In-Vitro Study of the Binding of Atorvastatin with Adenine using Multi-Spectroscopic Approaches.

Atorvastatin-an oral lipid regulating drug is a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), which is the rate determining enzyme for cholesterol synthesis. Adenine is a purine nucleobase that is found in deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) to generate genetic information. The binding mechanism of atorvastatin and adenine was studied for the first time utilizing various techniques, including UV-visible spectrophotometry, spectrofluorimetry, synchronous fluorescence spectroscopy (SF), Fourier transform infrared (FTIR), fluorescence resonance energy transfer (FRET), and metal ion complexation. The fluorescence spectra of the complex indicated that atorvastatin is bound to adenine via hydrophobic interaction through a spontaneous binding process, and the fluorescence quenching mechanism was found to be static quenching with a binding constant of 1.4893 × 104 Lmol-1 at 298 K. Various temperature settings were used to investigate thermodynamic characteristics, such as binding forces, binding constants, and the number of binding sites. The interaction parameters, including the standard enthalpy change (ΔHο) and standard entropy change (ΔSο) were calculated using Van't Hoff's equation to be 42.82 kJmol-1 and 208.9 Jmol-1K-1, respectively. The findings demonstrated that the adenine- atorvastatin binding was endothermic. Furthermore, the results of the experiments revealed that some metal ions (K+, Ca+2, Co+2, Cu+2, and Al+3) facilitate the binding interaction between atorvastatin and adenine. Slight changes are observed in the FTIR spectra of adenine, indicating the binding interaction between adenine and atorvastatin.

Fast concurrent determination of guaifenesin and pholcodine in formulations and spiked plasma using first derivative synchronous spectrofluorimetric approach.

Guaifenesin and pholcodine are frequently co-formulated in certain dosage forms. A new fast first derivative synchronous spectrofluorometric method has been used for their simultaneous analysis in mixtures. Here, first derivative synchronous spectrofluorometry enabled the successful simultaneous estimation of guaifenesin at 283 nm and pholcodine at 275 nm using a wavelength difference (Δλ) of 40 nm. The method was fully validated following International Council of Harmonization guidelines. For guaifenesin and pholcodine, linearity was determined within the corresponding ranges of 0.05-0.30 and 0.10-6.0 µg/ml. The two drugs were effectively analyzed using the developed approach in their respective formulations, and the results showed good agreement with those attained using reference methods. The method demonstrated excellent sensitivity, with detection limits down to 0.007 and 0.030 µg/ml and quantitation limits of 0.020 and 0.010 µg/ml for guaifenesin and pholcodine, respectively. Therefore, the procedure was successful in determining these drugs simultaneously in vitro in spiked plasma samples and syrup dosage form. The developed methodology also offered an environmentally friendly advantage by utilizing water as the optimal diluting solvent throughout the whole work. Different greenness approaches were investigated to ensure the method's ecofriendly properties.

Determination of omeprazole via fluorescence-quenching methodology; application to content uniformity testing.

A new, proven, economical spectrofluorimetric approach has been used to determine the proton pump inhibitor omeprazole (OMP). This innovative technique is based on the ability of OMP to quench the native fluorescence of the mercurochrome dye in an acidic (pH 3.6) solution. Because it was discovered that quenching is proportional to the drug concentration, this dye was used as a sensor for OMP detection. The fluorescence intensity was measured at 518/540 nm, and its linear response ranged from 0.2-10.0 µg/mL with a linear coefficient of 0.9999. The computation yielded a limit of quantification (LOQ) of 0.20 µg/mL and a limit of detection (LOD) of 0.07 µg/mL. Every circumstance and element impacting the reaction product was examined in detail. Pharmacopeial standards carried out the validation. The approved method investigated several commercial preparations and formulations, and the results were favorably compared with those provided by a reference method. According to United States Pharmacopeia (USP) rules, content consistency for two distinct formulations was evaluated.

A sustainable sensitive spectrofluorimetric approach for the determination of the multipotent protein lactoferrin in different pharmaceuticals and infant milk formula: Compliance with greenness metrics.

