Development Of Analytical Methods Research Articles

Analytical method development and validation for simultaneous estimation of seven markers in polyherbal formulation JKC by using RP-HPLC

BackgroundThe RP-HPLC method has been established to simultaneous estimation of seven markers in polyherbal formulation JKC using the C18 (25 × 0.46 cm, i.d,5 µm) column. The mobile phase consisted of methanol: water (80:20) at a flow rate of 1.0 mL/min and observed retention time at 2 to 11 min with sharp points. The marker compounds viz. Andrographolide (AG), Piperine (PP), Picroside-I (P-I), Picroside-II (P-II), α-Cyprone (AC), 6-Shogaol (6S), and 6-Gingerol (6G) were quantified in JKC formulations by HPLC method. Detection was performed at the wavelength (λ) of 229 nm for AG, 343 nm for PP, 279 nm for P-I, 264 nm for P-II, 254 nm for AC, and 280 nm for both 6S and 6G by HPLC–PDA detector.ResultsThe marker compounds in JKC formulations were observed in different retention times (Rt) i.e. AG at 3.060 ± 0.01 min, PP at 5.460 ± 0.03 min, P-I at 2.789 ± 0.02 min, P-II at 2.553 ± 0.03 min, AC at 10.951 ± 0.02 min, 6S at 6.302 ± 0.03 min, and 6G at 4.111 ± 0.02 min respectively. The proposed method was validated with acceptable linearity (r2 0.9995–0.9999), precision, robustness, ruggedness, and accuracy (RSD < 2%) under optimum conditions. The limit of detection and quantification of bioactive markers were as: AG (1.386; 4.200 ppm), PP (2.033; 6.161 ppm), P-I (2.822; 8.553 ppm), P-II (2.538; 7.691 ppm), AC (0.269; 0.815 ppm), 6G (0.158; 0.480 ppm), 6S (0.188; 0.569 ppm). The amount (mg/g) of bioactive markers detected and estimated in plants and formulation were as: AG (41.282 ± 0.48; 10.06 ± 0.18), PP (53.81 ± 0.25, 13.82 ± 0.37 in PN, PL; 4.27 ± 0.07), P-I (15.97 ± 0.01; 0.48 ± 0.003), P-II (63.24 ± 0.35; 2.31 ± 0.006), AC (0.42 ± 0.01; 0.36 ± 0.006), 6G (0.71 ± 0.03; 0.16 ± 0.001), and 6S (2.64 ± 0.09; 0.12 ± 0.004) respectively. Method was found to be rugged and robust. The results found for all the validation parameters were within the limits according to ICH guidelines.ConclusionThe proposed method is fast, precise, economic, and specific and used for the simultaneously quantifiable analysis of seven major bioactive markers in the ingredients (herbs) and the JKC formulations.

A new certified reference material IAEA-465 for radionuclides in Baltic Sea sediment

Certified reference material (CRM) for natural (40K,210Pb,210Po,226Ra,228Ra,228Th,230Th,232Th,234U,235U, and238U) and anthropogenic (137Cs,239+240Pu, and241Am) radionuclides in marine sediment from the Baltic Sea (IAEA-465) has been developed. Information values are given for 238Pu,239Pu and240Pu. Altogether 27 laboratories participated in this exercise. Radiometric (alpha-spectrometry, gamma-spectrometry and beta counting, as well as mass spectrometry (ICP-MS and AMS) techniques were applied in measurements. The CRM is intended to be used for Quality Assurance/Quality Control of radionuclide analyses, for the development and validation of analytical methods, for the development of reference methods and for training purposes.

Development of a simultaneous analytical method for the dietary exposure determination of β-Carboline alkaloids in foods

As β-carboline (βC) alkaloids, posing potential health risks, are present in a wide variety of foods, determining the exposure degrees of food to these alkaloids from dietary activity is key to ensuring food safety. Here, we developed a rapid and sensitive simultaneous analytical method for six βC alkaloids in food. We optimized the buffered QuEChERS method, which includes a clean-up process through dispersive solid phase extraction, to extract the target compounds from food matrices; then, these compounds were detected via liquid chromatography–tandem mass spectrometry. We established calibration ranges for each target compound and matrix within the range of 0.05–250 μg/kg, and verified linearity (R2 ≥ 0.99) and limit of quantitation (≤1.63 μg/kg). Furthermore, we validated trueness (85.8%–118.8%) and precision (≤18.7%) at three levels within the calibration range, including the lowest and highest concentrations. Finally, we employed the developed method to determine the βC alkaloid contents in 304 samples of 41 food items and dietary exposure of six βC alkaloids resulting from daily intake. Although βC alkaloids were detected in 86.2% of the samples, exposure level to the 41 food items was insufficient to cause toxicity.

