Combined Dosage Form Research Articles

Simultaneous micro-determination of eplerenone and torsemide in their combined tablets using HPTLC-Dual wavelength spectrodensitometric and spectrophotometric methods

Sudden death is common in patients with Congestive heart failure, occurring at a rate of six to nine times that of the general population. Day by Day, new combined therapies of more advanced drug generations are involved in treatment. Recently, the Eplerenone/ Torsemide binary therapy is displacing the older Spironolactone/ Frusemide treatment trend of heart disease because it proved better tolerability and potency. This necessitates fast development of analytical methods that are capable of simultaneous determination of such important drug mixture. In this study, validated analytical methods have been established for simultaneous quantitation of eplerenone (EPL) and torsemide (TOR). One of the methods represents the first high-performance thin-layer chromatographic (HPTLC) attempt for EPL-TOR simultaneous estimation. A mixture of ethyl acetate -methanol- chloroform (8:2:1, v/v/v) was applied as mobile phase. EPL and TOR were well resolved and scanned at 242 nm (at hRF 77) and 288 nm (at hRF 36), respectively. Other proposed methods include solvent-induced difference, derivative ratio and ratio subtraction (complemented with isosbetstic point) spectrophotometric methods that were adopted as faster, simpler and more economic alternatives for the routine analysis of the increasingly used binary EPL-TOR medication, especially in the developing countries. Both HPTLC and spectrophotometric methods successfully determined both drugs without interference from one another. All methods were validated as per ICH guidelines. In the HPTLC method, calibration plots were linear between 50–600 and 35–800 ng/band for EPL and TOR, respectively. The spectrophotometric methods proved excellent linearity in the range of 3–25 and 3–30 μg/mL for EPL and TOR, respectively. Within all the proposed methods, ratio subtraction spectrophotometric method was the most sensitive one for EPL (LOD = 0.75 μg/mL). Regarding TOR, direct spectrophotometry achieved the lowest LOD (0.71 μg/mL). The high accuracy, simplicity and low cost of the proposed methods recommend their application in the industrial analysis of EPL and TOR combined dosage forms. A full comparison with the reported techniques shows the privileges of the suggested analytical procedures.

METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF LAMIVUDINE AND ZIDOVUDINE IN TABLET BY REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

Objective: The objective of the study was to develop and validate reverse-phase high-performance liquid chromatography (RP-HPLC) method and apply method to tablet dosage form.
 Methods: A simple, rapid, economical, precise, and accurate RP-HPLC method for simultaneous estimation of lamivudine and zidovudine in their combined dosage form has been developed.
 Results: A RP-HPLC method was developed for the simultaneous estimation of lamivudine and zidovudine. In their combined dosage form has been developed. The separation was achieved by LC-C18 column (150 mm ×4.6 mm, 5 μm) and water: methanol (65:35v/v) as mobile phase, at a flow rate of 0.8 ml/min. Detection was carried out at 272 nm. Retention time of lamivudine and zidovudine was found to be 3.007 min and 4.647, respectively. The method has been validated for linearity, accuracy, and precision. The assay method was found to be linear from 50% to 150% for lamivudine and zidovudine.
 Conclusion: Developed method was found to be accurate, precise, and rapid for simultaneous estimation of lamivudine and zidovudine in their combined dosage form.

USB multiplex analyzer employing screen-printed silver electrodes on paper substrate; A developed design for dissolution testing

Multiplex ion analyzers have been introduced recently for the assay of several inorganic ions, whilst electrochemists have extensively employed screen printed sensors for pharmaceutical analyses. This work aims to develop a USB pluggable sensor with a user-friendly design for multiplex analysis of oppositely charged co-formulated organic ions. The miniaturized screen-printed electrode was developed using silver ink on paper substrate. A compact sensor design was attained by including three electrodes, a single reference electrode along with an indicator electrode for each of the determined ions. Optimized PVC membranes were drop-casted over each of the indicator electrodes for the determination of phenylephrine HCl (PHE) and ibuprofen (IBU). The proposed multiplex potentiometric sensors exhibit Nernstian slopes of 59.2 ± 0.26 and -56.8 ± 0.16 mV/decade for PHE and IBU, respectively, with respective detection limits of 1.6 × 10-7 and 6.53 × 10-8 mol L-1. The fast and stable response of the developed sensor enabled the real-time monitoring of the combined dosage form dissolution. The dissolution profiles obtained by this potentiometric analyzer and an off-line separation technique were compared favourably, albeit our proposed in-line sensor reduced waste and time of analysis. The developed method successfully complies with the most demanding stipulations of green analytical chemistry.

