Favipiravir (SARS-CoV-2) degradation impurities: Identification and route of degradation mechanism in the finished solid dosage form using LC/LC–MS method

Favipiravir finished dosage was approved for emergency use in many countries to treat SARS-CoV-2 patients. A specific, accurate, linear, robust, simple, and stability-indicating HPLC method was developed and validated for the determination of degradation impurities present in favipiravir film-coated tablets. The separation of all impurities was achieved from the stationary phase (Inert sustain AQ-C18, 250 × 4.6 mm, 5-μm particle) and mobile phase. Mobile phase A contained KH₂PO₄ buffer (pH 2.5 ± 0.05) and acetonitrile in the ratio of 98:2 (v/v), and mobile phase B contained water and acetonitrile in the ratio of 50:50 (v/v). The chromatographic conditions were optimized as follows: flow rate, 0.7 mL/min; UV detection, 210 nm; injection volume, 20 μL; and column temperature, 33°C. The proposed method was validated per the current International Conference on Harmonization Q2 (R1) guidelines. The recovery study and linearity ranges were established from the limit of quantification to 150% optimal concentrations. The method validation results were found to be between 98.6 and 106.2% for recovery and r² = 0.9995–0.9999 for linearity of all identified impurities. The method precision results were achieved below 5% of relative standard deviation. Forced degradation studies were performed in chemical and physical stress conditions. The compound was sensitive to chemical stress conditions. During the study, the analyte degraded and converted to unknown degradation impurities, and its molecular mass was found using the LC–MS technique and established degradation pathways supported by reaction of mechanism. The developed method was found to be suitable for routine analysis of research and development and quality control.     

基于甲基苯丙胺和吗啡的胶体金免疫层析试剂盒检测法的可靠性评价

通过固相萃取-液相色谱-多级质谱(SPE-LC-MS/MS)联用技术和毒品胶体金免疫层析试剂盒检测法对13种中药及调味品样品中甲基苯丙胺及吗啡分别进行定量分析,依据LC-MS/MS检测结果,对毒品胶体金免疫层析试剂盒检测法进行可靠性评价。实验结果表明:型号1试剂盒对甲基苯丙胺和吗啡的特异性均不高,检测准确率分别为57.7%与78.8%;型号2试剂盒对甲基苯丙胺的特异性不足,准确率为73.1%,但对吗啡的检测准确率达到100%。在利用毒品胶体金免疫层析试剂盒进行毒品快速筛查时,应注重排除干扰因素以提高免疫胶体金层析试剂盒的检测准确度。     

Development and validation of high-performance liquid chromatography method for analysis of β-CCT and its related substances

Compound 2β-carbomethoxy-3β-(4-chlorophenyl)tropane (β-CCT) is a key intermediate for the synthesis of some clinical dopamine transporter (DAT) imaging agents. Potential impurities from synthesis process of β-CCT and degradation during storage might have detrimental effect on the final imaging agents. Thus, it is necessary to guarantee the quality of β-CCT. In this study, a rapid, sensitive and accurate high-performance liquid chromatography (HPLC) method was developed and validated for the analysis of β-CCT and its related substances. The chromatographic separation was achieved on a reverse-phase phenomenex? Gemini C18 column with an isocratic mobile phase consisted of methanol, water and TFA (30:70:0.1 v/v/v). The flow rate was 1.0 mL/min at 30 °C and samples were monitored at 220 nm. The method was validated concerning system suitability, linearity, accuracy, precision, specificity, robustness and stability. The limit of detection (LOD) and the limit of quantification (LOQ) of β-CCT were 0.5 and 1.5 μg/mL, respectively. The linearity range of β-CCT was 1.5–450 μg/mL with a good linear correlation coefficient (R² = 0.9999) between the peak response and concentration. Specificity investigation through forced degradation experiments displayed that β-CCT was stable in acidic, thermal and photolytic degradation conditions, but significantly unstable in alkaline and oxidative conditions. With the developed chromatographic method, possible impurity α-CCT from synthetic process and potential degradation products could be well separated from β-CCT. Good recovery and precision were manifested in the assay method. These results indicated that the present method would be suitable for not only the quality assurance of β-CCT in regular production sample assays but also the monitoring and determination of its related substances.     

Analysis of aquatic‐phase natural organic matter by optimized LDI‐MS method

The composition and physiochemical properties of aquatic‐phase natural organic matter (NOM) are most important problems for both environmental studies and water industry. Laser desorption/ionization (LDI) mass spectrometry facilitated successful examinations of NOM, as humic and fulvic acids in NOM are readily ionized by the nitrogen laser. In this study, hydrophobic NOMs (HPO NOMs) from river, reservoir and waste water were characterized by this technique. The effect of analytical variables like concentration, solvent composition and laser energy was investigated. The exact masses of small molecular NOM moieties in the range of 200–1200 m/z were determined in reflectron mode. In addition, spectra of post‐source‐decay experiments in this range showed that some compounds from different natural NOMs had the same fragmental ions. In the large mass range of 1200–15 000 Da, macromolecules and their aggregates were found in HPO NOMs from natural waters. Highly humic HPO exhibited mass peaks larger than 8000 Da. On the other hand, the waste water and reservoir water mainly had relatively smaller molecules of about 2000 Da. The LDI‐MS measurements indicated that highly humic river waters were able to form large aggregates and membrane foulants, while the HPO NOMs from waste water and reservoir water were unlikely to form large aggregates. Copyright © 2014 John Wiley & Sons, Ltd.     

