Eco-friendly synchronous fluorescence spectrofluorimetric method for simultaneous determination of montelukast sodium and fexofenadine hydrochloride in their antiallergic rhinitis fixed-dose combination tablets.

An innovative, simple, accurate, sensitive, and eco-friendly synchronous fluorescence spectrofluorimetric method has been developed for the simultaneous analysis of montelukast sodium (MON) and fexofenadine hydrochloride (FEX). The method relies on measuring the relative synchronous fluorescence intensity of both drugs using Δλ of 60 nm in methanol at 405 nm for MON and 288 nm for FEX. The experimental parameters influencing the developed method were investigated and optimized. The method was linear over the ranges 0.1-2.0 and 2.0-20.0 µg/ml for MON and FEX, respectively. The limits of detection were 0.018 and 0.441 µg/ml, and the limits of quantitation were 0.055 and 1.336 µg/ml for MON and FEX, respectively. The developed method was applied successfully for the determination of the two drugs in their newly released fixed-dose combination prescribed for the treatment of allergic rhinitis. The mean per cent recoveries were found to be 100.680 ± 0.890 and 100.110 ± 0.940 for MON and FEX, respectively. Furthermore, the method was found to be eco-friendly green as was evaluated according to the Green Analytical Procedure Index tool guidelines and analytical eco-scale.

Green and cost-effective voltammetric assay for spiramycin based on activated glassy carbon electrode and its applications to urine and milk samples.

A simple, cost-effective, and efficient differential pulse voltammetric (DPV) assay for monitoring spiramycin adipate (SPA) in its dosage forms, urine, and milk samples at an activated glassy carbon electrode (GCE) was developed. GCE was electrochemically activated by anodization at a high positive voltage (2.5 V). The activated glassy carbon electrode (AGCE) was surface characterized, optimized, and utilized for the electrochemical assay of SPA. The electrochemical behavior of the AGCEs was investigated using cyclic voltammetry (CV) which shows a remarkable increase in the anodic peak of SPA in comparison with GCE. This behavior reflects a remarkable increase in the electrocatalytic oxidation of SPA at AGCE. The impacts of various parameters such as scan rate, accumulation time, and pH were investigated. The analytical performance of the activated glassy carbon electrodes was studied utilizing DPV. Under optimum conditions, the oxidation peak current exhibited two linear ranges of 80 nm to 0.8 µM and 0.85-300 µM with a lower limit of detection (LOD) of 20 nM. The developed assay exhibited high sensitivity, excellent repeatability, and good selectivity. Additionally, the developed SPA-sensitive modified GCE was successfully applied for SPA assay in its pharmaceutical dosage form and diluted biological fluids as well, with satisfactory recovery results which correlated well with the results obtained using spectrophotometry.

Hydrophilic gold nanospheres: influence of alendronate, memantine, and tobramycin on morphostructural features.

Turkevich gold nanospheres are the original nanospheres that have been modified over time. Its combination with targeting medications such as alendronate, memantine, and tobramycin will provide additional benefits in targeting specific areas in the bone, brain, and microorganisms, respectively. Hence, The reactivity and stability of nanospheres with various drug concentrations (milli-,micro-, and nano-levels) have been studied. With alendronate, the absorbance spectra of nanospheres at [Formula: see text] 520 nm were always stable and no redshifts occurred. In contrast, the spectra with memantine and tobramycin were stable at the nano-level and redshifts occurred at the milli- and micro-levels. HRTEM and DLS revealed that the core diameter was relatively stable in all cases, whereas the hydrodynamic diameter and zeta potential varied with varying drug concentrations. Increasing concentration increased hydrodynamic diameter slightly with memantine (from 64.99 to 98.41 nm), dramatically with tobramycin (from 135.3 to 332.16 nm), and almost negligibly with alendronate (from 52.08 to 58.94 nm ). Zeta Potential, conversely, is reduced as concentration increases. Memantine had the greatest reduction in negativity, followed by tobramycin, but alendronate had a slight increase in negativity. Benefits from this research would be in targeted drug delivery, where stability and reactivity of gold nanospheres are critical.

Glycerol as a new mobile phase modifier for green liquid chromatographic determination of ascorbic acid and glutathione in pharmaceutical tablets.

