Method Development and Validation

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Important Aspects of the Design of Experiments and Data Treatment in the Analytical Quality by Design Framework for Chromatographic Method Development
by Bianca F. G. Passerine andMárcia C. Breitkreitz *ORCID
Institute of Chemistry, University of Campinas (UNICAMP), Campinas 13083-970, SP, Brazil
*
Author to whom correspondence should be addressed.
Molecules 2024, 29(24), 6057; https://doi.org/10.3390/molecules29246057
Submission received: 1 November 2024 / Revised: 9 December 2024 / Accepted: 16 December 2024 / Published: 23 December 2024
(This article belongs to the Section Analytical Chemistry)
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Abstract
In the analytical quality by design (AQbD) framework, the design of experiments (DOE) plays a very important role, as it provides information about how experimental input variables influence critical method attributes. Based on the information obtained from the DOE, mathematical models are generated and used to build the method operable design region (MODR), which is a robust region of operability. Data treatment steps are usually carried out in software such as Fusion QbD, Minitab, or StaEase 360, among others. Although there are many studies in the literature that use the DOE, none of them address important aspects of data treatment for optimization and MODR generation and compare different software calculations. The purpose of this study is to contribute to a better understanding of data treatment aspects that are frequently misread or not fully understood, such as model selection, ANOVA results, and residual analysis. The discussion will be guided by the separation of curcuminoids, using ultra-high performance liquid chromatography and eight quality attributes as responses. This study highlights the importance of carefully selecting and evaluating models, as they significantly influence the generation of the MODR. Moreover, the findings emphasize that it is essential to incorporate uncertainties into the contour plots to accurately determine the MODR in compliance with the ICH Q14 guidelines and USP General Chapter <1220>.

Simple, accurate, precise, and sensitive reverse-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous measurement of ARI and ESC in pharmaceutical products by using a C18 column with dimensions of 250 mm x 4.6 mm and a particle size of 5 μm. The mobile phase consisted of a mixture of acetonitrile, methanol, and water in the ratio of 80:05:15 v/v/v. Orthophosphoric acid was added to adjust the pH to 7.00 ± 0.1. The detection wavelength used for measuring the absorbance of the components was 246 nm, and the Mobile Phase flow rate was set at 1.0 mL/min. The retention times of ARI and ESC were 5.63 and 3.65 minutes, respectively. The ARI and ESC correlation coefficients (R 2 -values) were observed at 0.9995 and 0.9997, respectively, demonstrating a robust linear relationship between the analyte concentration and the observed absorbance. The linearity range for ARI was 20 to 120 μg/mL, while ESC was 60 to 360 μg/mL. The recovery percentages for ARI and ESC ranged from 99.43% to 100.65% and 99.58% to 100.15%, respectively. Additionally, all parameter uncertainty levels were below 2. The method was validated following ICH guideline Q2 (R1). The analytical method validated will be successfully applied to the pharmaceutical dosage form.

Objective: A new and validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and used for the quantitative analysis of the anticancer drug Lurbinectedin in plasma samples. Methods: Liquid-liquid extraction method was used to extract the Lurbinectedin from plasma, and Ledipasvir was used as an internal standard (IS), Chromatographic separation of Lurbinectedin was accomplished with Zorbax SB-C18 column (250 mm×4.6 mm, 5 µm) using a mobile phase comprises of 0.1% v/v formic acid and acetonitrile (10:90, v/v) pumped at a flow rate of 0.80 ml/min. The total run time for Lurbinectedin and internal standard elution was 3.5 min. The linearity was attained for a concentration range of 1.50-5000 ng/ml with a correlation coefficient (r²) of 0.9994 by the optimized method. The lower limit of quantification (LLOQ) was established at 1.50 ng/ml, which resembles the sensitivity of the method toward Lurbinectedin. Results: The method showed high accuracy, ranging from 95.31% to 104.06% of nominal values, with intra-and inter-day precision within a 4.32% coefficient of variance (%CV). Matrix effects at the low-quality control (LQC) level varied between 94.25% and 104.85% (%CV: 4.61), while at the high-quality control (HQC) level, they ranged from 94.62% to 103.88% (%CV: 4.02). The mean recovery across all control standards was 95.58%. Conclusion: Stability assessments confirmed the robustness of the method. The developed LC-MS/MS approach is reliable for routine bioanalytical applications, including quality control, pharmacokinetic studies, and bioequivalence assessments of Lurbinectedin in biological matrices.

A simple, sensitive, and accurate chromatography (RP-HPLC) method for simultaneous estimation of Ciprofloxacin, Ofloxacin, and Marbofloxacin in their combined pharmaceutical dosage form or individually. The HPLC separation was achieved on a Hypersil (C18, 150 mm × 4.6 mm, 5 μm particle size) analytical column or equivalent. A mixture of triethanolamine (1 %), acetonitrile (80 %), and water was used as the mobile phase, with a flow rate of 1.2 mL /min and a detector wavelength of 280 nm at ambient temperature. In the HPLC method, the retention times of Ciprofloxacin, Ofloxacin, and Marbofloxacin were found to be 1.854, 2.480, and 4.688 min, respectively, and linearity was obtained in the range of 25-80 µg/mL for Ciprofloxacin, Ofloxacin, and Marbofloxacin. The correlation coefficient for the method was greater than 0.999. The RP-HPLC method validation parameter lay within its acceptance criteria as per the ICHq2 (R1) guideline. Hence, it can be successfully used for routine analysis of Ciprofloxacin, Ofloxacin, and Marbofloxacin in raw material or pharmaceutical dosage forms.

A simple, sensitive, and accurate chromatographic (RP-HPLC) method for simultaneous estimation of norfloxacin, levofloxacin, and moxifloxacin in their combined. Pharmaceutical dosage forms or individually. The HPLC separation was achieved on a Hypersil (C18, (150 mm x 4.6 mm,5 μm particle size) analytical column or equivalent. A Mixture of triethanolamine 1%: acetonitrile 80 % was used as the suitable mobile phase, at a flow rate of 1.2 mL/min and detector wavelength at 280 nm at ambient temperature (25 °C). In the HPLC method, the retention time of norfloxacin, levofloxacin, and moxifloxacin was found to be 1.854, 2.480, and 4.688 min, respectively, and the linearity of the method was obtained in the range from 25 to 80 µg/mL for norfloxacin, levofloxacin, and moxifloxacin. The Correlation coefficient for the method was greater than 0.999 RP-HPLC method validation parameter lay within its acceptance criteria as per the ICHq2 (R1) guideline Validation of analytical procedures. Hence it can be successfully used for routine analysis of norfloxacin, levofloxacin, and moxifloxacin in raw material or pharmaceutical dosage forms.

