Clonazepam Research Articles

Electrochemical classification of benzodiazepines: A comprehensive approach combining insights from voltammetry and liquid chromatography − mass spectrometry

The growing non-medical use of benzodiazepines (BZs) has led to the emergence of counterfeit BZ pills and new psycho-active substances (NPS) in the BZ class on the illicit market. Comprehensive analytical methods for BZ identification are required to allow law enforcement, first aid responders and drug-checking services to analyze a variety of sample types and contents to make timely decisions on the spot. In this work, the electrochemical behavior of diazepam (DZ), clonazepam (CZ) and alprazolam (AP) is studied on graphite screen-printed electrodes, both with and without dissolved oxygen in the solution, to link their redox signals to their chemical structure. After elucidation of their reduction mechanisms using liquid chromatography coupled to high-resolution mass spectrometry, three structural classes (Class 1, Class 2 and Class 3) were defined, each with different redox centers and electrochemical behavior. Subsequently, 22 confiscated pills containing 14 different BZs were correctly assigned to these three structural classes, with the deoxygenated conditions displaying the highest class selectivity. Finally, the three classes were successfully detected after being spiked into five alcoholic beverages in the context of drug-facilitated sexual assault. For analysis in red wine, which complicated the analysis by interfering with Class 1, a dual test strategy in pH 2 and pH 7 was proposed for accurate detection. Its rapid measurements, broad scope and lack of interference from diluents or colors makes this method a promising approach for aiding various services in combating problematic BZ use.

A versatile ratiometric fluorescence nanoprobe for the determination of clonazepam in patients' plasma samples.

Despite the necessity of the study of therapeutic drug monitoring of clonazepam (CLZ), there are only a few fast detection methods available for determining CLZ in biological media. This study aims to develop a cost-effective and ratiometric probe for the quantification of CLZ in plasma samples. Fluorescent polydopamine nanoparticles were produced through a self-polymerization process at a pH of 8.5. Rhodamine B molecules were employed as a fluorescent reference material, emitting stable fluorescence in the visible range. The fabricated probe exhibited a specific detection capability for CLZ. The fluorescence emission of the probe was enhanced in two concentration ranges: from 50 ng/mL to 1.0 μg/mL and from 1.0 to 15.0 μg/mL with a lower limit of quantification of 50 ng/mL, indicating the sensitivity of the probe for detecting CLZ plasma levels. The accuracy of the probe is favorable which could be recommended for CLZ monitoring in the biological media. Furthermore, this probe is highly specific towards CLZ in the presence of various interfering agents which is mainly caused by its ratiometric nature. The developed platform showed high reliability in quantifying CLZ concentrations in patients' plasma samples. Hence, the fabricated probe could be recommended as a reliable method for the routine detection of CLZ in clinical settings.

Designing Molecularly Imprinted Polymer-Modified Boron-Doped Diamond Electrodes for Highly Selective Electrochemical Drug Sensors.

Drug detection in biological solutions is essential in studying the pharmacokinetics of the body. Electrochemical detection is an accurate and rapid method, but measuring multiple drugs that react at similar potentials is challenging. Herein, we developed an electrochemical sensor using a boron-doped diamond (BDD) electrode modified with a molecularly imprinted polymer (MIP) to provide specificity in drug sensing. The MIP is a polymer material designed to recognize and capture template molecules, enabling the selective detection of target molecules. In this study, we selected the anticancer drug doxorubicin (DOX) as the template molecule. In the electrochemical measurements using an unmodified BDD, the DOX reduction was observed at approximately -0.5 V (vs Ag/AgCl). Other drugs, i.e., mitomycin C or clonazepam (CZP), also underwent a reduction reaction at a similar potential to that of DOX, when using the unmodified BDD, which rendered the accurate quantification of DOX in a mixture challenging. Similar measurements conducted in PBS using the MIP-BDD only resulted in a DOX reduction current, with no reduction reaction observed in the presence of mitomycin C and CZP. These results suggest that the MIP, whose template molecule is DOX, inhibits the reduction of other drugs on the electrode surface. Selective DOX measurement using the MIP-BDD was also possible in human plasma, and the respective limits of detection of DOX in PBS and human plasma were 32.10 and 16.61 nM. The MIP-BDD was durable for use in six repeated measurements, and MIP-BDD may be applicable as an electrochemical sensor for application in therapeutic drug monitoring.