History of toxicology at the Technical University of Munich (TUM).

Toxicology at the TUM is mainly associated with the Faculty of Medicine at the Klinikum rechts der Isar (MRI). The Department of Clinical Toxicology has been founded in 1963. Max von Clarmann, the head, focused his activities on the treatment of intoxications and the development of analytical methods and established a poison information center. His successors, Thomas Zielker and Florian Eyer, further developed this department to an internationally renown institution.In 1967, the MRI became the TUM faculty of medicine with its Institute of Pharmacology andToxicology. The director Melchior Reiter, formerly Institute of Pharmacology of the Ludwig Maximilians University (LMU), in 1970 initiated the foundation of the Department of Toxicology at the Gesellschaft für Strahlen- und Umweltforschung (GSF) with the director Gerhard Lange. The research focused on the neurotoxic effects of heavy metals and the metabolism and hepatoxicity of persistent chemicals. After Lange's unexpected death in 1973, he was succeeded in 1975 by Helmut Greim from the University of Tübingen. The now Institute of Toxicology rapidly expanded developing and standardizing in vitro test methods, investigating the mechanism of carcinogens and mutagens and heavy metal toxicity. Training courses in the 15 major areas of toxicology have been organized at the GSF and competent centers in Germany. In 1987, Greim became the director of the newly founded Institute of Toxicology and Environmental Hygiene of the TUM, with expanded research and teaching activities, especially in toxicology at the faculties of Chemistry of the TUM and LMU, which thereafter became mandatory for students of chemistry at German universities.

Analytical Method Development and Validation for the Simultaneous Estimation of Candesartan and Simvastatin by UV- Spectrophotometry Method

Analytical Method Development and Validation for the Simultaneous Estimation of Candesartan and Simvastatin by UV- Spectrophotometry Method

Analytical Method Development and Validation for Quantitative Estimation of Tebentafusp Bulk Dosage by RP-HPLC

Analytical Method Development and Validation for Quantitative Estimation of Tebentafusp Bulk Dosage by RP-HPLC

Analytical Method Development and Validation of Faricimab Injection by RP-HPLC

A robust and reliable Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the quantitative analysis of Faricimab in Vabysmo injections. Faricimab, an IgG1-based bispecific antibody targeting VEGF-A and Ang-2, is used for treating retinal vascular disorders like age-related macular degeneration and diabetic macular edema. The method development involved the use of an Inertsil C18 column (250 x 4.6 mm, 5 µm) with a mobile phase composed of KH2PO4 (pH 4.6) and methanol in a 1:1 ratio, sonicated for 20 minutes. The developed method was evaluated for various validation parameters as per ICH guidelines, including linearity, precision, accuracy, and sensitivity. Linearity was demonstrated within the concentration range of 50-150 µg/ml with a correlation coefficient (R²) of 0.999. The limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.050 µg/ml and 0.166 µg/ml, respectively, indicating high sensitivity. Precision, expressed as relative standard deviation (RSD), was below 0.1%, confirming the method's reliability. Accuracy was assessed, with mean assay values ranging between 97-101%, and RSD below 0.2%, indicating the method's precision. Robustness tests confirmed the method's resilience against slight variations in analytical conditions such as acetonitrile ratio, flow rate, detection wavelength, and pH. Stability studies showed that Faricimab remains stable under various stress conditions, including exposure to 0.1N HCl, 0.1N NaOH, 30% peroxide, heat, and sunlight, with assay values ranging from 88.93% to 94.07%. The optimized RP-HPLC method ensures good peak shape and resolution, making it suitable for routine quality control of Faricimab in pharmaceutical formulations. The validated method adheres to ICH guidelines, ensuring its reliability for consistent and accurate quantification of Faricimab

Analytical Method Development and Validation for the Simultaneous Estimation of Evinacumab Injection Formulation by RP-HPLC

This paper describes in detail the RP-HPLC method used to evaluate the content of evinacumab (EVC) in combination with evkeeza injections. The mobile phase used in the assay is a mixture of potassium dihydrogen phosphate and methanol in an isocratic ratio of 70:30 (v/v) on a column Shimpac solar C18, 150*4.6, 5μm column. The flow rate was 1.0 ml/min, isocratic. At 226 nm, the quantification and detection of EVC were done concurrently. It was discovered that the response obtained was linear over the concentration range of 75-225 μg/ml, and the estimated RT in minutes for EVC was 4.063. 1.805 μg/ml was the LOD and 6.017 μg/ml was the LOQ. For the EVC assay, precision was 0.3% RSD and accuracy was 100%.