Objective: Metformin hydrochloride and sitagliptin are the oral anti-hyperglycemic medications used to treat type 2 diabetes and are used in combination to treat patients. In this work, we have developed a bioanalytical method for simultaneous estimation of both the drugs form some formulation and subsequently the validation of the developed method metformin and sitagliptin in human plasma. Methods: The stability studies were done as per USFDA and EMA guidelines. The sample extraction approach presented here was a straightforward liquid extraction. The linearity range of metformin was 11.72 ng/ml to 3000 ng/ml, and sitagliptin was 4.68 ng/ml. to 1200 ng/ml. For metformin, the LOD was 1.0 ng/ml, and LLOQ was 11.72 ng/ml. and for sitagliptin, the LOD was 0.75 ng/ml, and LLOQ was 4.68 ng/ml. LC-ESI-MS/MS was used to develop and validate this method using the Phenomenex Kinetex C18 column. Milli-Q water containing 10 mmol Ammonium Acetate (pH =3.6) and Acetonitrile containing 0.1% Formi...

Nitrosamines impurities have strong carcinogenic potential which are making drug safety monitoring crucial. Recent detection of nitrosamines impurities in various drugs has led regulatory agencies to increase surveillance and almost all the regulatory bodies issue guidelines to control these impurities. Gabapentin, being one of the most widely prescribed drugs, has been one of the candidate for possible presence of nitrosamine impurities. In this article an analytical method developed to detect N-Nitrosodibutylamine (NDBA) in Gabapentin Capsules (100 mg, 300 mg, and 400 mg) using Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) has been validated as per the requirements of International Council for Harmonization (ICH) and United States Pharmacopeia (USP) guidelines. The LC-HRMS method was validated for its system suitability, specificity, system precision, method precision, quantification limits, and detection limits. System precision showed % RSD (n=6) of 5.0% with no blank interference, The method is specific for determination of NDBA Impurity as there was no interference from the blank and placebo at the retention time of NDBA impurity, while the system precision showed % RSD (n=6) of retention time of 0.7% and % RSD (n=6) of Area response was 5.0%. Method precision had % RSD (n=6) of 2.2%. The method achieved a limit of detection (LOD) of 0.05342 ng/mL (0.000742 ppm) and a limit of quantitation (LOQ) of 0.16188 ng/mL (0.002250 ppm). These results confirm the method's accuracy, sensitivity, and reliability for routine quality control analysis.

Objectives: To develop two (titrimetric and spectrophotometric) simple, rapid, sensitive and cost-effective methods for the determination of metoclopramide (MCP) in pharmaceutical dosage forms. Materials and Methods: The titrimetric method (A) was based on the N-oxidation reaction involving the use of potassium hydrogen peroxymonosulfate and subsequent iodometric back titration of a known residual reagent. The spectrophotometric method (B) was based on derivatization of MCP with potassium hydrogen peroxymonosulfate in the presence of iodide to produce a chromogen (triiodide) with a wavelength of maximum absorption at 350 nm. Results: Method "A" was applicable over the concentration range of 0.25-3.5 mg to end volume 10 mL. In method "B", Beer's law was obeyed over the concentration range of 0.3-3.5 μg/mL with molar absorptivity of 24600 L/mol cm. The limits of quantification were calculated to be 0.25 mg/10 mL (A) and 0.2 μg/mL (B), respectively. Conclusion: The proposed methods were suitable for determination of MCP as a pure substance in tablets and injection.

Terazosin hydrochloride is indicated in the symptomatic relief of benign prostatic hyperplasia and it is also used as an antihypertensive agent. A rapid, simple, specific, and economic UV-spectrophotometric method has been developed to determine the stability indicating method for terazosin using water as a solvent. The absorption spectrum of terazosin showed maximum absorbance at 250 nm and obeyed Beer's law in concentration range 1.5-9 µg/ml. Linear regression of absorbance on concentration gave the equation y=0.1103x+0.0101 with correlation coefficient 0.999. The method developed is validated for parameters like specificity, linearity, range, accuracy, precision, limit of detection (LOD), limit of quantitation (LOQ), robustness as per Q2A specifications of the ICH guidelines. The drug sample was exposed to acid, alkali, and oxidation by hydrogen peroxide, thermal and photolytic conditions to study the forced degradation of the drug. The obtained results proved that this method can be employed for the routine analysis of terazosin in bulk as well as in the commercial formulations.

Objective: The research work unveils the use of nuclear magnetic resonance (NMR) technique for quantitative determination and method validation of obeticholic acid. As standard expository methodology for more up to date medications or formulations may not be available in pharmacopeias, hence it is fundamental need to create novel analytical procedures which should be precise and accurate. Methods: Proton (1H) and carbon (13C) NMR analysis were initially performed to confirm the preliminary authenticity of obeticholic acid API. Method validation was accomplished on the basis of standard guidelines for the parameters, in which tetramethylbenzene as an internal standard and deuterated dimethyl sulfoxide as a diluent were used to assess the obeticholic acid. Results: For the quantification of the drug, the proton nuclear magnetic resonance signals at 0.602 ppm and 6.86 ppm corresponding to the analyte proton of drug and internal standard respectively were used. The curve equation calcul...

A simple, specific, accurate, and precise reverse phase high performance liquid chromatography (RP-HPLC0 method was developed and validated to analyse Rivaroxaban bulk dosage form. Using Sun Q C18 HPLC column separation was carried out. This was maintained at ambient temperature. During separation mobile phase consist of Acetonitrile: Buffer(sodium acetate buffer) 80:20 was delivered at a flow rate of 1mL/min. Using UV detector analysis was carried out at the wavelength 249 nm. RP-HPLC method was validated by using various parameter like system suitability, linearity, specificity, precision, limit of detection, limit of quantification and robustness. The RP-HPLC method was found to be linear over the concentration range from 5-30 µg/mL (r 2 =0.994). Retention time for bulk rivaroxaban was found to be 3.69 min. LOQ of method was 1.331 µg/mL and LOD 0.439 µg/mL. Thus the developed RP-HPLC method 14689 was found to be robust which can be applied for the regular analysis of Rivaroxaban in the bulk as well as pharmaceutical dosage form.

