Validation Of Analytical Method Research Articles

Analytical method development and validation for the simultaneous estimation of montelukast and Acebrophylline by RP- HPLC method

High performance liquid chromatography is at present one of the most sophisticated tool of the analysis. The estimation of Montelukast and Acebrophylline was done by RP-HPLC. The Phosphate buffer was pH 3.0 and the mobile phase was optimized with consists of Methanol: Phosphate buffer mixed in the ratio of 70:30 % v/ v. Inertsil C18 column C18 (4.6 x 150mm, 5mm) or equivalent chemically bonded to porous silica particles was used as stationary phase. The detection was carried out using UV detector at 260 nm. The solutions were chromatographed at a constant flow rate of 0.8 ml/min. the linearity range of Montelukast and Acebrophylline were found to be from 100-500 mg/ml of Montelukast and 1-5mg/ml of Acebrophylline . Linear regression coefficient was not more than 0.999.The values of % RSD are less than 2% indicating accuracy and precision of the method. The percentage recovery varies from 98-102% of Montelukast and acebrophylline. LOD and LOQ were found to be within limit. The results obtained on the validation parameters met ICH and USP requirements .it inferred the method found to be simple, accurate, precise and linear. The method was found to be having suitable application in routine laboratory analysis with high degree of accuracy and precision.

Development and Validation of Analytical Method for Simultaneous Estimation of Mometasone Furoate Topical Formulation by HPLC-DAD

Mometsone furoate is a corticosteroid (glucocorticoid) having an anti-inflammatory activity. Corticosteroids showed a wide range of effects on various cell types such as mast cells, eosinoplills, neutrophills, macrophages, and lymphocytes.

Comparison of Analytical Method validation guidelines used for release, stability in Biosimilar Manufacturing process

Comparison of Analytical Method validation guidelines used for release, stability in Biosimilar Manufacturing process

A Comprehensive Review on Advantages of Ultra Performance Liquid Chromatography (UPLC) for Analytical Method Development, Validation, and Stability Studies

Ultra Performance Liquid Chromatography (UPLC) represents a paradigm shift in analytical separation science, offering enhanced speed, sensitivity, and resolution while retaining the foundational principles of traditional liquid chromatography. This comprehensive review explores the principles, instrumentation, advantages, and applications of UPLC in analytical method development, validation, and stability studies, with a focus on pharmaceutical analysis. UPLC leverages finer particles and intensified flow rates to achieve unprecedented performance gains, facilitating rapid and precise separations without compromising analytical rigor. The discussion delves into the intricate instrumentation involved, including sample injection, column selection, and detector operation, highlighting the critical role of each component in ensuring optimal analytical performance. Advantages of UPLC include drastically reduced run times, streamlined method optimization, and increased sample throughput, while limitations such as higher back pressures and limited column regenerability are also discussed. Furthermore, the review outlines the diverse applications of UPLC across pharmaceutical, environmental, food and beverage, clinical, forensic, bioanalytical, and polymer analysis domains, underscoring its versatility and significance in advancing research, development, and quality control in various industries. In summary, UPLC represents a transformative tool in analytical chemistry, offering unparalleled capabilities for expedited and accurate analysis of complex sample matrices, ultimately driving innovation and scientific progress. Key Words: Ultra Performance Liquid Chromatography (UPLC), analytical method development, validation, stability studies, pharmaceutical analysis, instrumentation, advantages, applications, sample injection, column chemistry, detector technology, back pressures, method optimization, sample throughput, environmental analysis, food and beverage analysis, clinical diagnostics, forensic analysis, bioanalytical research, polymer analysis, analytical chemistry.

