estudo demonstra a necessidade de fortalecer nas instituições hospitalares a cultura de monitoramento de parâmetros de segurança e eficáciano uso de medicamentos, inclusive em terapias experimentais com a utilização de medicamentos off-labelpara minimizar riscos e ampliar possíveis benefícios (AU).

Even in health emergencies, when experimental therapies are employed, it is important to ensure the safety and efficacy of medicines, and the analysis of medical prescriptions is one of the ways to monitor safety aspects.Objective: Quantify and rank potential drug interactions with hydroxychloroquine according to risk in prescriptions of COVID-19 patients taking hydroxychloroquine admitted to an intensive care unit of a TeachingHospital.Methodology: This cross-sectional study was based on the analysis of 162 prescriptions of 38 patients admitted to an intensive care unit of a teaching hospital between April and June 2020. Micromedex® and UpToDate® were the clinical practice support databases used to establish potential drug interactions. Results: The mean number of days of hospitalization was 16.1 ± 14.0 and the mean number of days of days on hydroxychloroquine was 4.26 ± 1.74. 87.14% of the prescriptions presented potential drug interactions and the most common was between hydroxychloroquine and azithromycin. 76.4% of the analyzed prescriptions had potential drug interactions with hydroxychloroquine. 73.5% of prescriptions had at least one potential drug interaction between drugs that prolong the QT interval. Conclusions: In view of the risks of exposure of critically ill patients to drug interactions, this study interactions, this study demonstrates the need to strengthen in hospital institutions the culture of institutions the culture of monitoring safety and efficacy parameters in the use of medicines, including experimental therapies with the use of off-label drugs to minimize risks and increase possible benefits (AU).

Aunque en médio aemergencias sanitarias, cuando son empleadas terapias experimentales, es importante estimar la seguridad y eficacia en el uso de los medicamentos, y el análisis de prescripciones es una de las formas de acompanhar los aspectos de seguridad. Objetivo:Cuantificar y clasificar las interaciones farmacologicas potenciales con hidroxicloroquina de acuerdo com el riesgo em prescripciones de pacientes com Covid-19 em uso de hidroxicloroquina andmitidos em unidad de terapia intensiva de um Hospital Docente. Metodología: Este estudio transversal se asienta en el análisis de 162 prescripciones de 38 pacientes admitidos em uma unidad de terapia intensiva de um Hospital Docente entre abril y junio de 2020. El Micromedex®ï¸y el UpToDate®ï¸fueron las bases de datos de apoyo a la actuación clínica utilizadas para establecer las interacciones farmacológicas potenciales. Resultados:El promedio de días de internamiento fue de 16,1 ± 14,0 y el promedio de días en uso hidroxicloroquina fuede 4,26 ± 1,74. 87,14% de las prescripciones presentaron interacciones farmacológicas potenciales y la más común fue entre hidroxicloroquina y azitromicina. 76,4% de las prescripciones analizadas presentaron interaciones farmacológicas com hidroxicloroquina. 73,5% de las prescripciones tuvierion por lo menos uma interacción farmacológica potencial entre medicamentos que prolongam el intervalo QT. Conclusiones:Tenendo a la vista los riesgos de la exposición de pacientes críticos a las interaciones farmacológicas, este estudio demuestra la necesidad de reforzar em las instituiciones hospitalarias la cultura de monitoreo de parâmetros de seguridade y eficacio em el uso de medicamentos, incluso en terapias experimentales con utilización de medicamentos off-label, para minorar riesgos y ampliar los posibles beneficios (AU).

COVID-19 pharmaceuticals in aquatic matrices: The threatening effects over cyanobacteria and microalgae.

