ANALYSIS OF APPROACHES TO THE DEVELOPMENT AND VALIDATION OF THE METHODS OF ANALYSIS OF SOME ACTIVE PHARMACEUTICAL INGREDIENTS FROM THE GROUP OF ANGIOTENSIN CONVERTING ENZYME INHIBITORS IN DRUGS AND BIOLOGICAL LIQUIDS

The quantity of medication brought into the marketplace is growing each year. Analytical method development is increasingly being introduced into fundamental pharmaceutical research and pharmaceutical analysis practice, taking into account their high sensitivity, accuracy, specificity and expressiveness. Search criteria was analytical method development for medicines from group of ACE inhibitors. Literature survey has been done in range of years 1990-2018 to make the review updated and comprehensive and to show the new approacheches to the development of the methods of analysis ACE inhibitors. The sources were world recognized journals and key words used as filter were angiotensin-converting enzyme inhibitors, captopril, enalapril, method development, spectrophotometry, HPLC, UHPLC. The current review is created with an intended to focus on the advantage of HPLC. Literature survey revealed that a number of methods have been reported for estimation of ACE inhibitors individually or in combination with other drugs. However, there is very few analytical methods reported for the simultaneous analysis of these drugs in a combined dosage formulation by HPLC. In additional, analysis of approaches to the development of the methods of analysis of ACE inhibitors in drugs and biological liquids has been shown that HPLC is the most suitable method for analyses ACE inhibitors in substances, drugs, biological liquids to performe routine analysis of medicines, pharmacokinetic (bioequivalence in vivo), dissolution test for final dosages forms (bioequivalence in vitro, biowaiver procedure)

A HPLC-MS/MS METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS DETERMINATION OF NIFEDIPINE AND ENALAPRIL IN HUMAN PLASMA

Objective: The main purpose of this study was to develop a simple, precise, rapid and accurate method for the simultaneous quantification of nifedipine and enalapril in human plasma.Methods: Chromatography was achieved on Discovery C18, 50 × 2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A (acetonitrile-water–formic acid, 5: 95: 0.1 v/v), eluent B (acetonitrile–formic acid, 100: 0.1 v/v)). The initial content of the eluent B is 0%, which increases linearly by 1.0 min to 100% and to 1.01 min returns to the initial 0%. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 5 μl.Results: The total chromatographic run time was 2.5 min and the elution of nifedipine, enalapril and IS (verapamil) occurred at ~1.83, 1.57 and 1.61 min, respectively. A linear response function was established at 1-100 ng/ml for nifedipine and 2-200 ng/ml for enalapril maleate in human plasma. The % mean recovery for enalapril in LQC, MQC and HQC was 114.0 %, 112.9 % and 113.2 %, for nifedipine in LQC, MQC and HQC was 104.1 %, 105.0 % and 108.7 % respectively. The lowest concentration with the RSD<20% was taken as LLOQ and was found to be 2.16 ng/ml for enalapril, 1.01 ng/ml for nifedipine. The % accuracy of LLOQ samples prepared with the different biological matrix lots was found 108.2 % for enalapril and 100.5 % for nifedipine, which were found within the range of 80.00-120.00 % for the seven different plasma lots. % CV for LLOQ samples was observed as 3.2 % and 7.4 % respectively, which are within 20.00% of the acceptance criteria.Conclusion: A rapid method was developed for simultaneous determination of nifedipine and enalapril in human plasma. The method was strictly validated according to the ICH guidelines. Acquired results demonstrate that proposed strategy can be effortlessly and advantageously applied for routine examination of nifedipine and enalapril in human plasma.Â

EFFICIENT VALIDATED METHOD OF HPLC TO DETERMINE ENALAPRIL IN COMBINATED DOSAGE FORM CONTAINING ENALAPRIL AND BISOPROLOL AND IN VITRO DISSOLUTION STUDIES