Mesoporous silica nanoparticles decorated with C3N4 framework as a novel electrocatalyst for the design of a selective clonazepam sensor

In this study, a new chemical modifier based on copper nitrate hydroxide-containing mesoporous silica decorated with a C3N4 framework (MSN/C3N4/CNH) was developed to design a modified sensor for the determination of clonazepam (CZP). The resulting composite was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), energy-dispersive X-ray spectroscopy (EDS), simultaneous thermal analysis (STA), Brunauer-Emmett-Teller (BET) and Barrett Joyner–Halenda (BJH) analyses. The fabricated electrochemical sensor which was designed with a new modified glassy carbon electrode (GCE) for CZP analysis, exhibits outstanding electrocatalytic activities toward the reduction of CZP. The results showed that significant sensitivity compared to existing methods improved with an excellent detection limit of 2.50 nmol L−1 for the CZP detection, and the modified electrode has high activity toward the oxidation and reduction of CZP, with an oxidation peak around 0.32 V, and a lower reduction peak around −0.78 V; at pH = 7.0. The effect of experimental and instrumental factors on the electrochemical sensor response was examined, and the cyclic voltammetric results confirmed the 4H+/4e− process in the electrochemical studies. Moreover, in differential pulse voltammetry, the reduction peak current was proportional toward the CZP concentration from 0.82 to 76.90 μmol L−1, at optimum conditions. The modified electrode showed good reproducibility, and repeatability with relative standard deviation values of 2.7% and 1.2%, respectively. The proposed method can be applied as a simple, selective, and precise method to determine CZP drug in real samples, with satisfactory results.

Seizure control and complications in patients switching from clobazam to clonazepam due to drug shortage

BackgroundAntiseizure medication (ASM) shortages are a global problem that have a negative impact on outcomes such as seizure control in patients with epilepsy (PWE). In the case of clobazam (CLB) shortage, there is no study regarding the management strategy. This study aims to investigate the alteration in seizure frequency and the occurrence of side effects in PWE undergoing an abrupt switch from clobazam (CLB) to clonazepam (CLZ), during CLB shortage. Material and methodsA retrospective study was conducted from electronic health records at our neurology outpatient clinic from January to July 2022. Change in seizure frequency and percentage of CLZ-associated side effects were determined as primary and secondary outcomes, respectively. Potential drug-drug interactions (Level C and above) were evaluated by using Lexicomp Drug Interaction Checker. ResultsThe analysis included a total of 29 adult patients (15F, median age: 29). The switching ratio was 10 mg CLB for every 1 mg CLZ (10:1). Seizure frequency was higher during the CLZ period compared to the CLB period (p < 0.05), but no status epilepticus cases were observed. All patients exhibited potential drug-drug interactions, leading to reduced CLZ levels in 12 cases. A total of 36 CLZ-associated side effects were identified, with fatigue (19.4 %), drowsiness (16.6 %), and somnolence (13.8 %) being the most prevalent. A positive and strong correlation was found between CLZ dose and the number of side effects (r: 0.556; p: 0.002). ConclusionThe abrupt switch from CLB to CLZ was observed to increase seizure frequency without leading to status epilepticus in PWE. CLZ-associated side effects were found to be tolerable despite the abrupt switch. Future studies may explore the effect of alternative switching ratios.

Polysomnographic features associated with clonazepam and melatonin treatment in isolated REM sleep behavior disorder: Time for new therapeutic approaches?

Although clonazepam (CLO) and melatonin (MLT) are the most frequently used treatments for REM sleep behavior disorder, the polysomnographic features associated with their use are little known. The aim of this study was to evaluate polysomnographic and clinical parameters of patients with idiopathic/isolated REM sleep behavior disorder (iRBD) treated chronically with CLO, sustained-release MLT, alone or in combination, and in a group of drug-free iRBD patients. A total of 96 patients were enrolled: 43 drug-free, 21 with CLO (0.5-2 mg), 20 with sustained-release MLT (1-4 mg), and 12 taking a combination of them (same doses). Clinical variables and polysomnography were collected. Although clinical improvement was reported in all groups, MLT impacted sleep architecture more than the other treatments, with significant and large increase in N3 stage, moderate reduction in N2 and REM sleep, and moderate increase in REM latency. CLO moderately increased the percentage of both REM sleep and especially N2, while reducing N1 and wakefulness. Patients treated with both CLO and MLT did not show major changes in sleep architecture. These results suggest that the administration of MLT or CLO impacts (positively) on sleep parameters of iRBD patients. However, there is a need to better stratify patients, in order to treat them in a targeted manner, depending on the patient's individual sleep architecture and expected differential effects of these agents.