Water contamination by pharmaceuticals is a global concern due to their potential negative effects on aquatic ecosystems and human health. This study examined the presence of three repositioned drugs used for COVID-19 treatment: azithromycin (AZI), ivermectin (IVE) and hydroxychloroquine (HCQ) in water samples collected from three urban rivers in Curitiba, Brazil, during August and September 2020. We conducted a risk assessment and evaluated the individual (0, 2, 4, 20, 100 and 200 µg.L-1) and combined (mix of the drugs at 2 µg.L-1) effects of the antimicrobials on the cyanobacterium Synechococcus elongatus and microalga Chlorella vulgaris. The liquid chromatography coupled to mass spectrometry results showed that AZI and IVE were present in all collected samples, while HCQ occurred in 78 % of them. In all the studied sites, the concentrations found of AZI (up to 2.85 µg.L-1) and HCQ (up to 2.97 µg.L-1) represent environmental risks for the studied species, while IVE (up to 3.2 µg.L-1) was a risk only for Chlorella vulgaris. The hazard quotients (HQ) indices demonstrated that the microalga was less sensitive to the drugs than the cyanobacteria. HCQ and IVE had the highest values of HQ for the cyanobacteria and microalga, respectively, being the most toxic drugs for each species. Interactive effects of drugs were observed on growth, photosynthesis and antioxidant activity. The treatment with AZI + IVE resulted in cyanobacteria death, while exposure to the mixture of all three drugs led to decreased growth and photosynthesis in the cells. On the other hand, no effect on growth was observed for C. vulgaris, although photosynthesis has been negatively affected by all treatments. The use of AZI, IVE and HCQ for COVID-19 treatment may have generated surface water contamination, which could increased their potential ecotoxicological effects. This raises the need to further investigation into their effects on aquatic ecosystems.

Additively manufactured electrodes for the electrochemical detection of hydroxychloroquine.

Although studies have demonstrated the inactivity of hydroxychloroquine (HCQ) towards SARS-CoV-2, this compound was one of the most prescribed by medical organizations for the treatment of hospitalized patients during the coronavirus pandemic. As a result of it, HCQ has been considered as a potential emerging contaminant in aquatic environments. In this context, we propose a complete electrochemical device comprising cell and working electrode fabricated by the additive manufacture (3D-printing) technology for HCQ monitoring. For this, a 3D-printed working electrode made of a conductive PLA containing carbon black assembled in a 3D-printed cell was associated with square wave voltammetry (SWV) for the fast and sensitive determination of HCQ. After a simple surface activation procedure, the proposed 3D-printed sensor showed a linear response towards HCQ detection (0.4-7.5 µmol L-1) with a limit of detection of 0.04 µmol L-1 and precision of 2.4% (n = 10). The applicability of this device was shown to the analysis of pharmaceutical and water samples. Recovery values between 99 and 112% were achieved for tap water samples and, in addition, the obtained concentration values for pharmaceutical tablets agreed with the values obtained by spectrophotometry (UV region) at a 95% confidence level. The proposed device combined with portable instrumentation is promising for on-site HCQ detection.

Development of a chiral HPLC method for the separation and quantification of hydroxychloroquine enantiomers.

Hydroxychloroquine (2-[[4-[(7-Chloroquinolin-4-yl) amino]pentyl](ethyl) amino]-ethanol, HCQ), an effective anti-malarial drug, has been tested in the clinics for potential treatment of severe coronavirus disease 2019 (COVID-19). Despite the controversy around the clinical benefits of HCQ, the existence of a chiral center in the molecule to possess two optical isomers suggests that there might be an enantiomeric difference on the treatment of COVID-19. Due to their poor resolution and the inability of quantification by previously reported methods for the analysis of HCQ enantiomers, it is necessary to develop an analytical method to achieve baseline separation for quantitative and accurate determination of the enantiomeric purity in order to compare the efficacy and toxicity profiles of different enantiomers. In this study, we developed and validated an accurate and reproducible normal phase chiral high-performance liquid chromatography (HPLC) method for the analysis of two enantiomers of HCQ, and the method was further evaluated with biological samples. With this newly developed method, the relative standard deviations of all analytes were lower than 5%, and the limits of quantification were 0.27 µg/ml, 0.34 µg/ml and 0.20 µg/ml for racemate, R- and S-enantiomer, respectively. The present method provides an essential analytical tool for preclinical and clinical evaluation of HCQ enantiomers for potential treatment of COVID-19.