The current study presents the first spectrofluorimetric approach for the estimation of lactoferrin, depending on the measurement of its native fluorescence at 337 nm after excitation at 230 nm, without the need for any hazardous chemicals or reagents. It was found that the fluorescence intensity versus concentration calibration plot was linear over the concentration range of 0.1-10.0 µg/mL with quantitation and detection limits of 0.082 and 0.027 µg/mL, respectively. The method was accordingly validated according to the ICH recommendations. The developed method was applied for the estimation of lactoferrin in different dosage forms, including capsules and sachets with high percent recoveries (97.84-102.53) and low %RSD values (<1.95). Lactoferrin is one of the key nutrients in milk powder and a significant nutritional fortifier. In order to assess the quality of milk powder, it is essential to rapidly and accurately quantify the lactoferrin content of the product. Therefore, the presented study was successfully applied for the selective estimation of lactoferrin in milk powder with acceptable percent recoveries (96.45-104.92) and %RSD values (≤3.607). Finally, the green profile of the method was estimated using two assessment tools: Green Analytical Procedure Index (GAPI) and Analytical GREEnness (AGREE), which demonstrated its excellent greenness.

Insights on multi-spectroscopic approaches for estimating the in vitro binding of favipiravir with adenine nucleotide.

Favipiravir (FVP) is an oral antiviral drug approved in 2021 for the treatment of COVID-19. It is a pyrazine derivative that can be integrated into anti-viral RNA products to inhibit viral replication. While, adenine is a purine nucleobase that is found in deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) to generate genetic information. For the first time, the binding mechanism between FVP and adenine was determined using different techniques, including UV-visible spectrophotometry, spectrofluorimetry, synchronous fluorescence (SF) spectroscopy, Fourier transform infrared (FTIR), fluorescence resonance energy transfer (FRET), and metal ion complexation. The fluorescence spectra indicated that FVP is bound to adenine via Van der Waals forces and hydrogen bonding through a spontaneous binding process (ΔGο < 0). The quenching mechanism was found to be static. Various temperature settings were used to investigate thermodynamic characteristics, such as binding forces, binding constants, and the number of binding sites. The reaction parameters, including the enthalpy change (ΔHο) and entropy change (ΔSο), were calculated using Van't Hoff's equation. The findings demonstrated that the adenine-FVP binding was endothermic. Furthermore, the results of the experiments revealed that some metal ions (K+, Ca+2, Co+2, Cu+2, and Al+3) might facilitate the binding interaction between FVP and adenine. Slight changes are observed in the FTIR spectra of adenine, indicating the binding interaction between adenine and FVP. This study may be useful in understanding the pharmacokinetic characteristics of FVP and how the drug binds to adenine to prevent any side effects.

A Design-assisted Spectrofluorometric Method Utilizing a One-pot Fluorescent Probe for the Quantitation of some Calcium Channel Blockers.

Based on their reaction with highly fluorescent carbon quantum dots (CQDts), a precise and reliable spectrofluorometric approach was developed for the determination of three calcium channel blockers. The studied drugs are: lercanidipine, nimodipine and nifedipine. (CQDts) were produced using a one-step hydrothermal method with ascorbic acid as the carbon source. The produced CQDts were capped by alcohol to create yellow emitters displaying a high fluorescence emission at 524 nm when excited at 325 nm. The fluorescence intensity of CQDts was noticeably quenched by each of the three calcium channel blockers. The relation between their concentrations and fluorescence quenching is linear over the concentration range of 0.5-20 µg/mL for each of the three drugs. A full factorial design was used to optimize the effect of variable factors. Therefore, under optimum experimental design conditions, the detection limits for lercanidipine, nimodipine, and nifedipine were 0.11 ± 1.09, 0.10 ± 0.25 and 0.12 ± 0.71 µg/mL, respectively. The LOQ was 0.33, 0.30, and 0.37 µg/mL respectively. The quenching of fluorescent CQDts occurred through the inner filter effect (IFE) for nimodipine, while it was mixed with dynamic quenching for lercanidipine and nifedipine. The proposed method was effectively used to determine the cited drugs in their pharmaceutical products and had an acceptable level of precision. The selectivity of the CQDts system towards the studied drugs was examined indicating no interference from interfering species.