Objective: A rapid and reproducible HPLC method has been developed for the determination of enalapril in experimental combined dosage forms containing bisoprolol and enalapril and for drug dissolution studies. Methods: The separation was done using a column Hi Qsil C18 (4.6х250 mm, 5 μm particle size) and a mobile phase of methanol: phosphate buffer solution (65:35, v/v), flow-rate of 1.0 ml/min. The injection volume was 300 μL and the ultraviolet detector was set at 225 nm. Results: The method was validated as per ICH guidelines. Under these conditions, enalapril was eluted at 5.33 min. Total run time was shorter than 6 min. A linear relationship between the concentration and the area of ​​chromatographic peaks of enalapril in the range of 1.250 mg/ml–10.000 mg/ml (7.500 mg/ml at pH 1.2) was established. Requirements for linear dependency parameters are performed in this case throughout the range of application of the technique. Linearity studies were conducted in a wide range of concentrations (25-200% at pH 4.5 and 6.8, 25-150% at pH 1.2). In the medium with pH 1.2 release of enalapril from tablets in 5 min is 83.38 %, after 15 min-94.11%, after 30 min-97.17 %; in medium with pH 4.5 the release of enalapril from tablets in 5 min makes 47.13 %, after 15 min-88.34 %, after 30 min-95.86 %; in a medium with pH 6.8, the release of enalapril from tablets in 5 min is 71.04 %, and after 15 min-88.88 %, after 30 min-92.11 %. Conclusion: A simple and sensitive HPLC method was developed for the estimation of enalapril in tablets containing enalapril and bisoprolol. The proposed method was applied successfully for quality control assay of enalapril in experimental tablets and in vitro dissolution studies

ANALYSIS OF APPROACHES TO THE DEVELOPMENT AND VALIDATION OF THE METHODS OF ANALYSIS OF SOME ACTIVE PHARMACEUTICAL INGREDIENTS FROM THE GROUP OF CALCIUM CHANNEL BLOCKERS IN DRUGS AND BIOLOGICAL LIQUIDS

Calcium channel blockers prevent calcium from entering cells of the heart and blood vessel walls, resulting in lower blood pressure. Calcium channel blockers, also called calcium antagonists, relax and widen blood vessels by affecting the muscle cells in the arterial walls. Physico-chemical analysis methods are increasingly being introduced into fundamental pharmaceutical research and pharmaceutical analysis practice, taking into account their high sensitivity, accuracy, specificity and expressiveness. Analytical method development is increasingly being introduced into fundamental pharmaceutical research and pharmaceutical analysis practice, taking into account their high sensitivity, accuracy, specificity and expressiveness. Search criteria was analytical method development for medicines from group of calcium channel blockers. Literature survey has been done in range of years 1990-2018 to make the review updated and comprehensive and to show the new approacheches to the development of the methods of analysis of calcium channel blockers. The sources were  world recognized journals and key words used as filter were calcium channel blockers, amlodipine, nifedipine, verapamil, validation, method development, spectrophotometry, HPLC,  UHPLC. Chromatographic methods of analysis amongst others have the greatest specificity and objectivity and allow for qualitative and quantitative determination of API in combinated dosage forms and biological fluids without prior separation of the components. We can conclude that analysts are constantly working on developing new methods of analysis and their optimization in order to save time and consumables, which also ensures the efficiency of the developed method. The main disadvantage of the described methods of API analysis can be considered long term from the beginning of chromatography to API release and specific solvents used as the mobile phase in HPLC. It is necessary to develop methods and to select such chromatographic conditions that will provide high speed and high efficiency at lower pressure of the system. This reduces the amount of used mobile phase, which reduces cost analysis accordingly, while at the same time providing the necessary specificity, accuracy and reproducibility of the results of the analysis during quality control. Also, the reduction of analysis time is achieved by simplifying the conditions for sample preparation

A HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY–MASS SPECTROMETRY/MASS SPECTROMETRY METHOD DEVELOPMENT FOR THE QUANTITATIVE DETERMINATION OF BISOPROLOL FROM CACO-2 CELL MONOLAYERS