Liquid Chromatographic Methods for COVID-19 Drugs, Hydroxychloroquine and Chloroquine.

COVID-19 has been a threat throughout the world since December 2019. In attempts to discover an urgent treatment regime for COVID-19, hydroxychloroquine (HCQ) and chloroquine (CQ) have been on solidarity clinical trial. However, many countries have pulled HCQ and CQ from their COVID-19 treatment regimens recently, some countries still continue using them for patients who have previously started HCQ and CQ and they may complete their course under the supervision of a doctor. HCQ and CQ are 4-aminoquinoline drugs and it is safe to use them for autoimmune diseases, rheumatoid arthritis, systemic lupus erythematosus and malaria as well. Determination of CQ, HCQ and their metabolites in biologic fluids and in pharmaceuticals has great importance, especially for pharmacokinetics, pharmacodynamics and epidemiological studies. In this review, liquid chromatographic methods developed in the last 10 years were summarized focusing on sample preparation and detection methods for HCQ and CQ determination in biological fluids and pharmaceutical preparations. It is hoped that this article could be helpful to facilitate the use of these drugs in clinical trials or drug research studies as it provides comprehensive information on the reported analytical methods.

A metal organic framework-functionalized monolithic column for enantioseparation of six basic chiral drugs by capillary electrochromatography.

Poly(glycidyl methacrylate)-co-(ethylene dimethacrylate) [poly(GMA-co-EDMA)] monoliths were used as a support to grow a zeolitic imidazolate framework-8 (ZIF-8) via layer-by-layer self-assembly. Pepsin, acting as as chiral selector, was covalently linked to the surface of the amino-modified ZIF-8 through the Schiff base method. The material was characterized by scanning electron microscopy, thermogravimetric analysis, X-ray diffraction, Fourier transform infrared spectroscopy and elemental analysis. The pepsin-ZIF-8-poly(GMA-co-EDMA) column was utilized to the enantioseparation of the racemic forms of hydroxychloroquine (HCQ), chloroquine (CHQ), hydroxyzine (HXY), nefopam (NEF), clenbuterol (CLE) and amlodipine (AML). In comparison with a pepsin-poly(GMA-co-EDMA) monolithic column (without self-assembled ZIF-8 nanoparticles), the resolution is strongly enhanced (HCQ: 0.34 → 2.50; CHQ: 0.45 → 1.97; HXY: 0.39 → 1.43; NEF: 0.27 → 0.81; CLE: 0 → 0.81; AML: 0.16 → 0.72). Effects of self-assembly layers of ZIF-8, pepsin concentration, buffer pH values and applied voltage were investigated with hydroxychloroquine as the model analyte. The reproducibility of run-to-run, day-to-day and column-to-column were explored, and found to be satisfactory. Graphical abstractSchematic representation of capillary electrochromatography (CEC) systems with a pepsin-zeolitic imidazolate framework-8 (ZIF-8) modified poly(glycidyl methacrylate)-co-(ethylene dimethacrylate) [poly(GMA-co-EDMA)] monolithic column as stationary phases for separation of basic racemic drugs. ZIF-8 modified column was prepared via layer-by-layer self-assembly.

The role of photodegradation in the environmental fate of hydroxychloroquine.