Validation of HPLC and TLC analytical methods to determine radiochemical purity of 99mTc-cAbVCAM1-5, a new experimental radiotracer

Cardiovascular diseases, including fatal myocardial infarctions from atheromatous plaques, are the primary global mortality cause. Detecting stenotic atheromatous plaques is possible through coronary angiography, but vulnerable plaques with eccentric remodeling are undetectable with current diagnostic methods. Addressing this challenge, our group developed a radiopharmaceutical drug targeting vascular cell adhesion molecule 1 (VCAM-1), radiolabeled with technetium-99m. Given the absence of a monograph in the European Pharmacopoeia, and in order to draft the investigational medicinal product documentation, analytical methods had to be validated by high performance liquid chromatography (HPLC) and thin layer chromatography (TLC) to determine the radiochemical purity (RCP) of 99mTc-cAbVCAM1–5. This study therefore presents the results of the validation of analytical methods obtained in this context. The method validation followed the European Association of Nuclear Medicine (EANM) recommendations adapted from ICH Q2(R1), ensuring conformity with specificity, accuracy, repeatability and intermediate precision, linearity, robustness, quantification limit (LoQ), and range criteria. Regarding the results of specificity, both HPLC and TLC methods demonstrated excellent separation of 99mTc-cAbVCAM1–5 from impurities 99mTcO4-. Accuracy results indicated recovery percentages within the range of 99.52–101.40% for the HPLC and 99.51–101.97% for TLC, ensuring reliable measurements for each concentration of 99mTcO4-. Precision of the methods was validated by assessing repeatability and intermediate precision. Linearity was determined over the usual concentrations range and the correlation coefficient was greater than 0.99 for both methods. The limit of quantification was measured by diluting the 99mTcO4- to obtain a signal-to-noise ratio of around 10:1. Under these conditions, we obtained an LOQ of 2.10 MBq/mL for HPLC and 2Mbq/mL for TLC. In conclusion, the analytical methods developed in this study comply with EANM recommendations. This therefore allows us to correctly assess the radiochemical purity of 99mTc-cAbVCAM1–5, a new radiotracer targeting inflammation in vulnerable plaques.

ИЗУЧЕНИЕ ВАЛИДАЦИОННЫХ ХАРАКТЕРИСТИК МЕТОДИК ИСПЫТАНИЯ НА ПОДЛИННОСТЬ И КОЛИЧЕСТВЕННОГО ОПРЕДЕЛЕНИЯ АЦИЗОЛА В СТОМАТОЛОГИЧЕСКОМ ГЕЛЕ

In the quality assurance system for medicinal products, an important role is played by the procedure for validation of analytical methods at the stages of development, testing, technology and serial production. The need for validation at the stage of drug development is due to the fact that it is possible to promptly identify the shortcomings of the method and significantly improve it. The purpose of this study is to validate methods for testing «The authenticity» and «Quantitative determination» of acyzole in dental gel. Materials and methods. To achieve this goal, an active pharmaceutical substance of pharmacopoeial quality was used. When developing methods for testing for “Authenticity,” a reaction with oxalic acid to the zinc cation determined in the composition of acyzole was used. For the quantitative determination of acyzole, the complexometric method was used. The objects of the study were three series of gel samples. Results and discussion. As a result of the research, a method for testing «Authenticity» was developed; it was found that the proposed method is characterized by a negative analytical signal on model mixtures free of the analyte component and placebo, and a positive analytical signal on model mixtures of various compositions containing the analyte component. The validation characteristics of methods for the quantitative determination of acyzole in the gel were studied. positive results were obtained. The proposed methods can be used for inclusion in the regulatory documentation for the developed gel. Conclusion. When assessing the validation characteristics of the proposed methods, it was established that the methods for testing the «Authenticity» and «Quantitative determination» of acyzole in the gel are specific, the methods of quantitative determination are characterized by accuracy and repeatability, a linear dependence in the analytical range of ±20% of the declared amount of acyzole, which allows them to be used for a reliable assessment quality of the developed dental gel.

Methodological validation of sedimentation velocity analytical ultracentrifugation method for Adeno-Associated Virus and collaborative calibration of system suitability substance.

