Quality Control Laboratories Research Articles

“Eco-friendly HPLC method for analysis of dipyrone and hyoscine in different matrices with biomonitoring”

A selective, precise, and accurate reversed HPLC method has been developed and validated for simultaneous separation and determination of two veterinary drugs, dipyrone and hyoscine, in their combined dosage form in the presence of their official impurities, namely 4-aminoantipyrine and tropic acid, in addition to the formulated preservative: phenol. The linearity range was found to be (1.00–35.00 µg/mL) for dipyrone and (2.50–50.00 µg/mL) for hyoscine. It exhibited a satisfactory linearity regression R (0.9999) for both drugs with LOD 0.22 µg/mL and 0.72 µg/mL and LOQ 0.65 µg/mL and 2.19 µg/mL for dipyrone and hyoscine, respectively. Additionally, the two cited drugs were also determined in the presence of dipyrone active metabolite 4-aminoantipyrine using diclofenac as an internal standard in bovine urine. The linearity range was found to be (15–75 µg/mL) for dipyrone, (2.5–60 µg/mL) for hyoscine, and (2.5–60 µg/mL) for 4-aminoantipyrine with linearity regression R (0.9999–0.9998). The LLOQ (15, 2.5, 2.5 µg/mL), LQC (45, 7.5, 7.5 µg/mL), MQC (55, 25, 25 µg/mL), and HQC (60, 50 50 µg/mL) were determined for dipyrone, hyoscine and 4-aminoantipyrine, respectively. UV detection was carried out at 220 nm. The method was validated according to the ICH guidelines, as well as according to FDA guidelines for determining both drugs in bioanalytical matrices and both proved accuracy and precision. A statistical comparison was made between the results obtained and those obtained by the reported method, showing no significant difference in accuracy and precision at p = 0.05. The suggested method was proved eco-friendly through a greenness assessment using two different tools (The analytical eco-scale scored 83, and the AGREE-Analytical Greenness Metric approach scored 0.83). The suggested method can be used in the routine work of quality control labs, screening for drug abuse, and ensuring clean sport for horse racing.

Unravelling red deer (Cervus elaphus) meat adulteration in gourmet foods by quantitative real-time PCR

Red deer (Cervus elaphus) meat is highly regarded as an exclusive gourmet food due to its sensory quality, nutritional value and premium price, thus being susceptible to adulteration practices by its substitution with inferior meat species. This study proposes a novel real-time PCR method targeting the troponin I gene for detecting and quantifying red deer meat in processed food products. The assay showed acceptable performance parameters, achieving absolute and relative limits of quantification of 0.01 ng of DNA and 0.5% (w/w) of red deer meat in both raw and thermally processed pâté. The assay parameters, using raw and autoclaved reference mixtures, regarding PCR efficiency (89.3% and 100.0%, respectively), slope of the calibration curve (−3.6067 and −3.3224, respectively) and R2 (0.9889 and 0.9893, respectively) highlight the high performance of the calibration models for the determination of red deer meat. The method was successfully validated using blind mixtures and applied to 46 commercial meat products from Poland, Portugal and Spain. Data suggested the adulteration of 48% of the samples by the total or partial substitution of red deer species and the suitability of the method for the routine testing of game meat products in quality control laboratories.

Establishment of genomic RNA reference materials for BCR-ABL1 P210 measurement.

Quantitation of BCR-ABL1 with thequantitative reverse transcriptase polymerase chain reaction (RT-PCR) is very important in monitoring chronic myeloid leukemia (CML), which relies on an RNA reference material. A genomic RNA reference material (RM) containing the BCR-ABL1 P210 fusion mutation was developed, and an absolute quantitative method based on one-step reverse transcription digital PCR (RT-dPCR) was established for characterizing the RM. The proposed dPCR method demonstrates high accuracy and excellent analytical sensitivity, as shown by the linear relationship (0.94 < slope < 1.04, R2≧0.99) between the measured and nominal values of b2a2, b3a2, and ABL1-ref within the dynamic range (104-101 copies/reaction). Homogeneity and stability assessment based on dPCR indicated that the RM was homogeneous and stable for 24months at -80°C. The RM was used to evaluate inter-laboratory reproducibility in eight different laboratories, demonstrating that participating laboratories could consistently produce copy concentrations of b3a2 and ABL1-ref, as well as the BCR-ABL1/ABL1 ratio (CV < 2.0%). This work suggests that the RM can be employed in establishing metrological traceability for detecting mutations in the BCR-ABL1 fusion gene, as well as in quality control for testing laboratories.