A simple, specific, accurate, and precise reverse phase high performance liquid chromatography method was developed and validated to analyse Dolutegravir Sodium bulk dosage form. Using Princeton C18 HPLC column separation was carried out. This was maintained at 25ºC temperature. Shimadzu LC -20A prominence with LC solution softwarering separation mobile phase consist of Methanol: Water (0.1% OPA) in ratio of 80:20 v/v was delivered at a flow rate of 1mL/min. Using UV detector analysis was carried out at the wavelength 254 nm. RP-HPLC method was validated by using various parameter like system suitability, linearity, specificity, precision, limit 14664 of detection, limit of quantification and robustness. The RP-HPLC method was found to be linear over the concentration range from 10-60 µg/mL (r 2 =0.9998). Retention time for bulk Dolutegravir Sodium was found to be 4.383 min. LOQ of method was 1.3274 µg/mL and LOD 0.4380 µg/mL. Thus the developed RP-HPLC method was found to be robust which can be applied for the regular analysis of Dolutegravir Sodium in the bulk as well as pharmaceutical dosage form.

Dissolution analysis by HPLC for Magnesium Oxide Tablets 400 mg with Conductivity Suppression (SCD) Detector and/or by Atomic Absorption Spectroscopy as per current USP monograph method is quite difficult and non-feasible due to noneconomical availability of SCD detector and Atomic Absorption Spectroscopy. Hence, Dissolution analysis by Titration method for Magnesium Oxide Tablets has been developed using 0.025M Edetate disodium VS as titrant for titration method. The Dissolution Tolerance is NLT (Q) 75% of the labelled amount of Magnesium Oxide is dissolved. The method was validated as per ICH Quality guidelines Q2(R2) Validation of Analytical Procedure with specificity, method precision, linearity and accuracy. The study findings obtained the coefficient correlation of 0.9998 for linearity and %RSD of 2.9 with 91% of average dissolution for precision. The recovery results of 50%, 100% and 120% ensures the accuracy of the method with 100.1%, 98.1% and 97.0% respectively.

A straightforward method for the determination of histamine and other biogenic amines in Vietnamese traditional fish sauce was developed, avoiding a time-consuming derivatisation step. This resulted in the application of hydrophilic interaction in combination with liquid chromatography-tandem mass spectrometry (HILIC-MS/MS). The characteristics of the analytical method, such as linearity, limit of detection (LOD), limit of quantification (LOQ), repeatability, stability, and matrix effects were systematically investigated. The validated method was successfully applied to analyse 8 biogenic amines in 15 Vietnamese traditional fish sauce samples, as well as 10 broth samples directly collected from the fermentation tanks. Among the eight investigated biogenic amines, histamine was found in all analysed samples, in some at high levels. However, in traditional fish sauce samples collected from supermarkets histamine levels were lower than the maximum level as set by European Union regulations.

This study aims to develop and validate novel UV spectrophotometric methods for the simultaneous estimation of atenolol and hydrochlorothiazide in pharmaceutical dosage forms and bulk. The proposed methods are Simultaneous Equation and Q-Absorbance Ratio methods offer a cost-effective, eco-friendly alternative to chromatographic techniques. The λ max of atenolol and hydrochlorothiazide were identified at 224.00 nm and 234.00 nm, respectively, with an isoabsorptive point at 229.00 nm for the Q-Absorbance Ratio method. The validation followed ICH Q2 (R1) guidelines, demonstrating excellent linearity (R² > 0.99 for both drugs), accuracy (98.5 %-101.2 %), precision (%RSD < 2 %), and sensitivity (LOD: 1.61 µg/mL for atenolol, 0.78 µg/mL for hydrochlorothiazide). The developed methods were successfully applied to a marketed formulation, showing recovery rates within acceptable limits. The environmental sustainability of these methods was assessed using ComplexGAPI, AGREE, Analytical Eco-Scale, ChlorTox WHN, and CHEMS-1 tools. The AGREE score (0.71) and ComplexGAPI assessment (eight green attributes, two red) confirmed superior eco-friendliness over reported HPLC methods (AGREE: 0.62). ChlorTox scores further validated minimal toxicity risks. This research highlights an efficient, accurate, and environmentally benign alternative for atenolol and hydrochlorothiazide analysis, aligning with green analytical chemistry principles.

Peanut is one of the most widely used legumes due to its nutrition and taste. During the storage, this product is susceptible to pest infestations and microbial contamination. X-ray Ionizing radiation is a non-thermal, safe and effective technology used to inactivate pathogens and may positively affect peanut preservation. The European regulatory framework included the legumes matrices into the list of food and food ingredients which may be treated with ionizing radiation up to 1.0 kGy, therefore control methods to
identify any illicit treatments are essential for Member States. The UNI EN 13784:2002 DNA comet test is a rapid and sustainable screening tool for analyzing irradiated vegetables in terms of DNA breaks due to X-rays.
However, thermal, mechanical and irradiation DNA damage can be similar, therefore a confirmatory method is mandatory. HS-SPME/GC-MS is a green alternative to standard method UNI EN 1785:2003 for the qualitative determination of 2-dodecylcyclobutanone (2-DCB) known as radiation marker. 2-DCB is specific compounds generated from triglycerides containing palmitic acid by irradiation of fat-rich foods. In this work, shelled raw peanuts were irradiated in plastic bags using a low-energy X-ray irradiator at dose levels of 0.5, 1.0 and 1.5 kGy. The comet DNA was appropriately validated using 1.0 g of crushed peanuts and optimized
sedimentation and lysis times to 45 minutes for both. The HS-SPME/GC-MS analysis, performed using headspace extraction with PDMS fibre and SIM mode acquisition, was carried out on 5.0 g of ground samples in a 20 mL glass headspace vial containing 5 ml of deionized water. The identification criteria of 2-DCB were compliant with EN ISO 1785:2003. In not treated peanuts, used as control, intact cells without comets were observed while the irradiated samples showed comets with a homogeneous pattern and the fluorescence intensity is proportional to the treatment dose. The screening results were confirmed by the presence of 2-DCB only in the irradiated samples. Moreover, interference was not highlighted and the area of 2-DCB was increased directly to dose treatment, confirming the selectivity and linearity of HS-SPME/GC-MS method for the selected matrix. The outcomes confirmed the possibility to use the UNI EN 13784:2002 DNA comet assay
together with HS-SPME-GC/MS for vegetables products analysis within official laboratories for routinely control activity.

In order to accurately determine valsartan (API), this work aims to build a straightforward and exact UV spectroscopic approach. To improve sensitivity and accuracy, the procedure entails methodically optimizing experimental parameters, including solvent choice and wavelength selection. The produced stock solutions were tested for absorbance at 249 nm using a UV spectrophotometer. With %RSD values below 2% for both intra-and inter-day, the devised approach was determined to be quite accurate. At each additional concentration, the approach was shown to be accurate, yielding good drug recoveries ranging from 98 to 101%. The process was validated for linearity, specificity, accuracy, precision and limit of detection (LoD) and limit of quantification (LoQ) as per International Council of Harmonization (ICH) criteria. Its dependability for quantitative analysis of valsartan in pharmaceutical formulations was demonstrated. This UV spectroscopic approach offers a cost-effective and efficient alternative for routine quality control in pharmaceutical industries.

