Background Chlorate is an effective herbicide, but also a by-product of chlorinating agents used to disinfect water, one of the reasons why it is regularly found in food. Perchlorate is a ubiquitous contaminant, which is naturally occurring in the environment but also released from anthropogenic sources such as the industrial use of certain natural fertilizers. Chlorate affects the hematological system, and perchlorate the thyroid. Objective Implement and validate a simple and robust analytical method for the accurate determination of chlorate and perchlorate in baby food, infant and adult formulas, and ingredients thereof, which is suited for its application in routine environments where a broad variety of food commodities must be analyzed simultaneously. Method Typically, analytes are extracted with a mixture of water, acidified methanol and dichloromethane. Optionally, for dairy products and byproducts, extraction can be performed with water, acidified methanol and EDTA, followed by two steps clean-up (freezing-out and dispersive solid phase extraction with C18 in acetonitrile). Quantitative determination is carried out by isotopic dilution liquid chromatography tandem mass spectrometry (LC-MS/MS). Results The method was single laboratory validated in five Nestlé Quality Assurance Centers (NQACs) in a comprehensive range of representative matrices of different categories such as baby foods, infant/adult formulas and ingredients, with results generally in agreement with the acceptance criteria of the Standard Method Performance Requirements (SMPR) 2021.001 defined by AOAC INTERNATIONAL, in terms of representative matrices validated, LOQs, trueness and precision.

Background Propylene chlorohydrins (PCH) are formed as byproducts in the reaction between starch and propylene oxide (PO). For hydroxypropylated starch (HP-starch) applications in food, JECFA set the maximum allowed total propylene chlorohydrin (PHC-t) residues level at 1 mg/kg. Objective To develop an improved analytical method for the determination of the PCH-t content in starches in the low mg/kg range to replace the outdated JECFA method. Method A new GC-MS method that utilizes aqueous methanol as extraction medium for PCH. The GC-MS system is equipped with a programmable temperature vaporization injector and Stabilwax-DA column using helium as carrier gas. The quantitative detection is achieved in the selected ion monitoring mode. Results This single laboratory validation (SLV) study showed good linear calibrations for both 1-chloro-2-propanol (PCH-1) and 2-chloro-1-propanol (PCH-2) in the concentration range of 0.5-4 mg/kg in dry starch. The lower limit of quantitation of PCH-1 and PCH-2 was 0.2–0.3 mg/kg in dry starch, the relative standard deviation reproducibility (RSDR) at the concentration level of 1–2 mg/kg in dry starch was 3–5%, and the recovery values for both PCH-1 and PCH-2 were in the range of 78–112% at a concentration level of about 0.6 mg/kg in dry starch. Compared with the current, outdated JECFA method, the new GC-MS method is more sustainable, less laborious and therefore more economical. The analytical capacity of the new method is 4–5 times higher than the analytical capacity of the old JECFA method. Conclusion The GC-MS method is fit for a Multi Laboratory Trial (MLT). Highlight Based on the results of this SLV and the MLT (will be published in a second paper), the Joint FAO/WHO Expert Committee on Food Additives has recently decided to replace the outdated GC-FID JECFA method for the new GC-MS method for the determination of PCH-t content in starches.

Background Antibiotics are used in ethanol production to discourage undesirable bacteria growth. To determine if antibiotic residues remain in the distillers grain (DG) by-product, which is used as an animal food ingredient, the U.S. Food and Drug Administration/Center for Veterinary Medicine previously developed an LC-MS/MS method to detect residues of erythromycin A, penicillin G, virginiamycin M1, and virginiamycin S1 in DG to enable regulatory decision making. Objective Erythromycin and penicillin G were quantitated using the stable isotope dilution (SID) technique with their isotopically labeled compounds, which are considered optimal internal standards (ISTDs) for quantitative mass spectrometry. With the commercial availability of virginiamycin M1-d2 since then, the objectives of this study were to evaluate the feasibility of its use as it is only doubly deuterated, and to incorporate it in the method to enhance method performance. Methods Antibiotic residues were solvent-extracted from DG, the extract was cleaned up by a hexane wash and solid phase extraction, and analyzed by LC-MS/MS. Results We established suitability of virginiamycin M1-d2 as an ISTD and incorporated it in the method. For all analytes, accuracy and precision ranged 90 to 102% and 3.8 to 6.8, respectively. Conclusions We modified a previously developed LC-MS/MS method that uses virginiamycin M1-d2 as an ISTD to support surveillance studies to determine several drugs in DG. Highlights Virginiamycin M1-d2 was successfully incorporated into the method for better virginiamycin M1 quantitation. This addition also allowed calibration curves for all analytes to be constructed in solvent thereby simplifying the method.

