Environmentally benign first derivative synchronous spectrofluorimetry for the analysis of two binary mixtures containing duloxetine with avanafil or tadalafil in spiked plasma samples.

Antidepressants can cause sexual dysfunction side effects, necessitating the co-administration of phosphodiesterase type 5 inhibitors. The simultaneous determination of these drugs in biological fluids is critical for therapeutic drug monitoring. For the first time, two binary mixtures containing duloxetine with either avanafil or tadalafil were estimated utilizing simple green spectrofluorimetric methods without the need for a previous separation step. The study was based on first derivative synchronous spectrofluorimetry in ethanol using a change in wavelength difference (∆λ) of 20 and 25 nm for the first and second combinations, respectively. Duloxetine and avanafil were estimated at 297.7 and 331 nm in their binary mixture, while duloxetine and tadalafil were determined at 290.3 and 297.7 nm, respectively. The linearity was achieved over the ranges of 0.1-1.5 µg mL-1 for both duloxetine and avanafil and 0.01-0.40 µg mL-1 for tadalafil, with limits of detection of 0.013, 0.022, and 0.004 µg mL-1 for duloxetine, avanafil, and tadalafil, respectively. Successful application of the developed approaches was accomplished for the estimation of the two mixtures in dosage forms as well as human plasma with excellent percentage recoveries (96-103.75% in plasma), which supports their suitability for use in quality control laboratories and pharmacokinetic studies. Moreover, the adopted approaches' greenness was evidenced by applying three tools.

Self-ratiometric fluorescence approach based on plant extract-assisted synthesized silver nanoparticles for the determination of vanillin.

The current study designed and applied a novel self-ratiometric fluorescent nanosensor composed of green-synthesized silver nanoparticles (Ag-NPs) to determine vanillin in adult and infant foods and human plasma. A straightforward microwave-assisted approach is proposed for synthesizing Ag-NPs in less than 1 min using a reducing agent, tailed pepper seed extract. The synthesized Ag-NPs had a strong fluorescence with an intense emission band at 360 nm and a shoulder peak at 430 nm when excited at 265 nm. Upon interaction with vanillin, the fluorescence peak of Ag-NPs at 360 nm decreases in a concentration-dependent manner while being shifted to a longer wavelength, 385 nm. Meanwhile, the shoulder fluorescence peak at 430 nm is only slightly affected by vanillin addition. Thus, a new Ag-NP self-ratiometric probe was designed and validated for vanillin determination using the peak at 385 nm and the shoulder peak at 430 as two built-in reference peaks. The optimized system accurately measured vanillin with a detection limit of 9.0 ng/mL and a linear range of 0.05-8.0 µg/mL without needing pre-derivatization or high-cost instrumentation. The method successfully measured vanillin in adult and infant milk formula, biscuits, and human plasma samples with high percentage recoveries (95.3-104.6%) and excellent precision (relative SD; ≤3.85%).

Citrus/urea nitrogen-doped carbon quantum dots as nanosensors for vanillin determination in infant formula and food products via factorial experimental design fluorimetry and smartphone.

Vanillin is a flavouring agent that is prohibited for use in infant food products with ages lower than 6 months. Excessive vanillin usage could lead to eating disorders, nausea, headache, and vomiting. Therefore, it is essential to control the contents of vanillin in food samples, especially in infant formula. Here, we developed a highly sensitive nanosensor for vanillin based on using green synthesized highly fluorescent (QY = 29.5%) N-doped carbon quantum dots (N-CQDs) as a turn-off fluorescent nanoprobe. The N-doped CQDs synthesis was adopted using citrus bulb squeeze extract and the commonly used fertilizer, urea, as substrates. After mixing with vanillin, the fluorescence of the N-CQDs was largely quenched in a vanillin concentration-dependent manner. The sensing conditions were optimized by quality-by-design using a two-level full factorial design (22 FFD). The N-doped CQDs could detect vanillin in the range 0.1-12.0 µg/ml with a limit of detection of 0.013 µg/ml. Next, a smartphone imaging-based assay combined with a UV chamber was adopted and applied for vanillin determination. This simple detection technique showed sensitivity similar to that of the conventional fluorimetric method. Both conventional and smartphone-based methods were successfully applied for the determination of vanillin in infant milk formula and biscuits and could detect real vanillin concentrations in the analyzed samples with high % recoveries (94.5% to 105.5%). At last, the biocompatibility of the newly synthesized N-CQDs was tested, and it was found to be an excellent candidate for cancer cell imaging.