Stability-indicating HPTLC method development for determination of arterolane maleate and piperaquine phosphate in combined dosage form

Stability-indicating HPTLC method development for determination of arterolane maleate and piperaquine phosphate in combined dosage form

A Smart Green Spectrophotometric Method for Simultaneous Determination of Severely Overlapped Binary Mixtures Using Normalized Spectrum and Isosbestic Point as Resolving Tools

Background: The ability of the normalized spectra when used as a divisor and in combination with isosbestic point to resolve complex binary or ternary mixtures, Candesartan and Hydrochlorthiazide binary mixture was taken as a model. Introduction: A green simple smart and accurate method using ethanol as a solvent namely simultaneous derivative ratio (SIDD) was applied to prove the power of normalized spectra and isosbestic point as spectrophotometric resolving tools. Methods: In the proposed SIDD method, the zero order spectra of drugs were simply manipulated using the normalized spectra of CAN as divisor to obtain the ratio and first derivative spectra in two successive steps. Firstly, the total amplitude at isosbestic point 255.4 nm of the ratio spectra of the mixture was measured representing the total actual concentration of both drugs in the mixture. Then, the first derivative of the ratio spectra was obtained to determine Hydrochlorothiazide concentration at 233 nm. While the concentration of Candesartan was determined subsequently by subtracting the Hydrochlorothiazide concentration calculated after derivatization from the total concentration of both drugs obtained at the ratio spectra before the derivatization step. Results: The SIDD was successfully applied for simultaneous determination of both drugs in their pure form or in their binary mixture either in synthetic prepared mixtures or in combined dosage form the adopted method was validated according to the ICH guidelines and the results were found to be within the acceptable limits. Conclusion: The adopted method highlighted the important role of normalized spectrum when used as a divisor in addition to the importance of isosbestic point to resolve severely overlapped spectra. All the measurements were carried using ethanol which is considered one of the greenest solvents making the method an environmentally friendly one. the adopted method was validated according to the ICH guidelines and the results were found to be within the acceptable limits.

Application of analytical quality by design approach in method development for the simultaneous estimation of levocetrizine hydrochloride and montelukast sodium in their combined dosage form

Application of analytical quality by design approach in method development for the simultaneous estimation of levocetrizine hydrochloride and montelukast sodium in their combined dosage form

Advanced Derivative Spectroscopic Method for Estimation of Montelukast and Bilastine in Their Tablet Dosage Form

The present work aims to develop and validate the advanced derivative spectroscopic method for estimation of Montelukast (MON) and Bilastine (BIL) in their tablet dosage form. This tablet dosage form is used for anti-asthmatic and allergic rhinitis. The developed method can be applied for simultaneous estimation of Montelukast and Bilastine in their combined dosage form. For this advanced derivative method, the absorbance at 226.8 nm (ZCP of Montelukast) and 326.4nm (ZCP of Bilastine) was used to estimate Montelukast and Bilastine, respectively. The developed method is validated by ICH Q2R1 guidelines with validation parameters like linearity, LOD, LOQ, accuracy, precision, robustness, ruggedness and assay were performed using this guideline. The method was found to be linear in the concentration range of 2-14 µg/mL for Montelukast (R2 = 0.999) and 4–28 µg/mL for Bilastine (R2 =0.9997). LOD and LOQ found 0.1216 and 0.3686 for MON, and 0.3406 and 1.0320 for BIL. The precision study wasa carried out by comparing on 3 different concentrations and the result of their %RSD was is less than 2%. Robustness study carried out by the change in scanning speed and change in methanol manufacturer and Ruggedness study carried out by different analyst. Assay study was performed using tablet formulation. The developed method was utilized for simultaneous estimation of Montelukast and Bilastine for its tablet dosage form.