Currently, adeno-associated virus (AAV) is one of the primary gene delivery vectors in gene therapy, facilitating long-term in vivo gene expression. Despite being imperative, it is incredibly challenging to precisely assess AAV particle distribution according to the sedimentation coefficient and identify impurities related to capsid structures. This study performed the systematic methodological validation of quantifying the AAV empty and full capsid ratio. This includes specificity, accuracy, precision, linearity, and parameter variables involving the sedimentation velocity analytical ultracentrifugation (SV-AUC) method. Specifically, SV-AUC differentiated among the empty, partial, full, and High Sedimentation Coefficient Substance (HSCS) AAV particles while evaluating their sedimentation heterogeneity. The intermediate precision analysis of HE (high percentage of empty capsid) and HF( high percentage of full capsid) samples revealed that the specific species percentage, such as empty or full, was more significant than 50%. Moreover, the RSD (relative standard deviation) could be within 5%. Even for empty or partial less than 15%, the RSD could be within 10%. The accuracy recovery rates of empty capsid were between 103.9% and 108.7% across three different mixtures. When the measured percentage of specific species was more significant than 14%, the recovery rate was between 77.9% and 106.6%. Linearity analysis revealed an excellent linear correlation between the empty, partial, and full in the HE samples. The AAV samples with as low as 7.4×1011 cp/mL AAV could be accurately quantified with SV-AUC. The parameter variable analyses revealed that variations in cell alignment significantly affected the overall results. Still, the detection wavelength of 235nm slightly influenced the empty, partial, and full percentages. Minor detection wavelength changes showed no impact on the sedimentation coefficient of these species. However, the temperature affected the measured sedimentation coefficient. These results validated the SV-AUC method to quantify AAV. This study provides solutions to AAV empty and full capsid ratio quantification challenges and the subsequent basis for calibrating the AAV empty capsid system suitability substance. Due to the AAV structure and potential variability complexity in detection, we jointly calibrated empty capsid system suitability substance with three laboratories to accurately detect the quantitative AAV empty and full capsid ratio. The empty capsid system suitability substance could be used as an external reference to measure the performance of the instrument. The results could be compared with multiple QC (quality control)laboratories based on the AAV vector and calibration accuracy. This is crucial for AUC to be used for QC release and promote gene therapy research worldwide.

Fate of multi-residue insecticides and their metabolites in the process of vinification: Analytical method validation, dissipation kinetics, processing factor, and risk assessment

In viticulture, the use of synthetic chemical formulations introduces insecticide residues into harvested grapes and further into processed grape products, posing a safety concern to consumers. This study investigated the fate of ten insecticide residues and their metabolites from vine to wine. A rapid validated multi-residue approach using QuEChERS extraction and LC-MS/MS configuration was employed for targeted analysis in grape, pomace, and wine. The targeted insecticides showed satisfactory mean recoveries (76.03–111.95%) and precision (RSD = 0.75–7.90%) across the three matrices, with a matrix effect ranging from −16.88 to 35.18%, particularly higher in pomace. Preliminary grape washing effectively removed 15.52–61.31% of insecticide residues based on water solubility and systemic nature. Residue dissipation during fermentation ranged from 73.19% to 87.15% with a half-life spanning from 1 to 5.5 days. The mitigation rate was observed at 12.85–26.81% for wine and 17.76–51.55% for pomace, with the highest transfer rate for buprofezin (51.55%) to pomace and fipronil (25.72%) to wine. Calculated processing factors (PF) for final wine ranged from 0.16 to 0.44, correlating strongly with the octanol-water partition ratio of targeted insecticides. The reported PF, calculated hazard quotient (HQ) (0.003–5.800%), and chronic hazard index (cHI) (2.041–10.387%) indicate reduced residue concentrations in wine and no potential chronic risk to consumers, ensuring a lower dietary risk to wine consumers.

Analytical Method Development and Dermal Absorption of 4-Amino-3-Nitrophenol (4A3NP), a Hair Dye Ingredient under the Oxidative or Non-Oxidative Condition.