Quality by design (QbD) is modern and systematic approach for product development and quality of pharmaceuticals. In this concept of QbD it is essential to define desire product performance profile, analytical target Profile and identify critical quality attributes throughout designing and development of a process. This review aimed to identify implementation of QbD in analytical procedure validation. Integration of QbD is an essential step in developing a systematic method which effectively fit for purpose and assures quality. The outcomes of organized analytical process development and validation are understandings of critical quality attributes, risk assessment and outlining design space. Consequently, control strategy can be set for further investigation or other issues. Posaconazole is a approved lipophilic triazole antifungal agent that exhibits potent and broad-spectrum antifungal activity in vitro and in vivo against most Candida spp., Cryptococcus neoformans, Aspergillus spp., many Zygomycetes, endemic fungi and dermatophytes. It has been documented that posaconazole has potency and spectrum of activity similar to those of itraconazole and superior to those of fluconazole against clinically important isolates of Candida spp., C. neoformans and Aspergillus spp. A spectrophotometric method has been developed and validated for the determination of Posaconazole in pharmaceutical formulation. QbD approach was carried out by varying 19 parameters and critical parameters were extracted by using principal component analysis and by observation. The extracted critical parameters are summarized in Table .1. PCZ followed linearity in the concentration range of 10-50 g/mL. The proposed method was applied for pharmaceutical formulation and percentage label claim was found to be 99.05%. The amount of drug estimated by proposed method was in good agreement with the label claim. The method was found to be precise as indicated by the inter-day and intra-day analysis showing % RSD less than 2. There was no any interference of excipients showing that the method was specific. Limit of detection and limit of quantitation were 0.09 and 0.27 g/mL, respectively.

Candesartan cilexetil (CAN), a prominent angiotensin II receptor blocker (ARB), has gained significant attention in cardiovascular medicine. This comprehensive review article delves into various facets of CAN, ranging from its pharmacology and pharmacodynamics to drug interactions, contraindications, and adverse reactions. The pharmacological profile of this compound is explored, elucidating its mechanism of action and its role in modulating the renin-angiotensin-aldosterone system. A detailed description of CAN, its formulation, tablet names, brand names, and compositions, is presented in a tabular format for quick reference. A crucial aspect of this review is the discussion of drug interactions, highlighting potential combinations that require caution and vigilant monitoring. Robust analytical techniques are vital for quality control, bioavailability assessment, and bioequivalence studies. Moreover, this article goes beyond clinical aspects by encompassing analytical methods employed in the quantification of CAN. The incorporation of analytical methods provides a comprehensive overview of the scientific methodologies used to measure CAN concentrations in biological samples.

This article explores the integration of Artificial Intelligence (AI) in pharmaceutical quality control and manufacturing processes. We discuss how AI technologies, particularly Artificial Neural Networks (ANNs) and machine learning algorithms, are revolutionizing quality assurance in the pharmaceutical industry. The paper examines AI applications in process monitoring, batch consistency, Quality by Design (QbD) approaches, and data management. We also address ongoing challenges in AI adoption and potential solutions. The findings suggest that AI significantly improves accuracy, efficiency, and consistency in pharmaceutical manufacturing, aligning with regulatory initiatives like Process Analytical Technology (PAT) and QbD.

This study investigates the potential uses of palm shell, pretreated with sulfuric acid (APSP) for the adsorption of AOII, DSB, and AV7. The effects of different system variables like adsorbent dosage, pH, contact time, and temperature were studied. Optimum pH values for all the three dyes were determined as 1.0. Equilibrium was achieved within 30 min. Langmuir I, II, III, IV and Freundlich isotherm models were applied to describe the equilibrium isotherms at different temperatures, and the langmuir model was found to agree very well with the experimental data. The maximum adsorption capacity was found to be 2180.05 mg/g, 1199.99 mg/g, and 243.9 mg/g for AOII, DSB, and AV7, respectively. Thermodynamic parameters such as change in free energy (∆G0), enthalpy (∆H0), and entropy (∆S0) were also determined. Pseudo-first-order, pseudo-second order, and intraparticle diffusion models were used to fit the experimental data. The pseudo-second-order equation was able to fit well and provide a realistic description of the adsorption kinetics.

ABSTRACT In this present work very simple, highly rapid, highly accurate as well as more precise gradient RP-HPLC process is developed also validated to separate foreign substances (IMP) present in Erythromycin (ERT-A) tablets pharma dosage form. Chromatographic disengagement is drifting out on WatersX-Terra RP 18 (250 mm x 4.6 mm I.D., 3.5 µm particle size) at 65°C is recycled to this separation. Mobile Phase-A consists of Buffer solution which is prepared by taking 35.0000g of di-potassium hydrogen phosphate in 1000.00mL of triple distilled water. pH is noted at 7.0 by using dilute O-phosphoric acid, purified over 0.45 µm membrane filter) acetonitrile as well as water in ratio of 5v: 35v:60v Phosphate buffer pH 7.0, water and acetonitrile in ratio of 5v:45v:50v is used as mobile Phase-B. The rate of flow and injection volume was 1.00mL/min and 100.00μL respectively. Analysis is lugged out beneath gradient circumstances includes time in min./A (v/v): B(v/v); T0/100:00, T45/100:00, T47/0:100, T63/0:100, T65/100:00, and T70/100:00. 215 nm is the wavelength. The ERT-A degrades under different circumstances. Total degradation substances are well resolved by observed peaks from ERT. This process is observed as linear. The cumulative %RSD values are identified, found that they are within the range. By utilizing this analysis revealed that this process is more accurate, highly specific, more selective, highly precise, robust, and also useful in development of process as well as quality check in flawless dosage form manufacture.

This research aimed to design as well as evaluate reverse-phase high-performance liquid chromatography (RP HPLC) process to precise quantification for potential impurities in Mirabegron (MIRA) ER Tablets. The developed method utilized an Inertsil C8 - 3, 150 x 4.6 mm, 3µm column with a gradient elution program utilizing Buffer and Acetonitrile as mobile phases. The chromatographic separation achieved on a C8 stationary phase demonstrated robustness and reproducibility, with detection at a wavelength of 240 nm and a constant column temperature of 40°C. The research study was focused on MIRA and four impurities (MIR-1, Deshydroxy, Diamide-1, and Diamide 2). The method exhibited specificity, precision, linearity, accuracy, robustness, ruggedness, and stability indicating characteristics. Correlation coefficients exceeding 0.997 for MIRA and its foreign substances showcased the accuracy and sensitivity of the method. Low detection and quantization limits (ranging from 0.06ppm to 0.20ppm) highlighted its high sensitivity. Accuracy is finalized by rates of recovery as 96.1% to 100.3% for all foreign substances. The stability-indicating is validated by forced degradation studies, meeting peak purity circumstances also achieving mass balance. Robustness studies further established the method's reliability, demonstrating resilience to minor chromatographic variations. This proposed RP-HPLC method accommodates a dependable, reproducible, highly accurate, and more sensitive means to quantifying MIRA-related substances in MIRA ER Tablets, making it a valuable tool for pharmaceutical quality control.