 Objective: A simple, rapid high-performance liquid chromatography–mass spectrometry (MS)/MS method was developed for the determination of bisoprolol from confluent Caco-2 monolayers and from aqueous solution.Methods: Chromatography was achieved on discovery C 18, 50 mm×2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A [acetonitrile:water:formic acid, 5:95:0.1 v/v] and eluent B [acetonitrile:formic acid, 100:0.1 v/v]). The initial content of the eluent B is 0%, which increases linearly by 1.0 min to 100% and to 1.01 min returns to the initial 0%. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 5 μl.Results: Under these conditions, bisoprolol was eluted at 1.49 min. According to the Caco-2 test results, bisoprolol appeared to have moderate to high permeability. It should be noted that the recovery value for bisoprolol is 97.69%. Caco-2 permeability values for bisoprolol are in agreement with BCS Class I classification for these drugs and their known high bioavailability in humans.Conclusion: From results of analysis, it can be concluded that developed method is simple and rapid for the determination of bisoprolol from confluent Caco-2 monolayers and from aqueous solution. Acquired results demonstrate that proposed strategy can be effortlessly and advantageously applied for the examination of bisoprolol from Caco-2 cell monolayers

DEVELOPMENT OF METHODS FOR IDENTIFICATION OF DESLORATADINE IN MEDICINES

SUMMARY. Methods of identification of desloratadine in medicines by absorpt i on spectrophotometry and TLC have beendeveloped.KEY WORDS: desloratadine, identification, spectrophotometery, thin layer chromatography, validation

COMPUTER PROGNOSIS OF BIOLOGICAL ACTIVITY FOR A NUMBER OF NEW 7-R-8-SUBSTITUTED-1,3-DIMETHYL-XANTHINE

AbstractCreation of new potential active pharmaceutical ingredients of synthetic origin is an urgent problem of modern medical chemistry. With this purpose was obtained a number of original 7,8-disubstituted theophylline, and some molecular and pharmacological descriptors are calculated using public Web-resource Chemicalize.org. There was shown the influence of respective substituents in 7 and 8 positions of molecules of synthesized substances on druglike and was confirmed the prospects of chosen area of study. Keywords: virtual analysis, 7,8-disubstituted 1,3-dimethylxanthine, druglike, Ghosh, Maggie and Weber filters.Â

DEVELOPMENT OF THE METHODOLOGY OF THE CHROMATOGRAPHIC DETERMINATION OF NIFEDIPINE IN MEDICINES

ABSTRACTObjective: The aim was to develop a simple, rapid, less expensive, linear, precise, and accurate reverse phase high performance liquid chromatographymethod for determination of nifedipine in tablets.Methods: The chromatographic analysis of nifedipine was performed using liquid chromatograph Agilent 1290 Infinity II LC System. Selectedconditions were isocratic elution with binary mobile phase consisting of solution methanol and 0.1% trifluoroacetic acid (55:45). Detection wascarried out using spectrophotometric detector at 265 nm. The method was validated as per ICH guidelines.Results: The retention time for nifedipine by proposed high performance liquid chromatography (HPLC) method is observed as 3.5 minutes. Thecorrelation coefficients are 1.0000. The developed chromatographic method was found to be accurate with recovery 99.2-99.8% and was foundwithin the acceptance criteria (i.e. 98.0-102.0%) with acceptable % relative standard deviation of not more than 2% at each level. The assay results ofnifedipine in tablets by developed method are highly reproducible, reliable and are in good agreement with the label claim of the medicines (average99.62 %).Conclusion: Finally, it should be noted that a new simple, rapid, linear, precise, accurate HPLC method was developed and validated for thedetermination of nifedipine in medicines in accordance with the ICH guidelines. These results show the method are accurate, precise, sensitive,economic, and rugged. The proposed HPLC method is rapider (retention time is 3.5 minutes). This method can be useful for the routine analysis ofnifedipine in pharmaceutical dosage form.Keywords: Nifedipine, High-performance liquid chromatography, Validation, Linearity, Accuracy, Range of application