Background Flibanserin (FLB) was firstly synthesized as an antidepressant drug; however, due to its enhancing effects on sexual activity, it was approved for treatment of hypoactive sexual desire disorder in women in 2015. Objective The aim of this study was to develop a new and fully validated HPLC method for analysis of FLB in pharmaceutical formulations besides its degradation products, and identification of possible formation mechanisms by using HPLC-DAD-ESI-IT-TOF-MSn. Methods The HPLC separation was achieved in an Supelco Ascentis® Express series phenyl hexyl column (100 × 4.6 mm, ID 2.7µm). The mobile phase was acetonitrile-ammonium acetate solution (50:50, v/v, 10 mM, pH 5.4) mixture, which was pumped at the rate of 0.5 mL/min. Chromatography, detection and structural identification was performed by using LCMS-IT-TOF instrument (Shimadzu, Japan). Results 1-(2-(4-(3-hydroxy-5-(trifluoromethyl)phenyl)piperazine-1-yl)ethyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one is proposed as a novel degradation product, with a mass of 407.1695, and a formula of C20H21F3N4O2 with a margin of error about 0.001 ppm. The developed method is applicable with 98% accuracy within 2.5-50.0 µg/mL range. The LOD and LOQ was about 500 ng/mL and 1.50 µg/mL, respectively. The transferability and variation between laboratories were tested by inter-laboratory comparison and evaluated with one-way analysis of variance. Conclusions A novel FLB degradation product, which was produced under oxidative forced degradation condition was observed and identified for the first time; in addition, the formation kinetics of the degradation product, besides decomposition of FLB was studied. Furthermore, an inter-laboratory comparison was carried out and application of the proposed method on pseudo Addyi® sample was tested using both instrument configurations. Highlights A novel stability indicating assay method was developed and fully validated according to ICH (Q2)R1 for the analysis of FLB in the pharmaceutical preparations. A new degradation product was identified in the oxidative forced degradation condition and characterized using HPLC–DAD–ESI-IT-TOF-MS3. Moreover, the possible mechanism and the formation kinetic of the degradation product were revealed. In addition, the developed method was transferred to another LC-PDA instrument for inter-laboratory comparison. Finally, the current method was applied to pseudo formulation of Addy® in both instruments and ANOVA was applied for evaluation.

Background Schizonepetae Herba (SH, Jingjie) and Schizonepetae Herba Carbonisata (SHC, Jingjie Tan) are two different forms of the same herbal material, with SHC being the processed product of SH. The different clinical efficacies of SH and SHC may be caused by changes in their chemical compositions. Despite this, there have been few studies that have reported on the comparative identification of SH and SHC. Therefore, the aims of this experiment are to investigate the differential changes of non-volatile and volatile components before and after SH processing. Objectives To establish combination strategies for identifying the chemical markers in SH and SHC, ultra-high-performance liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-Q-TOF-MS) and headspace gas chromatography mass spectrometry (HS-GC-MS) to use. Methods An untargeted metabolomics approach using UHPLC-Q-TOF-MS and HS-GC-MS was utilized to comprehensively discriminate between SH and SHC. To identify chemical markers, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were performed on 14 batches of SH and SHC. Results A total of 71 non-volatile compounds and 81 volatile compounds were tentatively identified in SH and SHC. Among these, 14 non-volatile compounds and 18 volatile oils were found to be potential characteristic markers that can differentiate between SH and SHC. Conclusions The present work provides valuable information for understanding the chemical differences between SH and SHC. The results obtained from this research may serve as a scientific foundation for comprehensively revealing the mechanisms involved in the carbonizing processing method of stir-frying SH. Highlights The chemical changes that occur before and after carbonizing Schizonepetae Herba were investigated using integrated methods based on LC-MS and GC-MS, and chemical markers in SH and SHC were identified.