Fixed dose combinations of glutathione and ascorbic acid are marketed due to the synergetic effect of glutathione, which regenerates ascorbic acid, potentiates its action and prolongs its antioxidant potential. In this work, glycerol was introduced for the first time as a green mobile phase modifier for liquid chromatographic determination of glutathione and ascorbic acid in pharmaceutical tablets. The viscosity barrier was overcome by using water as a co-fluidifying solvents and by controlling the column temperature. Compared with methanol and acetonitrile, glycerol could result in better peak shapes and superior system suitability. To explore the retention process, free energy and enthalpy-entropy compensation were calculated for both glutathione and ascorbic acid. The chromatographic conditions were optimized using factorial design with the aid of Minitab 17® Software. The method was successfully validated according to the guidelines of the International Conference on Harmonization Q2 (R1) and was found linear in the range of 1 µg/mL to 300300 µg/mL for both drugs. The method was evaluated in terms of its greenness and then compared to the reported method. Glycerol has several advantages over commonly employed mobile phase modifiers, such as the high safety, the low UV cut-off point, low flammability, low volatility, the green features, and the biodegradability. This work paves the way for more applications of glycerol in green liquid chromatography.

An innovative spectrofluorimetric approach for determination of azelaic acid in bulk and pharmaceutical formulation through derivatization with 9-chloromethylanthracene.

A novel simple, selective and accurate spectrofluorimetric method has been developed for quantitation of azelaic acid (AZA) in bulk and cream dosage form. The proposed method depended on the reaction between dicarboxylic moiety of AZA and 9-chloromethylanthracence to produce fluorescent derivative that exhibited maximum fluorescence intensity at 413 nm after being excited at 365 nm. The numerous experimental parameters which affect the reaction product and stability have been carefully studied and optimized. The linearity of the calibration curve constructed has been 0.5-15 µg/ml (y = 26.864x + 31.793, r2 = 0.9999) with 0.143 and 0.434 µg/ml as LOD and LOQ values respectively. The method was used for quantitation of AZA in cream dosage form and the results showed that there was no interference from the cream excipients, the mean % recovery was 100.547% ± 0.775.

Carbon quantum dots as a sensitive fluorescent probe for quantitation of pregabalin; application to real samples and content uniformity test.

A novel optical nano-sensor for the detection of pregabalin (PG) in its pharmaceutical (Lyrica®) capsules and biological samples was reported. For the fabrication of highly fluorescent carbon quantum dots (CQDts), a simple green hydrothermal approach was described, and ascorbic acid (AA) was used as a carbon source. The obtained CQDts were confirmed by spectroscopic characterization such as transmission electron microscopy (TEM) and Fourier-transform infrared (FTIR) spectra. The synthesized CQDts were capped by alcohol to form yellow emitters, showing strong fluorescent emission at 524 nm, and excitation at 356 nm. The method is based on fluorescence quenching of CQDts in the presence of PG. The proposed analytical method was validated according to ICH guidelines. PG was successively assayed in the concentration range of 4.0 to 100 µg/ml). The detection and quantitation limits were 1.12 and 3.39 µg/ml, respectively. The proposed method could be used in both quality control and pharmacokinetic research for the studied drug.

UV spectrophotometric methods for quantitative determination of masitinib; extraction of qualitative information.

Masitinib is an orally administered selective tyrosine kinase inhibitor. It has emerged as a promising drug for multiple diseases including cancer and inflammation in either human or veterinary medicine. Five new and simple UV spectrophotometric methods were developed for its determination in bulk and in pharmaceutical tablets. These methods are based on measuring the absorbance of masitinib in either zero order or first, second, third or fourth derivative spectra. Measurements are optimized so as to minimize excipients' interferences. The methods are suitable for micro-analysis of masitinib. The proposed methods were validated according to the ICH-Q2(R1) guidelines and was successfully applied for determination of masitinib in laboratory prepared tablet. The presented methods are simple, fast, cost-effective and suitable for routine pharmaceutical analysis. Moreover, two derivative spectrophotometric-based methods were developed for identification of masitinib, the derivative ratio method and log-A derivative method. The impact of the developed methods on the environment was assessed by both analytical Eco-Sale and the Green Analytical Procedure Index (GAPI). The present work proves how derivative spectrophotometry could greatly extract qualitative and quantitative information from UV spectra.