Development and validation of stability indicating HPTLC method for estimation of Daclatasvir Dihydrochloride in pharmaceutical dosage form

A high-performance thin-layer chromatographic method was developed and validated for estimation of Daclatasvir Dihydrochloride in pharmaceutical dosage form. The proposed method was applied successfully to the pharmaceutical analysis of the recently approved dosage form of Daclatasvir Dihydrochloride which is available in market as a brand name of ‘NALDAC 60’ tablets. The drugs were satisfactorily show peak with RF 0.38 for Daclatasvir Dihydrochloride. Method was validated according to the ICH guidelines. The calibration plot was linear between 50-300 ng per band for Daclatasvir Dihydrochloride. The LOD and LOQ for Daclatasvir Dihydrochloride were found to be 0.171 μg per band and 0.521 μg per band, respectively. Accuracy and precision of the proposed method was evaluated by recovery studies (% recovery for Daclatasvir Dihydrochloride was 99.88%) and intra-day and inter-day precision studies (standard deviation value for precision studies was found to be 0.453). In stability testing, Daclatasvir Dihydrochloride was found susceptible to acid hydrolysis and alkaline degradation. Because the method could effectively separate the drugs from their degradation products, it can be used as a stability indicating method. The proposed validated stability indicating assay for the sensitive determination of the mentioned drugs is suitable for Quality control laboratories as a simple fast economic method. Degradation product of Daclatasvir Dihydrochloride in alkaline condition was carried out.

Chromatographic fingerprinting of ipratropium and fenoterol in their novel co-formulated inhaler treating major respiratory disorders; application to delivered dose uniformity testing along with greenness and whiteness assessment

Ipratropium bromide (IPR) and fenoterol hydrobromide (FEN) have recently been combined in a promising inhaler to treat two prevalent inflammatory illnesses of the airways: bronchial asthma and chronic obstructive pulmonary disease (COPD). The necessity for a single, sensitive, and trustworthy analytical approach to cover the diverse and necessary tests of in-vitro and in-vivo studies is greatly grown with the rising production of new fixed combinations. Two novel, selective and environmentally friendly LC techniques were developed in order to guarantee precise measurement of IPR and FEN in their challenging formulation. The initial technique involved high-performance thin-layer chromatography (HPTLC) in conjunction with densitometric quantification. Chromatographic separation was attained on HPTLC plates utilizing ethyl acetate - ethanol - acetic acid (5.0:5.0:0.1, by volume) as a developing system. Densitometric quantification of the separated bands was carried out at 220.0 nm over concentration ranges of 0.50–15.0 µg/band for IPR and 0.50–12.0 µg/band for FEN. High-performance liquid chromatography (HPLC) paired with diode array detection (DAD) was the core of the second technique. The optimized separation was achieved on a Zorbax SB C18 (150 × 4.6 mm, 5 μm) column with a combination of 10.0 mM potassium dihydrogen orthophosphate, pH 5.0 ± 0.1, adjusted with o-phosphoric acid and methanol (70:30, v/v) as the mobile phase and pumped at flow rate of 1.0 mL/min. The peaks were monitored at 220.0 nm using diode array detection, achieving linearity range of 5.0–200.0 µg/mL for both drugs. The ICH criteria have been verified and both methods have been confirmed to be valid, and successfully applied for assay the cited drugs in the Atrovent® comp HFA metered dose inhaler as well as delivered dose uniformity testing of the final product. Finally, whiteness appraisal and several state-of-the-art green evaluation metrics were applied to evaluate the sustainability of the proposed methods. The suggested approaches produced promising results and are the first simple and sustainable methodologies for the simultaneous quantification of both drugs in different real samples, all of which strongly suggest their application in quality control laboratories.