A Comprehensive Review on Analytical Methods of Rifampicin

Rifampicin, a cornerstone in the treatment of tuberculosis and other bacterial infections, necessitates robust analytical methods for its accurate determination in pharmaceutical formulations and biological matrices. This review aims to provide a comprehensive summary of the analytical methods developed and validated for rifampicin quantification, encompassing various chromatographic, spectroscopic, and other analytical techniques reported in the literature. The review begins with an overview of the physicochemical properties, pharmacological significance, and regulatory requirements pertinent to rifampicin analysis. Additionally, the review highlights the importance of method robustness, specificity, sensitivity, and stability-indicating capability in ensuring the quality and safety of rifampicin-containing formulations. Overall, this review serves as a comprehensive reference for researchers, analysts, and regulatory authorities involved in the development, validation, and quality control of analytical methods for rifampicin, facilitating the advancement of pharmaceutical sciences and therapeutic interventions aimed at combating infectious diseases effectively

Small-group originating model: Optimized individual-level GWAS simulation featured by SLiM and using open-access data

The development of analytical methods for Genome-wide Association Studies (GWAS) has outpaced the evolution of simulation techniques and pipelines. This disparity underscores the importance of innovative simulation methods that can keep pace with the rapidly increasing scale of GWAS. The median sample size of GWAS over the past ten years has exceeded 50,000 individuals, a trend that emphasizes the need for simulation tools capable of generating data on a similar or larger scale. This paper introduces a novel method, the small-group originating (SGO) model, utilizing the SLiM software for simulating individual-level GWAS data. Our standardized protocol facilitates the generation of tens of thousands of pseudo-individuals with millions of variants from small (30−90) open-access datasets.SGO stands out, especially when compared to the widely-used resampling method in HapGen, showcasing superior simulation efficiency for large sample sizes (> 13,000) of unrelated individuals. This capability is particularly relevant given the current trajectory towards larger GWAS, necessitating tools that can simulate datasets reflective of this growth. Additionally, SGO provides customization options and can model dynamic life cycles and mating across generations, positioning it as a highly promising alternative for GWAS simulations.In a case study, sensitivity analyses of chromosome-level principal component analysis and kinship coefficient estimation were conducted. The results highlighted the poor robustness of chromosome-level quality control (QC) indexes and the uneven distribution of population structure across chromosomes and ancestries, advocating for the caution against relying solely on chromosome-level QC statistics.With its flexible and efficient approach to generating pseudo GWAS data, our standardized SGO protocol emerges as a crucial asset for method development, power analysis, and benchmarking in GWAS research. It is especially vital in the context of accommodating the demands for large-scale simulations, aligning with the current and future scale of GWAS.

A tutorial on solid-phase analytical derivatization in sample preparation applications

Solid-phase analytical derivatization is a versatile sample preparation technique that enhances analysis accuracy, efficiency, reproducibility, and sensitivity. The technique combines the advantages of analytical derivatization and solid-phase extraction, providing a versatile approach for analysing analytes with various functional groups in complex matrices. Analytical derivatization is a technique used for functional group analysis that involves modifying the structure of an analyte to enhance sensitivity and specificity. Solid-phase analytical derivatization is a one-pot procedure that combines analytical derivatization and extraction. In the presence of acids, it can derivatize phenols, carboxylic acids, and other analytes. Furthermore, solid-phase analytical derivatization increases the reaction rate of carbonyl compounds, making it easier to extract aldehydes and ketones rapidly. This technique utilises electrophoresis, chromophores, fluorophores, and functional groups for detection and extraction. It initially began as a batch procedure but has now developed into an automated, microextraction, and derivatization method. Solid-phase analytical derivatization is highly efficient due to the significant excess of the derivatizing reagent relative to the analyte, along with the pre-impregnation of derivatizing reagents, which can speed up the process compared to traditional solution-based derivatization. This tutorial aims to provide detailed insights into the practical aspects of implementing solid-phase analytical derivatization in analytical method development and discuss the prospects and future trends.