Utility of a xanthene-based dye for determination of nilotinib using two spectroscopic approaches. Applications to bulk powder, capsules, and spiked human plasma.

Novel, selective, facile, and precise spectroscopic approaches were validated to determine nilotinib hydrochloride, a tyrosine kinase inhibitor used to treat patients with chronic myeloid leukemia. These approaches depend on the reaction of the tertiary amine group of nilotinib with erythrosine B in the Britton-Robinson buffer at pH 4. Method I, depends on measuring the absorbance of the formed complex at 551 nm. The absorbance concentration plot showed linearity over the concentration range of 1.0 to 9.0 µg/ml. Method II, involved the measurement of the quenching of the native fluorescence of erythrosine B by adding nilotinib in an acidic medium. The fluorescence quenching of erythrosine B was measured at 549 nm after excitation at 528 nm. This approach showed excellent linearity in the concentration range of 0.04 to 0.7 µg/ml. The limit of detection values for Method I and Method II were 0.225 and 0.008 µg/ml, respectively, while the limit of quantitation values for Method I and Method II were 0.68 and 0.026 µg/ml, respectively. To get the optimal conditions, factors that may affect the formation of the ion-pairing complex were thoroughly examined. The two approaches were carefully validated following the International Conference of Harmonization (ICH Q2R1) guidelines. Statistical assessment of the results achieved using the suggested and previously published comparison approaches showed no significant difference. The approaches were successful in determining nilotinib in a pharmaceutical dosage form as well as spiked human plasma samples. The eco-friendly properties of the methods were evaluated by three different tools.

Validated spectrofluorimetric assay of two co-administered drug mixtures containing hydroxychloroquine with either moxifloxacin or ofloxacin as a drug regimen for hospital-acquired pneumonia in patients with COVID-19.

Moxifloxacin and ofloxacin are two broad-spectrum quinolone antibiotics. They are among the most widely used antibiotics, at this time, applied to control the COVID-19 pandemic. Hydroxychloroquine is an FDA-approved drug for the treatment of COVID-19. This work describes a simple, green, selective, and sensitive spectrofluorimetric method for the assay of moxifloxacin and ofloxacin in the presence of hydroxychloroquine, two co-administered mixtures used in the treatment of hospital-acquired pneumonia in patients with COVID-19. Simultaneous assay of hydroxychloroquine and moxifloxacin was carried out in methanol using a direct spectrofluorimetric method (method I) at 375 and 550 nm, respectively, after excitation at 300 nm. The direct spectrofluorimetric assay was rectilinear over concentration ranges 50.0-400.0 and 300.0-2500.0 ng/ml for hydroxychloroquine and moxifloxacin, respectively, with limits of detection (LOD) of 6.4 and 33.64 ng/ml and limits of quantitation (LOQ) of 19.4 and 102.6 ng/ml, respectively, for the two drugs. The assay for hydroxychloroquine and ofloxacin was carried out by measuring the first derivative synchronous amplitude for hydroxychloroquine at the zero crossing point of ofloxacin and vice versa at Δλ = 140 nm (method II). Hydroxychloroquine was measured at 266 nm, while ofloxacin was measured at 340 nm over the concentration range 4-40 ng/ml for hydroxychloroquine and 200-2000 ng/ml for ofloxacin with LOD of 0.467 and 25.3 ng/ml and LOQ of 1.42 and 76.6 ng/ml, respectively, for the two drugs. The two methods were validated following International Conference on Harmonization guidelines and were applied to the analysis of the two drugs in plasma with good percentage recoveries (109.73-93.17%).

A Novel Quantum Dots-Based Fluorescent Sensor for Determination of the Anticancer Dacomitinib: Application to Dosage Forms.

