A simple and rapid determination of Al(III) in natural water samples using dispersive liquid-liquid microextraction after complexation with a novel antipyrine-based Schiff base reagent.

The purpose of this study is the development of a novel strategy for the determination of Al3+ ions using the combination of dispersive liquid-liquid microextraction (DLLME) and UV-Vis spectrophotometry. The method is grounded in the complexation between a novel antipyrine-based Schiff base reagent (EHMP) and Al3+ ions. Aluminum concentrations were detected using UV-Vis spectrophotometry at 260 nm and this technique was optimized using the absorbance value of EHMP-Al complex. pH, mixing period, type and volume of organic solvent, etc. were optimized stepwise in order to find out optimum experimental conditions. The limit of detection and the limit of quantification values for the improved analytical method were to be estimated 0.31 and 1.03 µmol.L-1, respectively. The new strategy was successfully performed to define Al3+ ions in natural water samples with RSD values (84.01-107.71%) and recovery values (0.01-0.09%).

Stress degradation of edaravone: Separation, isolation and characterization of major degradation products.

In the present study the International Conference on Harmonization-prescribed stress degradation was carried out to study the degradation profile of edaravone. To establish a Quality by Design (QbD)-assisted stability-indicating assay, the reaction solutions in which different degradation products were formed were mixed. Plackett Burman and central composite design were used to screen and optimize experimental variables to resolve edaravone and its impurities with good peak symmetry using an RP C18 column. The method was validated according to International Conference on Harmonization guidelines. Seven unknown and two known degradation products were identified and characterized by LC-MS/MS. Two major degradation products formed under thermal degradation were isolated and characterized as 4-(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl-4-(4,5-dihydro-5-hydroxy-3-methyl-1-phenyl-1H-pyrazol-4-yl)-3-methyl-1-phenyl-1H-pyrazol-5(4H)-one and 3-hydroxy-dihydro-thiazolo[1-(2-methyl-buta-1,3dienyl)-1-phenylhydrazine]5-one. The degradation pathways of degradants were proposed based on m/z values.

Pharmacokinetic profiles of metamizole (dipyrone) active metabolites in goats and its residues in milk.

Metamizole (dipyrone, MET) is a nonopioid analgesic drug commonly used in human and veterinary medicine. The aim of this study was to assess two major active metabolites of MET, 4-methylaminoantipyrin (MAA) and 4-aminoantipyrin (AA), in goat plasma after intravenous (IV) and intramuscular (IM) administration. In addition, metabolite concentration in milk was monitored after IM injection. Six healthy female goats received MET at a dose of 25 mg/kg by IV and IM routes in a crossover design study. The blood and milk samples were analyzed using HPLC coupled with ultraviolet detector and the plasma vs concentration curves analyzed by a noncompartmental model. In the goat, the MET rapidly converted into MAA and the mean maximum concentration was 183.97 µg/ml (at 0.08 hr) and 51.94 µg/ml (at 0.70 hr) after IV and IM administration, respectively. The area under the curve and mean residual time values were higher in the IM than the IV administered goats. The average concentration of AA was lower than MAA in both groups. Over 1 µg/ml of MAA was found in the milk (at 48 hr) after MET IM administration. In conclusion, IM is considered to be a better administration route in terms of its complete absorption with long persistence in the plasma. However, this therapeutic option should be considered in light of the likelihood of there being milk residue.

Titanium nanoparticles (TiO2)/graphene oxide nanosheets (GO): an electrochemical sensing platform for the sensitive and simultaneous determination of benzocaine in the presence of antipyrine.