Risk assessment based on spectrophotometric signals used in eco-friendly analytical scenarios for estimation of carvedilol and hydrochlorothiazide in pharmaceutical formulation

Special attention is given to the pharmacological treatment of combined medication of Carvedilol and hydrochlorothiazide which is the most effective and the most beneficial therapy for hypertensive patients with diabetes and various metabolic comorbidities. This work represents spectrophotometric platform scenarios based on factorized spectrum (FS) using interpoint data difference resolution scenarios (IDDRS) coupled with spectrum subtraction method (SS) for the concurrent quantification of carvedilol (CAR) and hydrochlorothiazide (HCT) when present together in a combination without the need for any initial physical separation steps. This IDD resolution scenario based on manipulating the zero-order spectra (D0) of both drugs in the mixture with various spectral features at different wavelength regions (200–400 nm), region I (220–250 nm), region II (240–300 nm) and region III (270–320 nm) via absorbance resolution (AR) and induced absorbance resolution (IAR) methods coupled with corresponding spectrum subtraction (SS). The calibration curves were established across the linearity ranges of 2.0–12.0 µg/mL at 242.50 nm and 4.0–40.0 µg/mL at 285.5 nm for CAR and 1.0–11.0 µg/mL at 226.10 nm and 2.0–20.0 µg/mL at 270.5 nm for HCT. Moreover, methods’ validation was confirmed via ICH guidelines. A Multicenter comparison between sensitivity, specificity in respect resolution sequence were applied using different wavelength regions with various concentration ranges was applied and finally spectral resolution recommendation is issued and cumulative validation score (CVS) is calculated as an indicator in the risk analysis. In quality control laboratories, the studied approaches are applicable for conducting analysis on the mentioned drugs. In addition, the selection of spectrophotometry aligns with the principles of green analytical chemistry, an approach that resonates with the overarching theme of minimizing environmental impact. Via four metric tools named: analytical greenness (AGREE), green analytical procedure index (GAPI), analytical eco-scale, and national environmental method index (NEMI), methods’ greenness profile was guaranteed.

Machine Learning for Patient-Based Real-Time Quality Control (PBRTQC), Analytical and Preanalytical Error Detection in Clinical Laboratory.

The rapidly evolving field of machine learning (ML), along with artificial intelligence in a broad sense, is revolutionising many areas of healthcare, including laboratory medicine. The amalgamation of the fields of ML and patient-based real-time quality control (PBRTQC) processes could improve the traditional PBRTQC and error detection algorithms in the laboratory. This narrative review discusses published studies on using ML for the detection of systematic errors, non-systematic errors, and combinations of different types of errors in clinical laboratories. The studies discussed used ML for detecting bias, the requirement for re-calibration, samples contaminated with intravenous fluid or EDTA, delayed sample analysis, wrong-blood-in-tube errors, interference or a combination of different types of errors, by comparing the performance of ML models with human validators or traditional PBRTQC algorithms. Advantages, limitations, the creation of standardised ML models, ethical and regulatory aspects and potential future developments have also been discussed in brief.

Carbon Nano-Dots based potentiometric sensor for the assay of the antiviral agent; Valacyclovir: In-Depth DFT Calculations, application to the pharmaceutical formulation and greenness assessment

Valacyclovir (VAL), as a prodrug to acyclovir antiviral agent, has recently held a lot of interest for its efficacy boosting owing to the significant improvement in bioavailability. This makes it a successful candidate in the management of herpes simplex strains as well as CORONA viruses. In this work a carbon nano dot-modified glassy carbon electrode (GCE) has been optimized as potentiometric sensor for the assay of VAL through a systematic approach. A mixture components of polyvinyl chloride (PVC) membrane have been selected and fine-tuned for optimum sensitivity and selectivity. The best results have been obtained by drop casting a blend in % w/w of PVC; 30.85, tricrasyl phosphate (TCP) as plasticizer; 64.32, phosphotungstic acid (PT) as cation exchanger; 1.61 and phosphorus – nitrogen-doped carbon nano-dots (PNCDs); 3.22 over the GCE. The density function theory (DFT) calculations have been applied to the optimized structures of VAL and the CDs and have proven high binding affinity of 46 kcal/mol dominated by π-π stacking and hydrogen bonding. The developed sensor has exhibited a swift response time of six seconds at a 5.00 pH value. The ICH guidelines have been applied to validate the optimized parameters. A good linearity range has been obtained from 1.18 × 10-3 down to 1.07 × 10-6 M for a correlation coefficient 0.9996 with a Nernstian slope of 33.16 mV/decade. The greenness has been assessed using the AGREE approach, giving a score of 0.8, which has indicated the environmental friendliness. An average % recovery of almost 99.00 % was obtained by application to the tablet formulations and furtherly it has been monitored by the standard addition method, giving recoveries varied from 98.00 and up to 102.00 % for relative standard deviation values (%RSD) less than 1.50 %, evidencing efficiency for application in the quality control laboratories.