Influence of amide linkages on acidity determinations of humic substances Testing with model-mixtures

A set of experiments, using model compounds, was carried out in order to evaluate the influence of amide linkages on acidity measurements of humic substances (HS). Three benzene-carboxylic acids (2,4-dihydroxybenzoic acid; 3,5-dihydroxybenzoic acid, and phthalic acid) and two peptides (dl-alanyl-dl-alanine, and glycil-l-leucine) were employed to simulate the major acidic functional groups present in humic material. The acidity of the samples was measured, using the Schnitzer and Gupta methods, for each compound separately, as well as for selected mixtures with different compound combinations. General results showed that, in the absence of peptides, phenolic groups with high pK_a values are not detected in the barium hydroxide reaction and the data so obtained do not represent the real concentrations of the acidic groups. For the mixtures containing peptides, the presence of amide linkages distorts the results relative to the total acidity (TA) mainly because under the extreme conditions of the reaction with Ba(OH)₂ hydrolysis of the peptide occurs, consuming extra hydroxyl groups and increasing artificially the phenolic content. Such a condition is minimized by the reduction of the reaction time.     

Impact of Extracellular Polymeric Substance in the Inactivation of Harmful Algae by Ag2O/g-C3N4 under Visible Light

Photocatalysis has attracted much attention as an emerging algae removal technology, but the inactivation performance is inevitably affected by the extracellular polymeric substance (EPS) produced by algae. In this study, a photocatalyst (Ag₂O/g-C₃N₄) with efficient algae inactivation is adopted to investigate the interactions with EPS, and the impact of EPS on photocatalytic algae removal is studied. The results show that EPS can adhere to the surface of Ag₂O/g-C₃N₄ by electrostatic force. The interaction with EPS decreases the surface zeta potential of the Ag₂O/g-C₃N₄ from 7.71 to −22.3 mV with the increase in EPS concentration, and the maximum ratio of particle size increases from 825 to 1281 nm. In addition, the interaction with EPS inhibits the release of Ag⁺ in Ag₂O/g-C₃N₄ by half, thus, the toxicity of metal ions will be alleviated. Meanwhile, EPS can also be degraded by Ag₂O/g-C₃N₄, indicating that EPS can work as a radical scavenger to protect the algae cells. Without the protection of EPS, 97.8% of algae cells are inactivated after 5 h photocatalysis. Therefore, more attention should be given to the interaction between EPS and photocatalyst to promote the design and application of the photocatalytic.     

Methyl methacrylate particles with amino groups on the surface: Colloid stability and sorption of biologically active substances

The present article investigates the influence of dispersion medium on the aggregate stability of cross-linked poly-(methyl methacrylate) particles on the surface layer of which aliphatic amino groups are localized. It is shown that particle size depends on the ionic strength of dispersion medium. The research determines the optimal content of cross-linked agent, ethylene glycol dimetharylate that results in the formation of cationic particles being stable in saline. Particle modification is performed by bovine serum albumin and luminophore fluorescein 5(6)-isothiocyanate. Protein sorption is observed not to influence luminescent properties of the particles. It is also determined that due to the aggregate stability of particles obtained from 5?wt% of ethylene glycol dimethacrylate in saline, the monodispersity and the absence of inversion of zeta potential in wide pH range is retained, such particles may be considered as perspective carriers of biologically active substances.     

Bienzyme Sensor Based on an Oxygen Reducing Bilirubin Oxidase Electrode

Based on an oxygen reducing electrode combining bilirubin oxidase and multiwalled carbon nanotubes modified gold (BOD‐MWCNT‐Au electrode) a bienzyme sensor is developed. Therefore the BOD‐MWCNT‐Au electrode is covalently coupled to enzymes catalysing oxygen‐consuming reactions (glucose oxidase and ascorbate oxidase) to result in a membrane‐free bienzyme electrode. The electrochemical characterisation of these bienzyme sensors reveals an enzyme substrate sensitivity down to 250 μM glucose and 100 μM ascorbate. In addition, the assembled sensor systems allow amperometric measurements in a potential range where the influence of interfering substances reacting directly at the transducing electrode is minimised. The results indicate that the BOD electrode provides a suitable platform for sensing analytes of medical and environmental interest for which oxidases of high activity are available.     

Uncertainty of measurement for summed masses: application to controlled substances

Because sentences for drug possession depend on the mass of the seized drug, testing laboratories must often determine the summed mass of numerous items submitted under a single case. One common practice for this purpose is to continue analyzing and weighing samples until a legal threshold is passed, at which point it is important to inform the court whether the summed mass is significantly above the threshold, or only marginally so. This paper develops a means for estimating the uncertainty of the summed mass for the common situation where the readability, d, of the balance dominates the uncertainty. It is seen that for all sample sizes the uncertainty, UM, is given by the remarkable simple expression UM = (d/2) × [N + SQRT(3N)] + N × CCE, where N is the number of items and CCE is the absolute value of the calibration check error. In most instances, this can be further simplified to UM = N × d.