An effective electrochemical sensing platform for the simultaneous determination of benzocaine (BEN) and antipyrine (ANT) based upon titanium dioxide nanoparticle (TiO2)/graphene oxide nanosheet (GO) bulk modified carbon paste electrodes (TiO2-GO/CPE) is reported. The TiO2-GO/CPE electrochemical sensing platform is found to exhibit linear ranges from 1.0 × 10-6 to 1.0 × 10-4 M and 1.2 × 10-8 to 8.0 × 10-5 M for BEN and ANT, respectively. The TiO2-GO/CPE sensor is explored towards the analysis of BEN and ANT in oral fluid (saliva) and pharmaceutical products. The synergy between the graphene oxide nanosheets and titanium dioxide nanoparticles results in a dramatic enhancement in the sensitivity of the sensor through a combination of increased surface area and improved electron transfer kinetics compared to other electrode alternatives. The fabricated TiO2-GO/CPE exhibits high sensitivity and good stability towards the sensing of BEN and ANT and has the potential to be utilised as a clinical assay and QA in pharmaceutical products.

Simultaneous determination of aminopyrine and antipyrine in porcine muscle, milk, and eggs using liquid chromatography with tandem mass spectrometry.

The concentrations of residual aminopyrine and antipyrine in porcine muscle, milk, and egg samples were analyzed using liquid chromatography with tandem mass spectrometry after undergoing a series of sample pretreatment steps. Owing to an ion suppression effect, matrix-matched calibrations were used for analyte quantitation with determination coefficients (R(2) ) ≥ 0.9931. The recovery rates for aminopyrine and antipyrine in various matrices at two spiking levels (5 and 10 ng/g) fell in the range of 60.96-68.87 and 61.87-66.99%, respectively. Meanwhile, the intra- and inter-day precisions (expressed as relative standard deviation) were 1.02-12.95 and 1.71-5.50%, respectively. The method's detection limit (1 ng/g) was very low, thus enabling the detection of low residue levels. The applicability of the developed method was demonstrated with actual market samples and none of the tested analytes was detected in any of the samples.

Green chromatography separation of analytes of greatly differing properties using a polyethylene glycol stationary phase and a low-toxic water-based mobile phase.

A simple, rapid, and environmentally friendly HPLC method was developed and validated for the separation of four compounds (4-aminophenol, caffeine, paracetamol, and propyphenazone) with different chemical properties. A "green" mobile phase, employing water as the major eluent, was proposed and applied to the separation of analytes with different polarity on polyethylene glycol (PEG) stationary phase. The chromatography separation of all compounds and internal standard benzoic acid was performed using isocratic elution with a low-toxicity mobile phase consisting of 0.04% (v/v) triethylamine and water. HPLC separation was carried out using a PEG reversed-phase stationary phase Supelco Discovery HS PEG column (15 × 4 mm; particle size 3 µm) at a temperature of 30 °C and flow rate at 1.0 mL min(-1). The UV detector was set at 210 nm. In this study, a PEG stationary phase was shown to be suitable for the efficient isocratic separation of compounds that differ widely in hydrophobicity and acid-base properties, particularly 4-aminophenol (log P, 0.30), caffeine (log P, -0.25), and propyphenazone (log P, 2.27). A polar PEG stationary phase provided specific selectivity which allowed traditional chromatographic problems related to the separation of analytes with different polarities to be solved. The retention properties of the group of structurally similar substances (aromatic amines, phenolic compounds, and xanthine derivatives) were tested with different mobile phases. The proposed green chromatography method was successfully applied to the analysis of active substances and one degradation impurity (4-aminophenol) in commercial preparation. Under the optimum chromatographic conditions, standard calibration was carried out with good linearity correlation coefficients for all compounds in the range (0.99914-0.99997, n = 6) between the peak areas and concentration of compounds. Recovery of the sample preparation was in the range 100 ± 5% for all compounds. The intraday method precision was determined as RSD, and the values were lower than 1.00%.

[Rapid identification 15 effective components of anti common cold medicine with MRM by LC-MS/MS].

This paper reports the establishment of a method for rapid identification 15 effective components of anti common cold medicine (paracetamol, aminophenazone, pseudoephedrine hydrochloride, methylephedrine hydrochloride, caffeine, amantadine hydrochloride, phenazone, guaifenesin, chlorphenamine maleate, dextromethorphen hydrobromide, diphenhydramine hydrochloride, promethazine hydrochloride, propyphenazone, benorilate and diclofenac sodium) with MRM by LC-MS/MS. The samples were extracted by methanol and were separated from a Altantis T3 column within 15 min with a gradient of acetonitrile-ammonium acetate (containing 0.25% glacial acetic acid), a tandem quadrupole mass spectrometer equipped with electrospray ionization source (ESI) was used in positive ion mode, and multiple reaction monitoring (MRM) was performed for qualitative analysis of these compounds. The minimum detectable quantity were 0.33-2.5 microg x kg(-1) of the 15 compounds. The method is simple, accurate and with good reproducibility for rapid identification many components in the same chromatographic condition, and provides a reference for qualitative analysis illegally added chemicals in anti common cold medicine.