A fast and green reversed-phase HPLC method with fluorescence detection for simultaneous determination of amlodipine and celecoxib in their newly approved fixed-dose combination tablets.

A fast, green, sensitive, and accurate analytical method using high-performance liquid chromatography couple with fluorescence detection was established and validated for the simultaneous determination of amlodipine besylate and celecoxib in their recently approved fixed-dose combination tablets (1:20). Separation of the two drugs was achieved on C18 reversed-phase column (Thermo ODS Hypersil, 4.6 × 250 mm, particle size 5 µm) using acetonitrile:potassium phosphate buffer (50 mM; pH 5.5, 60:40 v/v) as a mobile phase at 40°C, which eluted at a rate of 1 mL/min. Detection was carried out with excitation and emission wavelengths of 360 and 446 nm for amlodipine and 265 and 359 nm for celecoxib, respectively. The method was linear over a concentration range of 0.05-2 and 0.05-10 µg/mL and limit of detection reached to 0.017 and 0.0167 µg/mL for amlodipine and celecoxib, respectively. The developed method was successfully applied to assess the cited drugs in their newly FDA approved fixed-dose combination tablet dosage form. Furthermore, the method was found to be sensitive and eco-friendly green alternative to the reported methods as it was evaluated according to the green analytical procedure index tool guidelines and analytical Eco-Scale.

Simple spectrofluorimetric methods for determination of veterinary antibiotic drug (apramycin sulfate) in pharmaceutical preparations and milk samples.

This work describes the development of highly sensitive as well as simple two spectrofluorimetric methods for the determination of apramycin sulfate. The first method depends on measuring the inherent native fluorescence of the aqueous neutral solution of the drug at 388 nm (λex 335 nm). While the second method mainly based on enhancing the native fluorescence intensity of the drug using sodium dodecyl sulfate (SDS) micellar media by about 4 fold enhancement. The fluorescence intensity - concentration relationship for the two methods was found rectilinear over the concentration range 1.0-100.0 and 0.1-20.0 µg/mL for the first and second method respectively. The limit of detection for method I and II were 0.05 and 0.02 µg/mL respectively. The proposed methods can be effectively connected for the assurance of the medication without impedances from common normal excipients. Furthermore, the two methods were high sensitive enough for the assurance of the drug in spiked milk samples with high percentage recoveries.

Ultrasound-assisted dispersive liquid-liquid microextraction for determination of three gliflozins in human plasma by HPLC/DAD.

A novel, highly sensitive ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) high performance liquid chromatography with diode-array detection (HPLC/DAD) method was developed for the determination of empagliflozin, dapagliflozin and canagliflozin in human plasma using methanol as protein precipitating agent/disperser and 1-dodecanol as extracting solvent. The analytes were eluted with an isocratic mobile phase consisting of acetonitrile:aqueous 0.1% trifluoroacetic acid pH 2.5, (40:60, v/v), at a flow rate of 1 mL/min and UV detection at 210 nm. The microextraction conditions were optimized regarding type and volume of extractant, type of disperser, sample pH, extraction time and centrifugation time. Under the optimal conditions, the enrichment factors were 19 for empagliflozin, 27 for dapagliflozin and 50 for canagliflozin. Linearity ranges were 2-2500 ng/mL, 3.5-2500 ng/mL and 1.1-2500 ng/mL for empagliflozin, dapagliflozin and canagliflozin, respectively. The developed method employs very small volumes of organic solvents in sample extraction and allows determination of small concetrations of gliflozins in human plasma.

A UPLC/DAD method for simultaneous determination of empagliflozin and three related substances in spiked human plasma.

A simple, rapid and sensitive ultrahigh performance liquid chromatographic method was developed for the determination of the anti-diabetic drug: empagliflozin (EMPA) and three related substances in spiked human plasma, using dapagliflozin (DAPA) as an internal standard and tetrahydrofuran as a plasma protein precipitating agent. The chromatographic separation was achieved on an Acquity "UPLC® BEH" C18 column (50 mm × 2.1 mm i.d, 1.7 µm particle size), and a mobile phase consisting of aqueous trifluoroacetic acid (0.1%, pH 2.5): acetonitrile (60:40, v/v) at a flow rate of 0.5 mL/min. Upon using the UPLC system, the run time could be reduced to less than 1.2 min, and the solvents consumption decreased to 0.36 mL of acetonitrile per run. The response was linear over a concentration range of 50-700 ng/mL and 40-200 ng/mL (r2 = 0.9994-0.9999) with lower limits of detection and quantification (LOD/LOQ) of 15/50, 11.5/40, 12/40 and 12.5/40 ng/mL for EMPA and the three related substances, respectively. Good accuracy was obtained with mean percentage recoveries ≥ 96.97% for the studied compounds. The method was validated according to the ICH guidelines and was found suitable for routine analysis of EMPA and its related substances in human plasma.