Background Platycladus orientalis leaves (POL), as the sources of the traditional Chinese medicine Platycladi Cacumen, has frequently been found to be misused with five adulterants including Chamaecyparis obtusa leaves (COL), Cupressus funebris leaves (CFL), Juniperus virginiana leaves (JVL), Sabina chinensis leaves (SCL) and Juniperus formosana leaves (JFL). Objective The purpose of this study was to distinguish POL (fresh leaves) from its five adulterants (fresh leaves). Methods The micromorphological features in terms of transection and microscopic characteristics of POL and adulterants were captured and compared by the optical microscope. Both a high-performance liquid chromatography (HPLC) method and a thin layer chromatography (TLC) method for the simultaneous determination of six bioactive flavonoids (myricitrin, isoquercitrin, quercitrin, amentoflavone, afzelin and hinokiflavone) had been developed. Results There were significant differences in microscopic features of transverse section and powders. The TLC results suggested that the spots of myricitrin in POL were more obvious than those in five adulterants. The contents of myricitrin and quercitrin, or the total contents of flavonoids in POL, determined by HPLC, were significantly higher than those in adulterants. Conclusion POL was successfully distinguished from its five adulterants by the comparison of morphology, microscopic characteristics and chemical profiles. Highlights This research provided a comprehensive morphology, microscopic identification, TLC and HPLC anaylsis for authenticating POL and its five adulterants.

This review could be of significant importance in the area of ivermectin analytical method development studies. Background Ivermectin is one of the first, safe, broad spectrum, avermectin (AVM) class of antiparasitic agent, widely used to control parasitic growth in livestock. Ivermectin being highly lipophilic accumulates in fat tissues, causing its long-term existence in the body. Accordingly ivermectin residues are observed in various animal products such as milk and meat causing several health hazards. Therefore, monitoring of ivermectin residue levels in the various food products of animal origin is greatly important to ensure the safety of the consumers. Objective/Methods In this context, a critical review of analytical methods present in the literature for detection and quantification of ivermectin in pharmaceutical formulations, and biological materials including animal tissues. Results Several analytical methods based on HPLC FLD, UV-DAD, HPLC-MS/MS, UPLC-MS/MS, and capillary electrophoresis techniques have been utilized for the simultaneous determination of ivermectin singly or in the presence of other drugs with realistic retention times. Several derivatization strategies were used to introduce fluorophore followed by extraction into organic phase to remove the matrix interferences and enhance the sensitivity by pre-concentration. Methods such as HPLC linked to tandem-mass spectrometry were developed to lower the detection limit and quantification limit, with no requirement for derivatization. Conclusions More simple, selective, fast, sensitive and green chemistry oriented methods for ivermectin analysis are needed to be developed. Novel analytical devices based on pulsed electrochemical methods, voltammetry and amperometry can be developed for real time analysis of ivermectin in addition to biosensors based on nanotechnology including quantum dots and nanoparticles etc. Highlights Various methods described in the review include high-performance liquid chromatography techniques with ultraviolet or visible spectrophotometric, fluorescence and mass spectrometric detection techniques, capillary electrophoresis and immunological methods.