Assessment of groundwater pollution in Tokyo using PPCPs as sewage markers.

While the occurrence of pharmaceuticals and personal care products (PPCPs) in groundwater has typically been reported in bank filtration sites, irrigated fields, septic tanks, and sewage disposal practices, fewer studies have been conducted in highly urbanized areas, where infiltration of treated or untreated sewage is not supposed to be a source of groundwater recharge. Furthermore, little is known about the occurrence of various kinds of PPCPs in relation to microbial indicators in groundwater from different types of aquifers. Thus, we examined the city-wide occurrence of selected PPCPs (diethyltoluamide, crotamiton, ethenzamide, propyphenazone, carbamazepine, and caffeine) and E. coli in 50 groundwaters from unconfined aquifers (<30 m in depth) and confined aquifers (up to 500 m in depth) in Tokyo, where unintended groundwater contamination could take place due to decrepit sewer networks. PPCPs were detected in unconfined aquifers and springs (23/34 samples, 68%), and in confined aquifers (7/16 samples, 44%). Compared with published results for sewage influents, concentrations of PPCPs, excluding caffeine, were generally 1-2 orders of magnitude lower, while in some samples concentrations were quite comparable. The high occurrence rate of PPCPs, even in confined aquifers, indicated that such aquifers are not always protected from pollution by sewage near the land surface. Among the PPCPs analyzed, carbamazepine and crotamiton were most frequently detected, which would appear to be owing to their high persistence, combined with the high concentration of crotamiton in sewage. Crotamiton was detected in all four E. coli-positive groundwaters, and thus may potentially serve as a precautionary indicator of E. coli contamination. Using carbamazepine as a sewage marker, we estimated that 0.8%-1.7% of the dry-weather flow of sewage was leaking out into the unconfined aquifers.

Validation of an HPLC method for the simultaneous determination of diminazene diaceturate and phenazone in injectable veterinary granules and bulk powders.

A validated HPLC method for the simultaneous determination of diminazene diaceturate and phenazone has been established for the analysis of the compounds in veterinary granules for injection and in bulk powders. The compounds were separated using a Symmetry RP 18 analytical column and detected by UV absorbance at 250 nm. Linearity, accuracy as well as the intra-assay precision, inter-day precision and specificity of the method were established. The limits of detection and quantification were 3.2 and 9.7 µg mL-1 for diminazene diaceturate and 9.57 and 28.99 µg mL-1 for phenazone. Method had the potential to determine these drugs simultaneously from dosage forms without any interference with each other.

Placental transfer of the insulin analog glargine in the ex vivo perfused placental cotyledon model.

OBJECTIVE: The aim was to characterize the transfer of the insulin analog glargine across the placenta using the placental perfusion model. METHODS: Placentas were obtained and selected cotyledons were cannulated and dually perfused. Glargine, 50 mU/L (n = 2) and 200 mU/L (n = 1), and a reference marker, antipyrine (50 µg/mL), were added to the maternal circulation. Samples were taken from the maternal and fetal compartments. RESULTS: Glargine was not detected in the fetal compartment. In the maternal compartment, the steady state concentration was 50% lower than the starting concentration. CONCLUSIONS: Glargine probably does not cross the human placenta. Reduced maternal steady state concentrations may suggest insulin uptake by the placenta.

Reaction of phenazone-type drugs and metabolites with chlorine and monochloramine.