Efficacy of intravenous clonazepam for paediatric convulsive status epilepticus.

To compare the efficacy of intravenous clonazepam (CLZ) for the initial management of convulsive status epilepticus (CSE) in children as a function of the first-line in-hospital dose used. This monocentric retrospective study included children who received a first dose of CLZ for CSE at Montpellier University Hospital, France, between January 2016 and June 2019. Data from medical records (clinical, treatment, course) were collected and compared as a function of the first CLZ dose used. Among the 310 children treated for CSE, 105 received at least one CLZ dose (median age 3 years; quartile 1-quartile 3 [Q1-Q3] = 1 years 2 months-6 years 6 months). Among these 105 patients, 24 (22%) received a dose less than 0.03 mg/kg (low dose) and 69 (65%) received a dose of at least 0.03 mg/kg (high dose). Seizure cessation rate was not different between the low- and high-dose groups (62.5% vs 76%; odds ratio 0.53, 95% confidence interval [CI] 0.19-1.44, p = 0.29). The administration of a second dose of CLZ was more frequent in the low- than the high-dose group (37.5% vs 16%; odds ratio 3.2, 95% CI 1.1-9.1, p = 0.04). Our study did not find any difference in seizure termination rate as a function of CLZ dose in children with CSE. However, a second CLZ dose was more frequently needed in the group receiving low (less than 0.03 mg/kg) CLZ.

Feasibility of psychological screening in a tertiary epilepsy clinic

ObjectivePeople with epilepsy (PWE) have a high prevalence of developing depression and anxiety. The objective is to determine the feasibility of brief screening tools to screen for depression and anxiety in epilepsy, and the predictive factors. MethodThis is a cross-sectional study in the neurology clinic in a tertiary teaching hospital in Kuala Lumpur. The screening tools used were the Neurological Disorders Depression Inventory for Epilepsy (NDDI-E) and the General Anxiety Disorder Form (GAD-7). ResultsFive hundred and eighty-five patients were recruited in this study, and 50.8% of them were male, predominantly Chinese (46.7%), with a mean age of seizure onset of 21.8 ± 16.1 years. The majority had focal seizures (75.0%), and 41.9% had seizure remission. There were 15.5% who scored ≥15 in the NDDI-E, and 17.0% had moderate or severe anxiety (scored ≥10 in the GAD-7). In a regression model to predict the NDDI-E score, the age of seizure onset recorded a higher beta value (β = −0.265, p =< 0.001), followed by the duration of epilepsy (β = −0.213, p =< 0.001), use of levetiracetam (LEV) (β = 0.147, p = 0.002), clonazepam (CLZ) (β = 0.127, p = 0.011), and lamotrigine (LTG) (β = 0.125, p = 0.011), number of current antiseizure medications (β = −0.124, p = 0.049), seizure remission for ≥1 year (β = −0.108, p = 0.011), and female (β = 0.082, p = 0.049). For the GAD-7 score, the predictors included current age (β = −0.152, p = 0.001), the use of LEV (β = 0.122, p = 0.011), Indian ethnicity (β = 0.114, p = 0.006), and the use of carbamazepine (β = −0.090, p = 0.043). ConclusionImplementation of simple psychological screening using self-administered questionnaires was feasible in a busy tertiary epilepsy clinic.

Insights from molecular dynamics and DFT calculations into the interaction of 1,4-benzodiazepines with 2-hydroxypropyl-βCD in a theoretical study

This study delves into the interaction between benzodiazepine (BZD) drugs and 2-hydroxypropyl-β-cyclodextrin (2HPβCD), a cyclodextrin (CD) known to improve drug delivery and enhance therapeutic outcomes. We find that the 2HPβCD’s atoms become more rigid in the presence of chlordiazepoxide (CDP), clonazepam (CLZ), and diazepam (DZM), whereas they become more flexible in the presence of nordazepam (NDM) and nitrazepam (NZP). We also investigated the structure of 2HPβCD and found that loading these drugs increases both the area and volume of the 2HPβCD cavity, making it more suitable for drug delivery. Moreover, this research found that all drugs exhibited negative values for the binding free energy, indicating thermodynamic favorability and improved solubility. The binding free energy order of the BZDs was consistent in both molecular dynamics and Monte Carlo methods, with CDP and DZM having the highest affinity for binding. We also analyzed the contribution of different interaction energies in binding between the carrier and the drugs and found that Van der Waals energy is the primary component. Our results indicate that the number of hydrogen bonds between 2HPβCD/water slightly decreases in the presence of BZDs, but the hydrogen bond’s quality remains constant.

