Towards improved solubility of poorly water-soluble drugs: cryogenic co-grinding of piroxicam with carrier polymers.

Amorphous solid dispersions (SDs) open up exciting opportunities in formulating poorly water-soluble active pharmaceutical ingredients (APIs). In the present study, novel catalytic pretreated softwood cellulose (CPSC) and polyvinylpyrrolidone (PVP) were investigated as carrier polymers for preparing and stabilizing cryogenic co-ground SDs of poorly water-soluble piroxicam (PRX). CPSC was isolated from pine wood (Pinus sylvestris). Raman and Fourier transform infrared (FTIR) spectroscopy, X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC) were used for characterizing the solid-state changes and drug-polymer interactions. High-resolution scanning electron microscope (SEM) was used to analyze the particle size and surface morphology of starting materials and final cryogenic co-ground SDs. In addition, the molecular aspects of drug-polymer interactions and stabilization mechanisms are presented. The results showed that the carrier polymer influenced both the degree of amorphization of PRX and stabilization against crystallization. The cryogenic co-ground SDs prepared from PVP showed an enhanced dissolution rate of PRX, while the corresponding SDs prepared from CPSC exhibited a clear sustained release behavior. In conclusion, cryogenic co-grinding provides a versatile method for preparing amorphous SDs of poorly water-soluble APIs. The solid-state stability and dissolution behavior of such co-ground SDs are to a great extent dependent on the carrier polymer used.

Coformer screening using thermal analysis based on binary phase diagrams.

PURPOSE: The advent of cocrystals has demonstrated a growing need for efficient and comprehensive coformer screening in search of better development forms, including salt forms. Here, we investigated a coformer screening system for salts and cocrystals based on binary phase diagrams using thermal analysis and examined the effectiveness of the method. METHODS: Indomethacin and tenoxicam were used as models of active pharmaceutical ingredients (APIs). Physical mixtures of an API and 42 kinds of coformers were analyzed using Differential Scanning Calorimetry (DSC) and X-ray DSC. We also conducted coformer screening using a conventional slurry method and compared these results with those from the thermal analysis method and previous studies. RESULTS: Compared with the slurry method, the thermal analysis method was a high-performance screening system, particularly for APIs with low solubility and/or propensity to form solvates. However, this method faced hurdles for screening coformers combined with an API in the presence of kinetic hindrance for salt or cocrystal formation during heating or if there is degradation near the metastable eutectic temperature. CONCLUSIONS: The thermal analysis and slurry methods are considered complementary to each other for coformer screening. Feasibility of the thermal analysis method in drug discovery practice is ensured given its small scale and high throughput.

Liquid chromatographic method for the simultaneous determination of captopril, piroxicam, and amlodipine in bulk drug, pharmaceutical formulation, and human serum by programming the detector.

A highly sensitive LC method with UV detection has been developed for the simultaneous determination of coadministered drugs captopril, piroxicam, and amlodipine in bulk drug, pharmaceutical formulations, and human serum at the isosbestic point (235 nm) and at individual λmax (220, 255, and 238 nm, respectively) by programming the detector with time to match the individual analyte's chromophore, which enhanced the sensitivity with linear range. The assay involved an isocratic elution of analytes on a Bondapak C18 (10 µm, 25 × 0.46 cm) column at ambient temperature using a mobile phase of methanol/water 80:20 at pH 2.9 and a flow rate of 1.0 mL/min. Linearity was found to be 0.25-25, 0.10-6.0, and 0.20-13.0 µg/mL with correlation coefficient >0.998 and detection limits of 7.39, 3.90, and 9.38 ng/mL, respectively, whereas calibration curves for wavelength-programmed analysis were 0.10-6.0, 0.04-2.56, and 0.10-10.0 µg/mL with correlation coefficient >0.998 and detection limits of 5.79, 2.68, and 3.87 ng/mL, respectively. All the validated parameters were in the acceptable range. The recovery of drugs was 99.32-100.39 and 98.65-101.96% in pharmaceutical formulation and human serum, respectively, at the isosbestic point and at individual λmax . This method is applicable for the analysis of drugs in bulk drug, tablets, serum, and in clinical samples without interference of excipients or endogenous serum components.

