HPLC-Fluorescence Detection Method for Concurrent Estimation of Domperidone and Naproxen. Validation and Eco-Friendliness Appraisal Studies.

This work demonstrates a simple and reliable HPLC method with fluorimetric detection for simultaneous estimation of domperidone (DOM) and naproxen (NAP). Successful chromatographic separation was accomplished using Inertsil ODS C18 column (5 µm, 4.6 × 150 mm) with gradient elution of the mobile phase consisting of 0.01 M phosphate buffer (pH 5.5) solution and acetonitrile. The gradient elution started with 25% acetonitrile increased linearly to 65% in 5 min, then kept at this percentage till the end of the run. The mobile phase was pumped at a flow rate of 1.0 mL/min. The excitation wavelength at 284 nm was found suitable for both DOM and NAP since it corresponds to a maximum for the minor component DOM and measurable excitation for NAP, while using 316 and 355 nm as emission wavelengths for DOM and NAP, respectively. Peaks eluted with excellent resolution at retention times 4.4 and 6.3 min for DOM and NAP, respectively. Performance of the proposed method was tested according to ICH guidelines in regard to linearity, ranges, precision, accuracy, robustness, detection and quantitation limits. Calibration curves were linear in the ranges of 0.8-3.6 and 1.0-2.5 µg/mL for DOM and NAP respectively with correlation coefficients not less than 0.9996. The validated method was successfully applied to the analysis of DOM and NAP in their laboratory prepared tablets resembling the commercial dosage form, and assay results were favorably compared with a published reference HPLC method. The method's greenness was assessed using the Analytical Eco-Scale and the novel Analytical Greenness metric (AGREE).

Design and Optimization of 3D-Printed Gastroretentive Floating Devices by Central Composite Design.

This study aimed to optimize the size of capsule-shaped 3D-printed devices (CPD) using an experimental design by the response surface methodology to provide a gastroretentive drug delivery system (GRDDS) with optimal floating time. The CPD was fabricated using a fused deposition modeling (FDM) 3D printer. The central composite design was employed for the optimization of the devices. The morphology of the CPD was observed using a digital microscope and scanning electron microscope (SEM). The in vitro floating time and drug release were evaluated using a USP dissolution apparatus II. Appropriate total floating time (TFT) of the devices (more than 3 h) was obtained with the device's body, cap, and bottom thickness of 1.2, 1.8, and 2.9 mm, respectively. The release kinetics of the drug from the devices fitted well with zero-order kinetics. In conclusion, the optimization of CPD for GRDDS using the experimental design provided the devices with desirable floating time and ideal drug release characteristics.

Separation of Unprecedented Degradants of Domperidone by Ultra-Performance Convergence Chromatography and Their Structure Elucidation.

Domperidone, a gastroprokinetic agent, is a common drug to treat emesis. It was subjected to acid, base-mediated hydrolysis, peroxide-mediated oxidation, photolysis and thermal degradation according to ICH guidelines to observe stability of the selected drug under the stress conditions. Although the drug is resistant to base hydrolysis, photolysis and thermal stressors, two degradants (DP-ISO1 and DP-ISO2) were formed in acid mediated hydrolysis. Oxidation with hydrogen peroxide also resulted in one product (DP-OX). All three degradants were isolated from the crude reaction mixture by preparative high-performance liquid chromatography and supercritical fluid chromatography. Structures of isolated compounds were unambiguously characterized as 5-chloro-1-(1-(3-(6-chloro-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one (DP-ISO1), 5-chloro-1-(3-(4-(5-chloro-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidin-1-yl)propyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one (DP-ISO2), 4-(5-chloro-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)-1-(3-(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)propyl)piperidine 1-oxide (DP-OX) by analysis of mass spectrometry, 1D and 2D nuclear magnetic resonance spectra. To the best of our knowledge, DP-ISO1 and DP-ISO2 are new and DP-OX was previously reported as domperidone impurity.

Simultaneous voltammetric analysis of anti-ulcer and D2-antagonist agents in binary mixture using redox sensor and their determination in human serum.