For many organic pollutants present in surface waters, photolysis is considered as a major abiotic degradation process. The present study aimed to explore the role of photolysis in the environmental fate of hydroxychloroquine (HCQ) for the first time. The photolytic degradation of HCQ was investigated under simulated solar radiation (300-800 nm) in ultrapure, spring, river, and sea water. The effect of pH on the photodegradation rate was substantial and it was observed that degradation was faster at higher pH-values. Obtained half-lives ranged from 5.5 min at pH 9 to 23.1 h at pH 4. Humic acids, nitrate and iron(III) enhanced photodegradation of HCQ due to formation of hydroxyl radicals and its attack on HCQ molecule. In contrast, chloride, sulfate and bromide inhibited photodegradation. Additionally, the humic acids exhibited a dual role, photosensitization and inner filter effect. The study of the reaction kinetics was performed with HPLC-PDA, while the identification of degradation products formed during photolytic degradation was carried out using HPLC-MS/MS and NMR spectroscopy. The hydroxylation was recognized as the dominant path of photoproducts formation. The results of this research reveal the importance of photolytic degradation in environmental fate of HCQ and enable a better understanding of its behavior in the environment. Moreover, the results showing the significant effect of pH on the photodegradation of HCQ can be very useful in water treatment processes.

Simultaneous quantitation of hydroxychloroquine and its metabolites in mouse blood and tissues using LC-ESI-MS/MS: An application for pharmacokinetic studies.

Hydroxychloroquine (HCQ) has been shown to disrupt autophagy and sensitize cancer cells to radiation and chemotherapeutic agents. However, the optimal delivery method, dose, and tumor concentrations required for these effects are not known. This is in part due to a lack of sensitive and reproducible analytical methods for HCQ quantitation in small animals. As such, we developed and validated a selective and sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method for simultaneous quantitation of hydroxychloroquine and its metabolites in mouse blood and tissues. The chromatographic separation and detection of analytes were achieved on a reversed phase Thermo Aquasil C18 (50×4.6mm, 3µ) column, with gradient elution using 0.2% formic acid and 0.1% formic acid in methanol as mobile phase at a flow rate of 0.5mL/min. Simple protein precipitation was utilized for extraction of analytes from the desired matrix. Analytes were separated and quantitated using MS/MS with an electrospray ionization source in positive multiple reaction monitoring (MRM) mode. The MS/MS response was linear over the concentration range from 1 to 2000ng/mL for all analytes with a correlation coefficient (R2) of 0.998 or better. The within- and between-day precision (relative standard deviation, % RSD) and accuracy were within the acceptable limits per FDA guidelines. The validated method was successfully applied to a preclinical pharmacokinetic mouse study involving low volume blood and tissue samples for hydroxychloroquine and metabolites.

Sensitive and selective determination of hydroxychloroquine in the presence of uric acid using a new nanostructure self-assembled monolayer modified electrode: optimization by multivariate data analysis.

A highly sensitive electrochemical nanosensor was developed using covalent modification of a glassy carbon electrode (GCE) by self-assembly of a novel Schiff base. Scanning electron microscopy (SEM) and electrochemical techniques were used to investigate the immobilization of the self-assembled monolayer (SAM) on the GCE. The electrochemical behavior of hydroxychloroquine (HCQ) in the presence of uric acid (UA) at the surface of the modified electrode was studied using the differential pulse voltammetry (DPV) technique. Response surface methodology (RSM) is used to optimize the effects of various operating variables such as pH, immersion time, scan rate, step potential and modulation amplitude on the voltammetric response of HCQ. RSM formulates a mathematical model which correlates the independent parameters with the peak current of HCQ. The central composite rotatable design (CCRD) has been applied to conduct the experiments. Then, under the optimized conditions, HCQ was determined in the presence of UA. The electrochemical measurements demonstrated that this biosensor responded well to HCQ, confirming that the self-assembly immobilization method was effective. Also, the interference, the storage stability, and the reproducibility of the biosensor were studied and assessed. The developed nanosensor was economical and efficient, making it potentially attractive for application to real sample analysis.

Identification and characterization of process related impurities in chloroquine and hydroxychloroquine by LC/IT/MS, LC/TOF/MS and NMR.