DEVELOPMENT AND VALIDATION OF LC-MS/MS METHOD FOR ESTIMATION OF UROCARB IN HUMAN PLASMA

Objective: The present study was aimed to develop a rapid, specific and sensitive method based on LC-MS/MS method was developed for the determination of urocarb using etomidate as an internal standard. Methods: Chromatography was achieved on Discovery C18, 50 × 2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A (acetonitrile-water–formic acid, 5: 95: 0.1 v/v), eluent B (acetonitrile–formic acid, 100: 0.1 v/v)). The initial content of the eluent B of 8%, which increases linearly to 1.0 min to 100%, is maintained up to 1.5 min and returned to the original 8% to 1.51 min. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 4 μl. Results: The total chromatographic run time was 2.0 min and the elution of urocarb and IS (etomidate) occurred at ~1.53 and 1.67 min, respectively. A linear response function was established at 1-100 ng/ml for urocarb and etomidate in human plasma. The % mean recovery for urocarb in LQC, MQC and HQC was 104.1 %, 100.0 % and 97.4 %. The lowest concentration with the RSD<20% was taken as LLOQ and was found to be 1.03 ng/ml for urocarb. The within-run coefficients of variation ranged between 0.271 % and 0.478 % for urocarb. The within-run percentages of nominal concentrations ranged between 99.12 % and 100.21 % for urocarb. The between-run coefficients of variation ranged between 0.388 % and 0.601 % for urocarb. The between-run percentages of nominal concentrations ranged between 98.78 % and 101.11 % for urocarb. Conclusion: A highly sensitive, specific, reproducible, rapid and high-throughput LC-MS/MS assay was developed and validated to quantify urocarb in human plasma as per the regulatory guidelines. Due to the sensitivity of the developed method, it can be applied to the monitoring of plasma levels in the analysis of drug in preclinical and clinical pharmacokinetic studies. All the parameters and results were found within the acceptance limit as given in the validation protocol

A HPLC-MS/MS METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS DETERMINATION OF BISOPROLOL AND ENALAPRIL IN THE PRESENT OF ENALAPRILAT IN HUMAN PLASMA

Objective: A highly specific, sensitive and rapid HPLC-MS/MS method has been developed and validated for the simultaneous quantification of bisoprolol and enalapril in the present of enalaprilat in human plasma.Methods: Analytes were extracted from plasma using a protein precipitation extraction method. Chromatography was achieved on Discovery C18, 50 × 2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A (acetonitrile-water–formic acid, 5: 95: 0.1 v/v), eluent B (acetonitrile–formic acid, 100: 0.1 v/v)). The initial content of the eluent B is 0%, which increases linearly by 1.0 min to 100% and to 1.01 min returns to the initial 0%. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 5 μl.Results: The total chromatographic run time was 2.0 min and the elution of bisoprolol, enalapril, enalaprilat and IS (verapamil) occurred at ~1.01, 1.03, 0.96 and 1.09 min, respectively. A linear response function was established at 0.5-50 ng/ml for bisoprolol fumarate, 2-200 ng/ml for enalapril maleate, 1-100 ng/ml for enalaprilat dehydrate in human plasma. The intraday and interday accuracy and precisions were in the range of 0.311 %-0.647 % and 0.364 %-0.572 % for bisoprolol, 0.321 %-0.747 % and 0.390 %-0.673 % for enalapril, 0.221 %-0.547 % and 0.264 %-0.773 % for enalaprilat, respectively.Conclusion: A new rapid method was developed for simultaneous determination of bisoprolol and enalapril in the present of enalaprilat in human plasma. The method was strictly validated according to the ICH guidelines. The information thus obtained from the study can be used for the full pharmacokinetic profiling in individuals.Â