Chitosan coated niosomes for nose-to-brain delivery of clonazepam: Formulation, stability and permeability studies.

Crossing the Blood Brain Barrier constitutes a challenge in drug administration to the brain. In this context, nose-to-brain delivery is explored as an alternative route in the treatment of central nervous system disorders, and nanotechnology constitutes a promising tool for drug delivery to the brain. In this work, we explored niosomes and chitosan-coated niosomes (chitosomes) as possible tools for nose-to-brain delivery of clonazepam. The formulations have been optimised using different chitosan concentrations and different preparation methods as Thin Layer Evaporation-paddle (TLE-P), Evaporation (E), and Solvent Displacement Technique (SDT). The most suitable formulations were loaded with clonazepam (CLZ) and a full physicochemical characterization was performed. Chitosomes presented a size of around 200 nm, PDI < 0.3, a positive surface charge, spherical shape and a CLZ encapsulation above 60%. Chitosomes were stable for 12 weeks under storage conditions at 4ºC, in simulated nasal fluid for 24 h as well as after a lyophilization-sonication process. A CLZ release of 50% was also achieved after 4 h in this media. The mucoadhesive properties of chitosomes were also confirmed, with a 1.5-fold reduction of CLZ toxicity after encapsulation and a 10-fold increase of its permeability.

Clonazepam Loading Dose in Status Epilepticus: Is More Always Better?

Benzodiazepines are the first treatment line in status epilepticus (SE). Despite their well-established benefit, benzodiazepines are frequently underdosed with potential detrimental consequences. In some European countries, clonazepam (CLZ) is commonly used as the first line treatment. The aim of this study was to explore the correlation between CLZ loading doses and SE outcome. This study included a retrospective analysis of a prospective registry in Lausanne, Switzerland (CHUV Lausanne University Hospital), including all SE episodes treated between February 2016 and February 2021. Only adults (>16 years old) were included with CLZ used as the first treatment line. Post-anoxic SE were excluded because of significant differences in physiopathology and prognosis. Patient characteristics, SE features, the validated SE severity score (STESS), and treatment characteristics were prospectively recorded. We considered loading doses of 0.015 mg/kg or higher (following commonly recommended loading doses)as high doses. We analyzed outcome in terms of number of treatment lines after the CLZ, proportion of refractory episodes, intubation for airways protection, intubation for SE treatment, and mortality. We performed univariable analyses to investigate the association between loading doses and clinical response. A multivariable stepwise backward binary logistic regression was applied for adjusting for potential confounders. Multivariable linear regression was similarly used to analyze CLZ dose as a continuous variable. We collected 251 SE episodes in 225 adult patients. Median CLZ loading dose was 0.010 mg/kg. CLZ high doses were used in 21.9% of SE episodes (in 43.8% for >80% of the high dose). Thirteen percent of patients with SE were intubated for airways control, while intubation was needed in12.7% for SE treatment. High CLZ loading doses were independently associated with younger age (median 62 versus 68 years old, p = 0.002), lesser weight (65kg versus 75kg, p = 0.001) and more frequent intubation for airways protection (23% vs 11%, p = 0.013), but differing CLZ dose was not associated with any outcome parameter. CLZ high doses were more frequently used for SE treatment in younger patients with healthy weight and were more often associated with intubation for airways protection, probably as an adverse event. Varying CLZ dose did not alter outcome in SE, raising the possibility that commonly recommended doses are above what is needed, at least in some patients. Our results suggest that CLZ doses in SE may be individualized depending on the clinical setting.

Is anti-seizure medication the culprit of SUDEP?