The chemical 4-amino-3-nitrophenol (4A3NP) is classified as an amino nitrophenol and is primarily utilized as an ingredient in hair dye colorants. In Korea and Europe, it is exclusively used in non-oxidative or oxidative hair dye formulations, with maximum allowable concentrations of 1% and 1.5%, respectively. Despite this widespread use, risk assessment of 4A3NP has not been completed due to the lack of proper dermal absorption data. Therefore, in this study, both the analytical method validation and in vitro dermal absorption study of 4A3NP were conducted following the guidelines provided by the Korea Ministry of Food and Drug Safety (MFDS). Before proceeding with the dermal absorption study, analytical methods were developed for the quantitation of 4A3NP through multiple reaction monitoring (MRM) via liquid chromatography-mass spectrometry (LC-MS) in various matrices, including swab wash (WASH), stratum corneum (SC), skin (SKIN, comprising the dermis and epidermis), and receptor fluid (RF). These developed methods demonstrated excellent linearity (R2 = 0.9962-0.9993), accuracy (93.5-111.73%), and precision (1.7-14.46%) in accordance with the validation guidelines.The dermal absorption of 4A3NP was determined using Franz diffusion cells with mini-pig skin as the barrier. Under both non-oxidative and oxidative (6% hydrogen peroxide (H2O2): water, 1:1) hair dye conditions, 1% and 1.5% concentrations of 4A3NP were applied to the skin at a rate of 10 μL/cm2, respectively. The total dermal absorption rates of 4A3NP under non-oxidative (1%) and oxidative (1.5%) conditions were determined to be 5.62 ± 2.19% (5.62 ± 2.19 μg/cm2) and 2.83 ± 1.48% (4.24 ± 2.21 μg/cm2), respectively.

Development and validation of RP-HPLC method for simultaneous estimation of 5-fluorouracil and silibinin in corn-derived protein-polysaccharide nanocomplex: Forced degradation and mathematical modeling of drug release

In the current work, RP-HPLC method was developed and validated for simultaneous analysis of 5-Fluorouracil (5-FU) and silibinin from corn-derived protein polysaccharide nanocomplex. The method employs a mobile phase of water (A) and acetonitrile: methanol (60:40) (B) on a Waters C18 column (4.6 x 150 mm, 5 μm), with detection wavelengths set at 266 nm and 288 nm for 5-FU and silibinin, respectively. The runtime was 15 min at a flow rate of 0.7 mL/min. The analytical method validation adhered closely to ICH guidelines, exhibiting remarkable selectivity, specificity, linearity, and precision, all falling comfortably within acceptable thresholds. The analytical method was found to be linear from 1 to 150 µg/mL with R2 of 0.9998. Forced degradation studies were carried to assess the stability of 5-FU and silibinin within the nanocomplex across diverse conditions. Additionally, nanocomplexes loaded with 5-FU and silibinin were formulated and characterized, demonstrating a particle size of 209.83 ± 3.83 nm and a zeta potential of 57.93 ± 1.06 mV. The %entrapment efficiency for 5-FU and silibinin was found to be 88.73 ± 0.37 % and 97.00 ± 1.58 % with a %drug loading of 6.73 ± 0.92 % and 2.74 ± 0.61 % respectively. The %in vitro drug release was determined using developed and validated analytical methods. Further, various release kinetic models were applied to determine the release behavior from the nanocomplex system. To conclude, we ensured the eco-friendliness of the developed analytical method by assessing its greenness through Eco-scale, the green analytical procedure index (GAPI), and the Analytical GREEnness metric (AGREE).

Characterization and quantitation of a sulfoconjugated metabolite for detection of methyltestosterone misuse and direct identification by LC-MS

Methyltestosterone (MT) is one of the most frequently misused anabolic androgenic steroids detected in doping control analysis. The metabolism of MT in humans leads to several phase І metabolites and their corresponding phase Ⅱ conjugates. Previous studies have postulated the 3α-sulfoconjugate of 17α-methyl-5β-androstane-3α,17β-diol (S2) as principal sulfate metabolite of MT, with a detection window exceeding 10 days. However, a final direct and unambiguous confirmation of the structure of this metabolite is missing until now. In this study, we established an approach to detect and identify S2, using intact analysis by liquid chromatography hyphenated with tandem mass spectrometry (LC-MS/MS) without complex sample pretreatment. An in vitro study yielded the LC-MS/MS reference retention times of all 3-sulfated 17-methylandrostane-3,17-diol diastereomers, allowing for accurate structure assignment of potentially detected metabolites. In an in vivo excretion study with a single healthy male volunteer, the presence of the metabolite S2 was confirmed after a single oral dose of 10 mg MT. The reference standard was chemically synthesized, characterized by accurate mass mass spectrometry (MS) and nuclear magnetic resonance (NMR), and quantified by quantitative NMR (qNMR). Thus, this study finally provides accurate structure information on the S2 metabolite and a direct analytical method for detection of MT misuse. The availability of the reference material is expected to facilitate further evaluation and subsequent analytical method validation in anti-doping research.