Development and Validation for Simultaneous Estimation of Sofosbuvir and Daclatasvir Dihydrochloride in Pharmaceutical Dosage form by Ratio Derivative and Dual Wavelength Methods

Development and validation of new simple, sensitive, accurate and precise spectrophotometric method involving ratio derivative and dual-wavelength method, was done as per ICH Q2 (R1) for simultaneous estimation of Sofosbuvir (SOFO) and daclatasvir dihydrochloride (DACLA) in a combined dosage form. The overlapping in the spectra of both drugs was the reason for the selection of both methods. The absorbance difference (ΔA) value between 235.8 nm and 270.6 nm was selected for the quantitative determination of SOFO, where DACLA gives equal absorbance at the selected wavelength in the dual-wavelength method (Method A). The determination of DACLA is done quantitatively by measuring the difference in absorbance value at 249 nm and 268.6 nm where SOFO gives equal absorbance at a selected wavelength. Ratio spectra method (Method B) was based on dividing the spectra of a mixture with standard spectra of one of the analytes, and the first derivative spectra was recorded with Δλ = 8 nm and scaling factor 1. The amount of SOFO and DACLA was estimated in the binary mixtures by computing the first derivative signal at 247.0 nm and 341.0 nm, respectively. The calibration curve was linear in the concentration range of 10–90 μg/mL for SOFO and 4–20 μg/mL for DACLA for both the methods. The methods were successfully applied for the simultaneous determination of these drugs in combined dosage form with acceptable recoveries.

Analytical Method Development and Validation of First Order Derivative Spectrophotometric Method for Simultaneous Estimation of Telmisartan and Metformin Hydrochloride in their Combined Pharmaceutical Dosage Form

Ultra Violet (UV) Sectrophotomertic method has been developed for simultaneous estimation of telmisartan and metformin hydrochloride in bulk drug in their combined dosage form by first-order derivative. This method developed by using Methanol: Water (50: 50 v/v). Linearity was found near to 1, for telmisartan and Metformin hydrochloride. For Intraday, Interday, Intermediate precision, Robustness, %RSD was found less than 2. % Recovery was found to be between ranges 98-106% for both the drugs. These results indicate that the method is accurate, precise, and simple. All validation Parameter results comply with ICH guidelines.

METHODS OF METOPROLOL ANALYSIS IN DRUGS AND BIOLOGICAL FLUIDS: REVIEW AND SUGGESTIONS

Background. Analytical method is increasingly implemented into fundamental pharmaceutical chemistry and analysis, considering their high sensitivity, accuracy, specificity and expressiveness.
 Objective. Metoprolol’s analytical method development was the research goal.
 Methods. The sources were world recognized journals (1990-2019) and key words used as filter were “metoprolol”, “spectrophotometry” “high-performance liquid chromatography, HPLC”, “quantitative analysis”, “validation”.
 Results. Chromatographic methods of analysis have the highest specificity and objectivity and allow qualitative and quantitative determination of Active Pharmaceutic Ingredient (API) in combined dosage forms and biological fluids without prior components separation. The main disadvantage of the described API analysis methods is long terms from the beginning of chromatography to API release and specific solvents used as the mobile phase in HPLC. New methods development and selection such chromatographic conditions that provide high speed and high efficiency at lower pressure of the system are essential. Also, the reduction of analysis time is achieved by simplifying the conditions for sample preparation.
 Conclusions. Analysts are constantly working on developing new analysis methods and their optimization in order to save time and consumables, which also ensures the efficiency of the developed method. There is no monograph on the substance or dosage forms of metoprolol in SPhU. Therefore, some of the developed methods should be suggested for the SPhU monograph, which is important for ensuring pharmacopoeial quality control of medicines in Ukraine.

Synchronized stability indicating RP-LC methods for determination of metolazone with losartan potassium or spironolactone in presence of their degradation products