One of the most promising drugs recently approved for the treatment of various types of cancer is dacomitinib, which belongs to the tyrosine kinase inhibitor class. The US Food and Drugs Administration (FDA) has recently approved dacomitinib as a first-line treatment for patients suffering from non-small cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) mutations. The current study proposes the design of a novel spectrofluorimetric method for determining dacomitinib based on newly synthesized nitrogen-doped carbon quantum dots (N-CQDs) as fluorescent probes. The proposed method is simple and does not require pretreatment or preliminary procedures. Since the studied drug does not have any fluorescent properties, the importance of the current study is magnified. When excited at 325 nm, N-CQDs exhibited native fluorescence at 417 nm, which was quantitatively and selectively quenched by the increasing concentrations of dacomitinib. The developed method involved the simple and green microwave-assisted synthesis of N-CQDs, using orange juice as a carbon source and urea as a nitrogen source. The characterization of the prepared quantum dots was performed using different spectroscopic and microscopic techniques. The synthesized dots had consistently spherical shapes and a narrow size distribution and demonstrated optimal characteristics, including a high stability and a high fluorescence quantum yield (25.3%). When assessing the effectiveness of the proposed method, several optimization factors were considered. The experiments demonstrated highly linear quenching behavior across the concentration range of 1.0-20.0 µg/mL with a correlation coefficient (r) of 0.999. The recovery percentages were found to be in the range of 98.50-100.83% and the corresponding relative standard deviation (%RSD) was 0.984. The proposed method was shown to be highly sensitive with a limit of detection (LOD) as low as 0.11 µg/mL. The type of mechanism by which quenching took place was also investigated by different means and was found to be static with a complementary inner filter effect. For quality purposes, the assessment of the validation criteria adhered to the ICHQ2(R1) recommendations. Finally, the proposed method was applied to a pharmaceutical dosage form of the drug (Vizimpro® Tablets) and the obtained results were satisfactory. Considering the eco-friendly aspect of the suggested methodology, using natural materials to synthesize N-CQDs and water as a diluting solvent added to its greenness profile.

Insights on the Quantitative Concurrent Fluorescence-Based Analysis of Anti-COVID-19 Drugs Remdesivir and Favipiravir.

We hereby introduce a sensitive fast straightforward spectrofluorometric method for the estimation of remdesivir and favipiravir. The two drugs are prescribed in some regimens to treat COVID-19 pandemic disease, which is caused by SARS-CoV-2. The method is based on the first derivative synchronous spectrofluorimetry approach for the measurement of remdesivir and favipiravir. This was accomplished at 251 nm and 335 nm respectively using the first derivative order at delta lambda of 140 nm. A linear response with a correlation coefficient 0.9994 was achieved between the concentration and the derivative amplitudes in the ranges of 20.0-100.0 ng ml-1 and 40.0-100.0 ng ml-1 for remdesivir and favipiravir, respectively. The methods were validated for different parameters as stated by the pharmacopeial rules and were applied successfully for estimation of the studied drugs in their synthetic mixtures and in spiked human plasma samples. No significant difference was observed between the proposed and comparison methods as revealed from the analysis of data.

Novel approaches of erythrosine B as a food dye-derived spectroscopic probe for assessing trospium chloride in raw material and dosage form.

Two facile spectroscopic methodologies were designed for estimating trospium chloride (TPM) in raw material and tablets with high operational reliability and selectivity. The methods were based on using erythrosine B (EB) as a spectroscopic tool for ion-pair complex formation with the drug. In a mild acidic medium of Britton Robinson buffer (pH 4.0), the ionized hydroxyl group in the reagent interacted with the ionized amine in the studied drug. Method I was based on the spectrophotometric measuring of the absorbance of the reaction product at 557 nm. Method II was based on spectrofluorimetric measurement of the quenching effect of TPM on the inherent fluorescence of EB at 550 nm (λex. = 528 nm). The two methods showed linearity through ranges 1.0-10.0 and 0.5-10.0 µg/ml for Methods I and II, respectively. The suggested methods were exploited for analyzing TPM in Trospamexin® tablets and showed good applicability. The designed systems were validated as per International Conference on Harmonization guidelines. Experimental conditions were modulated to obtain the best sensitivities. The quenching mechanism was investigated and the quenching constant was computed relying on the Stern-Volmer equation. Environmental impact was appraised using novel metric green tools, GABI, and AGREE. The suggested systems excelled over other reported methods in terms of greenness, sensitivity, and cost-effectiveness.

