High-performance thin layer chromatography method for estimation of andrographolide in herbal extract and polyherbal formulations

A new, simple, sensitive, precise and robust high-performance thin layer chromatographic (HPTLC) method was developed for the estimation of andrographolide in herbal extracts and pharmaceutical dosage forms. Analysis of andrographolide was performed on TLC aluminium plates pre-coated with silica gel 60F-254 as stationary phase. Linear ascending development was carried out in twin trough glass chamber saturated with chloroform:toluene:methanol (66:26:8, v/v/v) at room temperature (25 ± 2 °C). The R_f value of andrographolide was found to be 0.49. Camag TLC scanner III was used for spectrodensitometric scanning and analysis in absorbance mode at 229 nm. The system was found to give compact spots for andrographolide (R_f value of 0.49). The data for calibration plots showed good linear relationship with r² = 0.9986 in the concentration range of 200 ng to 1000 ng with respect to peak area. The present method was validated by precision, recovery, robustness and reproducibility according to ICH guidelines. The limits of detection and quantification were determined and it was found to be 3.5 and 11.7 ng, respectively. Statistical analysis of the data showed that the method is reproducible and selective for estimation of andrographolide.     

Determination of Exemestane in bulk and pharmaceutical dosage form by HPTLC

An HPTLC method for analysis of Exemestane in bulk and pharmaceutical formulation has been established and validated. The analyte was separated on aluminium plates precoated with silica gel 60 F₂₅₄. The mobile phase was chloroform:methanol 9.2:0.8 (v/v). Quantification was done by densitometric scanning at 247 nm. Response was a linear function of Exemestane concentration in the range of 100–500 μg mL⁻¹. The limit of detection and quantification for Exemestane were 5.8 and 17.58 μg mL⁻¹, respectively. Average recovery of Exemestane was 100.1, which shows that the method was free from interference from excipients present in the formulation. The established method enabled accurate, precise, and rapid analysis of Exemestane in bulk as well as pharmaceutical formulation.     

Chiral stability-indicating HPLC method for analysis of arotinolol in pharmaceutical formulation and human plasma

An enantioselective stability-indicating high performance liquid chromatographic method was developed for the analysis of arotinolol in standard solution. The degradation behaviour of arotinolol was investigated under different stress conditions recommended by International Conference on Harmonization (ICH). Resolution of the drug and complete separation from its degradation products were successfully achieved on a Chirobiotic V column, using UV detector set at 315 nm, polar organic mobile phase (POM) consisting of methanol:glacial acetic acid:triethylamine, 100:0.02:0.03, (v/v/v), and a flow rate of 1 ml/min. The drug was subjected to oxidation, hydrolysis, photolysis, and heat to apply stress conditions. The drug was found to degrade in alkaline, acidic, oxidative conditions and when exposed to heat. The drug was stable to sunlight. The method reported here has also been successfully applied to pharmaceutical formulation and to human plasma that spiked with stock solutions of arotinolol enantiomers.Arotinolol enaniomers were recovered from plasma by using liquid–liquid extraction procedure with ethyl ether. The method was highly specific, where degradation products and coformulated compounds did not interfere, and was sensitive with good precision and accuracy and was linear over the range of 50–400 ng/ml (R² > 0.9981) with a detection limit of 20 ng/ml for each enantiomer. The mean extraction efficiency for arotinolol was in the ranges 96–104% for each enantiomer. The mean relative standard deviation (RSD) of the results of within-day precision and accuracy of the drug were ?7.1%. There was no significant difference between inter- and intra-day studies for each enantiomers which confirmed the reproducibility of the assay. The overall recoveries of arotinolol enantiomers from pharmaceutical formulations were in the ranges 97.6–101.8%.     

RP-HPLC-DAD method for the determination of phenylepherine,paracetamol, caffeine and chlorpheniramine in bulk and marketed formulation

A simple, specific and accurate isocratic RP-HPLC-DAD method was developed for the simultaneous determination of phenylephrine, paracetamol, caffeine and chlorpheniramine in bulk and tablet dosage form. The four contents are present in variable concentrations and have variable chromatographic behavior making the process of analysis very difficult. For present studies a reversed-phase C-18 column (150 mm × 4.5 mm i.d., particle size 5 μm) with mobile phase consisting of acetonitrile, methanol and 10 Mm phosphate buffer 16:22:62 (v/v) (pH of buffer 2.5 ± 0.02, adjusted with ortho phosphoric acid) was used. The flow rate was 1.0 ml/min and eluents were monitored at 280 nm. The mean retention times of phenylephrine, paracetamol, caffeine and chlorpheniramine were found to be 1.8, 3.1, 5.2 and 10.9 min, respectively. The method was validated in terms of linearity, range, specificity, accuracy, precision and robustness. The proposed method was successfully applied to the estimation of phenylephrine, paracetamol, caffeine and chlorpheniramine in combined tablet dosage form.     