Two precise and selective stability-indicating RP-LC methods have been developed and validated for simultaneous determination of metolazone in its binary mixture with losartan potassium (method 1) and spironolactone (method 2) in the presence of their degradation products. For method 1, the chromatographic separation was achieved on Kromasil C18 column, the mobile phase consisted of a mixture of 0.1% ortho-phosphoric acid in acetonitrile and 0.1% ortho-phosphoric acid in water (28:72, v/v) pumped at flow rate 2 mL/min and UV detection at 235 nm. Linearity was determined over the concentration range of 2–16µg/mL for metolazone and 40–320µg/mL for losartan potassium. For method 2, chromatographic separation of metolazone and spironolactone was achieved on a Symmetry C8 column using a mobile phase that consisted of acetonitrile, methanol, and 0.1% ortho-phosphoric in water in gradient mode pumped at a flow rate 1.5 mL/min with programed wavelength detection. Linearity was determined over the concentration range of 2–16µg/mL for metolazone and 20–160µg/mL for spironolactone. The suggested methods were proved to be highly selective, precise and accurate for simultaneous determination of the cited drugs in their combined pharmaceutical dosage form in the presence of their degradation products. The proposed methods were validated in compliance with ICH guidelines.HighlightsSynchronized determination of metolazone and co-formulated drugs in presence of their degradation products.Act as a method for screening of metolazone and co-formulated drugs in quality control laboratories.Validation of suggested methods according to ICH guidelines.The pathway of degradation of metolazone under different stress conditions was proposed.

A Sensitive HPTLC Method for the Estimation of Glibenclamide, Rosiglitazone Maleate and Metformin Hydrochloride from a Multicomponent Dosage Form.

A sensitive, rapid and cost-effective method based on HPTLC with UV detection was developed for the quantitation of Glibenclamide (GLIBEN), Rosiglitazone maleate (ROSI) and Metformin hydrochloride (MET) from a combined dosage form. Pre-coated RP-18 F254s aluminum sheets were used as the stationary phase. Methanol-tetrahydrofuran-water-glacial acetic acid (16: 3.6: 4: 0.4, v/v) used as the mobile phase, along with chamber saturation of 10min offered an optimum migration (Rf=0.54, 0.62 and 0.80 for GLIBEN, ROSI and MET, respectively). TLC Scanner 3 was used for densitometric evaluation of the chromatograms. DigiStore 2 Documentation System with winCATS software version 1.4.10 was used for the quantitation and photodocumentation. The LOD for GLIBEN, ROSI and MET was found to be 80 ng, 80 ng and 48ng, respectively. Moreover, the LOQ was 200 ng, 200 ng and 120ng for GLIBEN, ROSI and MET, respectively. The method was linear for GLIBEN (r=0.9991), ROSI (r=0.9993) and MET (r=0.9988) within the tested range (200-1000, 200-1000 and 120-600ng/band, respectively). The method was found to be precise and accurate for all the three drugs. The method was applied for the analysis of Triglucored tablets, and it proved to be a reliable quality control tool for the routine analysis of GLIBEN, ROSI and MET in a combined dosage form.

Assessments of Valsartan in the presence of Nebivolol or Amlodipine in solid formulations and its discriminative dissolution behavior via RP‑HPLC and RP‑UPLC methods.

The current work provides method development and validation of a rapid, robust, precise and accurate RP-HPLC and RP-UPLC for the assessment of Valsartan (VAL) in the presence of Nebivolol (NEB) or Amlodipine (AML) in their combined solid dosage form, following the ICH instructions. In RP-HPLC, Separation was achieved on an Inertsil column, (C18, 150 mm x 4.6 mm, 5μm) with (30:20:50 v/v%), mobile phase of (buffer pH 3.0: acetonitrile: methanol) in an isocratic mode with a flow rate 1.0 mL/min and using photodiode array (PDA) detection at 281 nm. However, in RP-UPLC, Mobile phase was phosphate buffer pH 3.0 and acetonitrile in a ratio of 55:45 v/v% at a flow rate 0.3 mL/min on an Eclipse Plus column, (C18, 1.8 µm, 50 mm × 2.1 mm), while detection at 240 nm on a PDA detector. Calibration curves were linear over the concentration ranges of (0.80-72.00,0.05-4.50) and (4.00-144.00, 0.125-4.50)µg/mL, for the assay and in-vitro drug release profile of (VAL, NEB) and (VAL, AML), respectively. In-Vitro dissolution studies revealed that not less than (NLT) 85% of the labeled amount of VAL and AML was released within 15 min and NLT 85% of the labeled amount of NEB was dissolved within 20 min from their fixed combination solid dosage forms.