Background Atractylodes chinensis (DC.) Koidz. (A. chinensis) is a perennial herbaceous plant that is widely used as a Chinese medicine herb to gastric diseases. However, the bioactive compounds of this herbal medicine have not been defined, and quality control is imperfect. Objective Although the method of quality evaluation method for A. chinensis by high performance liquid chromatography (HPLC) fingerprinting has been reported in related papers, it remains unknown whether the chemical markers selected are representative of their clinical efficacy. To develop methods for qualitative analysis and improved quality evaluation of A. chinensis. Methods In this study, HPLC was used to establish fingerprints and conduct similarity evaluation. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to reveal the differences of these fingerprints. Network pharmacology was used to analyze the corresponding targets of the active ingredients. Meantime, an active ingredient-target-pathway network was constructed to investigate the characteristics of the medical efficacy of A. chinensis and to predict potential Q-markers. Results Combining network pharmacological effectiveness and composition specificity with the Q-Marker concept, atractylodin (ATD), β-eudesmol, atractylenolide Ι (AT-I) and atractylenolide III (AT-III) were predicted to be potential Q-Markers of A. chinensis that showed anti-inflammatory, anti-depressant, anti-gastric and antiviral effects by acting on 10 core targets and 20 key pathways. Conclusions The HPLC fingerprinting method established in this study is straight forward, and the identified four active constituents can be utilized as Q-markers of A. chinensis. These findings facilitate effective quality evaluation of A. chinensis and suggest this approach could be applied to evaluate the quality of other herbal medicines Highlights The fingerprints of Atractylodis Rhizoma were organically combined with network pharmacology to further clarify its criteria for quality control.

Background Careful review of the scientific databases revealed that no stability-indicating analytical method is available for the binary mixture of Allopurinol (ALO) and Thioctic Acid (THA). Objective A comprehensive stability-indicating HPLC-DAD procedure has been executed for concurrent analysis of ALO and THA. Methods Successful chromatographic separation of the cited drugs was reached using Durashell C18 column (4.6 × 250 mm, 5 µm particle size). The mobile phase was comprised of a mixture of acidified water (pH 4.0) using phosphoric acid and acetonitrile pumped in gradient elution mode. For quantification of ALO and THA, their respective peak areas were recorded at 249 nm and 210 nm. A systematic validation of analytical performance was investigated in terms of system suitability, linearity, ranges, precision, accuracy, specificity, robustness, detection and quantification limits. Results ALO and THA peaks emerged at retention times 4.26 and 8.15 min, respectively. Linear ranges for ALO and THA were 5–100 µg/mL and 10–400 µg/mL, respectively with correlation coefficient values exceeding 0.9999. Both drugs were exposed to conditions of neutral, acidic and alkaline hydrolysis, oxidation and thermal decomposition. Stability-indicating features have been demonstrated by resolution of the drugs from their forced degradation peaks. For verification of peak identity and purity, the diode-array-detector (DAD) was utilized. In addition, degradation pathways for the cited drugs were postulated. Besides, separation of both analytes from about 13 medicinal compounds of different therapeutic classes disclosed optimum specificity of the proposed method. Conclusion Advantageous application of the validated HPLC method for the concurrent analysis of ALO/THA in their tablet dosage form was accomplished. Highlights So far, the described HPLC-DAD method is considered the first detailed stability-indicating analytical study for this pharmaceutical mixture.

Background Taurine is recognized as an essential growth factor and as being critical in the maintenance of functional tissue regulation. Objective To evaluate the analytical performance of a hydrophilic interaction liquid chromatography–tandem mass spectrometry (HILIC-MS/MS) method for compliance with AOAC Standard Method Performance Requirements (SMPR®) for taurine analysis described in SMPR 2014.013. Method Following protein precipitation with Carrez solutions, taurine is extracted and separated by HILIC with detection by triple quadrupole MS using multiple reaction monitoring. Stable isotope labelled taurine internal standard is used for quantification to correct for losses in extraction and variations in ionization in the ion source. Results The method was shown to meet the requirements specified in the SMPR with a linear range of 0.27–2700 mg/hg RTF (ready-to-feed), a limit of detection of 0.14 mg/hg RTF, acceptable recovery of 97.2–100.1%, and acceptable repeatability of 1.6–6.4% relative standard deviation. Additionally, the method was found to have no statistically significant bias compared with reference values for National Institute of Standards and Technology (NIST) 1849a certified reference material (CRM) (p-value = 0.95) and 1869 CRM (p-value = 0.31), and with results from AOAC 997.05 (p-value = 0.10). Conclusions A recent review of the method and validation data by the Stakeholder Program on Infant Formula and Adult Nutritionals (SPIFAN) Expert Review Panel (ERP) found that this method met all the criteria for analysis of taurine specified in SMPR 2014.013 and voted to adopt this method as First Action AOAC Official Method 2022.03. Highlights A method for the analysis of taurine in infant formulas and adult nutritionals by HILIC–MS/MS is described. A single laboratory validation study demonstrated the applicability of the method to meet requirements of SMPR 2014.013. In December 2022, the SPIFAN ERP voted to adopt this method as First Action AOAC Official Method 2022.03.