Utility and greenness appraisal of nuclear magnetic resonance for sustainable simultaneous determination of three 1,4-benzodiazepines and their main impurity 2-amino-5-chlorobenzophenone.

A robust, stability-indicating, and eco-friendly proton nuclear magnetic resonance (1H-qNMR) method was developed for the concurrent determination of three 1,4-benzodiazepines (BDZs), namely diazepam (DZP), alprazolam (ALP), and chlordiazepoxide (CDP) and their common impurity, synthesis precursor, and degradation product; 2-amino-5-chlorobenzophenone (ACB). In the present method, a novel approach was developed for composing a green and cost-efficient solvent system as an alternative to the common NMR organic solvents utilizing 0.3 M sodium dodecyl sulfate prepared in deuterated water. The conducted method is characterized by simplicity with no need for sample pretreatment or labeling. Phloroglucinol was used as an internal standard. The chosen signals for the determinations of ALP, CDP, DZP and ACB were at 2.35 ppm (singlet), 2.84 ppm (singlet), 3.11 ppm (singlet), and 6.90 ppm (doublet of doublet), respectively. The proposed method possessed linearity over the concentration range of 0.25-15.0 mg ml-1 for DZP, ALP, CDP and of 0.5-25.0 mg ml-1 for ACB with LOD values of 0.06, 0.03, 0.07 and 0.16 mg ml-1 respectively, and LOQ values of 0.18, 0.09, 0.21 and 0.49 mg ml-1, respectively. Accuracy of the method was evidenced by excellent recovery% (99.57-99.90%) and small standard deviation (≥ 1.10) for the three analyzed drugs. Intra- and inter-day precision were determined with coefficient of variation ranging from 0.12 to 1.14 and from 0.72 to 1.67, respectively. For the studied compounds, appraisal of the method greenness was achieved via four approaches: Analytical Eco-Scale, Green Analytical Procedure Index (GAPI), Analytical greenness metric (AGREE), and RGB Additive Color Model. The results proved that the proposed method has the privilege of being a green analytical method.

Sensitive determination of naftazone using carbon quantum dots nanoprobe by fluorimetry and smartphone-based techniques.

A simple, fast, and direct mix-and-read spectrofluorimetric method has been developed for the sensitive determination of naftazone (NFZ) utilizing graphene quantum dots (GQDs) as a greener and highly luminescent nanosensor. NFZ effectively quenches the strong fluorescence of GQDs at λex/λem of 350/440 nm via the inner filter effect mechanism. The nanosensor exhibits excellent linearity for NFZ over the concentration range of 0.46 to 186 µM with a limit of detection of 0.04 µM. The proposed method was validated for the successful determination of NFZ in tablets and on manufacturing equipment surfaces with good % recoveries of 98.4-101.6 and 96.3 - 102.2%, respectively. Furthermore, an integrated smartphone-based reader has been implemented and successfully applied for the determination of NFZ. The smartphone-based reader consists of a 365 nm UV torch as an excitation source, a smartphone for recording images, and smartphone-powered image analysis software for signal interpretation, together with a paper-based analytical device (PAD) utilizing filter paper as a substrate and correction fluid as a barrier for creation of detection zones. This smart platform showed excellent sensitivity with a limit of detection down to 0.12 nmol/zone, and it could be used for in-site determination of NFZ, especially for the limited resources laboratories.

Perception of the interaction behavior between pepsin and the antimicrobial drug secnidazole with combined experimental spectroscopy and computer-aided techniques.