Comparative study of HPLC-DAD and HPTLC for the simultaneous determination of a new multitarget antidiabetic ternary mixture in combined tablets

Qternmet XR® (FDA approval, May 2019) is a multitarget anti-diabetic drug combination composed of metformin (MET), saxagliptin (SAX) and dapagliflozin (DAP). To our present knowledge, no analytical reports were found in the scientific databases for the simultaneous quantification of MET, SAX and DAP in their ternary combined tablets, moreover, no articles have attempted the simultaneous estimation of the cited drugs in any matrix using high-performance liquid chromatography with diode-array detection (HPLC–DAD) or high-performance thin-layer chromatography (HPTLC) technique. The current work represents a comparative study on two developed and validated chromatographic methods for the simultaneous determination of the ternary mixture (MET, SAX and DAP) in pure form and in combined tablet dosage form. The first method is reversed-phase HPLC using Agilent C18 column (4.6 × 250 mm, 5 μm p.s.) with a mobile phase consisting of acetonitrile and acidic aqueous phase pH 3 with a photodiode array detection at 230 nm. The second method is HPTLC in which drug solutions were applied to Merck HPTLC silica gel plates developed with a mixture of chloroform:methanol:water:acetic acid (7.4:2.6:0.5:0.01, v/v) and scanned at 224 nm. Both methods were fully validated following the ICH guidelines in terms of linearity, accuracy, precision, selectivity and robustness.

Analytical quality risk management and DoE based development of robust chromatographic method for simultaneous estimation of tizanidine hydrochloride and nimesulide in their combined pharmaceutical dosage forms

Analytical quality risk management (ICH Q9 guideline) concept based robust high performance thin layer chromatographic method has been developed with help of design of experiment (DoE) tool for simultaneous estimation of tizanidine hydrochloride and nimesulide in their combined pharmaceutical dosage forms. Risk identification and assessment were done with brainstorming process with help of ishikawa diagram and experimental results based risk factor priority number (RPN). Critical risk factors which having RPN number more than sixty were further analysed for their criticality in method development by DoE based Taguchi screening design. From seven critical risk factors, volume of methanol in mobile phase and migration distance were identified as highly risky factors for development of HPTLC method. DoE based central composite design was applied for risk factors analysis and mitigation by optimisation of identified high risk factors. After implementation of risk minimisation operable design region and control strategy were set for development of robust HPTLC method for simultaneous estimation of tizanidine hydrochloride and nimesulide. Developed analytical method was validated for specificity, linearity, accuracy, precision and LOD-LOQ as per ICH guideline Q2R1. Validated HPTLC method was applied for assay of combined marketed pharmaceutical dosage forms of both drugs and results were found in good agreement with labelled claim of dosage forms.

Novel stability indicating RP-HPLC method for the simultaneous estimation of tobramycin and loteprednol in pharmaceutical dosage forms

A simple, accurate, rapid and precise isocratic stability indicating reversed-phase high-performance liquid chromatographic method has been developed and validated for simultaneous determination of Tobramycin and Loteprednol in combined tablet dosage form. The chromatographic separation was carried out on Zodiac C18 (150 x 4.6mm, 5µ) with a mixture of Phosphate buffer: acetonitrile (60:40%v/v) as a mobile phase at a flow rate of 1.0 mL/min. UV detection was performed at 243nm. The retention times were 2.442 and 3.269 min for Tobramycin and Loteprednol respectively. Calibration plots were linear (r2=0.999) over the concentration range of 3.75-22.5 µg/mL for Tobramycin 6.25-37.5µg/mL for Loteprednol. The method was validated for accuracy, precision, specificity, linearity, robustness, LOD and LOQ. The proposed method was successfully used for quantitative analysis of tablets. No interference from any component of pharmaceutical dosage form was observed. Validation studies revealed that developed method is specific, rapid, reliable, and reproducible. The high recovery and low relative standard deviation confirm the suitability of the method for routine determination of Tobramycin and Loteprednol in bulk and tablet dosage form.

Development and Validation of a Novel Stability-Indicating RP-HPLC Method for Simultaneous Determination of Tezacaftor and Ivacaftor in Fixed Dose Combination.