Pencil graphite electrode was successfully modified with a thin film of poly (eriochrome black T) and applied for the sensitive and selective voltammetric simultaneous determination of pantoprazole sodium and domperidone in a binary mixture. The preparation and basic electrochemical behavior of poly (eriochrome black T) film on the Pencil graphite electrode were investigated. The modified electrode has exhibited very high electro-catalytic activity towards the cited mixture. The anodic peaks of the both species were well defined with enhanced oxidation peak currents. Under the optimum conditions, the linearity ranges were 0.4-55×10-7M and 0.1-34×10-7M for pantoprazole sodium and domperidone, respectively with detection limits of 0.12×10-7M and 0.04×10-7M for pantoprazole sodium and domperidone, respectively. The proposed sensor has been successfully applied in the analysis of pantoprazole sodium and domperidone in synthetic binary mixtures and human serum.

Fourth-derivative synchronous spectrofluorimetry and HPLC with fluorescence detection as two analytical techniques for the simultaneous determination of itopride and domperidone.

Two simple, rapid and sensitive methods, namely, fourth-derivative synchronous spectrofluorimetry (method I) and HPLC with fluorescence detection (method II) were developed for the simultaneous analysis of a binary mixture of itopride HCl (ITP) and domperidone (DOM) without prior separation. The first method was based on measuring the fourth derivative of the synchronous fluorescence spectra of the two drugs at Δλ = 40 nm in methanol. The different experimental parameters affecting the synchronous fluorescence of the studied drugs were carefully optimized. Chromatographic separation was performed in < 6.0 min using a RP C18 column (250 mm × 4.6 mm i.d., 5 µm particle size) with fluorescence detection at 344 nm after excitation at 285 nm. A mobile phase composed of a mixture of 0.02 M phosphate buffer with acetonitrile in a ratio of 55 : 45, pH 4.5, was used at a flow rate of 1 mL/min. Linearity ranges were found to be 0.1-2 µg/mL for ITP in both methods, whereas those for DOM were found to be 0.08-2 and 0.05-1.5 µg/mL in methods I and II, respectively. The proposed methods were successfully applied for the determination of the studied drugs in synthetic mixtures and laboratory-prepared tablets.

Gold nanoparticles decorated poly-melamine modified glassy carbon sensor for the voltammetric estimation of domperidone in pharmaceuticals and biological fluids.

The electrochemical response of an unmodified glassy carbon (GCE), poly-melamine/GCE and gold nanoparticle (AuNP)/poly-melamine/GCE is compared in the present protocol for the sensitive and selective determination of domperidone (DOM). The AuNPs were synthesized in the laboratory and characterized using UV-visible spectroscopy and Transmission Electron Microscopy (TEM). Melamine was electropolymerized onto the glassy carbon surface using cyclic voltammetry and was investigated using Field Emission Scanning Electron Microscopy (FE-SEM) and Electrochemical Impedance Spectroscopy (EIS). The AuNP/poly-melamine/GCE exhibited the best electrochemical response among the three electrodes for the electro-oxidation of DOM, that was inferred from the EIS, cyclic and square wave voltammetry. The modified sensor showed a sensitive, stable and linear response in the concentration range of 0.05-100µM with a detection limit of 6nM. The selectivity of the proposed sensor was assessed in the presence of high concentration of major interfering molecules as xanthine, hypoxanthine, and uric acid. The analytical application of the sensor for the quantification of DOM in pharmaceutical formulations and biological fluids as urine and serum was also investigated and the results demonstrated a recovery of >95% with R.S.D of <5%.

Validation of four different spectrophotometric methods for simultaneous determination of Domperidone and Ranitidine in bulk and pharmaceutical formulation.

Four simple, specific, accurate and precise spectrophotometric methods were developed and validated for simultaneous determination of Domperidone (DP) and Ranitidine Hydrochloride (RT) in bulk powder and pharmaceutical formulation. The first method was simultaneous ratio subtraction (SRS), the second was ratio subtraction (RS) coupled with zero order spectrophotometry (D(0)), the third was first derivative of the ratio spectra ((1)DD) and the fourth method was mean centering of ratio spectra (MCR). The calibration curve is linear over the concentration range of 0.5-5 and 1-45 µg mL(-1) for DP and RT, respectively. The proposed spectrophotometric methods can analyze both drugs without any prior separation steps. The selectivity of the adopted methods was tested by analyzing synthetic mixtures of the investigated drugs, also in their pharmaceutical formulation. The suggested methods were validated according to International Conference of Harmonization (ICH) guidelines and the results revealed that; they were precise and reproducible. All the obtained results were statistically compared with those of the reported method, where there was no significant difference.