Drug-pepsin interaction possibly affects pepsin activity, leads to undesirable shift of its functionality, and likely induces adverse effects in the gastrointestinal tract. The present study aims at exploring the interaction of pepsin with the antiprotozoal/antibacterial drug secnidazole adopting a combination of experimental spectroscopy and computational techniques. For this purpose, different spectroscopic methods including fluorescence, synchronous fluorescence, UV-Visible absorption, and infrared spectroscopy were adopted and coordinated with in silico analysis via molecular docking. The employed synchronized approaches evidenced that; pepsin interacted with secnidazole via static mechanism at stomach pH inferring some consequent conformational changes in the structure of pepsin. Thermodynamic study of drug-pepsin interaction demonstrated that the interaction is spontaneous via van der Waals and hydrogen bonding interaction and the orientation of ligand within pepsin cavity was illustrated by molecular docking. The synchronous fluorescence study proved that tyrosine amino acid residues were involved in the interaction more than tryptophan amino acid residues. Eventually, the combined experimental and molecular docking approaches suggest that secnidazole interacts with pepsin and alter its structure, that finding correlates to gastrointestinal side effects related to secnidazole oral administration.

Fast one-pot microwave-assisted green synthesis of highly fluorescent plant-inspired S,N-self-doped carbon quantum dots as a sensitive probe for the antiviral drug nitazoxanide and hemoglobin.

In this study, we report a one-pot, green, cost-efficient, and fast synthesis of plant-based sulfur and nitrogen self-co-doped carbon quantum dots (S,N-CQDs). By 4-min microwave treatment of onion and cabbage juices as renewable, cheap, and green carbon sources and self-passivation agents, blue emissive S,N-CQDs have been synthesized (λex/λem of 340/418 nm) with a fluorescence quantum yield of 15.2%. A full characterization of the natural biomass-derived quantum dots proved the self-doping with nitrogen and sulfur. The S,N-CQDs showed high efficiency as a fluorescence probe for sensitive determination of nitazoxanide (NTZ), that recently found wide applicability as a repurposed drug for COVID-19, over the concentration range of 0.25-50.0 µM with LOD of 0.07 µM. The nanoprobe has been successfully applied for NTZ determination in pharmaceutical samples with excellent % recovery of 98.14 ± 0.42. Furthermore, the S,N-CQDs proved excellent performance as a sensitive fluorescence nanoprobe for determination of hemoglobin (Hb) over the concentration range of 36.3-907.5 nM with a minimum detectability of 10.30 nM. The probe has been applied for the determination of Hb in blood samples showing excellent agreement with the results documented by a medical laboratory. The greenness of the developed probe has been positively investigated by different greenness metrics and software. The green character of the proposed analytical methods originates from the synthesis of S,N-CQDs from sustainable, widely available, and cheap plants via low energy/low cost microwave-assisted technique. Omission of organic solvents and harsh chemicals beside dependence on mix-and-read analytical approach corroborate the method greenness. The obtained results demonstrated the substantial potential of the synthesized green, safe, cheap, and sustainable S,N-CQDs for pharmaceutical and biological applications.

Solid phase-fabrication of magnetically separable Fe3O4@graphene nanoplatelets nanocomposite for efficient removal of NSAIDs from wastewater. Perception of adsorption kinetics, thermodynamics, and extra-thermodynamics.

Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most detected pharmaceuticals in wastewater and likely pass to drinking water. Magnetic nanocomposite has been greenly fabricated for removal of different NSAIDs; ibuprofen (IP), ketoprofen (KP), naproxen (NX), and diclofenac sodium salt (DF); from wastewater. Solid-phase fabrication of the magnetically separable nanocomposite has been optimized using iron oxide (Fe3O4) and graphene nanoplatelets (GNP) precursors. The characterization showed the homogenous distribution of the magnetite over the GNP with a high specific surface area. Different parameters having a potential influence on the removal of the NSAIDs from wastewater by the Fe3O4@GNP nanocomposite were studied and optimized for efficient performance that achieves adsorption capacities of 8.76, 10.6, 14.3, and 7.63 mg/g for KP, NX, DF, and IP, respectively (296 K). The adsorption of the four NSAIDs by the Fe3O4@GNP nanocomposite followed pseudo-second-order kinetics, and it is mainly controlled by the intraparticle diffusion process. The thermodynamic and extra-thermodynamic studies demonstrated that the adsorption is endothermic and is affected by enthalpy-entropy compensation. The molecular interactions of the four drugs with the nanocomposite have been explored. Successful application of the nanocomposite for fast and efficient removal of the NSAIDs from wastewater has been accomplished with an average removal of 71.0%.

Facile Conversion of the Quinone-Semicarbazone Chromophore of Naftazone into a Fluorescent Quinol-Semicarbazide: Kinetic Study and Analysis of Naftazone in Pharmaceuticals and Human Serum.