Heart rate variability (HRV) reduction is a potential biomarker for sudden cardiac death. This study aimed to study the effects of anti-seizure medications (ASMs), adjusted with reported factors associated with sudden unexpected death in epilepsy (SUDEP) on HRV parameters. We recruited patients who were admitted in our epilepsy monitoring unit between January 2013 and December 2021. Two 5-min electrocardiogram epochs during wakefulness and sleep were selected in each patient. HRV analysis with Python® software was performed. The imputed datasets were used for linear regression analysis to assess association between each ASM item and all HRV parameters. The effects of ASM on HRV parameters were subsequently adjusted with the significant clinical characteristics and the concomitant use of other ASMs, respectively. Carbamazepine (CBZ), levetiracetam (LEV), lamotrigine (LTG), and clonazepam (CZP) were statistically significantly associated with changes of sleep HRV parameters. Only CBZ showed negative effects with reduction in HRV, evidenced as lower standard deviation of RR interval (SDNN), even when adjusted with concomitant use of other ASMs (p = 0.045) and had a trend of significance when adjusted with significant clinical characteristics of concurrent taking of beta-blocker drug (p = 0.052). LEV and CZP showed opposite effects with increased HRV even when adjusted with significant clinical characteristics and the concomitant use of other ASMs. CBZ showed negative effects on HRV. We proposed that CBZ should be cautiously used in patients with known risks for SUDEP. In addition, HRV assessment should be performed prior to commencing CBZ and re-performed in follow-up in cases of prolonged use.

An eco-friendly cellulose support functionalized with tin titanate nanoparticles for the fast removal of clonazepam drug from the drinking water: adsorption mechanisms

This research studied the performance of tin titanate (SnTiO3, SnT) and cellulose-based composites for the removal of clonazepam (CZP) drug by physical adsorption. The cellulose was extracted from a plant named tithonia tubaeformis, which is considered as weed in the crop fields of Mexico. The analysis by microscopy revealed that the SnTiO3 powders are formed by a mixture of coalesced grains and nanotubes with lengths in the range of 97-633 nm. Furthermore, the X-ray diffraction analysis indicated that the SnT powders present a mixture of cassiterite and rutile phases. Experiments for the CZP removal from drinking water were carried out, and several parameters such as initial drug concentration (1-10 mg/L), amount of SnT adsorbent per liter of contaminated solution (10-50 mg/L), and pH (3-10) were varied in order to study their influence on the CZP removal percentage. Essentially, we found that the SnT dosage of 50 mg/L produced the most efficient and fastest CZP removal, since 94.3% of CZP was removed after only 10 min of reaction. Moreover, a piece of cellulose (Cell) was decorated with 50 mg of SnT powder to form the Cell+SnT composite, and this was able to remove a maximum of 80.5% of CZP after 180 min of reaction. If the amount of SnT powder deposited on the Cell+SnT composite is raised up to 100 mg, the composite can remove 95.5% of CZP. The adsorption capacity was also calculated for the SnT powders and Cell+SnT composite and found that it was 6.3 times higher for the SnT powders. Furthermore, the Raman spectra recorded for the Cell+SnT composites demonstrated the presence of surface defects, which acted as adsorption centers for the CZP molecules. The results of this investigation demonstrate that eco-friendly and low-cost floatable composites can be used for the removal of pharmaceutical contaminants, which is an advantage over adsorbent powders.

Eco-friendly Analytical Method for Estimation for Benzodiazepine Drug in Pure and Pharmaceuticals Formulations by Oxidative Coupling Reaction with Phenylephrine Hydrochloride

A simple, fast, sensitive and selective spectrophotometric method has been developed based on the oxidative coupling reaction process, which depends on the determination of clonazepam (CZP) drug using an oxidizing agent sodium periodate (NaIO4) in the presence of a reagent, phenylephrine hydrochloride (PH-HCL), that uses the pink color of CZP at a wavelength of 495 nm. Where several factors aff ecting the color intensity and absorbance were studied, including the eff ect of color stability time, eff ect of the volume of the reagent, the eff ect of the volume of the oxidizing agent, order of addition and temperature. The calibration curve found to obey Lambert beer at range concentration (2–20 mg/L), while the limit of detection (LoD) (8. 9*10-2), limit of quantitation (LoQ) (2.6*10-2) and molar absorbance is 1.4 × 102 liters mol-1cm-1. The method was applied to pharmaceutical preparations (Tablet) and was found to characterize the best precision and accuracy. The standard methods not need any control of temperature and also not aff ected by interferences and henceforth successfully utilized to determine CZP in pharmaceutical preparations

Euterpe oleracea-based biochar for clonazepam adsorption: synthesis, characterization, adsorption properties, and toxicity assays.