The ultra-performance liquid chromatography determination of domperidone and its process-related impurities.

A rapid ultra-performance liquid chromatography (UPLC) method for the determination of domperidone in the presence of its process impurities and droperidol was developed and validated. The rapid chromatographic separation was achieved using a sub 2 µm Hypersil Zorbax eXtra Densely Bonded C18 column (30 × 4.6 mm, i.d., 1.8 µm). A gradient mobile phase consisting of Solvent A: 0.06 M ammonium acetate and Solvent B: methanol, with a flow rate of 1 mL/min was employed. The column temperature was set at 40°C, and the diode-array detector was set at 280 nm. An injection volume of 3 µL was used. The currently utilized European Pharmacopeia (Eur. Pharm.) method employed by Janssen Pharmaceuticals Ltd was run on a Hypersil Base-Deactivated Silica C18 column (100 × 4.6 mm, i.d., 3 µm) with a run time of 12.5 min. The developed UPLC method, with a run time of 7.5 min was determined to be accurate, precise, specific, robust and highly sensitive according to the International Conference on Harmonization guidelines. The method herein demonstrated a reduction in analysis time of 40%, allowing for a much higher sample throughput. A solvent consumption decrease of over 58% was also observed, which results in a dramatic reduction in running costs for Janssen Pharmaceuticals Ltd.

Spectrophotometric methods manipulating ratio spectra for simultaneous determination of binary mixtures with sever overlapping spectra: a comparative study.

Three simple, specific and accurate spectrophotometric methods manipulating ratio spectra were developed and validated for simultaneous determination of Rabeprazole sodium (RB) and Domperidone (DP) in their binary mixture without prior separation. Method A, is constant center spectrophotometric method (CC). Method B is a ratio difference spectrophotometric one (RD), while method C is a combined ratio isoabsorptive point-ratio difference method (RIRD). Linear correlations were obtained in range of 4-44µg/mL for both Rabeprazole sodium and Domperidone. The mean percentage recoveries of RB were 99.69±0.504 for method A, 99.83±0.483 for (B) and 100.31±0.499 for (C), respectively, and that of DP were 99.52±0.474 for method A, 100.12±0.505 for (B) and 100.16±0.498 for (C), respectively. Specificity was investigated by analysis of laboratory prepared mixtures containing the cited drugs and their combined tablet dosage form. The obtained results were statistically compared with those obtained by the reported methods, showing no significant difference with respect to accuracy and precision. The three methods were validated as per ICH guidelines and can be applied for routine analysis in quality control laboratories.

Liquid chromatography coupled with fluorimetric detection and third derivative synchronous fluorescence spectroscopy as two analytical methods for the simultaneous determination of rabeprazole sodium and domperidone after derivatization with 4-chloro-7-nitrobenzofurazan.

Two simple, sensitive, rapid, economic and validated methods, namely reversed phase liquid chromatography (method Ι) and third derivative synchronous fluorescence spectroscopy (method ΙΙ) have been developed for the simultaneous determination of rabeprazole sodium and domperidone in their laboratory prepared mixture after derivatization with 4-Chloro-7-nitrobenzofurazan. Reversed phase chromatography was conducted using a Zorbax® SB-Phenyl column (250.0 mm × 4.6 mm id) combined with a guard column at ambient temperature with fluorimetric detection at 540 nm after excitation at 483 nm. A mobile phase composed of a mixture of distilled water with methanol and acetonitrile in a ratio of 50:20:30 adjusted pH to 4 has been used at a flow rate of 1 mL/min. Sharp well resolved peaks were obtained for domperidone and rabeprazole sodium with retention times of 5.5 and 6.4 min respectively. While in method ΙΙ, the third-derivative spectra were estimated at 507 and 436 nm for rabeprazole sodium and domperidone respectively. Linearity ranges for rabeprazole sodium and domperidone respectively in both methods were found to be 0.15-2.0 and 0.1-1.5 µg/mL. The proposed methods were successfully applied for the analysis of the two compounds in their binary mixtures, and laboratory prepared tablets. The obtained results were favorably compared with those obtained by the comparison method. Furthermore, detailed validation procedure was also conducted.