Micellar-emphasized simultaneous determination of ivabradine hydrochloride and felodipine using synchronous spectrofluorimetry.

A sensitive and green micellar spectrofluorimetric approach was applied for the simultaneous estimation of ivabradine hydrochloride (IVB) and felodipine (FLD) in the ng/ml concentration range. The approach depended on measuring the first derivative synchronous peak amplitude (1 D) of both drugs at ∆λ = 60 nm in a Tween-80 micellar system. The method was rectilinear alongside the concentration ranges 0.02-0.4 µg/ml and 0.05-1.0 µg/ml at 269.5 nm and 378.5 nm for IVB and FLD, respectively. The proposed method was validated by following the International Council for Harmonization guidelines. The method was successfully applied without interference for laboratory-prepared synthetic mixtures, single pharmaceutical preparations, and within spiked biological fluids with acceptable percentage recoveries. A comparison of the performance of the suggested method with other methods, showed no discrepancy. The method's ecofriendly property evaluated using three different tools, confirming an excellent green method.

Green micellar solvent-free HPLC and spectrofluorimetric determination of favipiravir as one of COVID-19 antiviral regimens.

Quality control (QC) of pharmaceutical products requires fast, sensitive as well as economic methodologies in order to provide high through output at low cost which are the main aspects considered by such economic facilities. Meanwhile, the ecological impacts must be considered by researchers to minimize the hazardous effects of research laboratories. Favipiravir (FAV) is an antiviral agent recently approved for treatment of COVID-19 infections during 2020 pandemic crisis, so the size of its production by international pharmaceutical corporations evolved dramatically within the past few months. Two novel simple, sensitive, and green methods were developed and validated for FAV determination based on solvent-free micellar LC and spectrofluorimetry techniques. To improve FAV native fluorescence, several factors were studied including solvent type, buffering, pH and added surfactants. The best sensitivity for FAV fluorescence was obtained in Britton-Robinson buffer (pH 4) at 436 nm after excitation at 323 nm within concentration range of 20-350 ng mL-1. Another HPLC method was validated using C18-RP (5 µm, 250 × 4.6 mm) stationary phase and solvent-free mobile phase consisting of (0.02 M Brij-35, 0.15 M SDS, and 0.02 M disodium hydrogen phosphate, pH 5.0) isocratically eluted at a flow rate of 1 mL min-1 and detection wavelength of 323 nm. LC method was validated across concentration range of 10-100 µg mL-1 and FAV eluted in 3.8 min. The methods were validated according to the FDA guidelines and were applied successfully for determination of FAV in its marketed tablet dosage forms and in spiked human plasma samples. The proposed methods are eco-friendly since they are typically based on biodegradable reagents in aqueous solvent-free phases, which was proven by their assessment on two recent greenness metrics (GAPI and AGREE) to prove their eco-friendly properties.

New spectroscopic methods for determination of dexlansoprazole using mercurochrome.

New spectroscopic methods were developed for dexlansoprazole estimation in capsule formulation based on the formation of a reaction between dexlansoprazole and Mercurochrome (MER) at pH 3.7. The formed complex was measured spectrophotometrically (Method I) at 557 nm and spectrofluorometrically (Method II) at 300 nm/538 nm, because the drug caused quantitative quenching of the native fluorescence of Mercurochrome. The spectrophotometric method was linear over the concentration 25-55 µg/ml with a limit of detection (LOD) of 1.15 µg/ml and a limit of quantification (LOQ) of 3.48 µg/ml. The spectrofluorometric method had a linear range 20-45 µg/ml with an LOD of 1.13 µg/ml and an LOQ of 3.45 µg/ml. The suggested methods were used to analyze capsules to test the interference from excipients and the data indicated good selectivity. Data obtained were statistically analyzed and were favourably good. The new methods are environmentally benign and depend on distilled water mainly as the diluting solvent. This property was confirmed by assessing their greenness.