Background Linezolid (LNZ) is extremely prone to resistance. The development of resistance to LNZ should be taken into consideration when selecting this drug as a therapeutic option. By considering the hypothesis that reactive oxygen species (ROS) generated by iron oxide nanoparticles (MNPs) could kill the infecting bacteria. So, we hypothesized the synergistic antibacterial effect of iron oxide nanoparticles and LNZ. Objective To study the release and antibacterial effects of LNZ loaded superparamagnetic iron oxide nanoparticles (SPIONs) on Staphylococcus aureus and Streptococcus pneumoniae. Methods Ferrofluid containing SPIONs was synthesized via chemical co-precipitation method and stabilized by sodium lauryl sulphate (SLS). SPIONs were then loaded with LNZ and characterized for particle size, FT-IR, XRD, and entrapment efficiency. Further antibacterial activity of SPIONs and LNZ-loaded SPIONs was investigated. For the in-vitro release findings, a HPLC analytical method development and validation was performed. Results Isolation of LNZ was accomplished on a C-18 column with methanol: TBHS (Tetra butyl ammonium hydrogen sulphate, 50:50 v/v). The eluate was monitored at 247 nm with retention time of 4.175 min. The MNP’s DLS measurement revealed monodispersed particles with an average size of 16.81 ± 1.07 nm and PDI 0.176 ± 0.012. In optimized formulation, 25 ± 1.75% (w/w) of the drug was found to be entrapped. XRD revealed uniform coating of oleic acid covering the entire magnetic particles surface with no change in its crystallinity. An effective antimicrobial activity was observed at the lowered dose of drug. Conclusion A robust HPLC method was developed to quantify the LNZ in MNPs and outcomes showed that the reduced dose of LNZ incorporated in SPIONs was able to show similar activity as the marketed product. Highlights Successfully reduction of dose of LNZ was established with the aid of biocompatible MNPs to attain the equivalent antibacterial activity.

Background Due to its medicinal properties, Pistacia integerrima is in high demand and is extensively used as a key ingredient in various formulations. However, its popularity has led to IUCN threatened category list. In Ayurvedic texts, such as Bhaishajaya Ratnavali, Quercus infectoria is recommended as a substitute for P. integerrima in different formulations. Additionally, Yogratnakar highlights that Terminalia chebula shares similar therapeutic properties with P. integerrima. Objective The objective of the current study was to gather scientific data on metabolite profiling and markers-based comparative analysis of Q. infectoria, T. chebula and P. integerrima. Methods In present study, hydro-alcoholic and aqueous extracts of all three plants were prepared and standardized for the comparative evaluation of secondary metabolites. Thin-layer chromatography was carried out for the comparative fingerprinting of the extracts using chloroform: methanol: glacial acetic acid: water (60: 8: 32: 10, v/v/v/v) as a solvent system. A fast, sensitive, selective, and robust HPLC method was developed to determine gallic acid and ellagic acid from both extracts of all three plants. The method was validated for precision, robustness, accuracy, limit of detection and quantitation as per the International Conference on Harmonization guidelines. Results The TLC analysis revealed the presence of several metabolites, and the pattern of metabolites in the plants exhibited a certain degree of similarity. A highly precise and reliable quantification technique was created for gallic acid and ellagic acid, operating within the linear concentration range of 81.18–288.22 µg/mL and 3.83–13.66 µg/mL, respectively. The correlation coefficients for gallic acid and ellagic acid were 0.999 and 0.996, indicating strong relationships. The gallic acid content in all three plants ranged from 3.74% to 10.16% w/w, while the ellagic acid content ranged from 0.10% to 1.24% w/w. Conclusions This pioneering scientific approach highlights the phytochemical similarities between Q. infectoria, T. chebula and P. integerrima.