Naftazone is a quinone-semi carbazone drug that possesses a strong orange color, and hence it was usually analyzed colorimetrically or by HPLC-UV. However, these methods are not sensitive enough to determine naftazone in biological samples. Naftazone lacks intrinsic fluorescence and does not possess easily derivatizable functional groups. In this contribution, we introduced the first spectrofluorimetric method for naftazone assay through reduction-elicited fluorogenic derivatization through the reduction of its quinone-semicarbazone moiety to the corresponding quinol-semicarbazide derivative by potassium borohydride as a reduction probe. The solvent-dependent fluorescence of the reaction product was studied in various protic and aprotic solvents. Eventually, the fluorescence of the reduced naftazone was measured in 2-propanol at λemission of 350 nm after excitation at λecxitation of 295 nm. The relative fluorescence intensity was linearly correlated to the drug concentration (r = 0.9995) from 10.0 to 500 ng/mL with high sensitivity, where the lower detection limit was 2.9 ng/mL. Hence, the method was effectively applied for naftazone tablets quality control with a mean %recovery of 100.3 ± 1.5, and the results agreed with those of the comparison HPLC-UV method. Furthermore, a new salting-out assisted liquid-liquid extraction (SALLE) method was established for naftazone extraction from human serum, followed by its determination using the developed reduction-based fluorogenic method. The developed SALLE method showed excellent recovery for naftazone from human serum (92.3-106.5%) with good precision (RSD ≤ 6.8%). Additionally, the reaction of naftazone with potassium borohydride was kinetically monitored, and it was found to follow pseudo-first-order kinetics with an activation energy of 43.8 kcal/mol. The developed method's greenness was approved using three green analytical chemistry metrics.

Correction pen as a hydrophobic/lipophobic barrier plotter integrated with paper-based chips and a mini UV-torch to implement all-in-one device for determination of carbazochrome.

The design of a cheap, simple, and handy sensing system for rapid quantitation of pharmaceuticals becomes mandatory to ease drug development procedures, quality control, health care, etc. This work describes a simple, innovative, and easily manufactured paper-based device using a correction pen as a plotter for hydrophobic/lipophobic barriers and graphene quantum dots for recognition and quantification of the hemostatic drug carbazochrome, via fluorescence turn-off mechanism mediated by the inner filter effect. A smartphone-based all-in-one device fitted with an inexpensive 365 nm flashlight as a UV light source and a free image processing software was developed for rapid and reliable interpretation of the fluorescence change from the paper-based device upon introduction of the drug. The simple and convenient steps permit the analysis of many samples in a very short time. The smartphone-based all-in-one device featured excellent sensitivity for carbazochrome with a limit of detection equals to 12 ng/detection zone and good %recovery (100.0 ± 0.4). The reliability of the device was ascertained by favorable statistical comparison with the analogous optimized conventional fluorimetry method and a reference HPLC method. The device has been successfully applied for versatile quantitation of carbazochrome in tablets and on manufacturing equipment surfaces with excellent recoveries. The device offers many green aspects that definitely assist the implementation of the sustainability concept to analytical laboratories. The cost-efficiency, reliability, and ease of fabrication as well as the greenness and user friendship qualify the device for wide application in low-income communities.

High-temperature liquid chromatography for evaluation of the efficiency of multiwalled carbon nanotubes as nano extraction beds for removal of acidic drugs from wastewater. Greenness profiling and comprehensive kinetics and thermodynamics studies.