Improvement of drug loading onto ion exchange resin by cyclodextrin inclusion complex.

CONTEXT: Ion exchange resins have ability to exchange their counter ions for ionized drug in the surrounding medium, yielding "drug resin complex." Cyclodextrin can be applied for enhancement of drug solubility and stability. OBJECTIVE: Cyclodextrin inclusion complex of poorly water-soluble NSAIDs, i.e. meloxicam and piroxicam, was characterized and its novel application for improving drug loading onto an anionic exchange resin, i.e. Dowex® 1×2, was investigated. METHODS: ß-Cyclodextrin (ß-CD) and hydroxypropyl ß-cyclodextrin (HP-ß-CD) were used for the preparation of inclusion complex with drugs in solution state at various pH. The inclusion complex was characterized by phase solubility, continuous variation, spectroscopic and electrochemistry methods. Then, the drug with and without cyclodextrin were equilibrated with resin at 1:1 and 1:2 weight ratio of drug and resin. RESULTS AND DISCUSSION: Solubility of the drugs was found to increase with increasing cyclodextrin concentration and pH. The increased solubility was explained predominantly due to the formation of inclusion complex at low pH and the increased ionization of drug at high pH. According to characterization studies, the inclusion complex was successfully formed with a 1:1 stoichiometry. The presence of cyclodextrin in the loading solution resulted in the improvement of drug loading onto resin. CONCLUSIONS: Enhancing drug loading onto ion-exchange resin via the formation of cyclodextrin inclusion complex is usable in the development of ion-exchange based drug delivery systems, which will beneficially reduce the use of harmful acidic or basic and organic chemicals.

Investigation on the micelle-sensitized Ce(IV)-lornoxicam-Rh B chemiluminescence system and its application.

Based on the micelle synergism mechanism, a simple and sensitive flow injection chemiluminescence (FI-CL) method for the assay of lornoxicam was described. The CL signal generated from the reaction of Ce (IV) with lornoxicam in acidic solution was very weak, while the interfusion of sodium dodecyl benzene sulfonate (SDBS) resulted in a highly CL intensity. Under the optimum experimental conditions, the CL intensity was proportional to lornoxicam concentration over the range 1.0 × 10(-10)-7.3 × 10(-8) g/mL with a detection limit of 4.9 × 10(-11) g/mL (3σ). The relative standard deviation for 11 replicate measurements of 3.0 × 10(-9) g/mL of lornoxicam was 1.9%. The proposed method was successfully applied for the assay of lornoxicam in pharmaceuticals, human serum and urine with excellent recovery. The possible mechanism of CL reaction was also discussed briefly.

Electrokinetic supercharging focusing in capillary zone electrophoresis of weakly ionizable analytes in environmental and biological samples.

Five non-steroidal anti-inflammatory drugs, naproxen, fenoprofen, ketoprofen, diclofenac and piroxicam, were separated and analyzed by electrokinetic supercharging in CZE. Three different setups of the ITP technique were assayed for the separation and preconcentration of these five non-steroidal anti-inflammatory drugs. For the setup that gave the best results, we evaluated the influence of different parameters on separation and preconcentration efficiency such as sample pH, concentration of the leading stacker, BGE composition, electrokinetic injection time, composition and hydrodynamic injection of the solvent plug and of the terminating stacker. In the selected setup, the BGE (10 mM Na(2)B(4)O(7) + 50 mM NaCl in 10% of MeOH aqueous solution) contained the leading electrolyte while the terminating electrolyte, hydrodynamically injected after the sample (50 mbar x 12 s), was 50 mM of CHES. Prior to sample injection at (700 s at -2 kV) a short plug of MeOH (50 mbar x 3 s) was hydrodynamically injected. The results show that this strategy enhanced detection sensitivity 2000-fold compared with normal hydrodynamic injection, providing detection limits of 0.08 µg/L for standard samples with good repeatability (values of relative standard deviation, %RSD < 1.03%). Method validation with river water samples and human plasma demonstrated good linearity, with detection limits of 0.9 and 2 µg/L for river water samples and human plasma samples, respectively (as well as satisfactory precision in terms of repeatability and reproducibility).