Analytical Method Development and Validation of Tofisopam in Pure and Pharmaceutical formulation

Tofisopam is an anxiolytic used in treatment of anxiety like disorders, prescribed in the dosage of 50mg to 300mg. As the drug is insoluble in water it is necessary to develop an economical and streamlined liquid chromatography for determination of pure and commercial dosage of Tofisopam. The main objective of the work is to develop and validate an RP-HPLC method for determination of Tofisopam as per ICH guidelines. An RP- HPLC method was devised using 0.1% Orthophosphoric acid in water: methanol in the ration 10:90% v/v and validated for parameters such as system suitability, linearity, limit of detection, limit of quantitation, precision, accuracy, assay, robustness, and ruggedness, as per ICH Guidelines. Tofisopam showed the maximum absorbance at 238nm with good system suitability and linearity was entrenched within the range of 10-60µg/ml, with the regression coefficient of 0.9996. The limit of detection and limit of quantitation were found to be 2.75 µg/ml and 8.855µg/ml respectively. Assay was valuated to be 101%, Accuracy was valuated between 98-103%, and %RSD was less the 2% for precision, robustness and ruggedness, so the method is precise and accurate. The method robustness is demonstrated by insignificant variations in absorption values and mobile phase ratio with deliberate alterations. In accordance with the validation results, the procedure is simple, specific, precise, accurate, robust and suitable for routine analysis of Tofisopam in bulk and commercial products.

Organophosphate esters and their metabolites in marine fish from Vietnam: Analytical method development and validation

Organophosphate esters (tri-OPEs), which are a significant group of emerging organic pollutants, have been widely found in aquatic products worldwide. While tri-OPEs have been extensively detected in aquatic products and are known to represent potential risks to human health, there is limited information available regarding their metabolites (di-OPEs). Currently, the techniques for analyzing the composition of tri- and di-OPEs have been distinguished and have certain drawbacks in terms of their expense, time requirements, and reliability. The objective of the study is to develop, validate, and utilize a modified QuEChERS technique for extracting tri- and di-OPEs in marine fish. As a result, EMR-Lipid and WAX were selected as adsorbents for the QuEChERS method to purify the samples and a combination of centrifugal vacuum and nitrogen at a temperature of 1 °C was selected as a technique to enhance the sensitivity of the developed method. The parameters of the method validation are in accordance with the criteria specified in SANTE/11312/2021 (a recovery ranging from 84.0% to 107.4% and RSD of precision below 20%). The aforementioned methodology was employed to assess the tri-OPEs and their metabolites in a total of 72 fish muscle samples obtained from the presence of OPEs in all recorded instances. The average concentration of ΣOPEs was determined to be 87.4±55.0 ng/g dw, with TBOEP and BEHP identified as the predominant compounds. Furthermore, no significant variation in the content of tri- and di-OPEs was observed in fish samples, regardless of species or geographical location.

Analytical Method Development and Validation of Brivaracetam in API and Marketed Formulation by RP-HPLC

A rapid and precise reverse phase high performance liquid chromatographic method has been developed for the validated of brivaracetam in its pure form as well as in tablet dosage form. Chromatography was carried out on a Symmetry ODS RP C18,5mm, 15mm x 4.6mm column using a mixture of phosphate buffer: methanol: acetonitrile (30:35:35% v/v) as the mobile phase at a flow rate of 1.0ml/min, the detection was carried out at 285 nm. The retention time of the brivaracetam was 2.183. The method produces linear responses in the concentration range of 60-140μg/ml of brivaracetam. The method precision for the determination of assay was below 2.0%RSD. The method was found to be sensitive, accurate and precise useful in the quality control of bulk and pharmaceutical formulations.