The focus of this study is identification and characterization of major unknown impurities in chloroquine (CQ) and hydroxychloroquine (HCQ) bulk drug samples using liquid chromatography/ion trap mass spectrometry (LC/IT/MS) and liquid chromatography/time of flight mass spectrometry (LC/TOF/MS). The newly developed LC/MS method was employed for the analysis of both the drugs. The analysis revealed the presence of two impurities in each of the drugs. The impurities are designated as CQ-I, CQ-II (for chloroquine); HCQ-I and HCQ-II (for hydroxychloroquine). Three of the impurities, CQ-II, HCQ-I and HCQ-II were unknown have not been reported previously. Accurate masses of the impurities were determined by using Q-TOF mass spectrometer and fragmentation behavior was studied by an ion trap mass spectrometer. Based on the spectrometric data and synthetic specifics the structures of CQ-II, HCQ-I and HCQ-II were proposed as 1,4 pentanediamine, N(4)(7-chloro-4-quinolinyl), N(4)-chloromethyl, N(4)-ethylamine; 2-(4-(7-chloroquinolin-4-ylamino) pentylamino) ethanol and [[4-[(7-chloro-4-quinolyl) amino] N-pentyl] N-chloromethyl-N-ethylamino] ethanol respectively. The impurities were isolated by semi-preparative HPLC and structures were confirmed by NMR spectroscopy. The formation and through characterization of known CQ-I impurity is also discussed.

Enantioselective analysis of the metabolites of hydroxychloroquine and application to an in vitro metabolic study.

A one-step chiral method for the quantification of the enantiomers of two hydroxychloroquine (HCQ) metabolites, desethylchloroquine (DCQ) and desethylhydroxychloroquine (DHCQ) by HPLC is described, in addition to its application to the in vitro study of HCQ metabolism in rat liver microsomes. Liquid-liquid extraction was used to extract the enantiomers from microsome samples and the separation was performed on a Chiralpak AD-RH column protected with an RP-8 guard column using hexane:isopropanol (92:8, v/v) plus 0.1% diethylamine as the mobile phase, at a flow rate of 1.0 mL min(-1). The detection was carried out at 343 nm. The method proved to be linear in the range of 50-5000 ng mL(-1) for DCQ enantiomers and 125-2500ngmL(-1) for DHCQ enantiomers, with a quantification limit of 50 and 125 ng mL(-1), respectively. Precision and accuracy, demonstrated by within-day and between-day assays, were lower than 15%. The metabolic study demonstrated that metabolism is stereoselective for HCQ. The major metabolites formed in the incubation of racemic HCQ were (-)-(R)-DCQ and (-)-(R)-DHCQ with R/S ratios of 2.2 and 3.3, respectively.

Electrochemical study of hydroxychloroquine and its determination in plaquenil by differential pulse voltammetry.

Hydroxychloroquine (HCQ) is a halogenated aminoquinoline that presents wide biological activity, often being used as an antimalarial drug. The electrochemical reduction of HCQ was investigated by cyclic voltammetry and chronoamperometry using glassy carbon electrodes. By cyclic voltammetry, in acid medium, only the cathodic peak was observed. The electrochemical behavior of this peak is dependent on pH and the electrodic process occurs through an E(r)C(i) mechanism. The electron number (1e) consumed in the reduction of HCQ was obtained by chronoamperometry. A method for the electrochemical determination of HCQ in pharmaceutical tablets was developed using differential pulse voltammetry. The detection limit reached was 11.2 microg ml(-1) of HCQ with a relative standard deviation of 0.46%. A spectrophotometric study of HCQ has been also carried out utilizing a band at 343 nm. The obtained detection limit and the relative standard deviation were 0.1 microg ml(-1) and 0.36%, respectively. The electrochemical methods are sufficiently accurate and precise to be applied for HCQ determination, in laboratorial routine, which can be used to determine the drug at low level.