The aim of the present study is to assess the prescribing errors inhypertensive outpatients taking Antiplatelet drugswith other drugs and consequently the frequency and pattern of potential drug-drug interactions. A retrospective study was conductedfrom December 2017- May 2018.The data analysis was carried out by SPSS. Potential drug-drug interactions were determined by Micromedex 2. Severity of errors was evaluated according to NCCMERP (National Coordinating Council for Medication Error Reporting and Prevention). In current study 1142 medication errors (MEs) were trailed in 300 prescriptions. The utmost ubiquitous ME was the patient’s weight not mentioned in (95.3%) prescriptions continued by drug-drug interactions (95%). Diagnosis was missing in (73.3%) prescriptions. The average number of drugs per encounter was 5.64. According to NCCMERP highest number of the prescriptions (n=193, 64.33%) were fall in category D, followed by category C and I. Out of 300 prescriptions, majority o...

Turmeric is of huge importance in Indian food and traditional medicine that’s why it is also called as golden spice. It not only increases the taste of food but pharmacologically it helps to eliminate or reduce pain, oxidative stress and inflammation. Turmeric contains a chemical constituent known as curcumin which have scientifically proven analgesic, anti-inflammatory and antioxidant properties. Turmeric as a spice provides many health benefits and very useful ingredient of Ayurvedic medicines but if we came to know that this yellow spice is harmful, then what will happen and what can we do. Recent research says that, the levels of lead in samples of turmeric taken from different countries of the Asian continent were found 1,000 μg/g. This much level is too much high and responsible thing behind it was found Lead chromate. It is very shocking that although Lead chromate, a yellow colour pigment is not a food ingredient but it is used in food items (turmeric) to increase the colour appearance. According to the Indian standards, the lead level in turmeric (whole and powdered) cannot be more than 10 μg/g. Recent report of World health organization tells that people lead exposure makes a man sick. More than 0.15 crores deaths occurred all over the world due to lead exposure in the year 2021. Mostly the reason of death was found problem in the heart. In this article, we will go over some of the facts of turmeric adulteration, where lead comes from, analysis of lead and, the dangers of lead poisoning and lead chromate. The approaches for preventing and treating lead poisoning have also been covered.

Introduction
Laboratories offer use of biochemical tests on body fluids that are clinically relevant Body fluid biochemical tests provide valuable diagnostic information, guiding healthcare providers in making accurate diagnoses, monitoring disease progression, and evaluating treatment responses. Validating tests for off label use for unconventional fluids in chemistry poses challenges and requires thorough planning. Existing research on this topic offers partial guidance for clinical labs to create their own validations. This study aims to fill some of these gaps by providing evidences for performance specification using in house validation of these assays.
Methods
Unconventional types of body fluid were use such as pleural fluid, ascetic fluid (for glucose, albumin, protein, LDH, amylase, lipase, triglyceride, urea, creatinine, bilirubin total, cholesterol, ADA) where as cerebrospinal fluid was used for (albumin, LDH, ADA). A validation protocol was used to validate each analytical method for accuracy precision, linearity, sensitivity, specificity and interfering substances along with their acceptance criteria. Alternative calibrators having fixed assigned value were use to validated recovery. The equipment used was Beckman Coulter DxC 700 AU. Statistical analysis was done using Microsoft Excel (Version 2019, Microsoft, Seattle, WA)
Results
Precision for each assay and body fluid type had a CV <9%. Recovery for the various analytes in all fluid types was 93.5-108.6%. Biorad EQAS samples results comparisons were within 95.8 – 104.2%. Serial dilution with serum samples showed specificity of >95%. Sensitivity using specimen at lower limit of detection was confirmed with precision of less than 15% of CV. There were no interferences found for icterus, lipemia and haemolysis using spiked samples. The tests assays for various body fluids were found linear till the higher measurement range with and without dilutions
Conclusions
Beckman Coulter DxC 700 AU is validated for the off label use of biochemical analysis of various body fluids.

Gas Chromatography Isotope Ratio Mass Spectrometry (GC-C-IRMS) has traditionally been used in routine laboratories to measure the δ13C values of anabolic steroids in urine, enabling differentiation between endogenous and synthetic testosterone (T) in sports doping control. Liquid Chromatography Isotope Ratio Mass Spectrometry (LC-IRMS) has not been applied due to its limitation of not allowing organic solvents or modifiers in the mobile phase for δ13C analysis. Mid-to non-polar analytes like steroids can be analyzed in water heated to high temperatures (up to 200°C), where water's polarity becomes comparable to an 80/20 methanol/water mixture at ambient temperature. This study developed a method for analyzing steroids in urine, extending the application of LC-IRMS to non-polar analytes in complex matrices.

An HT-LC-IRMS method was successfully developed and validated to determine the δ13C values of four steroids: testosterone (T), 5α-androstane-3α,17β-diol (ααβ), 5β-androstane-3α,17β-diol (βαβ), and pregnanetriol (PT) in urine. Accuracy ranged from 0.23‰ (ααβ and βαβ) to 0.49‰ (T), with a detection limit of 10 ng/mL for T and ααβ+βαβ. Validation data and comparisons of authentic urine samples analyzed with HT-LC-IRMS and GC-C-IRMS showed comparable performance between the two methods.

HT-LC-IRMS is suitable for determining the δ13C values of anabolic steroids, expanding the applications of both HT-LC and LC-IRMS to non-polar substances in complex matrices in routine laboratory settings.

There appears to be a constantly incontrovertible, yet delicately inseparable cord, between land resource as an all-time invaluable asset, and meaningful socioeconomic growth and development. Much as this assertion holds, what enigmatically remains almost unsolvable is the seeming unending mystery that surrounds 'wilful' prevention of access to this very non-substitutable resource, in spite of its tangentiality to virtually all endeavours, around which the survival of all human beings revolves. Thence, it is compellingly tempting to reason that, some 'unseen' factors are decidedly making the delivery and accessibility of land, especially within urban milieu, extremely difficult, so much that, enviable growth and development might be so perennially elusive, particularly to the poor segment of the populace, which unfortunately forms the largest proportion of any country's citizenry, Nigeria inclusive. Although, the trend facades itself to be solely caused by economic and other attendant pecuniary factors, but as all the economically-defined strategies seem not too effective over the years, to finally nail several challenges associated with formal land delivery and accessibility; it behoves upon researchers and policy-makers alike, to look beyond the ordinary. Hence, the demystification exercise such as this, which is geared towards unravelling this mystery, reveals more factors that are indeed non-pecuniary, this is the crux of this study.