Background The Thermo Scientific™ SureTect™ Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus PCR Assay method is a real-time PCR method for the multiplex detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus in seafood. Objective The Thermo Scientific SureTect Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus Assay was evaluated for AOAC® Performance Tested Methods℠ certification. Methods Inclusivity/exclusivity, matrix, product consistency/stability and robustness studies were conducted to assess the method’s performance. For the matrix study, the method was validated using the Applied Biosystems™ QuantStudio™ 5 Real-Time PCR Food Safety Instrument and the Applied Biosystems™ 7500 Fast Real-Time PCR Food Safety Instrument against the U. S. Food and Drug Administration Bacteriological Analytical Manual, Chapter 9 (2004), Vibrio and ISO 21872–1:2017 Microbiology of the food chain—Horizontal method for the determination of Vibrio spp. – Part 1: Detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus reference methods. Results Matrix studies showed equivalent or superior performance of the candidate method compared to the reference method and overall, no difference between presumptive and confirmed results, except for one matrix due to high background flora. The inclusivity/exclusivity study correctly identified/excluded all strains analyzed. Robustness testing showed no statistically significant differences in assay performance under varied test conditions. Product consistency and stability studies demonstrated no statistically significant differences between assay lots with different expiration dates. Conclusions The data presented shows that the assay constitutes a rapid and reliable workflow for the detection of V. cholerae, V. parahaemolyticus and V. vulnificus in seafood matrixes. Highlights The SureTect PCR Assay method allows for fast, reliable detection of stipulated strains in seafood matrixes with results obtained in as little as 80 minutes post-enrichment.

Background The CompactDry “Nissui” BC is a ready to use dry media sheet using a chromogenic medium with selective agents for the detection and enumeration of Bacillus cereus in products after incubation at 30 ± 1 °C for 24 ± 2 h. Objective The CompactDry “Nissui” BC method was validated to achieve AOAC Performance Tested Methods SM certification. Methods The performance of the CompactDry “Nissui” BC was compared to that of ISO 7932:2004 for 10 matrixes including panna cotta, double cream, dried baby food, dried vegetable soup mix, seafood sticks, salmon pâté, sliced ham, pork liver pâté, ham and cheese sandwich, and Caesar pasta salad with chicken and bacon. Performance indicators included repeatability, difference of means (DOM), and inclusivity/exclusivity. Results After log10 transformation of the data, the relative standard deviation of repeatability (RSDr) was ≤9.2% for 28 of the 30 materials (10 matrixes each at 3 contamination levels) analyzed by the CompactDry “Nissui” BC method and ≤13% for 27 of the 30 matrix/level combinations analyzed by the reference method. Method equivalence was demonstrated in 28 of the 30 matrix/level combinations based on the 90% confidence interval of the DOM being within (-0.5, 0.5). For inclusivity, 47 of 50 strains tested showed typical colonies and confirmed positive. For exclusivity, 28 of 33 strains tested resulted in no growth or were negative, and 5 were positive. Inclusivity and exclusivity results were similar on the reference method agar. The method was shown to be robust to changes in sample volume, incubation temperature, and incubation time and data are presented supporting product consistency and 18-month shelf life. Conclusions The CompactDry “Nissui” BC method is validated for the determination of Bacillus cereus in a variety of matrixes. Highlights The CompactDry “Nissui” BC method is equivalent to the ISO 7932:2004 reference method and is suitable for Performance Tested MethodsSM certification for the matrixes tested.