Development and Validation of HPLC Fluorescence and UPLC/DAD Stability-Indicating Methods for Determination of Hepatitis C Antiviral Agent Daclatasvir.

Background: Few stability-indicating chromatographic methods were published for determination of daclatasvir. All used UV detection. Objective: This work aimed to develop rapid, specific, and novel stability-indicating methods using HPLC with fluorescence detection and ultra performance liquid chromatography (UPLC) with UV detection for the determination of daclatasvir in bulk powder and in its dosage form. Methods: The drug was subjected to hydrolysis (acidic and alkaline) as per International Conference on Harmonization (ICH) guidelines. The fragmentation pattern of the drug was studied using LC-MS. Separation was carried out first by HPLC using Thermo BDS Hypersil Phenyl (300 mm × 4 mm, 5 µm) column and a mobile phase consisting of ammonium phosphate buffer (0.02 M) pH 3-methanol (40+60, v/v) at flow rate 1 mL/min. Quantitation was achieved by fluorescence detection at 305 and 457 nm for excitation and emission, respectively. The second method used UPLC equipped with diode-array detector. Acquity BEH C18 (50 mm × 2.1 mm, 1.7 µm) column was used with the same mobile phase at flow rate 0.4 mL/min and detection wavelength at 305 nm. ICH guidelines were used for validation. Results: The mean percentage recovery ± SD values for tablet assay were found to be 101.12 ± 0.655 and 99.78 ± 0.632 by HPLC and UPLC methods, respectively. The assay results showed a good agreement with the reported method. Conclusions: The developed HPLC and UPLC methods provide simple, accurate, and reproducible analysis of daclatasvir without interference from degradates. Highlights: This is the first research using fluorescence detection in a stability-indicating chromatographic method for determination of daclatasvir.

Simultaneous determination of amlodipine and metoprolol in their combined dosage form using a synchronous fluorescence spectrofluorimetric method.

Highly sensitive, rapid, accurate and precise synchronous fluorescence spectrofluorimetric method has been developed for simultaneous analysis of a mixture of amlodipine (AMD) and metoprolol (MET). The method relies on measuring the relative synchronous fluorescence intensity of both drugs at Δλ of 90 nm in acetate buffer solution at pH 5. The experimental parameters influencing the developed method were investigated and optimized. The method was linear over the ranges 0.2-2 µg/ml and 0.5-10 µg/ml for AMD and MET, respectively. The limits of detection were 50 ng/ml for AMD and 130 ng/ml for MET while the limits of quantitation were 150 ng/ml for AMD and 390 ng/ml for MET. The developed method was applied successfully for the determination of the two drugs in their co-formulated tablet. The mean percent recoveries were found to be 100.51 and 99.57 for AMD and MET, respectively.

Development and Validation of Chemometric Spectrophotometric Methods for Simultaneous Determination of Simvastatin and Nicotinic Acid in Binary Combinations.

BACKGROUND: The development and introduction of combined therapy represent a challenge for analysis due to severe overlapping of their UV spectra in case of spectroscopy or the requirement of a long tedious and high cost separation technique in case of chromatography. Quality control laboratories have to develop and validate suitable analytical procedures in order to assay such multi component preparations. METHODS: New spectrophotometric methods for the simultaneous determination of simvastatin (SIM) and nicotinic acid (NIA) in binary combinations were developed. These methods are based on chemometric treatment of data, the applied chemometric techniques are multivariate methods including classical least squares (CLS), principal component regression (PCR) and partial least squares (PLS). In these techniques, the concentration data matrix were prepared by using the synthetic mixtures containing SIM and NIA dissolved in ethanol. The absorbance data matrix corresponding to the concentration data matrix was obtained by measuring the absorbance at 12 wavelengths in the range 216 - 240 nm at 2 nm intervals in the zero-order. The spectrophotometric procedures do not require any separation step. The accuracy, precision and the linearity ranges of the methods have been determined and validated by analyzing synthetic mixtures containing the studied drugs. CONCLUSION: Chemometric spectrophotometric methods have been developed in the present study for the simultaneous determination of simvastatin and nicotinic acid in their synthetic binary mixtures and in their mixtures with possible excipients present in tablet dosage form. The validation was performed successfully. The developed methods have been shown to be accurate, linear, precise, and so simple. The developed methods can be used routinely for the determination dosage form.