Validated ecofriendly chromatographic method for quantitative determination of anti‐migraine quaternary mixture

A novel ecofriendly, cost and time saving high‐performance thin‐layer chromatographic method was developed and validated for simultaneous determination of metoclopramide, ergotamine, caffeine, and paracetamol in bulk and pharmaceutical formulation. The separation was carried out on silica gel plates, using ethyl acetate:ethanol:ammonia (9:1:0.1, v/v/v) as a developing system. Ultraviolet detection was carried out at 272 nm. The resulting retention times were 0.15, 0.36, 0.49, and 0.74 min for metoclopramide, ergotamine, caffeine, and paracetamol, respectively. The greenness profile assessment was achieved to the proposed method to evaluate its greenness characters to the environment with acceptable results. Validation parameters were checked according to International Conference of Harmonization guidelines to achieve the international requirements for quality control analysis of the proposed drugs.     

Stability-indicating liquid chromatography for determination of clopidogrel bisulfate in tablets: Application to content uniformity testing

The present study describes a simple stability-indicating reversed-phase HPLC assay for antiplatelet drug, clopidogrel bisulfate. Separation of the drug and the degradation products, under stress conditions was successfully achieved on a C-18 column utilizing 0.01 M Na₂HPO₄ (pH 4): acetonitrile in the ratio 80:20 v/v, pumped at a flow rate of 0.5 ml min^?1 with UV detection at 235 nm. The retention time of clopidogrel was 6.84 min. The method was satisfactorily validated with respect to linearity, precision, accuracy, selectivity, sensitivity and ruggedness. The response was linear in the range of 0.2–3.5 μg ml^?1 with detection limit 0.079 μg ml^?1. The suggested method was successfully applied for the analysis of clopidogrel in bulk and in commercial tablets. The results were favorably compared to those obtained by a reference method. The proposed method was successfully applied to the content uniformity testing of tablets and for determination of clopidogrel in presence of its co-administered drug, acetyl salicylic acid.     

New,simple and validated kinetics spectrophotometric method for determination of moxifloxacine in its pharmaceutical formulations

The objective of this research was to develop a kinetic spectrophotometric method for determination of moxifloxacine (MOXF) in pure form and pharmaceutical formulations. The method was based on the formation of a colored N-vinyl chlorobenzoquinone derivative of MOXF by its reaction with 2,3,5,6-tetrachloro-1,4-benzoquinone in presence of acetaldehyde.The formation of the colored product was monitored spectrophotometrically by measuring the absorbance at 652 nm. Factors affecting the reaction were studied and optimized. The stoichiometry of the reaction was determined, and the reaction pathway was postulated. The activation energy of the reaction was calculated and found to be 6.65 kJ mol^?1. Under the optimized conditions, the initial rate and fixed time (at 5 min) methods were utilized for constructing the calibration graphs. The graphs were linear in concentration ranges 5–100 and 15–150 μg ml^?1 with limit of detection of 2.0 and 5.0 μg ml^?1 for the initial rate and fixed time methods, respectively. The analytical performance for both methods was fully validated, and the results were satisfactory. No interference was observed from the excipients that are commonly present in the pharmaceutical formulations. The proposed method was successfully applied to the determination of MOXF in its pharmaceutical formulations. The label claim percentages were 101.25 ± 0.73% and 100.92 ± 0.65% for the initial rate and fixed time method, respectively. Statistical comparison of the results with those obtained by a reference spectrophotometric method showed excellent agreement between the accuracy and precision of the two methods. The proposed method has great value in its application to the analysis of MOXF in quality control laboratories.     

Silicon surface texturing by electro-deoxidation of a thin silica layer in molten salt

A new method of silicon surface texturing is reported, which is based on thin silica layer electrochemical reduction in molten salts. A thermal silica layer grown on p-type silicon was potentiostatically reduced in molten calcium chloride at 850 °C. Typical nano–micro-formations obtained at different stages of electrolysis were demonstrated by SEM. X-ray diffraction measurements confirmed conversion of the amorphous thermal silica layer into crystalline silicon. The proposed approach shows promise in photovoltaic applications, for instance, for production of antireflection coatings in silicon solar cells.     