Simultaneous determination of Cinnarizine and Domperidone by area under curve and dual wavelength spectrophotometric methods.

Accurate, selective and sensitive spectrophotometric methods have been developed and validated for simultaneous determination of Cinnarizine and Domperidone, a binary mixture with overlapping spectra, without preliminary separation. These methods include area under the curve (AUC) and dual wavelength spectrophotometry. For the AUC method, the area under curve of mixture solutions in the wavelength ranges 241-258 nm and 280-292 nm were selected for determination of Cinnarizine and Domperidone and by applying Cramer's rule, concentration of each drug was obtained. In dual wavelength method, two wavelengths were selected for each drug in a way so that the difference in absorbance is zero for another drug. Domperidone shows equal absorbance at 240.2 nm and 273.2 nm, where the differences in absorbance were measured for the determination of Cinnarizine. Similarly, differences in absorbance at 230.8 nm and 259.2 nm were measured for determination of Domperidone. The proposed methods were applied for determination of Cinnarizine and Domperidone over the concentration ranges of 2-20 and 2-22 µg mL(-1), respectively. The suggested methods were validated as per USP guidelines and the results revealed that they are reliable, reproducible and precise for routine use with short analysis time. The results obtained by the proposed methods were statistically compared to the reported method, and there was no significant difference between them regarding both accuracy and precision.

Capillary electrophoretic determination of antimigraine formulations containing caffeine, ergotamine, paracetamol and domperidone or metoclopramide.

A novel, fast, sensitive and specific technique using capillary electrophoresis coupled to a diode array detector has been developed for the separation and simultaneous determination of two antimigraine mixtures in tablet formulation. The two combinations are ergotamine tartrate (ERG), caffeine (CAF) and paracetamol (PAR) with either domperidone (DOM), combination (I) or metoclopramide (MET), combination (II). The proposed method utilized a fused silica capillary (55 cm × 75 µm i.d.) and background electrolyte composed of phosphate buffer (25 mM, pH 9.8). The separation was achieved at 20 KV applied voltage and at 25°C. The described method was linear over the range of 1-80 and 2-100 µg/mL for CAF and MET, respectively, and 1-80 µg/mL for DOM, ERG and PAR. Intra-day and inter-day relative standard deviation (n = 5) was ≤1.10%. The limits of detection of CAF and PAR were 0.20 and 0.10 µg/mL, respectively, and 0.50 µg/mL for MET, DOM and ERG. Other aspects of analytical validation were also evaluated. The proposed method was successfully applied to the analysis of the two combinations in their tablets. Therefore, the proposed method is suitable for the routine control of these ingredients in multicomponent dosage forms.

Stability-indicating TLC-densitometric method for estimation of dexrabeprazole and domperidone in pharmaceutical dosage form.

A stability-indicating thin layer chromatographic (TLC) method for determination of dexrabeprazole (DEX) and domperidone (DOM) in combined dosage form has been developed and validated. The mobile phase selected was acetone:toluene:methanol (4.5:4.5:0.5, v/v/v) with ultraviolet (UV) detection at 285 nm. The retention factors for DEX and DOM were found to be 0.49 ± 0.02 and 0.24 ± 0.03, respectively. The method was validated with respect to linearity, accuracy, precision, and robustness. The linearity range for DEX was 50-350 ng/band (r² = .9960) and for DOM was 100-700-n /band (r² = .9982), respectively. The method was successfully applied for the analysis of drugs in pharmaceutical formulation.

High-performance thin-layer chromatographic determination of rabeprazole sodium and domperidone in combined dosage form.

A new, simple, high-performance thin-layer chromatographic method for determination of rabeprazole sodium (RAB) and domperidone (DOM) in combined tablet dosage form has been developed and validated. The mobile phase was toluene-acetone-methanol (4.5 + 4.5 + 0.5, v/v/v) with UV detection at 285 nm. The retention factors for RAB and DOM were found to be 0.53 +/- 0.12 and 0.32 +/- 0.20. The method was validated with respect to linearity, accuracy, precision, and robustness. Beer's law was obeyed in the concentration range of 50-800 ng/band for both RAB and DOM. The method has been successfully applied for the analysis of drugs in a pharmaceutical formulation.