The retention behavior of a series of acidic drugs, namely ketoprofen (KET), naproxen (NAP), diclofenac (DIC), and ibuprofen (IBU), on the heat-resisting ZORBAX 300SB-C18 column, was studied thermodynamically using high-temperature liquid chromatography (HTLC). A perfect correlation of the compounds' lipophilicity and the calculated thermodynamic indicators evidenced its contribution to the retention behavior. Besides, the steric fitting has a subsidiary effect on IBU retention. Isocratic HTLC separation of the four compounds was achieved using an aqueous mobile phase containing 30% acetonitrile-0.2% acetic acid-0.2% triethylamine at 60 °C. This method has been utilized to monitor the adsorption efficiency of multiwalled carbon nanotubes (MWCNTs) for the removal of the four NSAIDs from water. Different variables affecting the remediation process have been optimized such as the time of contact, pH, ionic strength, temperature, and the mass of MWCNTs. The kinetics and thermodynamics of the adsorption were investigated. The adsorption was evidenced to take place via pseudo-second-order kinetics and the intraparticle diffusion is the rate-controlling step. The thermodynamic investigation showed that the adsorption process is exothermic and enthalpy-driven, and the adsorption is more extensive at a lower temperature. The MWCNTs showed excellent adsorption efficiency of about 76.4 to 97.6% at the optimum conditions. The obtained results are promising and encouraging for the full-scale application of MWCNTs for remediation of NSAIDs-related water pollution. The green analytical chemistry metric "AGREE" and the analytical eco-scale score tool confirmed that the developed protocol is greener and more favorable to the environment and user than most of the reported literature.

Pentabromobenzyl-RP versus triazole-HILIC columns for separation of the polar basic analytes famotidine and famotidone: LC method development combined with in silico tools to follow the potential consequences of famotidine gastric instability.

The competence of hydrophilic interaction (HILIC) and reversed phase liquid chromatography (RPLC) modes, employing two new stationary phases: triazole- and pentabromobenzyl-bonded silica (PBr), respectively, was inspected for separation of two polar basic analytes: famotidine (FAM) and its acidic degradant famotidone (FON). Comparison of the chromatographic efficiency, greenness, and economy aspects showed that the RPLC is superior to the HILIC. Hence, the RPLC method was adopted and validated adhering to the FDA guidelines showing excellent linearity for FAM (1.0-20.0 µg/mL) with a detection limit of 0.14 µg/mL. The method was applied to study the behavior of FAM in simulated gastric juice (SGJ), where it exhibited rapid degradation yielding FON. This degradation pathway is a probable major reason for the poor bioavailability of FAM. The kinetic study of the gastric degradation of FAM in SGJ demonstrated pseudo-first order reaction with a rate constant of 8.1 × 10-3 min-1. Moreover, FAM degradation has been proven to be pH-dependent and catalyzed by the gastric juice components. Hence, in situ buffered dosage form is recommended to overcome or decrease this problem. Molecular docking study shows that FON is missing a crucial stabilizing interaction with the key amino acid Asp98 causing a reduced activity at hH2R receptor relative to FAM. Moreover, ADMET properties prediction revealed some differences in the toxicity, pharmacokinetics, metabolism, and solubility profiles of FAM and FON.

Green conventional and first-order derivative fluorimetry methods for determination of trimebutine and its degradation product (eudesmic acid). Emphasis on the solvent and pH effects on their emission spectral properties.

In this report, the fluorescence properties of the antimuscarinic drug trimebutine maleate (TRB) were fully studied and characterized. TRB exhibited intrinsic fluorescence that is greatly dependent on the local environmental factors including the solvent nature and the pH. Yet, its fluorescence was not significantly influenced by the existence of some surface active agents and polymer. The outcomes of this investigation verified that TRB fluorescence emission is intense in ethanol: 1.0 M aqueous acetic acid (9:1, v/v) with emission maxima at 357 nm and excitation maxima at 270 nm. Whereas, going towards higher pH causes fluorescence quenching. These conditions permitted ultrasensitive fluorimetric determination of TRB over the concentration range of 2.00-1500.0 ng/mL with a lower detection limit of 0.40ng/mL Application for the determination of TRB in tablets, ampoule and suspension was successfully achieved with %recoveries ranged between 98.21-100.17%. Furthermore, a first order derivative fluorimetric method was validated for resolving and simultaneous determination of TRB and its degradation product and impurity, eudesmic acid (EUA) making use of the pH-mediated fluorescence spectral shift of EUA. An ethanolic solution containing acetate buffer (pH 5.3) was used for this goal with excitation at 255 nm and measurement of the first order derivative peak amplitudes at respective zero-crossing points of 375 and 351 nm over the corresponding concentration ranges of 20.00-500.00 and 10.00-300.00 ng/mL for TRB and EUA, respectively. The two methods were assessed regarding greenness and eco-friendship by the National Environmental Methods Index and analytical eco-scale score approaches which confirmed their excellent greenness and safety.