Quality evaluation of selected expired fluoroquinolones medicines obtained from the public hospitals in Jimma zone, Oromia regional state, Ethiopia.

The problem of medicine expiration presents a notable obstacle, resulting in considerable financial losses. Nevertheless, there is currently limited data indicating that certain medications do not experience a significant decrease in effectiveness after their expiration date. Therefore, the aim of the study was to assess the physico-chemical quality of expired fluoroquinolone antibiotics. The expired samples of fluoroquinolone antibiotics were purposively collected from public hospitals in the Jimma zone of the Oromia regional state, Ethiopia. A World Health Organization quality evaluation sampling strategy was employed. Then, simple random sampling techniques were utilized for the selection of tablets for the laboratory quality control test. The assay, identification, and dissolution were performed in accordance with the United States Pharmacopeia (USP) guidelines, as well as failure mode and effect analysis (FMEA) techniques. The finding revealed that about 100% (7/7) expired samples passed pharmacopeia quality specifications for identity and assay tests. However, of the seven expired brands, about 14.3% (1/7) of the sample (Code-002) was unable to release its API content within the USP criteria of 30 min. The risk-based quality evaluation revealed that assay was the most critical quality attributed to ciprofloxacin tablets (RPN = 189), followed by identity (RPN = 100). Assay was also the most critical quality attribute (RPN = 378), followed by identity (RPN = 100) for Norfloxacin tablets. The risk-based desirability function approach showed that 75% (3/4) of ciprofloxacin products were of good quality, and 25% (1) were found to be of acceptable quality, while the desirability function of norfloxacin tablets was found to be excellent 1 (33.3%), good 1 (33.3%), and acceptable 1 (33.3%). The study revealed that medications can maintain their quality beyond their labeled expiration date. By combining pharmacopeial standards with risk-based approaches like failure mode and effect analysis (FMEA), the study provides a comprehensive evaluation framework. This approach not only confirms the continued effectiveness of expired fluoroquinolone antibiotics but also underscores the potential waste reduction and cost-saving benefits. This could significantly contribute to addressing healthcare challenges in low-resource settings, promoting more efficient pharmaceutical resource utilization.

Whole-genome sequencing for antimicrobial surveillance: species-specific quality thresholds and data evaluation from the network of the European Union Reference Laboratory for Antimicrobial Resistance genomic proficiency tests of 2021 and 2022.

As antimicrobial resistance (AMR) surveillance shifts to genomics, ensuring the quality of whole-genome sequencing (WGS) data produced across laboratories is critical. Participation in genomic proficiency tests (GPTs) not only increases individual laboratories' WGS capacity but also provides a unique opportunity to improve species-specific thresholds for WGS quality control (QC) by repeated resequencing of distinct isolates. Here, we present the results of the EU Reference Laboratory for Antimicrobial Resistance (EURL-AR) network GPTs of 2021 and 2022, which included 25 EU national reference laboratories (NLRs). A total of 392 genomes from 12 AMR-bacteria were evaluated based on WGS QC metrics. Two percent (n = 9) of the data were excluded, due to contamination, and 11% (n = 41) of the remaining genomes were identified as outliers in at least one QC metric and excluded from computation of the adjusted QC thresholds (AQT). Two QC metric correlation groups were identified through linear regression. Eight percent (n = 28) of the submitted genomes, from 11 laboratories, failed one or more of the AQTs. However, only three laboratories (12%) were identified as underperformers, failing across AQTs for uncorrelated QC metrics in at least two genomes. Finally, new species-specific thresholds for "N50" and "number of contigs > 200 bp" are presented for guidance in routine laboratory QC. The continued participation of NRLs in GPTs will reveal WGS workflow flaws and improve AMR surveillance data. GPT data will continue to contribute to the development of reliable species-specific thresholds for routine WGS QC, standardizing sequencing data QC and ensure inter- and intranational laboratory comparability.IMPORTANCEIllumina next-generation sequencing is an integral part of antimicrobial resistance (AMR) surveillance and the most widely used whole-genome sequencing (WGS) platform. The high-throughput, relative low-cost, high discriminatory power, and rapid turnaround time of WGS compared to classical biochemical methods means the technology will likely remain a fundamental tool in AMR surveillance and public health. In this study, we present the current level of WGS capacity among national reference laboratories in the EU Reference Laboratory for AMR network, summarizing applied methodology and statistically evaluating the quality of the obtained sequence data. These findings provide the basis for setting new and revised thresholds for quality metrics used in routine WGS, which have previously been arbitrarily defined. In addition, underperforming participants are identified and encouraged to evaluate their workflows to produce reliable results.