Development and Validation of Chemometric-Assisted Spectrophotometric Methods for Simultaneous Determination of Phenylephrine Hydrochloride and Ketorolac Tromethamine in Binary Combinations.

The present work describes new spectrophotometric methods for the simultaneous determination of phenylephrine hydrochloride and ketorolac tromethamine in their synthetic mixtures. The applied chemometric techniques are multivariate methods including classical least squares, principal component regression, and partial least squares. In these techniques, the concentration data matrix was prepared by using the synthetic mixtures containing these drugs dissolved in distilled water. The absorbance data matrix corresponding to the concentration data was obtained by measuring the absorbances at 16 wavelengths in the range 244-274 nm at 2 nm intervals in the zero-order spectra. The spectrophotometric procedures do not require any separation steps. The accuracy, precision, and linearity ranges of the methods have been determined, and analyzing synthetic mixtures containing the studied drugs has validated them. The developed methods were successfully applied to the synthetic mixtures and the results were compared to those obtained by a reported HPLC method.

Stability-indicating HPLC-DAD methods for determination of two binary mixtures: Rabeprazole sodium-mosapride citrate and rabeprazole sodium-itopride hydrochloride.

Two selective stability-indicating HPLC methods are described for determination of rabeprazole sodium (RZ)-mosapride citrate (MR) and RZ-itopride hydrochloride (IO) mixtures in the presence of their ICH-stress formed degradation products. Separations were achieved on X-Bridge C18 column using two mobile phases: the first for RZ-MR mixture consisted of acetonitrile: 0.025 M KH2PO4 solution: TEA (30:69:1 v/v; pH 7.0); the second for RZ-IO mixture was at ratio of 25:74:1 (v/v; pH 9.25). The detection wavelength was 283 nm. The two methods were validated and validation acceptance criteria were met in all cases. Peak purity testing using contrast angle theory, relative absorbance and log A versus the wavelengths plots were presented. The % recoveries of the intact drugs were between 99.1% and 102.2% with RSD% values less than 1.6%. Application of the proposed HPLC methods indicated that the methods could be adopted to follow the stability of their formulations.

Role of alpha-lipoic acid in the management of anemia in patients with chronic renal failure undergoing hemodialysis.

INTRODUCTION: Anemia associated with chronic kidney disease is a serious complication necessitating expenditure of huge medical efforts and resources. This study investigates the role of alpha-lipoic acid (ALA) in end stage renal disease patients undergoing hemodialysis. By the virtue of its antioxidative effects, ALA is expected to act as an erythropoietin (EPO) adjuvant, and also has extended beneficial effects on endothelial dysfunction. METHODS: Forty-four patients undergoing hemodialysis and receiving EPO were randomized into two groups: the first group received ALA 600 mg once daily for 3 months; while the other group represented the control group. Parameters measured at baseline and at end of study were hemoglobin, EPO doses, EPO resistance index (ERI), iron store indices, malondialdehyde, oxidized low-density lipoprotein (ox-LDL), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and asymmetric dimethylarginine (ADMA), as well as routine laboratory follow-up. RESULTS: EPO doses and ERI were significantly decreased in the treatment group, while they did not change in the control group. Hemoglobin, iron store indices, malondialdehyde, oxidized ox-LDL, IL-6, TNF-α, and ADMA were similar in both treatment and control groups at baseline, and did not change by the end of study period. Likewise, routine laboratory measures were not affected by the treatment. CONCLUSION: ALA could be used in hemodialysis patients to reduce requirements for EPO. However, larger and longer term studies are required to clarify the exact role of ALA in hemodialysis as well as in pre-hemodialysis patients.

Effect of ketoprofen and indomethacin on methotrexate pharmacokinetics in mice plasma and tumor tissues.