Novel spectrofluorimetric technique for determination of amoxicillin and ethopabate in chicken tissues, liver, kidney, eggs, and feed premix.

A new smart spectrofluorometric method was developed for the quantitation of amoxicillin and ethopabate simultaneously for the first time. The method is based on measuring their first derivative synchronous amplitudes in water at Δλ = 80 nm. The peak amplitudes were recorded at their crossing points; 240 nm for amoxicillin and 280 nm for ethopabate. The method is linear over the concentration ranges of 100.0-1,000.0 ng/ml for amoxicillin and 2.0-20.0 ng/ml for ethopabate. The limits of detection were 20.0 ng/ml and 0.58 ng/ml and limits of quantitation were 60.0 ng/ml and 1.92 ng/ml for amoxicillin and ethopabate, respectively. The method sensitivity permitted the determination of the two drugs below their maximum residue limit stated by the federal regulations. The developed method was applicable to the analysis of both drugs in the veterinary powders, feed premix, chicken tissues, liver, kidney, and eggs samples with percentage recoveries ranging 93.72-104.71%.

Solvent-free mixed micellar mobile phases: An advanced green chemistry approach for reversed-phase HPLC determination of some antihypertensive drugs.

Minimizing the amount of organic solvents without loss in chromatographic performance has been an important step toward greening analytical methodologies. Mobile-phase composition is the key for maintaining separation efficiency in liquid chromatography while decreasing the procedure hazardousness. If sodium dodecyl sulfate is mixed with Brij-35 in the mobile phase, they could be used as a green alternative for using organic modifiers. In this research, the effect of changing the relative amounts of both surfactants was studied on the chromatographic performance and separation efficiency of ten antihypertensive drugs belonging to different categories. The use of surfactants has many advantages including low cost and toxicity, safe environmental disposal, unique selectivity besides high solubilization capabilities. The optimum separation was maintained using a mobile phase (0.01 M Brij-35, 0.08 M sodium dodecyl sulfate and 0.01 M sodium dihydrogen phosphate/pH 5) on reversed-phase C18 core-shell column at flow rate 1.5 mL/min and temperature 30°C. The method was successfully applied for the determination of the drugs in various marketed dosage forms. International Conference of Harmonization guidelines were followed to validate the developed method. Additionally, the method was verified on the Green Analytical Procedure Index in regards to the greenness and found to be an excellent green alternative method.

Use of eosin for green spectroscopic determination of mebendazole.

New, sensitive, and reliable spectroscopic methods were constructed for the fast determination of the anthelmintic drug mebendazole. The methods depended on the reaction of the amino group in mebendazole with eosin in acidic medium forming an ion pair complex. The first method, Method I, relied on quenching of the native fluorescence of eosin after reaction with mebendazole at pH 3.7 using acetate buffer. Fluorescence quenching was measured at 538 nm after excitation at 518 nm. This method showed a linear response over the concentration range 5.0-20.0 µg/ml. The second method, Method II, was based on measuring the absorbance of the formed complex at 554 nm; the method showed good linearity from 7.0 to 22.0 µg/ml. Different parameters that influenced the formation of the reaction product were carefully investigated to reach the optimized conditions. A comparison between the proposed methods and a previous spectrophotometric method was carried out and there was no significant difference between them. The methods could be applied successfully to determine mebendazole in its tablet form. Moreover, the methods used water as diluting solvent, which made them compatible with the 'green' analytical chemistry principles. No organic solvents were used throughout the study.

Facile green spectrophotometric approaches for the determination of three natural edible antioxidant polyphenols in different matrices.