Capacity Assessment of the National Medicines Regulatory Authority in a Low -Income Country.

Access to medical products of the required efficacy, quality and safety is essential for everyone's health and wellbeing. To achieve this milestone, every country needs a robust and strong performing National Regulatory Authority (NRA) that is independent and outcome oriented. With the help of the World Health Organization (WHO), the global benchmarking tool is the gold standard used to assess the regulatory capacity of NRAs. This study assessed the capacity of the National Medicines Regulatory Authority in Sierra Leone to perform its regulatory functions. This descriptive cross-sectional study used both qualitative and quantitative approaches. A self-administered questionnaire was used for the quantitative approach, and the qualitative aspect consisted of a desk review looking at key regulatory documents such as laws, regulations, policies, guidelines, standard operating procedures and reports. The data collection tool used was the WHO global benchmarking tool (GBT) for "Evaluation of National Regulatory System of Medical Product Version VI. The majority of the participants had a postgraduate degree (60%), and 72% had over 10 years of experience working at the NRA. Out of 251 sub-indicators assessed, 85 (34%) sub-indicators were fully implemented. Of the eight (8) functions assessed, sub-indicators related to clinical trial oversight and vigilance were the most implemented, with 67% and 62%, respectively. Of the 9 indicators assessed, 79% of the sub-indicators that are related to quality and risk management were implemented. The results of this study showed that PBSL operates at maturity level 1. The absence of laws and regulations that give PBSL the mandate to perform its regulatory functions was a major challenge even though other indicators were met. The study reported other challenges toward effective functioning, including but not limited to a lack of sufficient staff, weak enforcement of the sale of medicines and a poorly equipped quality control laboratory.

Box-Behnken design in optimization of the green liquid chromatographic method for the quantification of Afatinib in drug product: AQbD approach

The aim of this research work was to quantitatively determine the tyrosine kinase inhibitor, Afatinib, in tablets using an eco-friendly chromatographic method. The development of a green liquid chromatographic method using analytical quality-by-design (AQbD) approach principles to estimate Afatinib via Box-Behnken design. A simple and effective green liquid chromatographic method was developed and validated. The analysis was done on the Zodiac C18 column by isocratic elution mode. Mobile phase composed of pH 3.0 potassium dihydrogen phosphate buffer and ethanol in the ratio of 65:35%V/V. The flow rate was set to 0.6 mL/min. with an injection volume of 10 µL and at detection wavelength of 254 nm. The optimized method was found to be specific, linear, precise, accurate, and robust. Established linearity in the range of 24.6 to 73.9 µg/mL. The developed method exhibited excellent greenness. Based on the greenness assessment tools, the analytical eco-score value of 94 and the pictograms of NEMI, Modified NEMI, GAPI, Complex GAPI, AGREE, and AGREE prep expressed the method’s greenness. Hence, the AQbD approach based green method and the validation data indicate that the method can be employed in routine analysis for the determination of Afatinib in pharmaceutical dosage form in quality control laboratory use.