Determination of nicotinamide by stopped-flow injection method in pharmaceutical formulations

A simple stopped-flow injection system with spectrophotometric detection was proposed for the determination of nicotinamide (NAM) in pharmaceutical formulations. In this system cyanogen chloride formed from the combination of an acidic KSCN with the NaClO streams reacts with injected NAM to form glutaconic aldehyde. Then the product of these three components was coupled with another buffered (pH 3.5) stream of barbituric acid and directed towards the detector. A 45 s after sample injection the pump was stopped for 130 s. During this time the reactants in the flow cell were provided with the required temperature (40 °C) by placing the cell in a home made cell jacket to increase the yield of the polymethine dye product. Eventually, the absorbance of the formed pink color dye was monitored spectrophotometrically at 560 nm and NAM in the concentration range of 1.0–25.0 μg/mL (R = 0.9974 and D.L = 0.5 μg/mL) was determined. The results obtained by this method were compared statistically and agree with those obtained by the method described in the British Pharmacopoeia.     

Simultaneous HPLC-UV Determination of Lactic Acid,Glycolic Acid,Glycolide, Lactide and Ethyl Acetate in Monomers for Producing Biodegradable Polymers

A simple, rapid high-performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of glycolic acid, lactic acid, glicolide, lactide and ethyacetate in monomers for obtaining biopolymers. The separation was effected on the reversed-phase C18 column 250mm×4.6 mm with particle size 5 μ using a mobile phase mixture buffer and acetonitrile in a ratio 88:12 v/v and elution was isocratic at a flow-rate of 1.0 mL/min. The determinations were performed with a UV-Vis detector at 200 nm. The volume of the injected sample was 20 μL. Detection limits for acids and its dimers (glycolic acid, lactic acid, glicolide, lactide) and ethylacetate range between 82 and 182 ng/mL. The analytes are separated in 13 min. Recovery studies showed good results for all solutes (99–102%). The method is linear for all compounds over the concentration range tested, and shows good precision and accuracy, making it suitable for quantitation of acids and its dimers (glycolic acid, lactic acid, glicolide, lactide) and ethyl acetate in monomers.     

Stability-indicating HPLC method for the determination of nicardipine in capsules and spiked human plasma. Identification of degradation products using HPLC/MS

In this stability-indicating, reversed-phase high-performance liquid chromatographic method for nicardipine (NIC), forced degradation has been employed and the formed degradants were separated on a C18 (150 mm × 3.9 mm, 5 μm) analytical column using a mobile phase consisted of 70% methanol: acetic acid containing 0.01 M triethylamine with pH 4. The flow rate was 1.0 mL/min and the photodiode array detection wavelength was 353 nm. Forced degradation of the drug was carried out under acidic, basic, photolytic, and oxidative stress conditions. Chromatographic peak purity data indicated no co-eluting peaks with the main peaks. This method resulted in the detection of seven degradation products. Among these, two major degradation products from basic hydrolysis, one from oxidation by H₂O₂ and four from photolytic stress were identified by mass spectral data. A good linear response was achieved over the range of 0.5–40 μg/mL with a limit of detection (LOD) of 0.011 μg/mL and limit of quantification (LOQ) of 0.036 μg/mL. The suggested method was successfully applied for the analysis of NIC in its commercial capsules, with mean% recovery value of 100.11 ± 2.26%. The method was extended to the in vitro determination on NIC in spiked human plasma samples with mean% recovery of 99.04 ± 5.67%. The suggested method was utilized to investigate the kinetics of photolytic induced degradation.     

Kinetic spectrophotometric determination of hyoscine butylbromide in pure form and in pharmaceutical formulations

A simple and sensitive kinetic method was described for the determination of hyoscine butylbromide in pharmaceutical preparations. The method is based upon a kinetic investigation of the oxidation reaction of the drug with alkaline potassium permanganate at room temperature for a fixed time of 15 min. The absorbance of the colored manganate ion was measured at 610 nm. The absorbance–concentration plot was rectilinear over the range of 1.0–10 μg mL^?1 (r = 0.9999) and detection limit of 0.092 μg mL^?1. The concentration of hyoscine butylbromide was calculated using the corresponding calibration equation for the fixed-time method. The determination of hyoscine butylbromide by the fixed-concentration and rate constant methods is also feasible with the calibration equations obtained but the fixed-time method has been found to be more applicable. The different experimental parameters affecting the development and stability of the colors were carefully studied and optimized. The proposed method was applied to the determination of hyoscine butylbromide in pharmaceutical formulations. The results obtained were in good agreement with those obtained using the official British Pharmacopeial method (2004).     