Recent solid-phase microextraction-based analytical approaches for the profiling of biliary bile acids in pre-transplant assessments of liver grafts subjected to normothermic ex vivo liver perfusion

BackgroundLiver transplantation is the definitive treatment for end-stage liver failure, but the scarcity of donor organs remains a significant challenge. Leveraging organs from extended criteria donors (ECD) offers a potential avenue to address worldwide shortages, though these organs are more susceptible to post-reperfusion injury. This study explores the use of normothermic ex vivo liver perfusion (NEVLP) as a method for organ preservation – an approach that sustains liver metabolism and facilitates pre-transplant assessments of organ viability via bile analysis. The focal point of this study revolves on the development of analytical methods for determining the bile acid profile throughout the peritransplantation period as a potential indicator of liver function and viability. ResultsThe study optimized and validated a high-throughput analytical method to quantify selected bile acids in bile samples using a thin-film microextraction-liquid chromatography-mass spectrometry (TFME-LC-MS) platform. Furthermore, it introduced a solid-phase microextraction-microfluidic open interface-mass spectrometry (SPME-MOI-MS) method for rapid direct analysis of bile acid isobar groups. In the animal study, discernible variations in the concentrations of specific bile acids were observed between donors after circulatory death (DCD) and heart-beating donors (HBD), particularly following normothermic perfusion and reperfusion. Noteworthy fluctuations in individual bile acid concentrations were observed throughout the entire organ transplantation process, with taurocholic acid (TCA), glycocholic acid (GCA), and glycochenodeoxycholic acid (GCDCA) emerging as promising indicators of organ quality. The efficacy of the SPME-MOI-MS platform in corroborating these trends highlights its potential for real-time bile acid analysis during liver transplantation procedures. SignificanceOur findings underscore the efficacy of NEVLP in tandem with advanced bile acid analysis methods as a reliable strategy for pre-transplant assessments of organ viability, potentially increasing the use of ECD organs and reducing organ shortages. The ability to monitor bile acid profiles in real-time provides crucial insights into liver function and ischemic injury, making significant strides in improving transplant outcomes and patient survival rates.

Sensitive determination of Gabapentin in plasma sample using dispersive liquid-liquid microextraction coupled with ion mobility spectrometry

Gabapentin (GBP) is an anticonvulsant agent and its determination using a fast and simple method is challenging. In this research, a novel sensitive, and economical method for the fast determination of GBP in biological samples was developed. This method benefits from the advantages of ion mobility spectrometry (IMS) as a fast detection and separation system and dispersive liquid-liquid microextraction (DLLME) for the preconcentration/extraction of GBP. In the developed method (DLLME-IMS), Ag nanoparticles serve as modifiers. Moreover, the quantum theory of atoms in molecules (QTAIM) analysis was employed to investigate the interaction mechanism between GBP and Ag nanoparticles. Also, a delay time after the injection of the sample was applied to improve the method sensitivity. Under the optimum condition, the developed method showed a linear response over the concentration range of 0.3–12 μg mL−1 and a detection limit of 0.12 μg mL−1. Eventually, DLLME-IMS was successfully employed in plasma sample, and was validated against the HPLC method with UV detection as a reference method. The developed method will open up a new avenue for the development of effective, rapid, and simple analytical methods.

Wearable solid-phase microextraction sampling for enhanced detection of volatile analytes in human ears

BackgroundInvestigating ear at molecule level is challenging task, since there is a lack of molecular detection by traditional diagnosis techniques such as otologic endoscopy, ear swab culture, and imaging diagnostic technique. Therefore, new development of noninvasive, highly sensitive, and convenient analytical method for investigating human ears is highly needed. ResultsWe developed a wearable sampling device for extracting trace analytes in ear by fixing solid-phase microextraction fibers into modified earmuffs (SPME-in-earmuffs). After sampling, SPME fiber was coupled with gas chromatography-mass spectrometry (GC-MS) for identification and quantification of extracted analytes. Enhanced detection of various analytes such as volatile metabolites, exposures, and therapeutic drugs of ears were demonstrated in this work. Particularly, sport-induced metabolic changes such as fatty acids, aldehyde compounds and oxidative produces were found from human ears using this method. Acceptable analytical performances were obtained by using this newly developed method for detecting ear medicines, e.g., low limit of detection (LOD, 0.005–0.021 ng/mL) and limit of quantification (LOQ, 0.018–0.071 ng/mL), excellent linear dynamic responses (R2 > 0.99, ranging from 0.050–8.00 ng/mL), good relative standard deviations (RSDs, 13.19 % ∼ 21.40 %, n = 6) and accuracy (84.43–150.18 %, n = 6) at different concentrations. SignificanceFor the first time, this work provides a simple, convenient, and wearable microextraction method for enhanced detection of trace volatiles in human ears. The enclosed space between ear and earmuff allows headspace SPME sampling of volatile analytes, and thus provides a new wearable method for monitoring ear metabolites and human exposures, showing potential applications in human health, disease diagnosis, and sport science.