Estimation of nizatidine gastric nitrosatability and product toxicity via an integrated approach combining HILIC, in silico toxicology, and molecular docking.

The liability of the H2-receptor antagonist nizatidine (NZ) to nitrosation in simulated gastric juice (SGJ) and under WHO-suggested conditions was investigated for the first time. For monitoring the nitrosatability of NZ, a hydrophilic interaction liquid chromatography (HILIC) method was optimized and validated according to FDA guidance. A Cosmosil HILIC® column and a mobile phase composed of acetonitrile: 0.04 M acetate buffer pH 6.0 (92:8, v/v) were used for the separation of NZ and its N-nitroso derivative (NZ-NO) within 6 min with LODs of 0.02 and 0.1 µg/mL, respectively. NZ was found highly susceptible to nitrosation in SGJ reaching 100% nitrosation in 10 min, while only 18% nitrosation was observed after 160 min under the WHO-suggested conditions. The chemical structure of NZ-NO was clarified by ESI+/MS. In silico toxicology study confirmed the mutagenicity and toxicity of NZ-NO. Experiments evidenced that ascorbic acid strongly suppresses the nitrosation of NZ suggesting their co-administration for protection from potential risks. In addition, the impacts of the HILIC method on safety, health, and environment were favorably evaluated by three green analytical chemistry metrics and it was proved that, unlike the popular impression, HILIC methods could be green to the environment.

Green micellar HPLC analysis of three angiotensin-converting enzyme inhibitors in their mixtures with hydrochlorothiazide and modeling of their retention behavior by fitting to Foley's model.

We present an environmentally friendly method for the analysis of three angiotensin-converting enzyme inhibitors and hydrochlorothiazide simultaneously using a green micellar eluent for the first time. The chromatographic separation of enalapril maleate, lisinopril dihydrate, benazepril hydrochloride, and hydrochlorothiazide was implemented on an octadecyl silica column with a solution containing sodium dodecyl sulfate (0.12 M), 1-propyl alcohol (10% v/v), triethylamine (0.3% v/v), and H3 PO4 (0.02 M) at pH 3.6 as the mobile phase and UV detection at 210 nm. Validity of the method was confirmed and it exhibited good linearity within the ranges of 5.0-50.0 µg/mL for hydrochlorothiazide and 10.0-60.0 µg/mL for the three angiotensin-converting enzyme inhibitors with a limit of detection of 0.39 to 1.15 µg/mL for all the studied drugs. The developed micellar high-performance liquid chromatography method enables the quantification of the targeted angiotensin-converting enzyme inhibitors in combined tablets with hydrochlorothiazide by isocratic elution. There is no need for special precautions to prevent broadening and splitting of their chromatographic peaks. The method fulfills the society rights for safe and green analytical methods. The retention behavior of the four studied drugs was fitted to Foley's model and their association equilibria to the micelles (KAM ) and to the surface-modified stationary phase (KAS ) were calculated.

Fast separation and quantification of three anti-glaucoma drugs by high-performance liquid chromatography UV detection.

In this study, a simple and accurate high-performance liquid chromatography method was developed and validated for fast separation of three anti-glaucoma drugs: timolol maleate (TM), brimonidine tartrate (BM), and latanoprost (LP). Separation of the three drugs was achieved in < 6 minutes using a BDS Hypersil phenyl column and a mobile phase consisting of acetonitrile: 25mM phosphate buffer, pH 4.0 (50: 50, v/v) at 1.2 mL/min with UV detection at 210 nm. The method was linear over the concentration ranges of 5.0-200.0 µg/mL, 2.0-80.0 µg/mL and 1.0-25.0 µg/mL with lower detection limits of 0.21 µg/mL, 0.10 µg/mL and 0.11 µg/mL for TM, BM and LP, respectively. The method was applied for the determination of two fixed-dose combination eye drops for the treatment of glaucoma, containing TM together with either BM or LP. Commercial samples of single-ingredient ophthalmic solutions containing the studied drugs were also successfully analyzed. The results obtained by the proposed method were favorably compared with those obtained by the comparison methods using Student's t test and the variance ratio F test.

Development and Validation of a New HPLC Method for the Simultaneous Determination of Paracetamol, Ascorbic Acid, and Pseudoephedrine HCl in their Co-formulated Tablets. Application to in vitro Dissolution Testing.