Preparation and evaluation of solid dispersions of piroxicam and Eudragit S100 by spherical crystallization technique.

OBJECTIVE: To improve the dissolution rate of piroxicam (PX), enteric-release microparticles having solid dispersion (SD) structure were prepared in one step. METHODS: SD of PX and Eudragit S100 (Eu S100) with the aid of silicon dioxide (Aerosil 200), as an antiadhesion agent, were prepared by spherical crystallization technique. The microparticles were characterized by differential scanning calorimetry, X-ray powder diffraction, and Fourier transform infrared spectroscopy and were evaluated for yield, encapsulation efficiency, flowability, packability, and drug release (at pH 1.2 and pH 7.4). The samples were stored at severe condition [40 degrees C, 75% relative humidity (RH)] for 3 months to investigate their stability. The effects of the polymer-drug and polymer-Aerosil ratios on the characteristics of the microparticles were also investigated. RESULTS: PX microparticles exhibited significantly improved micromeritic properties in comparison to the crystalline pure drug. The dissolution of drug from microparticles in phosphate buffer (pH 7.4) indicated a significant increase in dissolution of PX when dispersed in Eu S100. The results of X-ray powder diffraction and differential scanning calorimetry analysis indicated that in microparticles at 2:1 Eu S100:PX ratio the crystalline form of PX was disordered, suggesting that PX was highly dispersed in microparticles, as that in the amorphous state. Fourier transform infrared spectroscopy analysis demonstrated the presence of intermolecular hydrogen bonding between PX and Eu S100 in SD. In stability test, the release profiles of the microparticles were unchanged as compared with the freshly prepared SDs; amorphous PX in the SD particles did not crystallize under storing at 40 degrees C, 75% RH for 3 months.

Failure Mode Critical Effect Analysis and Design of Experiment-Based Robust Chromatographic Method for Simultaneous Estimation Lornoxicam and Eperisone Hydrochloride.

Failure mode critical effect analysis and design of experiment-based high performance thin layer chromatography (HPTLC) method has been developed for simultaneous estimation of lornoxicam (LOC) and eperisone hydrochloride (EPR). Failure modes were identified on the basis of prior knowledge and experimental data with the help of Ishikawa diagram for the development of method. The criticality of failure mode was assessed by giving risk priority number and criticality rank on the basis of preliminary experimental trials. The identified critical failure modes were analyzed for their effect by design of experiment (DoE)-based Plackett-Burman screening design. From 11 critical factors, the volume of methanol and modifier in mobile phase composition were found as critical failure modes. Critical failure mode was further analyzed by DoE based on central composite design for study of their relationship with resolution of both drugs. Quadratic model suggested by design was further used for failure mode risk control and navigation of design space for a resolution of both drugs more than 1.5. Failure mode risk control strategy was set for robust HPTLC method for simultaneous estimation of both drugs in laboratory mixture. Developed and validated HPTLC method was applied for assay of LOC and EPR in their laboratory mixture and assay values were found in good agreement with a spiked amount of drugs.

Soft-modeling based spectrofluorimetric study of simultaneous equilibria.

A two-way soft resolution method will fail when applied to a simultaneous equilibria system due to rank deficiency in its concentration profiles. Increasing the dimensionality of measurements from two-way to three-way data can be used to overcome this problem. Simultaneous dissociation of two weak acids is considered as a model for simultaneous equilibria. Three-way data obtained from excitation-emission spectrofluorimetric monitoring of a pH-metric titration is analyzed using a proper combination of well-known soft-modeling methods. Multivariate curve resolution-alternating least squares is used for calculating the excitation and emission spectral profiles of involved species and rank annihilation factor analysis for obtaining the contribution of each species in measured excitation-emission matrices at different pHs. The results of simulated and real simultaneous acids dissociation equilibria showed that the proposed combined method performs well even in situation when the equilibrium constants are close to each other. The applicability of method for study of an acidic dissociation is also shown.

Chemiluminescence method for the determination of piroxicam by the enhancement of the tris-(4,7-diphenyl-1,10-phenanthrolinedisulphonic acid) ruthenium(II) (RuBPS)-cerium(IV) system and its application.