Background Coated Wire Electrodes are considered the most effective and selective type of ion selective electrodes because of the modified surface with different materials according to the substance to be analyzed. Objective This study aims to construct the first potentiometric method for analysis of Levocetirizine (LVZ) in its combination form with Montelukast drug. Method A novel potentiometric sensor which consists of a silver wire coated with Zinc Oxide (ZnO) nanorod modified with a polymeric membrane combining β-Cyclodextrin and tetraphenyl borate, plasticized with di-butyl phthalate was constructed for the determination of Levocetirizine.2HCl in its pure form and its combination dosage form. Results The fabricated sensor exhibited a linearity range (5x10 −6- 1x10−2) mol/L with a Nernstian slope 57.88 mV/decade, over pH range (2–4.5), the effect of the temperature on the constructed sensor was studied and it was found that the electrode works effectively over (10–50) C°. The electrode showed a fast response time and the lifetime of the electrode was found to be 72 days without significant change in the Nernstian slope value. The selectivity of the electrode toward levocetirizine was estimated in the presence of some obstructive ions. Conclusions The method was validated according to ICH rules and applied to the determination of levocetirizine in its pure and combined pharmaceutical dosage forms. Highlights This paper introduces the synthesis of the first modified Coated Wire Electrode with ZnO Nanorods for determination the Levocetirizine drug. The obtained results demonstrate the ability of the electrochemical methods to analyze drugs in their combination. The presented method excels over another analytical methods in terms of sensitivity, selectivity and simplicity.

Background This Azelastine HCl (AZ) and Fluticasone propionate (FL) nasal spray drug product is commonly used in the treatment of allergic rhinitis worldwide. The impurity profiling of this product is not reported till date. Objective The present study is aimed to develop and validate a novel stability indicating analytical method for the estimation of impurities from Azelastine Hydrochloride and Fluticasone Propionate nasal spray drug product. Methods A mixture of octane sulfonic acid sodium salt and trifluroacetic acid is used as a mobile phase A. Acetonitrile is used as a mobile phase B. Good separation was achieved on Baker bond phenyl hexyl, 250 x 4.6, 5 µm column at 1 mL/min flow rate in gradient elution mode. The chromatograms were monitored at 239 nm. Results The limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.006 and 0.019 µg/mL for AZ and 0.010 and 0.030 µg/mL for FL, respectively. The correlation coefficient for all the known impurities and principle analytes was observed 0.999 from LOQ level to 150% of standard concentration. The recovery for all the known impurities was found to be between 90 to 110%. During stress study, 15% degradation was observed in basic condition and 8.7% in acidic condition. No significant degradation was observed in thermal and oxidative conditions. Conclusion The impurity profiling method for AZ and FL combination nasal spray product was successfully developed, validated and demonstrated to be accurate, precise, specific, robust and stability indicating. The method can be routinely used for the impurity testing of commercial batches in quality control laboratories in the pharmaceutical industry. Highlights No impurity study has been reported for this combination product till date.

Background In April 2020, the BIOMILK 300 LAC method for the quantification of lactose in lactose-free and low lactose dairy products was adopted as First Action Official Method of Analysis 2020.09 by the AOAC INTERNATIONAL. In order to test the reproducibility of the method, as the last step towards the Final Action status, a collaborative study was organized by BIOLAN Microbiosensores. Objective Fifteen collaborators within the European Union took part in this study, where nine different samples were sent as duplicate blind test portions for lactose determination. The data obtained were used to determine method repeatability and reproducibility and also to validate the new version of the biosensor. Method The test method is based on the direct enzymatic recognition-electrochemical detection of trace levels of lactose over a wide range of dairy samples by means of the BIOMILK 300 biosensor, in less than five minutes, and without intricate sample pre-treatments. Results All samples resulted in a repeatability relative standard deviation (RSDr) of < 8.1% and a reproducibility relative standard deviation (RSDR) of 14.0% maximum, meeting requirements from SMPR 2018.09. Conclusions On the basis of these results, the enzymatic amperometric biosensor method developed by BIOLAN Microbiosensores was adopted as Final Action Official MethodSM in July 2022.