Direct electrochemistry of hemoglobin and glucose oxidase in electrodeposited sol–gel silica thin films on glassy carbon

This work points out that electrogeneration of silica gel (SG) films on glassy carbon electrodes (GCEs) can be applied to immobilize biomolecules – hemoglobin (Hb) or glucose oxidase (GOD) or both of them in mixture – without preventing their activity. These proteins were physically entrapped in the sol–gel material in the course of the electro-assisted deposition process applied to form the thin films onto the electrode surface. SG films were prepared from a precursor solution by applying a suitable cathodic potential likely to induce a local pH increase at the electrode/solution interface, accelerating thereby polycondensation of the silica precursors with concomitant film formation. Successful immobilization of proteins was checked by various physico-chemical techniques. Both Hb and GOD were found to undergo direct electron transfer, as demonstrated by cyclic voltammetry. GCE–SG–Hb gave rise to well-defined peaks at potentials E_c = −0.29 V and E_a = −0.17 V in acetate buffer, corresponding to the Fe^III/Fe^II redox system of heme group of the protein, while GCE–SG–GOD was characterized by the typical signals of FAD group at E_c = −0.41 V and E_a = −0.33 V in phosphate buffer. These two redox processes were also evidenced on a single voltammogram when both Hb and GOD were present together in the same SG film. Hb entrapped in the silica thin film displayed an electrocatalytic behavior towards O₂ and H₂O₂ in solution, respectively in the mM and μM concentration ranges. Immobilized GOD kept its biocatalytic properties towards glucose. Combined use of these two proteins in mixture has proven to be promising for detection of glucose in solution via the electrochemical monitoring of oxygen consumption (decrease of the oxygen electrocatalytic signal).     

Development of off-line layer chromatographic and total reflection X-ray fluorescence spectrometric methods for arsenic speciation

Rapid and low cost off-line thin layer chromatography–total reflection X-ray fluorescence spectrometry and overpressured thin layer chromatography–total reflection X-ray fluorescence spectrometry methods have been developed for separation of 25 ng of each As(III), As(V), monomethyl arsonic acid and dimethylarsinic acid applying a PEI cellulose stationary phase on plastic sheets and a mixture of acetone/acetic acid/water = 2:1:1 (v/v/v) as eluent system. The type of eluent systems, the amounts (25–1000 ng) of As species applied to PEI cellulose plates, injection volume, development distance, and flow rate (in case of overpressured thin layer chromatography) were taken into consideration for the development of the chromatographic separation. Moreover, a microdigestion method employing nitric acid for the As spots containing PEI cellulose scratched from the developed plates divided into segments was developed for the subsequent total reflection X-ray fluorescence spectrometry analysis. The method was applied for analysis of root extracts of cucumber plants grown in As(III) containing modified Hoagland nutrient solution. Both As(III) and As(V) were detected by applying the proposed thin layer chromatography/overpressured thin layer chromatography–total reflection X-ray fluorescence spectrometry methods.     

A study on solution deposited CuSCN thin films: Structural,electrochemical, optical properties

A cost-effective successive ionic layer adsorption and reaction (SILAR) method was used to deposit copper (I) thiocyanate (CuSCN) thin films on glass and steel substrates for this study. The deposited thin films were characterized for their structural, morphological, optical and electrochemical properties using X-ray diffraction (XRD), scanning electron microscopy (SEM), UV–visible spectroscopy and VersaSTAT potentiostat. A direct band gap of 3.88 eV and 3.6 eV with film thickness of 0.7 μm and 0.9 μm was obtained at 20 and 30 deposition cycles respectively. The band gap, microstrain, dislocation density and crystal size were observed to be thickness dependent. The specific capacitance of the CuSCN thin film electrode at 20 mV/s was 760 F g⁻¹ for deposition 20 cycles and 729 F g⁻¹ for deposition 30 cycles.     

Sorption of iron(II) and ruthenium(III)-triazine complexes on silica gel and its analytical applicability

2,4,6-Tri(2′-pyridyl)-s-triazine (TPTZ) complexes with iron(II) and ruthenium(III) were prepared. Their sorption and desorption features on silica gel have been investigated. Both complexes were strongly adsorbed. This has been utilized for separating and preconcentrating iron(II) and ruthenium(III) using TPTZ-impregnated silica gel. The chromatographic behavior of TPTZ on silica gel column was examined and found to be effective modifier for silica gel surface. The sorption capacity of silica gel for those metal-triazine complexes has been determined under static conditions and was found to be 5.28 × 10^–3 mM (Fe(TPTZ)₂²⁺) and 2.9 × 10^–3 mM (Ru(TPTZ)₂³⁺). Saturated methanolic solutions of KI or 25% NaClO₄ solutions desorbed both complexes quantitatively from the silica gel surface.     