A simple, rapid chemiluminescence (CL) method was described for the determination of piroxicam, a commonly used analgesic agent drug. A strong CL signal was detected when cerium(IV) sulphate was injected into tris-(4,7-diphenyl-1,10-phenanthrolinedisulphonic acid) ruthenium(II) (RuBPS)-piroxicam solution. The CL signal was proportional to the concentration of piroxicam in the range 2.8 x 10(-8)-1.2 x 10(-5) mol/L. The detection limit was 2 x 10(-8) mol/L and the relative standard deviation (RSD) was 3.7% (c = 7.0 x 10(-7) mol/L piroxicam; n = 11). The proposed method was applied to the determination of piroxicam in pharmaceutical preparations in capsules, spiked serum and urine samples with satisfactory results.

MXene with Great Adsorption Ability toward Organic Dye: An Excellent Material for Constructing a Ratiometric Electrochemical Sensing Platform.

Ratiometric electrochemical sensors coupled with an intrinsic built-in correction have received much attention in biochemical analysis, which can effectively avoid potential impacts from both intrinsic and extrinsic factors. However, the complex modification procedure and the unstable reference signal limit development and application of ratiometric sensing. To address these issues, we proposed a novel ratiometric electrochemical platform based on MXene. Introduction of built-in correction was realized via simple one-step incubation of MXene in solution containing the reference molecule methylene blue (MB), and their firm electrostatic interaction ensures the strong adsorption capability of MXene toward MB. Remarkable enhancement in repeatibility and stability compared with nonratio sensor was proved by detecting the model analyte piroxicam. Furthermore, compatibility of the ratio sensor was demonstrated by integrating copper nanoparticles (CuNPs) into the platform. It turned out that sensing performance of the hybrid electrochemical sensor was significantly improved owing to synergistic effect of MXene and CuNPs, where the former affords a large specific surface area as well as quick electron transport, and the latter possess decent electrical catalytic ability. In all, the proposed ratiometric sensor based on MXene features easy preparation, superb reproducibility, robustness, and broad applicability, affording the platform highly competitive and reliable in the determination of a wide range of substances.

With or Without Internal Standard in HPLC Bioanalysis. A Case Study.

The mandatory strategy of using internal standard in HPLC is still controversial. Despite that the introduction of internal standard methodology in the early stage of development of HPLC technology was used to improve method accuracy and precision, there are still practical situations in which a simple external standard quantification is adequate. The aim of the study is to compare the determination of meloxicam (MXC) in human plasma by HPLC with or without using an internal standard according to some key points related to the reason of introducing the internal standardization such as the reducing of sample preparation errors or variability for low injection volumes. The HPLC analysis was performed on reversed phase with UV detection after protein precipitation. Piroxicam (PXC) was used as an internal standard. The two methods are compared in terms of accuracy and precision over the same concentration range. The stability of the analyte has been proved. According to the results, the quantitative determination of MXC in human plasma after simple protein precipitation by using PXC as an internal standard does not bring any significant improvement of accuracy and precision of the experimental measurements.

Smart bi-metallic perovskite nanofibers as selective and reusable sensors of nano-level concentrations of non-steroidal anti-inflammatory drugs.

A strategy for trace-level carbon-based electrochemical sensors is investigated via exploring the interesting properties of BaNb2O6 nanofibers (NFs). Utilizing adsorptive stripping square wave voltammetry (ASSWV), an electrochemical sensing platform was developed based on BaNb2O6 nanofibers-modified carbon paste electrode (CPE) for the sensitive detection of lornoxicam (LOR). Different techniques were used to characterize the fabricated BaNb2O6 perovskite NFs. The obtained data show the feasibility to electro-oxidize LOR and paracetamol (PAR) on the surface of the fabricated sensor. The amount of nanofiber and testing conditions were optimized using response surface methodology and ASSWV technique. The optimized BaNb2O6/CPE sensor exhibits low detection limit of 6.39 × 10-10 mol L-1, even in the presence of the co-formulated drug paracetamol (PAR). The sensor was successfully applied for biological applications.

Determination of non-steroidal anti-inflammatory drugs in pharmaceuticals and human serum by dual-mode gradient HPLC and fluorescence detection.