A simple and precise novel stability-indicating method for the simultaneous estimation of tezacaftor and ivacaftor in combined tablet dosage form was developed and validated using reversed-phase high-performance liquid chromatography (RP-HPLC). The method is being reported for the first time and includes an estimation of degradation products produced post-stress conditions without any extraction or derivatization. The chromatographic separation of the drugs was achieved with a Symmetry Shield RP18 Column (100Å, 5μm, 4.6mm × 250mm) using a mixture of buffer, methanol and acetonitrile (42:27:31v/v/v) as mobile phase. The buffer used in mobile phase contained 35mM potassium dihydrogen phosphate, and its pH was adjusted to 7.0±0.02 with 20% orthophosphoric acid. The instrument was set at flow rate of 1.2 mL min-1 at ambient temperature and the wavelength of UV-visible detector at 275nm. The developed method could be suitable for the quantitative determination of these drugs in pharmaceutical preparations and also for quality control in bulk manufacturing. Stress testing was performed to prove the specificity. No interference was observed from its stress degradation products. The statistical analysis was done by using F-test and t-test at 95% confidence level.

Method Development and Validation for the Simultaneous Estimation of Clonazepam and Paroxetine in Combined Dosage Form using FT-IR

An analytical methodology was developed for the simultaneous estimation of Clonazepam and Paroxetine in combined dosage form. The method involves the use of N-naphthyl ethylene diamine (NED) as reagent. In this method the absorbance was measured at 535nm for Clonazepam and 398nm for Paroxetine. Linearity was observed in the range of 1–9μg/ml for Clonazepam 25–225μg/ml. The regression equation for the calibration data was y=0.001x + 0.653 with correlation coefficient of 0.99 for Clonazepam and y=0.047x + 0.130 with correlation coefficient of 0.981for Paroxetine. The limits of detection were 0.2μg/ml for Clonazepam and 2μg/ml for Paroxetine and the limit of quantitation were 0.6μg/ml and 16μg/ml for Clonazepam and Paroxetine respectively. The precision of the method was good. The values of the relative standard deviation did not exceed 2%. The proposed method was successfully applied for the simultaneous estimation of Clonazepam and Paroxetine in its combined tablet dosage form with good accuracy and precision. All the analytical validation parameters were determined and found in the limit as per ICH guidelines, which indicates the validity of the method. The developed method was found to be precise, accurate, rugged and robust for the simultaneous estimation of Clonazepam and Paroxetine in its combined dosage forms.

A Validated RP-HPLC Method for the Simultaneous Estimation of Atorvastatin calcium and Clopidogrel Bisulphate in Combined Dosage Forms

The present work describes an isocratic, simple, accurate and precise RP-HPLC method for the simultaneous estimation of Atorvastatin calcium and Clopidogrel Bisulphate in combined capsule dosage forms. Waters HPLC System with 515-HPLC Pump, Spherisorb 5μ silica (4 × 250mm i.d.) column and PDA-2998 detector was used for the analysis. The mobile phase consisting of Acetonitrile and 0.1% ortho-phosphoric acid in the ratio of 65: 35% v/v was used for the separation of drugs. The mobile phase flow rate was 1ml/min and the eluents were detected at 245nm. Using the optimized chromatographic conditions, resolved sharp peaks corresponding to Atorvastatin Calcium and Clopidogrel Bisulphate could be obtained at retention time of 2.007 and 5.977min respectively. The method was validated in terms of linearity, precision and accuracy.

METHOD DEVELOPMENT FOR SIMULTANEOUS ESTIMATION OF ATORVASTATIN AND NATEGLINIDE IN COMBINED DOSAGE FORM BY UV SPECTROSCOPY.

Keywords: Simultaneous estimation, combined dosage forms, Atorvastatin, Nateglinide. In the present work, the quantitative estimation of both the drugs in combined dosage forms was carried out. A new, simple, reliable, sensitive, rapid, and economical procedure for simultaneous estimation of atorvastatin calcium and nateglinide in a combined dosage form by UV spectroscopy using the simultaneous equation method was developed. Native ultraviolet absorbance maxima of the two chemotherapeutic agents were used. As both compounds do not interact chemically in methanol, the two wavelengths 246.15 nm for atorvastatin calcium and 206.6 nm for nateglinide were used. Both the drugs obeyed Beer's law in the concentration range (1-10 µg/ml) that was employed in this method. The method developed was validated to determine its linearity (r2=0.996 for atorvastatin and r2=0.997 for nateglinide), precision, reproducibility, and sensitivity. .