Kyrgyz Republic production-resource and export potential state regulation mechanism

The study addressed the mechanism of state regulation of the production, resource and export potential of the Kyrgyz Republic. The study aimed to improve the system of state regulation of the agricultural sector to increase national food security. The analysis revealed a steady increase in production of the main types of agricultural products over the period 2014-2024: cereals by 11.6%, potatoes by 12.5%, vegetables by 18.2%, meat by 24.5% and milk by 17.1%. However, the study also showed uneven levels of self-sufficiency across different types of products: the high levels of self-sufficiency in potatoes (167.4%), vegetables (164.5%) and milk (116.5%) contrast with the low levels of self-sufficiency in fruit and berries (18.2%), vegetable oil (44%) and sugar (69%). An analysis of the existing state regulatory mechanisms revealed a comprehensive legal framework, including the Law of the Kyrgyz Republic No. 320 “On Food Security” and the “Food Security and Nutrition Programme for 2019-2023”, but their implementation is limited by insufficient funding and administrative barriers. Based on the analysis, recommendations were developed to optimise the state regulatory mechanism, including the introduction of a differentiated subsidy system, optimisation of the structure of exports and imports of agricultural products, increasing the efficiency of agricultural production, development of agricultural processing and improving the food quality control system. Proposed measures include the creation of the brand “Ecologically clean products of Kyrgyzstan”, the introduction of the Global G.A.P. certification system, the creation of a network of regional agricultural-consulting centres, the development of agro-industrial parks and the modernisation of quality control laboratories. The implementation of these measures aims to overcome the identified problems create more favourable conditions for the development of agriculture and ensure food security in the Kyrgyz Republic

ON THE ISSUE OF QUALITY CONTROL OF MEDICAL LABORATORY TESTS

As healthcare systems around the world increasingly rely on laboratory testing, maintaining high quality diagnostic services is imperative. In Kazakhstan, the importance of quality control in medical laboratories is being recognized, but there is a need to evaluate and improve current practices against international standards. Purpose: The purpose of this review article is to assess the current state of quality control in medical laboratories in Kazakhstan and then compare it with quality control procedures in other countries. Search strategy: Perform a comprehensive search in electronic databases such as PubMed, Google Scholar, Scopus and Web of Science with relevant keywords including “international standards”, “medical laboratories”, “quality control”, “quality assurance” and other. The search was limited to articles published in English between 2001 and 2023 to ensure inclusion of recent research and current practice. Results: A total of 79 publications were selected for the study based on their significance for quality control in medical laboratories, especially in Kazakhstan. The articles covered a wide range of topics, including regulatory frameworks, accreditation requirements, training programs, equipment requirements, data management systems, and collaborative efforts. A comparative study of quality control procedures in Kazakhstan with other countries revealed its positive and negative aspects.

SIMPLE AND SENSITIVE SPECTROPHOTOMETRIC ASSAYS FOR THE DETERMINATION OF TERBINAFINE HCL ANTIFUNGAL DRUG IN PHARMACEUTICALS

Objective: Two highly sensitive, accurate, inexpensive, and simple spectrophotometric assays were developed and validated for the determination of an anti-fungal drug, Terbinafine HCl (TBH), in pure drug and tablets using potassium permanganate (PP) and disodium 2-(1,3-dioxo-2,3-dihydro-1H-inden-2-yl)quinoline-6,8-disulfonate (DSOQ). Material and Method: In the present study, Sebifin and Terbiforce 250 mg tablets were used as pharmaceuticals, potassium permanganate KMnO4 and disodium 2-(1,3-dioxo-2,3-dihydro-1H-inden-2-yl)quinoline-6,8-disulfonate in water were used as reagents, and DR 3900 spectrophotometer equipped with 1cm matched quartz cells was used for absorbance measurements. Result and Discussion: The amount of terbinafine hydrochloride reacting with permanganate and disodium 2-(1,3-dioxo-2,3-dihydro-1H-inden-2-yl)quinoline-6,8-disulfonate in an acidic medium has been determined. The colored reaction products in both cases were measured at the maximum absorptions of 540 nm and 440 nm, respectively. The absorbance measured in each assay as a function of TBH concentration was related to TBH concentrations. Different experimental and variable conditions of assays were done carefully, accurately studied, and optimized. The validation of two assays also was done by following the current guidelines of the International Conference on Harmonization (ICH). Beer’s law for the two methods is obeyed over the concentration ranges 1-15 µg/ml (Correlation coefficient = 0.9983) and 1-18 µg/ml (Correlation coefficient = 0.9989) for methods PP and DSOQ, respectively. Molar absorptivity, limits of detection, and quantification (LOD &amp;amp; LOQ) values were (1.38×104 l/ mol cm, 0.92 &amp;amp; 2.78 µg/ml) for PP assay, and (1.73×104 l/ mol cm, 0.09 &amp;amp; 0.27 µg/ml) for DSOQ assay, respectively. The two assays were successfully applied for the determination of TBH in commercial tablets with reliable and satisfactory results, and hence the proposed assays can be applied in pharmaceutical laboratories of quality control.