A Review of Machine Learning and QSAR/QSPR Predictions for Complexes of Organic Molecules with Cyclodextrins.

Cyclodextrins are macrocyclic rings composed of glucose residues. Due to their remarkable structural properties, they can form host-guest inclusion complexes, which is why they are frequently used in the pharmaceutical, cosmetic, and food industries, as well as in environmental and analytical chemistry. This review presents the reports from 2011 to 2023 on the quantitative structure-activity/property relationship (QSAR/QSPR) approach, which is primarily employed to predict the thermodynamic stability of inclusion complexes. This article extensively discusses the significant developments related to the size of available experimental data, the available sets of descriptors, and the machine learning (ML) algorithms used, such as support vector machines, random forests, artificial neural networks, and gradient boosting. As QSAR/QPR analysis only requires molecular structures of guests and experimental values of stability constants, this approach may be particularly useful for predicting these values for complexes with randomly substituted cyclodextrins, as well as for estimating their dependence on pH. This work proposes solutions on how to effectively use this knowledge, which is especially important for researchers who will deal with this topic in the future. This review also presents other applications of ML in relation to CD complexes, including the prediction of physicochemical properties of CD complexes, the development of analytical methods based on complexation with CDs, and the optimisation of experimental conditions for the preparation of the complexes.

Analytical method development and validation of residual solvents in ethosuximide by GC – headspace technique

Analytical method development and validation of residual solvents in ethosuximide by GC – headspace technique

Analytical method development and validation for estimation of indapamide in bulk drug and dosage form by UV-Spectroscopy

Analytical method development and validation for estimation of indapamide in bulk drug and dosage form by UV-Spectroscopy

Utilization of the FTIR spectroscopic method for the quantitative determination of the narrow therapeutic index levothyroxine sodium in pharmaceutical tablets

Levothyroxine sodium is a narrow therapeutic index drug used for the treatment of hypothyroidism. The medication is marketed in tablet form with very low doses ranging from 25 to 150 µg, which requires the development of a sensitive quantitative analytical method to ensure a safe and effective pharmacological response. In the present work, a Fourier transform infrared method has been developed and validated for levothyroxine sodium determination in various pharmaceutical formulations. The proposed method involves selectively extracting levothyroxine sodium from the studied tablets using chloroform as solvent, then depositing it on a KBr pellet, followed by infrared measurements and spectra analysis. The peak band area corresponding to the C=C centered at 1409 cm-1 has been chosen for the quantification. The method has been validated according to ICH guidelines and was found to be simple, precise, accurate, and specific. The linearity, detection, and quantitation limits are 25–800, 8.121, and 24.545 µg/pellet, respectively. These values confer the method’s sensitivity and applicability for the determination of different pharmaceutical tablets with various dosages. A statistical comparison with a reference HPLC method showed no significant difference. Accordingly, the developed method can be employed for quality control testing of levothyroxine sodium due to its simplicity and the absence of sophisticated instrumentation and procedures. Graphical abstract:

Chemical spaces of taste and odor

AbstractThe thesis provides a draft map of the chemical space of chemosensory active molecules in foods and enables, by application of cheminformatics approaches, the deduction of general principles of raw material influence and processing on the sensory perception of complex food systems. The summarized data supports the development of analytical methods targeting the efficient quantitation of key compounds, provides a database for screening of chemosensory receptor agonists and facilitates the optimization of in‐silico tools for prediction of receptor activation determining sensory quality and biological function.

A review of analytical method development and validation of labetalol hydrochloride

A review of analytical method development and validation of labetalol hydrochloride