Background Azo dyes are among the most widely used dyes in the textile industry, releasing a series of carcinogenic aromatic amines that can the through skin absorption. Objective This work aims to show that 22 azo dye amines in a textile matrix may be quantified using a GC-MS method. Method Based on the notion of total error and β-content, γ-confidence statistical intervals (β,γ CCTI), a chemometric approach known as the “Uncertainty Profile” has been used to completely validate a gas chromatography coupled with mass spectrometry (GC-MS) method for the simultaneous assay of 22 azo amines in fabrics. According to ISO 17025 guidelines, analytical validation and measurement uncertainty estimates have evolved to be two main principles for ensuring the accuracy of analytical results and controlling the risk associated with their use. Results The calculated tolerance intervals allowed for the determination of the uncertainty limits at each concentration level. These limits when compared to the acceptable limits show that a significant portion of the expected outcomes is in conformity. Additionally, the relative expanded uncertainty values, calculated with a proportion of 66.7% and a 10% risk, do not exceed 27.7%, 12.2%, and 10.9% for concentration levels 1 mg/L, 15 mg/L, and 30 mg/L respectively. Conclusions The capability and flexibility of the intervals β-content, γ-confidence have been established through the use of this innovative approach to carrying out a qualimetry of the GC-MS method depending on the behavior, required conformity proportion, and acceptable tolerance limits of each amine. Highlights An efficient GC-MS technique for the simultaneous determination of 22 azo amines in a textile matrix has been completed. Analytical validation using a new strategy based on the uncertainty concept is reported, uncertainty associated to measurement results was estimated and the applicability of our approach to the GC-MS method is investigated.

Background Spectrophotometric resolution of a mixture of several drugs is considered a cheaper, simpler and more versatile alternate compared to the costly chromatographic instruments. Objective The work aims to resolve the interfering spectra of ephedrine hydrochloride, naphazoline nitrate and methylparaben in nasal preparations using smart spectrophotometric methods. Methods In our work, derivative and dual wavelength methods were combined to eliminate this interference, under the name of derivative dual wavelength method. Other methods, namely successive derivative subtraction and chemometric analysis were also able to eliminate this interference. The methods have proven their applicability as they follow the ICH requirements regarding repeatability, precision, accuracy, selectivity, and linearity. Eco- scale, GAPI and AGREE tools were used to estimate the possible environmental effects of the methods. Results Acceptable results of repeatability, precision, accuracy, selectivity, and linearity were obtained. LOD values were 2.2 for ephedrine and 0.3 for naphazoline. The correlation coefficients were above 0.999. The methods were proven to be safe for application. Conclusion The introduced methods are cheap and easily implemented compared to chromatographic techniques. They can be used in purity checking of raw material and estimation of concentrations in market formulations. The replacement of the published chromatographic techniques with our developed methods is usefulwhen saving money, effort and time is needed. Highlights The three components of a decongestant nasal preparation were determined using cheap green versatile spectrophotometric methods that keep the advantages of chromatographic techniques including accuracy, reproducibility and selectivity.

Background Computationally-designed molecular imprinted polymers (MIP) incorporation into electrochemical sensors has many advantages to the performance of the designed sensors. The innovative self-validated ensemble modeling (SVEM) approach is a smart machine learning-based technique that enabled the design of more accurate predictive models utilizing smaller data sets. Objective The novel SVEM experimental design methodology is exploited here exclusively to optimize the composition of four eco-friendly PVC membranes augmented by a computationally designed magnetic molecularly imprinted polymer to quantitatively determine drotaverine hydrochloride in its combined dosage form and human plasma. Besides, the application of hybrid computational simulations such as molecular dynamics and quantum mechanical calculations (MD/QM) is a time-saving and eco-friendly provider for the tailored design of the MIP particles. Methods Here, for the first time, the predictive power of machine learning is assembled with computational simulations to develop four PVC-based sensors decorated by computationally designed MIP particles utilizing four different experimental designs known as central composite, SVEM-LASSO, SVEM-FWD, and SVEM-PFWD. The pioneering Agree approach further assessed the greenness of the analytical methods, proving their eco-friendliness. Results The proposed sensors showed decent Nernstian responses towards drotaverine hydrochloride in the range of (58.60—59.09 mV/decade) with a linear quantitative range of (1 x 10−7—1 x 10−2 M) and limits of detection in the range of (9.55 x 10−8 – 7.08 x 10−8 M). Moreover, the proposed sensors showed ultimate eco-friendliness and selectivity for their target in its combined dosage form and spiked human plasma. Conclusion The proposed sensors were validated as per IUPAC recommendations proving their sensitivity and selectivity for drotaverine determination in dosage form and human plasma. Highlights This work presents the first ever application of both the innovative SVEM designs and MD/QM simulations in the optimization and fabrication of drotaverine-sensitive and selective MIP-decorated PVC sensors.