The combination of Curcumin (CRN), resveratrol (RSV), and quercetin (QRN) has significant antioxidant effects and is found to be more effective than a single polyphenol. Spectrophotometric methods are considered one of the most common analytical techniques for the determination of the drugs due to their sensitivity, rapidness, low cost, and reproducibility. Therefore, the presence of new, and simple methods for the determination of such compounds will be highly valuable, specially in the presence of spectral overlap. In this research, five different facile spectrophotometric methods were investigated for the simultaneous determination of that ternary mixture for the first time, including zero order (I), first derivative (II), ratio difference double divisor (III), first derivative ratio spectra (IV), and mean centering (V) methods. The designed approaches were linear over the concentration ranges of (1.0-10.0), (0.5-8.0), and (1.0-14.0) µg/mL, respectively for curcumin, resveratrol, and quercetin. The different methods were then validated as stated by the International Council of Harmonization. The accuracy and precision have been evaluated by statistical analysis including student t-test, variance ratio F-test, and ANOVA. Moreover, the greenness and whiteness of the proposed methods were assessed to ensure the adherence to the greenness characters.

Stability indicating eco-friendly quantitation of terbutaline and its pro-drug bambuterol using quantitative proton nuclear magnetic spectroscopy.

Quantitative 1H-NMR became an increasingly important issue in pharmaceutical analytical chemistry. This study used NMR spectroscopy to assay the bronchodilator drug terbutaline sulfate and its pro-drug bambuterol hydrochloride in pure form and pharmaceutical preparations. The technique proceeded using deuterium oxide (D2O) as an 1H-NMR solvent and phloroglucinol anhydrous as an internal standard (IS). Comparatively, to the phloroglucinol signal at 5.9 ppm, the resulting quantitative signals of the studied drugs were corrected. The terbutaline singlet signal at 6.3 ppm was chosen for quantification, while the bambuterol quantitative singlet signal was at 2.9 ppm. The two drugs were rectilinear over the concentration range of 1.0-16.0 mg/mL. LOD values were 0.19 and 0.21 mg/mL while LOQ values were 0.58 and 0.64 mg/mL for terbutaline and bambuterol respectively. The developed method has been validated according to the International Conference of Harmonization (ICH) regarding linearity, accuracy, precision, specificity, and robustness. A greenness profile assessment was applied, and the method proved to be green. The method enables the assay of the two drugs in pure drug and pharmaceutical preparations. The method also enables the assay of the two drugs in the presence of each other; thus, it is considered a stability-indicating method where terbutaline is an acid degradation product of bambuterol.

Investigation of green synchronous spectrofluorimetric approach for facile sensitive estimation of two co-administered anti-cancer drugs; curcumin and doxorubicin in their laboratory-prepared mixtures, human plasma, and urine.

Recently, phytochemicals play an important role in cancer management. Curcumin (CUR), a natural phytochemical, has been co-administered with widespread chemotherapeutic agents such as doxorubicin (DOX) due to its excellent antitumor activity and the ability to lower the adverse reactions and drug resistance cells associated with DOX use. The present study aims to determine DOX and CUR utilizing a label-free, selective, sensitive, and precise synchronous spectrofluorimetric method. The obvious overlap between the emission spectra of DOX and CUR prevents simultaneous estimation of both analytes by conventional spectrofluorimetry. To solve such a problem, synchronous spectrofluorimetric measurements were recorded at Δλ = 20 nm, utilizing ethanol as a diluting solvent. Curcumin was recorded at 442.5 nm, whereas DOX was estimated at 571.5 nm, each at the zero-crossing point of the other one. The developed method exhibited linearity over a concentration range of 0.04-0.40 µg/mL for CUR and 0.05-0.50 µg/mL for DOX, respectively. The values of limit of detection (LOD) were 0.009 and 0.012 µg/mL, while the values of limit of quantitation (LOQ) were 0.028 and 0.037 µg/mL for CUR and DOX, respectively. The adopted approach was carefully validated according to the guidelines of ICH Q2R1. The method was utilized to estimate CUR and DOX in laboratory-prepared mixtures and human biological matrices. It showed a high percentage of recoveries with minimal RSD values. Additionally, three different tools were utilized to evaluate the greenness of the proposed approach.