Sensitivity analysis for matched observational studies with continuous exposures and binary outcomes

Summary Matching is one of the most widely used study designs for adjusting for measured confounders in observational studies. However, unmeasured confounding may exist and cannot be removed by matching. Therefore, a sensitivity analysis is typically needed to assess a causal conclusion’s sensitivity to unmeasured confounding. Sensitivity analysis frameworks for binary exposures have been well-established for various matching designs and are commonly used in various studies. However, unlike the binary exposure case, there still lacks valid and general sensitivity analysis methods for continuous exposures, except in some special cases such as pair matching. To fill this gap in the binary outcome case, we develop a sensitivity analysis framework for general matching designs with continuous exposures and binary outcomes. First, we use probabilistic lattice theory to show our sensitivity analysis approach is finite-population- exact under Fisher’s sharp null. Second, we prove a novel design sensitivity formula as a powerful tool for asymptotically evaluating the performance of our sensitivity analysis approach. Third, to allow effect heterogeneity with binary outcomes, we introduce a framework for conducting asymptotically exact inference and sensitivity analysis on generalized attributable effects with binary outcomes via mixed- integer programming. Fourth, for the continuous outcomes case, we show that conducting an asymptotically exact sensitivity analysis in matched observational studies when both the exposures and outcomes are continuous is generally NP-hard, except in some special cases such as pair matching. As a real data application, we apply our new methods to study the effect of early-life lead exposure on juvenile delinquency. An implementation of the methods in this work is available in the R package doseSens.

Validation of an efficient analytical method for quantification of tetracycline antibiotic residues in egg samples along with investigating their decontamination using ultraviolet waves

ABSTRACT This study describes the development and application of a dispersive solid-phase extraction procedure combined with dispersive liquid – liquid microextraction for the extraction of tetracycline antibiotics from egg samples. In this method, a bio-metal organic framework was prepared and used in extraction of the analytes from sample solution. In this method, the sorbent synthesis was relatively easy and its crosslinker was vitamin B3 which is a green chemical compound. The sorbent provides particles can extract the analytes via interactions such as surface adsorption, occlusion van der waals and π–π interactions. The extraction procedure was done by mixing trichloroacetic acid with the egg sample to remove the sample proteins and then the synthesised sorbent at mg – level was added to the supernatant phase and the mixture was vortexed. After centrifuging, the supernatant phase was removed and sorbent particles containing the extracted analytes were eluted using acetonitrile. After that, this phase containing the analytes was taken and used in the following dispersive liquid – liquid microextraction process that was done to more enrichment the analytes using a deep eutectic solvent. After optimisation of the extraction process, sensitive limits of detection and quantification were found in the ranges of 0.21–0.28 and 0.70–0.93 ng g−1, respectively. Acceptable extraction recoveries of the analytes ranged from 60 to 73%. The method was selective regarding the analytes, as there were no interfering peaks eluted at the retention times of the analytes. After the validation step, the offered method was employed for the determination of the tetracycline antibiotics in various egg samples and the effect of ultraviolet waves in decontaminating them from egg samples was investigated.

Comparative study and validation of a new analytical method for hydraulic modelling of bidirectional low temperature networks

Bidirectional Low Temperature Networks (BLTNs) are an innovative technology of district heating and cooling systems. Because of their intrinsic feature of supplying simultaneous heating and cooling either directly from an energy source/sink or through distributed heat pumps and chillers, circulation pumps installed at each building extract water from the network and a distinct pressure cone is subsequently created. Variations in the amount (and direction) of the extracted water call for accurate hydraulic design and analysis to ensure safe and adequate network operation. This paper proposes a new analytical solution method for fast hydraulic modelling of BLTNs. The analytical method was compared against three numerical solution methods and validated using different current and future operating scenarios of a real-world bidirectional network. In all modelled cases, the calculated mass flows and pressure drops using the new analytical method yielded very small Normalised Mean Bias Error (NMBE) and Coefficient of Variation of the Root Mean Square Error (CV(RMSE)), suggesting excellent agreement between the analytical and numerical solution methods. The new analytical method offers better performance over its numerical counterparts due to its explicit nature, ease of implementation, and quick modelling time.