The consumption of açaí fruit (Euterpe oleracea) has largely increased worldwide, resulting in a significant increase in the demand for its pulp. As a result, the small producing communities end up with large amounts of açaí endocarp residues, creating local environmental pollution problems. Therefore, chemical and physical routes were investigated for producing açaí endocarp adsorbents to propose a locally viable solution for this problem. The adsorption properties of the produced biochars were tested for clonazepam (CZM) removal, and the toxicity of the final solutions was evaluated. The results revealed that the chemical route generated biochar with about twice the surface area and pore volume (762 m2 g-1 and 0.098 cm3 g-1) than the physical route (498 m2 g-1 and 0.048 cm3 g-1). Furthermore, the Sips isotherm better described the CZM adsorption equilibrium for both biochars, with qs values of 26.94 and 61.86 mg g-1 for the physical- and chemical-activated adsorbents. Moreover, recycling studies were performed, and the chemical-activated biochar was stable for up to three cycles, reaching removal rates superior to 80%. Besides, the final toxicity decreased after the adsorptive treatment. Therefore, chemical activation can be used as a simple and effective method for producing stable and compelling adsorbents as an elegant way of adding value to the residues from açaí production, helping solve local environmental problems.

Electrical Status Epilepticus during Sleep: Risk Factors, Clinical Course, and Treatment Approaches

Abstract Background The efforts of clinicians are focused on determining the predictors for electrical status epilepticus in sleep (ESES) manifestation, due to the negative effect of ESES on cognition. Treatment approaches remain a leading problem because of therapeutic resistance. Objective We looked for potential risk factors for ESES manifestation and summarize the clinical course and therapeutic approaches in patients with idiopathic and symptomatic ESES. Patients and Methods We retrospectively reviewed the medical data of 51 children with idiopathic ESES and 20 children with symptomatic ESES. Results We observed an earlier age of seizure onset (p = 0.0002) and a higher percentage of cases with multiple seizures (p &lt; 0.00001) and with postictal paralysis (p &lt; 0.00001) in idiopathic ESES compared with childhood epilepsy with centrotemporal spikes. In the idiopathic ESES, the treatment consisted of corticosteroids in patients with permanent ESES remission and transient remission, levetiracetam (LEV) children with permanent ESES remission and transient, clonazepam (CZP) children with permanent ESES remission and transient, ethosuximide (ESM), and sulthiame. The patients with symptomatic ESES had more unfavorable evolution, as 19 children had persistent or relapsing ESES course. Conclusion We consider the earlier age of seizure onset (below 5 years) and the presence of multiple seizures and postictal paresis as risk factors for ESES manifestation. ESES is characterized by a significant therapeutic resistance, especially in the group of symptomatic cases. Good results are observed with LEV, ESM, CZP, and steroids.

Bimodal Release Two-In-One Clonazepam Matrix Lozenge Tablets for Managing Anxiety-Related Disorders: Formulation, Optimization and In Vivo Evaluation

Clonazepam (CLZ), an antipsychotic drug reported for its efficiency in managing anxiety-related disorders, is being marketed only as conventional tablets. Some patients have abstention to swallow the conventional tablets; therefore, the proposed study was aimed at developing a buccal lozenge tablet by direct compression of two types of optimized granules. Conazepam’s water solubility was first enhanced by a solid dispersion technique for a fast and better dissolution of type 1 granules, while the impact of gelling polymers was investigated on controlled-release type 2 granules. The optimized formulae met the acceptable pharmacopeial limits for tablets’ evaluation. A differential scanning calorimetry study revealed the compatibility between the drug and used excipients. All formulae gave a burst release of CLZ in the first hour of investigation, followed by a sustained release over 24 h. The formula that showed the highest prolonged in vitro release (99.0 + 0.1%), following the Higuchi diffusion model (R2 = 0.99), was then selected for further study. The formula succeeded in controlling the induced stress in a rat model with a significant impact on the behavioral tests throughout the experiment. The results were further confirmed by a pharmacokinetic study that showed a significant increase in Cmax, Tmax, and AUC (1.5, 2, and 3.9 folds), respectively, compared to oral suspension. The newly proposed delivery system has proven a better efficacy with a reduced dosing frequency.

Simultaneous Estimation of Escitalopram and Clonazepam in Tablet Dosage Forms Using HPLC-DAD Method and Optimization of Chromatographic Conditions by Box-Behnken Design.