This work studies the chlorination and monochloramination reaction kinetics of two phenazone-type drugs (phenazone - Phe and propyphenazone - PrPhe) and three metabolites of phenazone-type drugs (4-formylaminoantipyrine - FAA, 4-aminoantipyrine - AA and 4-acetoamidoantipyrine - AAA). Kinetics were faster with chlorine (apparent second-order constants between 100 and 66,500 times higher) than with monochloramine. For FAA and AAA, no significant reaction was observed during monochloramination. Further, apparent rate constants decreased as the pH increased from pH 5.7 to 8.3, except during chlorination of AA. The transformation products (TPs) formed were also elucidated by liquid chromatography-high resolution mass spectrometry. The main transformation pathway for Phe and PrPhe consisted of halogenations, hydroxylations and dealkylations, while AAA and FAA were firstly transformed to AA, then followed by pyrazole ring opening and hydroxylations. The extend of the reaction was also tested in real water samples, where, in general, slower reaction kinetics were obtained during monochloramination, while the chlorination reaction showed similar half-lives to ultrapure water. Finally, acute and chronic toxicity of the TPs were estimated using two quantitative structure-activity relationship (QSAR) software (ECOSAR and TEST), showing that some TPs could be more toxic than their precursor compounds.

Simultaneous Determination of Paracetamol, Propyphenazone and Caffeine in Presence of Paracetamol Impurities Using Dual-Mode Gradient HPLC and TLC Densitometry Methods.

Two chromatographic methods were validated for the determination of the widely prescribed analgesic and antipyretic drug combination of paracetamol (PC) (recently integrated into the supportive treatment of COVID-19), propyphenazone (PZ) and caffeine (CF) in the presence of two PC impurities, namely 4-aminophenol and 4-nitrophenol. A "dual-mode" gradient high-performance liquid chromatography method was developed, where the separation was achieved via "dual-mode" gradient by changing both the ternary mobile phase composition (acetonitrile: methanol: water) and the flow rate. This enables a good resolution within a relatively shorter analysis time. The analysis was realized using Zorbax Eclipse XDB column C18, 5 µm (250 × 4.6 mm) and the UV detector was set at 220 nm. The other method is a thin-layer chromatography densitometry method, where the separation was achieved using a mobile phase composed of chloroform: toluene: ethyl acetate: methanol: acetic acid (6: 6: 1: 2: 0.1, by volume). Densitometric detection was performed at 220 nm on silica gel 60 F254 plates. The developed methods were fully validated as per the ICH guidelines and proved to be accurate, robust, specific and suitable for application as purity indicating methods for routine analysis of PC in pure form or in pharmaceuticals with PZ and CF in quality control laboratories.

Degradation of selected pharmaceuticals in aqueous solution with UV and UV/H(2)O(2).

The degradation of four pharmaceutical compounds (PhACs), ibuprofen (IBU), diphenhydramine (DP), phenazone (PZ), and phenytoin (PHT) was investigated via ultraviolet (UV) photolysis and UV/H(2)O(2) process with a low-pressure (LP) UV lamp. For each PhAC tested, direct photolysis quantum yields at 254 nm were found to be ranging from 6.32 x 10(-2) to 2.79 x 10(-1)molE(-1) at pH 7. The second-order rate constants of the reaction between the PhACs and OH were determined to be from 4.86 x 10(9) to 6.67 x 10(9)M(-1)s(-1) by using a competition kinetic model which utilized para-chlorobenzoic acid (pCBA) as a reference compound. The overall effect of OH radical scavenging from humic acid (HA) and anions HCO(3)(-), NO(3)(-) was measured utilizing R(OH,UV) method through examining the aqueous photodegradation of pCBA as a probe compound. Moreover, these fundamental direct and indirect photolysis parameters were applied in the model prediction for oxidation rate constants of the PhACs in UV/H(2)O(2) process. It was found that the predicted oxidation rate constants approximated the observed ones. The results indicated that the new R(OH,UV) probe compound method was applicable for measuring background OH radical scavenging effects in water treatment process of UV/H(2)O(2). Furthermore, by GC-MS analysis, most of the intermediates created during the photodegradation of the selected PhACs in UV/H(2)O(2) process were identified. For the photodegradation of PZ, a competition mechanism existed between the direct UV photolysis and the oxidation of OH. An appropriate dosage of H(2)O(2) could hinder the occurrence of the direct photolysis.