Column reversed-phase high-performance liquid chromatographic method for simultaneous determination of rabeprazole sodium and domperidone in combined tablet dosage form.

A new, simple column reversed-phase high-performance liquid chromatographic (HPLC) method for simultaneous determination of rabeprazole sodium (RAB) and domperidone (DOM) in a combined tablet dosage form has been developed and validated. Determination was performed using a Jasco HPLC system with a HiQ SiL octadecylsilane (C18) column (250 x 4.6 mm id), acetonitrile-0.1 M ammonium acetate (50 + 50, v/v) mobile phase, and paracetamol as an internal standard. The detection was performed using a UV detector set at 280 nm. The method was validated with respect to linearity, accuracy, precision, and robustness. Beer's law was obeyed in the concentration range of 1.0-10.0 and 0.5-5.0 microg/mL for RAB and DOM, respectively. The method has been successfully applied for the analysis of drugs in a pharmaceutical formulation.

Second-derivative synchronous fluorometric method for the simultaneous determination of cinnarizine and domperidone in pharmaceutical preparations. Application to biological fluids.

A rapid, simple and highly sensitive second derivative synchronous fluorometric method has been developed for the simultaneous analysis of binary mixture of cinnarizine (CN) and domperidone (DOM). The method is based upon measurement of the native fluorescence of these drugs at Deltalambda=80 nm in aqueous methanol (50% V/V). The different experimental parameters affecting the native fluorescence of the studied drugs were carefully studied and optimized. The fluorescence-concentration plots were rectilinear over the range of 0.1 to 1.3 microg mL(-1) and 0.1-3.0 microg mL(-1) for CN and DOM, respectively with lower detection limits of 0.017 and 5.77 x 10(-3) microg mL(-1) and quantification limits of 0.058 and 0.02 microg mL(-1) for CN and DOM. The proposed method was successfully applied for the determination of the studied compounds in synthetic mixtures and in commercial tablets. The results obtained were in good agreement with those obtained with reference methods. The high sensitivity attained by the synchronous fluorometric method allowed the determination of CN in real and spiked human plasma. The mean % recoveries in case of spiked human plasma (n=3) were 96.39+/-1.18 while that in real human plasma (n = 3) was 104.67+/-4.16.

An improved HPLC assay with fluorescence detection for the determination of domperidone and three major metabolites for application to in vitro drug metabolism studies.

Domperidone is currently used in Canada and Europe for the treatment of intestinal motility disorders as well as for its antiemetic properties. Recent drug metabolism studies have indicated that domperidone is a substrate of different subtypes of CYP3A family and consequently, the drug requires complete characterization of its metabolism for the identification of major drug-drug interactions. Therefore, the purpose of our studies was to develop a simple, sensitive and rapid HPLC assay for the determination of domperidone and its major metabolites. This assay had to be suitable for the conduct of in vitro drug metabolism study with human liver microsomes. Baseline resolution of internal standard, domperidone and three of its major metabolites was achieved in a run time of less than 15 min using an Ultrasphere ODS column (250 mm x 4.6 mm x 5 microM) and a mobile phase consisting of disodium citrate buffer (10 mM, pH 3.4):methanol:acetonitrile:trietylamine, 54.6:34.7:9.9:0.8 at a flow rate of 1.0 mL/min. Chromatographic separation was executed at room temperature. Quantification was performed by tandem fluorescence (excitation lambda=282 nm and emission lambda=328 nm) and ultraviolet detectors (lambda=254 nm for the quantification of encainide, internal standard). Calibration curves were constructed and showed linearity in the range of 0.1-20 micromol/L and 10-250 micromol/L. Intra- and interday coefficients of variation were less than 8% and 11%, respectively. Mean accuracy was 100.5+/-9.9% and limit of quantification was established at 0.06 micromol/L for domperidone and its metabolites. The assay allows estimation of enzymatic parameters (K(m) and V(max)) of domperidone for the formation of its various metabolites and sensitivity is sufficient for the conduct of inhibition studies with potent CYP3A inhibitors.

Simultaneous estimation of pantoprazole and domperidone in pure powder and a pharmaceutical formulation by high-perfomance liquid chromatography and high-performance thin-layer chromatography methods.