Objective: A unique liquid chromatography-mass spectrometry technique is essential for determining the concentration of asciminib in biological matrices, and its development is of the utmost importance.

Methods: The samples that were processed were separated using a Reversed Phase-Phenomenex (100 mm x 4.6 mm, 5 µm) C18 analytical column. The column was equipped with an isocratic moveable phase that consisted of 0.1% (v/v) HCOOH and acetonitrile at a ratio of 18:82% (v/v). The flow rate of the phase was 0.70 ml/min. For asciminib, the multiple reaction monitoring mode was used at m/z 450.23/257.3, while for canagliflozin, it was used at m/z 445.13/267.31.

Results: With a correlation coefficient of 0.9998, the method was linear for asciminib throughout the concentration range of 1.0-2100.00 ng/ml. Each day's accuracy percentage relative standard deviation was within 5.74%. For analytes at the low-quality control level, the mean matrix factors ranged from 96.34 to 104.85% with a % Coefficient of Variance (CV) of 4.21; at the high-quality control level, the range was from 94.62 to 103.88% with a %CV of 3.67.

Conclusion: The method that has been developed has the potential to be used to examine the pharmacokinetics and toxicokinetics of asciminib in various biological samples for both forensic and clinical purposes.

Background Zinc helps with cell division, growth, wound healing, and carbohydrate breakdown. Humanbeings have to obtain zinc from food or supplements because our bodies do not produce it naturally. In view of the greater advantages (such as low cost, time of analysis, and stability-indicating) compared to other quantification methods (titration, ion chromatography, Atomic absorption spectroscopy) proposed in the literature, a refractive index detector coupled with HPLC has been used in quantification of zinc. Objective The goal of this research is to develop and validate a sensitive, low-cost, high-resolution, and stability- indicating method for detecting and quantifying zinc levels in zinc supplement pharmaceutical products (injectables, tablets, and capsules). Methods A novel isocratic reverse-phase HPLC method with a refractive index detector (with sensitivity 64, detector temperature 35°C, and positive polarity) was developed using a carboxyl functional group packed column and ...

Background: Zinc helps with cell division, growth, wound healing, and carbohydrate breakdown. Humanbeings have to obtain zinc from food or supplements because our bodies do not produce it naturally. In view of the greater advantages (such as low cost, time of analysis, and stability-indicating) compared to other quantification methods (titration, ion chromatography, Atomic absorption spectroscopy) proposed in the literature, a refractive index detector coupled with HPLC has been used in quantification of zinc. Objective: The goal of this research is to develop and validate a sensitive, low-cost, high-resolution, and stability-indicating method for detecting and quantifying zinc levels in zinc supplement pharmaceutical products (injectables, tablets, and capsules). Methods: A novel isocratic reverse-phase HPLC method with a refractive index detector (with sensitivity 64, detector temperature 35 C, and positive polarity) was developed using a carboxyl functional group packed column and 0.8% (v/v) formic acid as the mobile phase to detect and quantify zinc content. Results: The reported method has a good optimal sensitivity (LOQ: 0.006 mg/mL and LOD: 0.002 mg/mL). The correlation coefficient (r) obtained from the zinc calibration plot was greater than 0.998, indicating that the method was linear and that there was a strong correlation between zinc concentration (0.006 mg/mL to 0.375 mg/mL) and peak response. The accuracy at LOQ level was found to be 95-105% and 97-103% at the remaining levels (50%, 100%, and 150%). Conclusion: The proposed method was successfully developed and validated as per International council for harmonisation (ICH) guidelines. Therefore, this method can be used for the quantitative testing of zinc in the QC laboratory. Highlights: A novel method was developed for zinc levels determination in pharmaceutical products using HPLC with a refractive index detector. The present approach has a quick run time of 10 min and is inexpensive.

To quantify the sulfamoxole content in bulk as well as tablet formulation, a new isocratic RP-HPLC method was developed and then validated. Kromasil C18 column with a dimension of 250 X 4.6mm was used which was filled with a particle size of 5μ. A mixture containing CH3OH, CH3CN and H2O in the volume ratio of 55, 30 and 15 was used as a mobile phase at room temperature with an optimized flow rate of 1ml/min. UV detector was set at 241nm for sulfamoxole determination. The run time of the current method is ten minutes. Accomplished the forced degradation studies to understand the stability indicating nature of the current method. As per the current ICH guides (Q2R1), validation of the method was conducted.

To quantify the sulfamoxole content in bulk as well as tablet formulation, a new isocratic RP-HPLC method was developed and then validated. Kromasil C18 column with a dimension of 250 X 4.6mm was used which was filled with a particle size of 5μ. A mixture containing CH3OH, CH3CN and H2O in the volume ratio of 55, 30 and 15 was used as a mobile phase at room temperature with an optimized flow rate of 1ml/min. UV detector was set at 241nm for sulfamoxole determination. The run time of the current method is ten minutes. Accomplished the forced degradation studies to understand the stability indicating nature of the current method. As per the current ICH guides (Q2R1), validation of the method was conducted.

Background: There are 400 licensed pharmaceutical companies in Pakistan; one third of Pakistan total consumption of pharmaceuticals is imported. Market potential is good for antibiotics, vaccines, hormones etc.  Introduction: High performance liquid chromatography is an important tool for routine determination of antimicrobial drugs with specific emphasis on fluoroquinolones. The quality testing of norfloxacin was also done by Liquid chromatography coupled with mass spectrometry. Antibacterial activity of norfloxacin is good for aerobic gram negative bacteria, including Enterobacteriaceae and Pseudomonas aeruginosa. It has moderate antibacterial activity against gram positive microorganisms, such as staphylococci; anaerobic bacteria are generally resistant. Method: Our investigation focuses on the quality testing of brands of Norfloxacin available in market with variety of trade names. Reverse phase chromatographic technique RP-HPLC was used for the quality testing of norfloxacin at...

Il metodo chimico per l’identificazione del marker radiolitico 2-dodecilciclobutanone (2-DCB), già impiegato nei nostri laboratori per l’analisi di matrici animali, è stato ottimizzato per la matrice uova intere, non irradiate e irradiate a 0.5 kGy. Inoltre sono state effettuate le fasi preliminari della validazione al fine di ampliare il campo di applicazione del metodo.
Mediante HS-SPME-GC/MS è stato possibile identificare correttamente sia i campioni non irradiati che i campioni irradiati alla dose di 0.5 kGy.