A novel fluorescent assay for edaravone with aqueous functional CdSe quantum dots.

Aqueous thiol-capped CdSe QDs with a narrow, symmetric emission were prepared under a low temperature. Based on the fluorescence enhancement of thiol-stabilized CdSe quantum dots (QDs) caused by edaravone, a simple, rapid and specific quantitative method was proposed to the edaravone determination. The concentration dependence of fluorescence intensity followed the binding of edaravone to surface of the thiol-capped CdSe QDs was effectively described by a modified Langmuir-type binding isotherm. Factors affecting the fluorescence detection for edaravone with thiol-stabilized CdSe QDs were studied, such as the effect of pH, reaction time, the concentration of CdSe QDs and so on. Under the optimal conditions, the calibration plot of C/(I-I(0)) with concentration of edaravone was linear in the range of (1.45-17.42) microg/mL (0.008-0.1 micromol/L) with correlation coefficient of 0.998. The limit of detection (LOD) (3sigma/kappa) was 0.15 microg/mL (0.0009 micromol/mL). Possible interaction mechanism was discussed.

Simultaneous determination of acrylamide, its metabolite glycidamide and antipyrine in human placental perfusion fluid and placental tissue by liquid chromatography-electrospray tandem mass spectrometry.

A rapid and sensitive method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the simultaneous determination of acrylamide (AA) and its genotoxic metabolite glycidamide (GA) with a test marker antipyrine (AP) in placental tissue and perfusion medium used in human placental perfusion studies. An internal standard ((13)C-acrylamide) was added to the samples which were then deproteinized with acetonitrile. Chromatographic separation was performed on a reversed phase column with a gradient elution of acetonitrile and 0.01% formic acid at a flow rate of 0.3 mL/min. Detection and quantification of the analytes were carried out with a triple quadrupole mass spectrometer using positive electrospray ionization (ESI) and multiple reaction monitoring (MRM). The method was validated and linear over a concentration range of 0.5-20 microg/mL for acrylamide and glycidamide and 5-200 microg/mL for antipyrine. The lower limit of quantification for acrylamide and glycidamide was 0.5 microg/mL and for antipyrine 5 microg/mL. The method was selective, and good accuracy, precision, recovery, and stability were obtained for concentrations within the standard curve. The method was successfully used to analyze the placental perfusion medium and tissue samples in a toxicokinetic study for transplacental transfer of acrylamide and glycidamide. This is the first time that acrylamide, glycidamide and antipyrine are measured simultaneously.

Behaviour and redox sensitivity of pharmaceutical residues during bank filtration - Investigation of residues of phenazone-type analgesics.

The behaviour of residues of phenazone-type pharmaceuticals during bank filtration was investigated at a field site in Berlin, Germany, where bank-filtered water is used for drinking water production. The concentrations of the pharmaceutical residues in the shallow, young bank filtrate (travel times

Determination of caffeine and associated compounds in food, beverages, natural products, pharmaceuticals, and cosmetics by micellar electrokinetic capillary chromatography.

A method is described for quantitating caffeine, theobromine, theophylline, paracetamol, propyphenazone, acetylsalicylic acid, salicylic acid, and codeine phosphate in corresponding real samples of food, beverages, natural products, pharmaceuticals, and cosmetic preparations by micellar electrokinetic capillary chromatography. The separation is carried out at 25 degrees C and 25 kV, using a 20 mM phosphate buffer (pH 9.0), 80 mM sodium dodecyl sulfate, and 7.5% (v/v) acetonitrile. UV detection is at 210 nm. The method is shown to be specific, accurate (recoveries over the range 98.9-101.2%), linear over the tested range (correlation coefficients>or=0.9993), and precise (relative standard deviation below 2.1%). The method is applied for the quantitative analysis of these compounds in different foods, beverages, natural products, pharmaceuticals, and cosmetic products.

Concurrent determination of topotecan and model permeability markers (atenolol, antipyrine, propranolol and furosemide) by reversed phase liquid chromatography: utility in Caco-2 intestinal absorption studies.