Smart Chemometric‑Assisted Spectrophotometric Approaches for Simultaneous Quantification of Tertiary Combination Recommended for COVID-19 Supportive Treatments with Their Greenness Assessment.

There is an increasing interest of scientific community in developing innovative methodologies for their analysis needs within green analytical chemistry framework. UV spectrophotometry is one of the most promising eco-friendly methods, which is integrated with advanced chemometric tools to enhance the selectivity of the analysis of complex mixtures with severe overlapped signals. Simultaneous determination of a triple-combination of pseudoephedrine hydrochloride (PSE), carbinoxamine maleate (CRX), and paracetamol (PAR) using an artificial intelligence system and multivariate calibration methods. This combination is recently recommended for COVID-19 home-treated patients as part of a symptomatic treatment. Namely, the suggested models are: Artificial Neural Networks, Partial Least Squares, and Principal Component Regression. The proposed algorithms were optimized and developed with the aid of a five-level, three-factor experimental design. The investigated methods were applied over the concentration range of 100-180 μg/mL, 18-16 μg/mL, and 4-12 μg/mL for PSE, CRX, and PAR, respectively. The models validation results demonstrated excellent recoveries (around 98 to 102%), signaling the approaches outstanding resolution capacity for the cited compounds in the presence of common excipients. The outcomes of the studied methods were statistically compared to the official approaches, and no significant difference was found. The suggested models were efficiently employed to determine the selected drugs in their combined tablets without any initial separation steps. The impact of these methods on the environment was evaluated via greenness tools, namely; National Environmental Method Index, Raynie and Driver's green assessment method, analytical Eco-Scale, Green Analytical Procedure Index, and Analytical Greenness Metric. Green chemometric quality assessment of PSE, CRX, and PAR in their pure and pharmaceutical dosage forms. The established approaches are innovative, sustainable, smart, fast, selective, and cost-effective. These models are potential green nominees for routine analysis of the investigated mixture in quality control laboratories.

Assessing the Effect of Shodhana (Purification) Process by Triphala Decoction on Shilajit Using High Performance Thin Layer Chromatography

Standardization of Shodhit Shilajit is a major challenge. Objective: Objective of the present study was to assessing the effect of Shodhana purification process on Shilajit as well as assessing the difference in fingerprint of raw Shilajit and Shodhit Shilajit by HPTLC and physicochemical tests. Methods: The standardization parameters such as appearance, identification of active marker compound Gallic acid and Fingerprint comparison by HPTLC, Physicochemical test parameters like loss on drying, pH value, ash value, water soluble extractive, ethanol soluble extractive and assay of humic acid and fulvic acid were evaluated during the study. Chromatographic separation was achieved by using Toluene: Ethyl acetate: Formic acid (5.0:4.0:1.0 v/v/v) as mobile phase followed by comparing the Rf values. Results: The presence of active marker compound gallic acid in Shodhit Shilajit was confirmed by comparing the Rf value with standard. HPTLC fingerprint also confirmed that constituents from raw Shilajit transferring to the Shodhit Shilajit after the process of Shodhana. Total Ash value and Acid insoluble ash value reduced from 52.95% to 21.67% and 2.35% to 0.39% respectively while Water soluble extractive value increased from 59.33% to 72.13% after Shodhana process. There is also an increase in percentages of humic acid and fulvic acid from 7.10% to 8.23% and 36.91% to 50.12 % in Shodhit Shilajit. Conclusion: All these results showed the significance and effectiveness of Shodhana process in reducing the impurities and ultimately enhancing the therapeutic activity of Shodhit Shilajit. These evaluation parameters can be used for the standardization of Shodhit Shilajit and also can be routinely employed for analysis in Quality Control Lab.