Analytical Method Development and Validation of a UV‐Spectrophotometric Technique for Pioglitazone HCL Estimation in Bulk and Polymeric Nanoparticles

AbstractThe type 2 diabetes drug pioglitazone is encapsulated in polymeric (Chitosan/PEG blended PLGA) nanoparticles utilizing solvent evaporation methods. The current research aimed to develop and validate an accessible, accurate, fast, and commercial UV spectrophotometric approach for assessing Pioglitazone in bulk and polymeric nanoparticles. In the estimation method that involves a pH 7.4 phosphate buffer, the maximum absorbance of Pioglitazone is reported to be 269 nm. With a correlation coefficient of 0.999, the drug exhibits linearity in the concentration series of 5–60 µg mL‐1. The proposed technique is validated as per ICH guidelines for linearity, accuracy, precision, and ruggedness. When the proposed technology is used in nano‐formulation, the percent amount of drug estimated to be 99.35% agreed well with the label claim. Recovery experiments at three distinct levels, 50%, 75%, and 100% are used to test the method's accuracy. The recovery rate is estimated to be between 98.91% and 99.11%. The low percent relative standard deviation (RSD) value indicates the method's accuracy and reproducibility. The method's repeatability, intraday, and interday precision are all examined. The approach is precise if the percent RSD value is less than 2. With the assistance of two analysts, the ruggedness of the suggested method is investigated. For the scheduled examination of Pioglitazone in bulk and polymeric nanoparticles, the above process has been developed and validated and proved to be a quick and cost‐effective quality‐control tool in pharmaceuticals.

Development of an analytical method for the determination of more than 300 pesticides and metabolites in the particulate and gaseous phase of ambient air

Pesticides can enter the atmosphere during spraying or after application, resulting in environmental or human exposure. The study describes the optimisation and validation of analytical methods for the determination of more than 300 pesticides in the particulate and gaseous phases of the air. Pesticides were sampled with high-volume air samplers on glass-fibre filters (GFFs) and glass columns filled with polyurethane foam (PUF) and XAD-2 resin. Comparing different extraction methods, a QuEChERS extraction with acetonitrile was selected for the GFFs. For the PUF/XAD-2 columns, a cold-column extraction with dichloromethane was used. Instrumental determination was performed using liquid chromatography/electrospray ionisation-time-of-flight mass spectrometry (LC/ESI-QTOF) and gas chromatography/electron impact ionisation-tandem mass spectrometry (GC/EI-MS/MS). Recovery experiments showed recovery rates between 70 and 120% for 263 compounds on the GFFs and 75 compounds on the PUF/XAD-2 columns. Semi-quantitative determination was performed for 39 compounds on the GFFs and 110 compounds on the PUF/XAD-2 columns. Finally, 27 compounds on the GFFs and 138 compounds on the PUF/XAD-2 columns could be determined only qualitatively. For the determination of the PUF/XAD-2 samples, signal suppression (LC) or signal enhancement (GC) due to matrix effects were determined. Method quantification limits of the optimised methods ranged from 30 to 240 pg/m3 for the target compounds on the GFFs, and from 8 to 60 pg/m3 on the PUF/XAD-2 columns. The applicability of the method was demonstrated by means of environmental air samples from an agricultural area in the Netherlands.

The Quality by design Approach for Analytical Method Development and Validation of the RP-HPLC Method for Estimation of Quercetin in Pure, Marketed, and Cream Formulation

ABSTRACT: A simple, precise reverse phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the flavonoid quercetin, isolated from Tridax procumbens L. The method was validated by using Phenomenex C18 (250 x 4.6mm i.d) Column. A simple, cost effective mobile phase consisting of (ACN and 10 m/moL Phosphate buffer as mobile phase in proportion of 50:50 v/v ) pH 3, 1.0 ml/min Flow rate at 370nm by using UV Visible detector. The retention time of Quercetin was found to be 3.392 minutes. The Quercetin linearity range was found to be 05 to 25 μg/mL. The accuracy and precision of commercially available preparations and in-house cream formulations were investigated using a one-way ANNOVA test. The Percentage recovery of both formulations was found to be 99.83%, 99.88%, 99.82% and 98.92%, 98.18%, 98.86%. Robustness of analytical method was studied by using 2/3 full factorial design by using Design expert software. The Percentage assay and % RSD of marketed capsule and in house cream was found to be 98.38 %, 97.40% and 0.05273, 0.02053 respectively. The Limit of Quantitation and Limit of detection were found to be 0.9053μg/mL and 2.5435μg/mL respectively. The development method of quercetin is simple, accurate, precise, sensitive, and robust