Methotrexate (MTX) has been used in combination with nonsteroidal anti-inflammatory drugs in the treatment of inflammatory diseases as well as malignancies. Severe adverse effects with this combination may occur, usually resulting from inhibition of renal transporters. Solid Ehrlich carcinoma was experimentally induced by implantation of Ehrlich ascites Carcinoma cells subcutaneously into the thigh of mice, and after 30 days, mice were divided into three groups: Group I that served as control group received MTX (50 mg/kg, i.p.); Group II received ketoprofen (100 mg/kg, i.p.) and then after half an hour received MTX (50 mg/kg, i.p.); Group III received indomethacin (10 mg/kg, i.p.) and then after half an hour received MTX (50 mg/kg, i.p.). Plasma and tissue samples were collected at different time points and then MTX concentrations were determined by HPLC. The injection of ketoprofen or indomethacin before MTX injection resulted in significant increase in the AUC and CPmax of MTX (p < 0.05) and significant decrease in CL/F and Vd/F of MTX (p < 0.05) in mice plasma. The effects were more significant after injection of indomethacin than in case of ketoprofen. The study showed that administration of ketoprofen or indomethacin prior to MTX caused significant decrease in MTX elimination and significant increase in MTX extent of absorption which may lead to severe adverse effects if coadministered in human.

Effect of clarithromycin and fluconazole on the pharmacokinetics of montelukast in human volunteers.

OBJECTIVE: Montelukast, a leukotriene receptor antagonist, is used in the treatment of asthma. The objective of the study reported here was to determine whether multiple doses of clarithromycin or fluconazole affect the pharmacokinetics of montelukast. METHODS: This was a four-phase cross-over study with a washout period of 2 weeks between phases. In phase 1, 12 volunteers received a single oral dose of 10 mg montelukast. In phase 2, the volunteers received a single, oral dose of 1,000 mg clarithromycin once daily for 2 days, followed by, on day 3, a single oral dose of 10 mg montelukast co-administered with clarithromycin. In phase 3, a single oral dose of 50 mg fluconazole was given once daily for 6 days, followed by, on day 7, a single oral dose of 10 mg montelukast co-administered with 50 mg fluconazole. In the last phase (phase 4), a single oral dose of 150 mg fluconazole was given once daily for 6 days, followed by, on day 7, a single oral dose of 10 mg montelukast co-administered with 150 mg fluconazole. The plasma concentration of montelukast was measured by high performance liquid chromatography for 24 h. RESULTS: Following clarithromycin co-administration, the area under the concentration-time curve from zero to infinity ( AUC(0-∞)) of montelukast increased by 144% [90% confidence interval (CI) 2.03-2.86]. The co-administration of a single oral dose of 150 and 50 mg fluconazole decreased the montelukast AUC(0-∞) by 30.7 (90% CI 0.53-0.81) and 38.8% (90% CI 0.57-0.69), respectively. CONCLUSIONS: Clarithromycin increased the plasma concentrations of montelukast whereas fluconazole reduced the plasma concentrations of montelukast. The mechanism of the interaction is probably due to interference of the interacting drugs with transporters mediating the uptake of montelukast.

Simultaneous determination of the two non-steroidal anti-inflammatory drugs ; diflunisal and naproxen in their tablets by chemometric spectrophotometry and HPLC.

Chemometric spectrophotometry and HPLC were applied to the simultaneous determination of the two non-steroidal anti-inflammatory drugs; diflunisal (I) and naproxen (II). The applied chemometric techniques are multivariate methods including classical least squares (CLS), principal component regression (PCR) and partial least squares (PLS); and the second derivative of the ratio spectra ((2)D(r)) method. To develop the multivariate methods, the UV absorption spectra of the standard solutions of the training and validation sets in methanol were recorded in the range of 242-274 nm at 2 nm intervals. The specificity of the studied multivariate methods has been tested. In the (2)D(r) method, analytical signals at 235 and 259 nm were selected for the determination of (I) and (II), respectively. The HPLC method depends on reversed-phase separation using C18 column. The mobile phase consists of a mixture of acetonitrile - acetate buffer (pH 4.2; 50 mM) (60:40, v/v). The UV detector was set at 255 nm. The developed methods were validated and successfully applied to the simultaneous determination of (I) and (II) in their tablets. The assay results obtained using the chemometric methods were statistically compared to those of the HPLC method and good agreement was observed.