Non-steroidal anti-inflammatory drugs (NSAIDs) such as piroxicam and mefenamic acid are commonly prescribed to treat inflammation, pain and fever. Similarly acetylsalicylic acid is used to prevent strokes and heart attacks. A rapid and selective method was developed for the simultaneous assay of three NSAIDs and salicylic acid via HPLC with fluorescence detection. The separation was performed using a "dual-mode" gradient (acetonitrile-0.1% aqueous orthophosphoric acid) and the analysis was completed within 7 min using an ACE column C18, 5 microm, 150 mm x 4.6 mm. Naproxen was used as internal standard. The proposed method is simple, selective as well as with a good sensitivity reaching LOD lower than 2 pmol (0.05 microM) and was applied for quantitative analysis in pharmaceuticals and in human serum samples. The mean recovery was more than 95% and the within-day and between-days precisions were found to be satisfactory having RSD within the acceptable limits (<10%).

Electrochemical monitoring of piroxicam in different pharmaceutical forms with multi-walled carbon nanotubes paste electrode.

The electrochemical behavior of piroxicam on a multi-walled carbon nanotubes electrode for the first time was investigated. A highly sensitive and fast responding sensor for determination of piroxicam was simply and conveniently fabricated. The constructed electrode exhibits efficiently catalytic activity for the electrooxidation of piroxicam at a reduced over potential with high sensitivity, stability, and long lifetime in the wide concentration rang of piroxicam. The oxidation process was found to be dependent on the pH of the supporting electrolyte. The behavior is further exploited as a sensitive detection method for piroxicam determination by differential pulse voltammetry. Under the optimized conditions the calibration plots are linear in the concentration range of 0.15-5 microg ml(-1). Application of the method for the determination of the drug in the dosage form (Feledene capsules and tablets and also piroxicam gel), without any interference, from the excipients, resulted in acceptable deviation from the stated concentrations. Recoveries were obtained in the range 96.35-104.16%. The detection limit of 0.1 microg ml(-1) was obtained for piroxicam determination.

Simultaneous spectrofluorometric determination of piroxicam and pyridoxine using generalized rank annihilation method.

The application of generalized rank annihilation method (GRAM) to the analysis of fluorescence excitation-emission matrices of mixtures of piroxicam and pyridoxine is described. The input of GRAM consists of two bilinear data matrices, i.e. one for unknown and one for the calibration sample. The excitation wavelength range was from 290 to 340 nm and the emission was recorded from 370 to 560 nm. Piroxicam and pyridoxine were determined in the concentration ranges 0.33 - 4.00 microg ml(-1) and 0.66 - 8.00 microg ml(-1), respectively. To check the accuracy of the proposed method, several binary synthetic mixtures and one real sample were analyzed successfully. No matrix effect was observed in mixture analysis, so a single external calibration sample was used for each analyte. The ability of GRAM to quantify the studied compounds and the comparability of GRAM results were evaluated by comparing them with those of PLS regression as a standard first-order calibration.

Indirect flow-injection spectrophotometric determination of meloxicam, tenoxicam and piroxicam in pharmaceutical formulations.

A simple and sensitive indirect spectrophotometric method for the assay of meloxicam (MX), tenoxicam (TX) and piroxicam (PX) in pure and in pharmaceutical formulations by flow injection analysis (FIA) has been proposed. The method is based on the oxidation of these drugs by a known excess of N-bromosuccinimide (NBS) in an acidic medium, followed by a reaction of excess oxidant with chloranilic acid (CAA) to bleach its purple color. The absorbance values increased linearly with increasing concentrations of the drugs. Variables, such as the acidity, reagent concentrations, flow rate of reagents and other FI parameters were optimized to produce the most sensitive and reproducible results. The system obeyed Beer's low over concentration ranges of 10 - 160, 20 - 200 and 10 - 160 microg/ml for MX, TX and PX, respectively. The common excipients and additives did not interfere with their determinations. The method was successfully applied to the determinations of MX, TX and PX in various pharmaceutical preparations. The results obtained by the proposed method were found to be in good agreement with those found by the official HPLC methods.

Miniaturized X-ray powder diffraction assay (MixRay) for quantitative kinetic analysis of solvent-mediated phase transformations in pharmaceutics.