Flow injection analysis system with electrochemical detection for the simultaneous determination of nanomolar levels of acetaminophen and codeine

A simple, rapid and low-cost electroanalytical method is proposed for the determination of acetaminophen (ACP) and codeine (COD) at nanomolar levels in pharmaceutical and biological samples. The analytical procedure is based on a flow injection analysis system coupled to electrochemical detection, which was multiple pulse amperometry (FIA-MPA). Boron-doped diamond was used as the working electrode for electrochemical detection. The electrode was subjected to a cathodic pretreatment and was selected in this work due its good electrochemical performance. By applying the FIA-MPA method, after a number of optimization assays, the analgesics were simultaneously determined at excellent linear concentration ranges. The analytical curves ranged from 80 nmol L⁻¹ to 100 µmol L⁻¹ for ACP and from 50 nmol L⁻¹ to 10 µmol L⁻¹ for COD, and the obtained limits of detection were 30 nmol L⁻¹ and 35 nmol L⁻¹ for ACP and COD, respectively. The practical applicability of the electroanalytical method was evaluated from the ACP and COD determination in two sample matrices: commercial pharmaceutical samples and biological fluids. In the case of pharmaceutical formulation samples, the obtained results were statistically similar to those obtained using a reference chromatographic method. In addition, these drugs were simultaneously quantified in biological fluid samples of urine and human serum with excellent recovery percentages.     

Development and validation of a UPLC method for quantification of antiviral agent,Acyclovir in lipid-based formulations

PurposeThe objective of the current study is to evaluate the Ultra Performance Liquid Chromatography (UPLC) method for quantification of Acyclovir in lipid-based formulations.MethodA simple, rapid, reliable and precise reversed phase UPLC method has been developed and validated according to the regulatory guidelines, which composed of isocratic mobile phase; 0.25% formic acid (FA) in Milli-Q water with a flow rate of 0.5 ml/min, and column BEH C18 (2.1 × 50 mm, 1.7 μm). The detection was carried out at 254 nm.ResultsThe developed UPLC method was found to be rapid (1.2 min run time), selective with well resoluted Acyclovir peak (0.89 min) from different lipid matrices and sensitive (Limit of Detection (LOD) was 0.3 ppm and Lower Limit of Quantification (LLOQ) was 1 ppm). The accuracy and precision were determined and were perfectly matching with the standard FDA limits.ConclusionThe study showed that the proposed UPLC method can be used for the assessment of drug purity, stability, solubility and lipid-formulation release profile with no interference of excipients or related substances of active pharmaceutical ingredient.     

Lithium storage in perovskite lithium lanthanum titanate

Perovskite lithium lanthanum titanate (LLTO) was synthesized using sol–gel method. It shows a reversible capacity of 145 mA h g^− 1 and moderate cycling performance between 0.01 and 2.00 V. Cyclic voltammetry and X-ray diffraction results demonstrate a two-step solid–solution reaction behavior in the voltage range of 0.00–3.00 V upon lithium insertion/extraction. A stable solid electrolyte interphase (SEI) layer is formed on the surface of LLTO after the initial discharge. Carbon coating by chemical vapor deposition improves its cycling performance significantly.     

Determination of phenylephrine hydrochloride and chlorpheniramine maleate in binary mixture using chemometric-assisted spectrophotometric and high-performance liquid chromatographic-UV methods

Four methods have been developed for the simultaneous determination of phenylephrine hydrochloride and chlorpheniramine maleate without previous separation. In the first method both drugs are determined using first derivative UV spectrophotometry, with zero-crossing measurement. The second method depends on first derivative of the ratios spectra. The third method describes the use of multivariate spectrophotometric calibration for the simultaneous determination of the analyzed binary mixture where the resolution is accomplished by using partial least squares (PLS) regression analysis. In the fourth method (HPLC), a reversed-phase column and a mobile phase of methanol:water:acetonitrile (80:12:8 v/v/v/) at 0.9 ml/min flow rate have been used to separate both drugs with a UV detection at 270 nm. All the proposed methods are extensively validated. They have the advantage to be economic and time saving. All the described methods can be readily utilized for analysis of pharmaceutical formulations. The results obtained using the proposed methods are statistically analyzed and compared with some reported methods.