Objective: Methyl-4-Chlorobutyrate (M4CB), a genotoxic impurity, was identified in the active pharmacological components of the fourthgeneration fluoroquinolone, moxifloxacin (MXFN). There has not yet been a report on the analysis of the M4CB impurity content in the MXFN molecule. Consequently, a Gas Chromatography-Electron Ionization-Mass Spectrometry (GC-EI-MS) method was established that has the ability to identify and measure M4CB impurity content at ppm level. Methods: The column exploited in M4CB impurity assay was a Dura Bond 624 (DB-624) type stationary column. Temperatures of 220 °C and 280 °C were consistently maintained at the injection and detection sites, respectively. The helium, as carrier gas, in split mode with ratio of 1:7 was used. The column's flow rate remained steady around 2.0 ml/min. The mass spectrometer was operated in Single Ion Monitoring (SIM) mode at m/z = 74. Results: The impurity M4CB is generated during the manufacturing process of cyclopropanamine, which is an intermediary molecule in the manufacturing process of MXFN. This new GC-EI-MS approach can measure the M4CB at 0.9452 ppm in MXFN samples with a 500 mg/ml concentration following International Council for Harmonisation (ICH) standards. Very low quantification limits (0.9452 ppm), high linearity (range=0.945 ppm to 5.625 ppm; regression coefficient= 0.9999), and a reasonable recovery range (94.60-94.63%) were all provided by this new validated GC-EI-MS approach. Three batches were analysed for M4CB content by new GC-EI-MS approach and found that none of the batches contained M4CB impurity. Conclusion: The GC-EI-MS approach has excellent applicability in the quality assurance testing of MXFN for M4CB content since it was adequate in terms of linearity, precision, sensitivity, accuracy, specificity, and robustness.

Objective: Development of RP-HPLC method for determination of Hydrochlorothiazide (HCTZ), Olmesartan medoxomil (OLM) and their related substances in combined tablet dosage form and validation of the developed method. Methods: Gradient mobile phase system was used for estimation of drug contents and their related substances. Mobile phase A contained the mixture of Acetonitrile and 15 mM Phosphate buffer (pH adjusted to 3.4 with orthophosphoric acid) in the ratio of 20:80. Mobile phase B contained the same mixture in the ratio of 80:20. Chromatographic separation was carried out at the mobile phase flow rate of 0.8 mL/min using C18 Results: The linearity of developed method was tested in the range of 62.5-187.5 μg/mL for Hydrochlorothiazide, 100-300 μg/mL for Olmesartan medoxomil, 1-1.8 μg/mL for Hydrochlorothiazide. The % recovery was found to be 99.88-100.67 % (HCTZ), 99.14-99.91 % (OLM), 99.11-100.71% (HCTZ-IMP) and 98.13-100.83% (OLM-IMP). The assay of marketed formulation was found to be 99.78% (HCTZ) and 99.26% (OLM). Phenomenax inplace of Enable (250 × 4.6 mm) 5 μm column and detection was made at 254 nm. Conclusion: A simple, precise and accurate RP-HPLC method was developed for determination of Hydrochlorothiazide, Olmesartan medoxomil and their related substances.

Objective: Development of RP-HPLC method for determination of Hydrochlorothiazide (HCTZ), Olmesartan medoxomil (OLM) and their related substances in combined tablet dosage form and validation of the developed method. Methods: Gradient mobile phase system was used for estimation of drug contents and their related substances. Mobile phase A contained the mixture of Acetonitrile and 15 mM Phosphate buffer (pH adjusted to 3.4 with orthophosphoric acid) in the ratio of 20:80. Mobile phase B contained the same mixture in the ratio of 80:20. Chromatographic separation was carried out at the mobile phase flow rate of 0.8 mL/min using C18 The linearity of developed method was tested in the range of 62.5-187.5 μg/mL for Hydrochlorothiazide, 100-300 μg/mL for Olmesartan medoxomil, 1-1.8 μg/mL for Hydrochlorothiazide. The % recovery was found to be 99.88-100.67 % (HCTZ), 99.14-99.91 % (OLM), 99.11-100.71% (HCTZ-IMP) and 98.13-100.83% (OLM-IMP). The assay of marketed formulation was found to be 99.78% (HCTZ) and 99.26% (OLM). Phenomenax inplace of Enable (250 × 4.6 mm) 5 μm column and detection was made at 254 nm. A simple, precise and accurate RP-HPLC method was developed for determination of Hydrochlorothiazide, Olmesartan medoxomil and their related substances.

Simple, accurate, precise, and sensitive reverse-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous measurement of ARI and ESC in pharmaceutical products by using a C18 column with dimensions of 250 mm x 4.6 mm and a particle size of 5 μm. The mobile phase consisted of a mixture of acetonitrile, methanol, and water in the ratio of 80:05:15 v/v/v. Orthophosphoric acid was added to adjust the pH to 7.00 ± 0.1. The detection wavelength used for measuring the absorbance of the components was 246 nm, and the Mobile Phase flow rate was set at 1.0 mL/min. The retention times of ARI and ESC were 5.63 and 3.65 minutes, respectively. The ARI and ESC correlation coefficients (R 2-values) were observed at 0.9995 and 0.9997, respectively, demonstrating a robust linear relationship between the analyte concentration and the observed absorbance. The linearity range for ARI was 20 to 120 μg/mL, while ESC was 60 to 360 μg/mL. The recovery percentages for ARI and ESC ranged from 99.43% to 100.65% and 99.58% to 100.15%, respectively. Additionally, all parameter uncertainty levels were below 2. The method was validated following ICH guideline Q2 (R1). The analytical method validated will be successfully applied to the pharmaceutical dosage form.

Land has been variously acknowledged as a perennial harbinger of socio-economic power that exudes prestige and class. It is the foundation for food and shelter, driving the very bedrock that significantly provides for most employment opportunities in the country side. Regrettably, as global urbanisation trends soars uncontrollably, land is rapidly becoming increasingly scarce assets in cities. Therefore, supplying of and demand for lands, especially within urban spheres, are though unfortunately never at equilibrium, but the present scenarios in formal window of urban land market, which is solely responsible for the supply of formal land for various developmental purposes, has been very unimpressive. This simply, has rendered delivery to be at its lowest ebb, such that accessibility, which is the measure of demand coefficients from the part of the various land users, to be hampered, the resultant effects of which is consistent decrease of the real estate stock. Hence, it is in astou...