Many pharmaceutical compounds exhibit polymorphism, which may result in solvent-mediated phase transformations. Since the polymorphic form has an essential influence on physicochemical characteristics such as solubility or dissolution rate, it is crucial to know the exact polymorphic composition of a drug throughout pharmaceutical development. This study addressed the need to perform quantitative X-ray analysis of polymorphic mixtures on a 96-well scale (MixRay). A calibration of polymorphic mixtures (anhydrate and hydrate) was performed with three model drugs, caffeine, piroxicam, and testosterone, and linear correlations were obtained for all compounds. The MixRay approach for piroxicam was applied to a solubility and residual solid screening assay (SORESOS) to quantify the amount of hydrate and anhydrate corresponding to kinetic bulk concentrations. Changes in these drug concentrations correlated well with the kinetic changes in the residual solid. The influence of excipients on the solid state and kinetic concentrations of piroxicam was also studied. Excipients strongly affected polymorphic transformation kinetics of piroxicam and concentrations after 24h depended on the excipient used. The new calibration X-ray method combined with bulk concentration analysis provides a valuable tool for both pharmaceutical profiling and early formulation development.

Quantification of piroxicam and 5'-hydroxypiroxicam in human plasma and saliva using liquid chromatography-tandem mass spectrometry following oral administration.

Saliva sampling used to quantify piroxicam and 5'-hydroxypiroxicam is a noninvasive and painless method when compared to sequential blood sampling. For that, a rapid, selective and sensitive liquid chromatography-tandem mass spectrometric method for simultaneous determination of piroxicam and 5'-hydroxypiroxicam in saliva and human plasma was developed and validated. Piroxicam and its major metabolite were separated using a LiChroCART 125-4 RP Select-B Sorbent C18 column using a mixture of methanol and 2% phosphoric acid (pH 2.7) (70:30, v/v) for the mobile phase with a flow injection of 1mL/min. The run time was 4min. Volunteers had saliva and blood sampled before, 1, 2, 3, 4, 5, 6, 8, 11, 24, 48 and 72h after taking a 20mg oral dose of piroxicam. The pharmacokinetic parameters of piroxicam in plasma samples were as follows: AUC0-72 (64819hng/mL), predicted clearance (0.2L/h), distribution volume (14.8L), elimination half-life (50.7h) and saliva/plasma concentration ratio (0.003). The estimation of all pharmacokinetic parameters for 5'-hydroxypiroxicam would require collections beyond 72h; however, it was possible to quantify the mean maximum concentration (133ng/mL), time to peak concentration (53.6h), mean AUC0-72 (6213hng/mL), predicted clearance (110.3L/h) and saliva/plasma concentration ratio (0.04). The developed methods proved effective and sensitive for determining the lower quantification limit of piroxicam in plasma (6.1ng/mL) and saliva (0.15ng/mL) and of 5'-hydroxypiroxicam in plasma (1.2ng/mL) and saliva (0.15ng/mL).

Determination of Degradation Products of Cyclobenzaprine Hydrochloride, Lidocaine and Piroxicam in a Semi-Topical Formulation: MS-MS Confirmation of Unknown Impurities.

Association of cyclobenzaprine hydrochloride, piroxicam and lidocaine in a topical formulation is one of the newest innovations in the pharmaceutical formulary field. In this study, a reversed-phase liquid chromatographic method was developed for the establishment of the impurities of cyclobenzaprine hydrochloride, lidocaine and piroxicam in the semisolid topical formulation. In this study, we not only determined 2,6-dimethylaniline, 2-pyrydilamine but also specified impurities of cyclobenzaprine hydrochloride (dibenzosuberenone, amitriptyline, carbinole, cyclobenzaprine N-oxide and anthrachinone). The target compounds were determined using a mobile phase that consisted of a mixture of phosphate buffer (0.025 M; pH 6.2)-acetonitrile-methanol (60 : 13 : 27, v/v/v). A minimum of three supplementary possible degradation products were determined. Using mass spectrometry, the unspecified impurities were identified and the use of correlation matrices permitted the association with the possible source compounds. The chromatographic conditions were qualified and validated according to ICH guideline requirements to confirm specificity, linearity, accuracy and precision.