A simple, sensitive, specific and high-resolution reversed-phase liquid chromatographic method utilizing ultraviolet detection has been developed and validated for simultaneous determination of topotecan and four intestinal permeability markers (atenolol, antipyrine, propranolol and furosemide) as suggested by US-FDA. Chromatography was carried out on C-18 column with mobile phase comprising water (pH 3.0) and acetonitrile gradient pumped at a flow rate of 1 ml min(-1). The validation parameters included specificity, accuracy, precision, sensitivity and stability studies. Topotecan, an anti-cancer drug widely used in metastatic carcinoma, is a P-glycoprotein substrate having oral bioavailability of 30% with large inter-patient variability. The present method was successfully applied for demonstrating P-gp mediated transport of topotecan and its inhibition using verapamil in Caco-2 cell monolayer. The method can be used in identification of novel P-gp inhibitors for topotecan and estimating the contribution of P-gp in affecting oral bioavailability of topotecan. The other applications of method include its use in validation of Caco-2 monolayer assay for getting biowaiver based on Biopharmaceutic Classification System and its extrapolation to in situ and/or in vivo studies.

Simultaneous determination of paracetamol, caffeine and propyphenazone in ternary mixtures by micellar electrokinetic capillary chromatography.

A new micellar electrokinetic capillary chromatographic method has been developed to analyze the pharmaceutical preparations containing ternary combination of paracetamol (PAR), caffeine (CAF) and propyphenazone (PRO). Best results were obtained by using 20mM pH 9.0 borate buffer containing 30mM sodiumdodecylsulphate as the background electrolyte. Diflunisal (DIF) was used as internal standard (IS). The separation was performed through a fused silica capillary (50microm internal diameter, 44cm total length, 35.5cm effective length) at 25 degrees C with the application of 3s of hydrodynamic injection at 50mbar pressure and a potential of 29kV. Detection wavelength was 200nm. Under these conditions, the migration times were found to be 5.174min for PAR, 5.513min for CAF, 7.195min for DIF, and 9.366min for PRO. Linearity ranges for the method were determined as 2-200microgmL(-1) for PAR and CAF and 3-200microgmL(-1) for PRO. Limit of detections were found as 0.6microgmL(-1) for PAR and CAF and 0.8microgmL(-1) for PRO. According to the validation study, the developed method was proved to be accurate, precise, sensitive, specific, rugged and robust. Three pharmaceutical preparations, which are produced by different drug companies in Turkey, were analyzed by the developed method. One of the same preparations was also analyzed by the derivative ratio spectro zero-crossing spectrophotometric method reported in literature. No significant differences were found statistically.

Development of a multicommuted flow-through optosensor for the determination of a ternary pharmaceutical mixture.

The combination of multicommutation and flow-through multioptosensing is presented in this work as a powerful strategy for the routine analysis of active principles in pharmaceuticals. By coupling methodologies, the selectivity and sensitivity of optosensors is maintained, while the use of the multicommutation approach provides additional advantages, such as low reagent consumption, low waste generation and reduced human supervision. The potential of this integration is enhanced when implemented with multiwavelength detection mode. An UV sensor is here developed for the simultaneous determination of three widely used active principles: salicylamide, caffeine and propyphenazone. The measuring wavelengths were 276 nm for caffeine and propyphenazone, and 302 nm for salicylamide. The five three-way solenoid valves used in the system are controlled by Java-written home-made software. The sensor is based on the on-line selective retention of two of the three analytes on a precolumn placed just before the sensing zone and filled with the same solid support than the flow-through cell (C(18) silica gel). This approach allows the sequential arrival of the analytes to the sensing zone, so allowing their determination with only one sample injection. So, the use of C(18) placed, in both the precolumn and the flow-cell combines the advantages of the increase of sensitivity and selectivity in the detection solid zone with the additional increase of the selectivity in the precolumn. The sensor was applied to the determination of the analytes in several pharmaceutical preparation of the Spanish Pharmacopoeia, obtaining satisfactory results.