The first HPLC method was developed for the simultaneous determination of paracetamol (PC), ascorbic acid (AA), and pseudoephedrine HCl (PE) in their co-formulated tablets. Separation was achieved on a C18 column in 5 min using a mobile phase composed of methanol-0.05 M phosphate buffer (35:65, v/v) at pH 2.5 with UV detection at 220 nm. Linear calibration curves were constructed over concentration ranges of 1.0 - 50.0, 3.0 - 60.0 and 3.0 - 80.0 µg mL(-1) for PC, AA, and PE, respectively. The method was validated and applied for the simultaneous determination of these drugs in their tablets with average % recoveries of 101.17 ± 0.67, 98.34 ± 0.77, and 98.95 ± 1.11%, for PC, AA, and PE, respectively. The proposed method was also used to construct in vitro dissolution profiles of the co-formulated tablets containing the three drugs.

Stability study of the antihistamine drug azelastine HCl along with a kinetic investigation and the identification of new degradation products.

The first stability-indicating HPLC method was developed and validated for azelastine HCl (AZL). The separation of AZL from its degradation products was achieved on a C18 column using acetonitrile-0.04 M phosphate buffer of pH 3.5 (32:68, v/v) as a mobile phase with UV-detection at 210 nm and naftazone as an internal standard. The method was rectilinear over the range of 0.2-20.0 µg mL(-1) with a detection limit of 7.05 ng mL(-1). The degradation behavior of AZL was studied under different ICH-recommended stress conditions along with a kinetic investigation; also, degradation products were identified by mass spectrometry. The method was applied for the quality control and stability assessment of AZL in eye drops and nasal spray. The obtained results were favorably compared with those obtained by a comparison method.

Simultaneous determination of floctafenine and its hydrolytic degradation product floctafenic acid using micellar liquid chromatography with applications to tablets and human plasma.

A stability-indicating micellar liquid chromatography (MLC) method was developed and validated for the assay of floctafenine (FLF) in the presence of its degradation product and main metabolite, floctafenic acid (FLA). The analysis was carried out on a CLC Shim-Pack octyl silane (C8) column (150 x 4.6 mm id, 5 microm particle size) using a micellar mobile phase consisting of 0.15 M sodium dodecyl sulfate, 10% n-propanol, and 0.3% triethylamine in 0.02 M orthophosphoric acid (pH = 3). The mobile phase was pumped at a flow rate of 1.0 mL/min with UV detection at 360 nm. The method showed good linearity for FLF and FLA over the concentration ranges of 0.5-25.0 and 0.4-10.0 microg/mL, with LODs of 0.16 and 0.12 microg/mL, respectively. The developed method was successfully applied to the determination of FLF in commercial dispersible tablets, with mean recovery of 98.87 +/- 1.37%. Also, the proposed method was specific for the analysis of FLF in presence of the co-formulated drug thiocolchicoside in laboratory-prepared tablets, with mean recovery of 100.50 +/- 1.07%. Statistical comparison of the results obtained by the proposed MLC method with those obtained by a comparison method showed good agreement. Moreover, the method was extended to study the degradation behavior of FLF under different International Conference on Harmonization recommended conditions such as alkaline, acidic, oxidative, thermal, and photolytic. The method was further applied for direct determination of FLA as the main metabolite of FLF in human plasma without prior extraction steps, with mean recovery of 110.50 +/- 6.5%.

Validated stability-indicating spectrofluorimetric method with enhanced sensitivity for determination of repaglinide in tablets.

A simple and highly sensitive spectrofluorimetric method was developed and validated for determination of the antidiabetic agent repaglinide (RG) in tablets. The proposed method is based on measurement of the native fluorescence of RG in 0.1 M H(2)SO(4)/methanol medium at 360 nm after excitation at 243 nm. The method showed a linear dependence of the relative fluorescence intensity on drug concentration over the range of 0.02-0.50 µg mL(-1) with lower detection limit of 6.0 ng mL(-1) and lower quantification limit of 18 ng mL(-1). The method was successfully applied for determination of RG in different tablets and the obtained results were in good agreement with those obtained by the official method. The proposed method was extended to investigate the kinetics of oxidative degradation of the drug. A proposal for the degradation pathway was postulated.