Design of point-of-care electrochemical sensor for therapeutic drug monitoring of ofloxacin in biological fluids

A disposable screen-printed sensor has been crafted specifically for therapeutic drug monitoring purposes, particularly for detecting ofloxacin in biological fluids. To enhance selectivity toward ofloxacin, a supramolecular calix [6] arene serves as the ionophore of choice. The sensor incorporates a graphene nanocomposite as an ion-to-electron transducer layer, which not only boosts potential stability but also mitigates potential drift. The developed ofloxacin sensor underwent rigorous characterization following IUPAC guidelines. The linearity range spans from 1 × 10–6 to 1 × 10–2 M, with a measured slope of 59.0 mV/decade. Impressively, it boasts a percentage recovery of 100.18 ± 1.60 and a low detection limit (LOD) of 6 × 10–7 M. Stability assessments indicate reliable performance over an extended period of 8 weeks. The versatility of this sensor extends to various applications, including the determination of ofloxacin in pharmaceutical formulations, bulk powder, and biological fluids. Notably, it has demonstrated efficacy post-bioanalysis validation, adhering to Food and Drug Administration regulations. This advancement holds promise for personalized therapeutic drug monitoring in clinical pharmacy studies and quality control laboratories, thereby optimizing patient care at the point-of-care.Graphical abstract

Evaluation and clinical application of a bracketing calibration-based isotope dilution liquid chromatography-tandem mass spectrometry candidate reference measurement procedure for serum theophylline.

A candidate reference measurement procedure (RMP) for serum theophylline via isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. With a single-step precipitation pretreatment and a 6-min gradient elution, the method achieved baseline separation of theophylline and its analogs on a C18-packed column. A bracketing calibration method was used to ensure repeatable signal intensity and high measurement precision. The intra-assay and inter-assay imprecisions were 1.06%, 0.84%, 0.72% and 0.47%, 0.41%, 0.25% at concentrations of 4.22 µg/mL (23.40 µmol/L), 8.45 µg/mL (46.90 µmol/L), and 15.21 µg/mL (84.43 µmol/L), respectively. Recoveries ranged from 99.35 to 102.34%. The limit of detection (LoD) was 2 ng/mL, and the lowest limit of quantification (LLoQ) was 5 ng/mL. The linearity range extended from 0.47 to 60 µg/mL (2.61-333.04 µmol/L). No ion suppression and carry-over (< 0.68%) were observed. The relative bias for this candidate RMP that participated in 2023 External Quality Control for Reference Laboratories (RELA) conducted by the International Federation of Clinical Chemistry (IFCC) was within a range of 0.17 to 0.93%. Furthermore, two clinical immunoassay systems were compared with this candidate RMP, demonstrating good correlations. The results of the Trueness Verification Plan indicate significant differences among routine systems, highlighting the need for standardization efforts. The developed candidate RMP for serum theophylline serves as a precise reference baseline for standardizing clinical systems and assigning values to reference materials.

Tuning fluorescence of fexofenadine by switching OFF intramolecular photoinduced electron transfer: Application to development of one-step green and high throughput microwell spectrofluorimetric assay for analysis of tablets and plasma.

Fexofenadine (FEX) is a non-sedating antihistamine commonly used for the treatment of allergic conditions such as seasonal rhinitis and chronic idiopathic urticaria. This study describes the tuning "ON" the intrinsic fluorescence of FEX by switching "OFF" its intramolecular photoinduced electron transfer (PET) through the protonation of the piperidinyl nitrogen atom using sulfuric acid. The resulting fluorescence was utilized as a basis for the development of a highly sensitive microwell spectrofluorimetric assay (MW-SFA) for the one-step determination of FEX in pharmaceutical tablets and plasma. The linear range of the assay was 10-500 ng ml-1, and its limit of quantitation was 25.9ng ml-1. The proposed MW-SFA was successfully applied to analyze FEX in pharmaceutical tablets and plasma samples, demonstrating good accuracy and precision. The greenness of the assay was confirmed using three metric assessment tools. In conclusion, the MW-SFA is a straightforward, single-step analysis that requires no experimental adjustments. It offers high sensitivity, efficient sample processing, and environmental sustainability. This assay is highly recommended for pharmaceutical quality control and clinical lab use, particularly for measuring FEX levels.