This paper describes validated high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) methods for the simultaneous estimation of pantoprazole (PANT) and domperidone (DOM) in pure powder and capsule formulations. The HPLC separation was achieved on a Phenomenex C18 column (250 mm id, 4.6 mm, 5 pm) using 0.01 M, 6.5 pH ammonium acetate buffer-methanol-acetonitrile (30 + 40 + 30, v/v/v, pH 7.20) as the mobile phase at a flow rate of 1.0 mL/min at ambient temperature. The HPTLC separation was achieved on an aluminum-backed layer of silica gel 60F254 using ethyl acetate-methanol (60 + 40, v/v) as the mobile phase. Quantification was achieved with ultraviolet (UV) detection at 287 nm over the concentration range 400-4000 and 300-3000 ng/mL with mean recovery of 99.35+/-0.80 and 99.08+/-0.57% for PANT and DOM, respectively (HPLC method). Quantification was achieved with UV detection at 287 nm over the concentration range 80-240 and 60-180 ng/spot with mean recovery of 98.40+/-0.67 and 98.75+/-0.71% for PANT and DOM, respectively (HPTLC method). These methods are simple, precise, and sensitive, and they are applicable for the simultaneous determination of PANT and DOM in pure powder and capsule formulations.

Analysis of domperidone in pharmaceutical formulations and wastewater by differential pulse voltammetry at a glassy-carbon electrode.

The redox characteristics of the drug domperidone at a glassy-carbon electrode (GCE) in aqueous media were critically investigated by differential-pulse voltammetry (DPV) and cyclic voltammetry (CV). In Britton-Robinson (BR) buffer of pH 2.6-10.3, an irreversible and diffusion-controlled oxidation wave was developed. The dependence of the CV response of the developed anodic peak on the sweep rate (nu) and on depolizer concentration was typical of an electrode-coupled chemical reaction mechanism (EC) in which an irreversible first-order reaction is interposed between the charges. The values of the electron-transfer coefficient (alpha) involved in the rate-determining step calculated from the linear plots of E (p,a) against ln (nu) in the pH range investigated were in the range 0.64 +/- 0.05 confirming the irreversible nature of the oxidation peak. In BR buffer of pH 7.6-8.4, a well defined oxidation wave was developed and the plot of peak current height of the DPV against domperidone concentration at this peak potential was linear in the range 5.20 x 10(-6) to 2.40 x 10(-5) mol L(-1) with lower limits of detection (LOD) and quantitation (LOQ) of 6.1 x 10(-7) and 9.1 x 10(-7) mol L(-1), respectively. A relative standard deviation of 2.39% (n = 5) was obtained for 8.5 x 10(-6) mol L(-1) of the drug. These DPV procedures were successfully used for analysis of domperidone in the pure form (98.2 +/- 3.1%), dosage form (98.35 +/- 2.9%), and in tap (97.0 +/- 3.6%) and wastewater (95.0 +/- 2.9%) samples. The method was validated by comparison with standard titrimetric and HPLC methods. Acceptable error of less than 3.3% was also achieved.

Stability indicating simultaneous determination of domperidone (DP), methylparaben (MP) and propylparaben by high performance liquid chromatography (HPLC).

A simple, specific and precise high performance liquid chromatographic method has been developed and validated for the simultaneous determination of methylparaben (MP), propylparaben (PP), and domperidone (DP) in oral suspension. Isocratic mobile phase consists of 0.5% w/v aqueous ammonium acetate buffer:methanol, 40:60 (v/v). Column containing octylsilyl chemically bonded to porous silica particles (Optimapak, OP C8, 150 mmx4.6 mm, 5 microm, stainless steel analytical column from RS tech) is used as stationary phase. The detection is carried out using variable wavelength UV-vis detector set at 280 nm. The solutions are chromatographed at constant flow rate of 1.0 mL/min. The method separates MP, PP, DP and droperidol (DR) impurity in less than 12 min with good resolution, peak shapes and minimal tailing. Retention times (RT) for MP, PP, DP and DR are about 3.4, 7.0, 9.0 and 10.9 min, respectively. Linearity range and percent recoveries for MP, PP and DP are 90-270, 10-30, 50-1500 microg/mL and 100.30%, 100.78% and 100.48%, respectively. Method was validated according to ICH guidelines and proved to be suitable for stability testing, homogeneity testing and quality control of these compounds in pharmaceutical preparations.