The study aimed to develop a new reverse-phase high-performance liquid chromatography (RP-HPLC) method with diode array detection (DAD) detection for simultaneous estimation of escitalopram (EST) and clonazepam (CZP) in tablet dosage forms with a quality by design (QbD) approach. The chromatographic conditions were optimized by Box-Behnken design (BBD) and developed method was validated for the linearity, system suitability, accuracy, precision, robustness, sensitivity, and solution stability according to International Council for Harmonization (ICH) guidelines. EST and CZP standard drugs peaks were separated at retention times of 2.668 and 5.046 min by C-18 column with dimension of 4.6 × 100 mm length and particle size packing 2.5 µm. The mobile phase was methanol: 0.1% orthophosphoric acid (OPA) (25:75, v/v), with a flow rate of 0.7 mL/min at temperature of 26 °C. The sample volume injected was 20 µL and peaks were detected at 239 nm. Using the standard calibration curve, the % assay of marketed tablet was founded 98.89 and 98.76 for EST and CZP, respectively. The proposed RP-HPLC method was able to detect EST and CZP in the presence of their degradation products, indicating the stability-indicating property of the developed RP-HPLC method. The validation parameter’s results in terms of linearity, system suitability, accuracy, precision, robustness, sensitivity, and solution stability were in an acceptable range as per the ICH guidelines. The newly developed RP-HPLC method with QbD application is simple, accurate, time-saving, and economic.

An electrochemical sensor based on cobalt oxyhydroxide nanoflakes/reduced graphene oxide nanocomposite for detection of illicit drug-clonazepam

Clonazepam (CNZ) is a commonly prescribed drug for the management of mental illness including insomnia, panic anxiety and status epilepsy. Due to the associated sedative effects, CNZ has also become a drug of abuse amongst addicted users, sexual assault cases, and suicide. Therefore, it becomes vital to analyze CNZ in human specimens for criminal investigational purposes. Herein, we report the construction of an electrochemical sensor based on self-assembled hybrid nanocomposite of 2-D transition metal oxide nanoflakes i.e., cobalt oxyhydroxide (CoOOH) nanoflakes and reduced graphene oxide (rGO) for determination of clonazepam (CNZ), especially in different beverages. The physico-chemical characterization of the as-synthesized hybrid nanocomposite was done by scanning electron microscopy (SEM), X-Ray diffraction (XRD), raman spectroscopy, X-ray photoelectron spectroscopy (XPS), fourier transform infrared spectroscopy (FTIR) and UV–Visible spectroscopy. Further, the surface of the screen printed carbon electrode (SPCE) was modified with CoOOH-rGO hybrid nanocomposite and its electrochemical characteristics were evaluated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques. A synergistic combination of CoOOH nanoflakes with abundant catalytic sites and highly supportive skeletons of rGO nanosheets that act as conductive bridges for charge transfer makes the nanocomposite sensor unique for electrochemical detection. Under optimized conditions, the specific interaction of CNZ with CoOOH-rGO modified SPCE was determined by differential pulse voltammetry (DPV) to measure the dose dependent increase in reduction current values at a potential of 0.11 V. The developed sensor achieved a detection limit of 38 nM with a sensitivity of 0.054 µA µM−1 cm−2 over a dynamic concentration range of 0.1–350 µM. Moreover, the sensor was applied for determination of CNZ in spiked beverages samples and showed satisfactory recovery in the range of 87–115%. This work provides a new avenue for the development of hybrid nanocomposites based electrochemical sensing platforms for on-site screening of illicit drugs, as necessitated for forensics and clinical settings.

Spectrophotometric Estimation of Clonazepam as Pure Form and in its Pharmaceutical Formulation (Tablet) Using Alizarin Red S

An easy and sensitive spectrophotometric method has been suggested for the assay of clonazepam (CLZM) as pure form and in its formulation as tablets. The present method based on the reaction of CLZM with alizarin red S reagent (ALRS) via proton transfer reaction to form a colored solution with a maximum absorption at wavelength of 528 nm and the optimal conditions for the reaction were studied. The linearity was within the concentration range from 1 to 30 µg/ml, with an R2 value (determination coefficient) of 0.9882 and the sensitivity express via the molar absorption of 7.04x103 l mol-1.cm-1, while Sandell's sensitivity index has been determined and equal to 0.0448 µg/ cm2.The percentage of relative standard deviation (RSD%) which express the precision of the method, percentage of relative error (RE%) express the accuracy, the LOD and LOQ also have been estimated. The application of the suggested method to the determination of CLZM in tablet gave satisfactory results.