The primary objective of this review is to provide a comprehensive overview of the challenges involved in detecting ancient ayahuasca, a traditional hallucinogenic drink from the Amazon region, which is prepared using Banisteriopsis caapi liana and other plants, by utilizing advanced analytical techniques. The presence of harmine and harmaline in Andean archaeological findings has led certain authors to speculate that the Banisteriopsis liana may have been the source plant responsible for their occurrence. Consequently, the utilization of this liana can be traced back to at least 500 CE. However, a combination of archaeological, ethnobotanical, biochemical and analytical chemistry considerations has rendered uncertain the true origins of the presence of harmine and harmaline in ancient mummies and artifacts. Thus, the archaeological evidence does not convincingly prove the ancient usage of ayahuasca.

Now days as the expense and complications involved in marketing new drug entities are increased, with concomitant recognition of the therapeutic advantages of controlled drug delivery, greater attention has been focused on development of sustained or controlled release drug delivery systems (DDS). Hence we will change the area of focusing it is suitable to designing sustained drug delivery is to reduce the frequency of dosing or to increase the effectiveness of the drug by localization at the site of action, reducing the dose required or providing uniform drug delivery. The design of oral sustained release DDS depends on various factors such as, physicochemical properties of drug, type of delivery system, disease being treated, and patient condition, and treatment duration, presence of food, gastrointestinal motility, and co-administration of other drugs.

An accurate, easy, detailed, selective and fast RP-HPLC stability representative technique was developed and validated for assessment of ivermectin in tablet dosage form. The Reverse Phase High Performance Liquid Chromatographic technique was developed for routine quantification of ivermectin in laboratory prepared mixtures as well as in combined dosage forms. The chromatographic separation was accomplished with INERTSIL C-18 ODS 250×4.6mm, 5µm particle size column along with acetonitrile and methanol as the mobile phase at a flow rate of 1ml/min. Quantification was completed by using a UV detector at 245 nm and the run time was 10 minutes. The retention time was found to be 4.198 min for ivermectin. The linearity was observed in the range of 1-32µg/ml with correlation coefficient r= 0.9798. The % RSD for intraday and interday precision was 1.352 and 1.589 respectively. The LOD and LOQ values were found to be 2.93 and 8.79, respectively. The system suitability parameters for ivermectin such as theoretical plates and tailing factor were found to be 129.949 and 2.0, respectively. Robustness was also studied and there was no significant variation in the system suitability of the analytical method by incorporating small changes in experimental parameters. The technique has been validated for linearity, precision, accuracy and other parameters as approved by ICH guidelines. The results obtained by RP-HPLC methods are found to be fast, detailed, selective and accurate. Therefore, proposed analytical method can be used for regular analysis of ivermectin in injection, tablet and other formulations.

A new simple, accurate, precise and reproducible gradient phase ultra-performance liquid chromatography method was developed and fully validated for the estimation of Amlodipine Besylate and Olmesartan Medoxomil in pharmaceutical tablet dosage form gradientlly using acetonitrile: triethylamine buffer(pH4.0+0.5) as mobile phase and Acquity BEH C8 column (4.6x 150 mm,2.6μg) as stationary phase and chromatogram was recorded at 237nm at a flow rate of 0.4ml/min. The retention time of AML were 1.5to 2.8 and OLM 3.2 to 5.5 min respectively and showed a good linearity in the concentration range of 5-25μg/ml with a concentration coefficient (R) of 0.99956 and 0.99985 respectively. The developed UPLC method was validated with respect to specificity, linearity, precision, accuracy, ruggedness (reproducibility), robustness and stability. The recovery data was in the range of 98.0%to 102.0%. the proposed method was validated as per ICH guidelines and successfully as per ICH guidelines and successfully applied to the development and validation of AML and OLM in tablet formulation.

Simple, accurate, precise, and sensitive reverse-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous measurement of ARI and ESC in pharmaceutical products by using a C18 column with dimensions of 250 mm x 4.6 mm and a particle size of 5 μm. The mobile phase consisted of a mixture of acetonitrile, methanol, and water in the ratio of 80:05:15 v/v/v. Orthophosphoric acid was added to adjust the pH to 7.00 ± 0.1. The detection wavelength used for measuring the absorbance of the components was 246 nm, and the Mobile Phase flow rate was set at 1.0 mL/min. The retention times of ARI and ESC were 5.63 and 3.65 minutes, respectively. The ARI and ESC correlation coefficients (R 2-values) were observed at 0.9995 and 0.9997, respectively, demonstrating a robust linear relationship between the analyte concentration and the observed absorbance. The linearity range for ARI was 20 to 120 μg/mL, while ESC was 60 to 360 μg/mL. The recovery percentages for ARI and ESC ranged from 99.43% to 100.65% and 99.58% to 100.15%, respectively. Additionally, all parameter uncertainty levels were below 2. The method was validated following ICH guideline Q2 (R1). The analytical method validated will be successfully applied to the pharmaceutical dosage form.

Simple, accurate, precise, and sensitive reverse-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous measurement of ARI and ESC in pharmaceutical products by using a C18 column with dimensions of 250 mm x 4.6 mm and a particle size of 5 μm. The mobile phase consisted of a mixture of acetonitrile, methanol, and water in the ratio of 80:05:15 v/v/v. Orthophosphoric acid was added to adjust the pH to 7.00 ± 0.1. The detection wavelength used for measuring the absorbance of the components was 246 nm, and the Mobile Phase flow rate was set at 1.0 mL/min. The retention times of ARI and ESC were 5.63 and 3.65 minutes, respectively. The ARI and ESC correlation coefficients (R 2-values) were observed at 0.9995 and 0.9997, respectively, demonstrating a robust linear relationship between the analyte concentration and the observed absorbance. The linearity range for ARI was 20 to 120 μg/mL, while ESC was 60 to 360 μg/mL. The recovery percentages for ARI and ESC ranged from 99.43% to 100.65% and 99.58% to 100.15%, respectively. Additionally, all parameter uncertainty levels were below 2. The method was validated following ICH guideline Q2 (R1). The analytical method validated will be successfully applied to the pharmaceutical dosage form.

A simple and sensitive UV spectrophotometric method has been developed and subsequently validated for the determination of Moxifloxacin in bulk and pharmaceutical formulations. 0.01N HCL was used as a solvent in the present investigation. Quantitative measurements were made at the maximum absorption of 294.4 nm. The method was validated over the range of 2 to 8 µg/mL with correlation coefficient r = 0.9999. The method was shown to be accurate and precise with inter-day and intra-day percent relative standard deviation values in the range of 0.176 to 0.356 and 0.448 to 1. 07. The percent recoveries of Moxifloxacin were found to be 99.31 to 100.13. The limit of detection was 0.011 µg/mL and limit of quantification was 0.038 µg/mL. The method has been successfully utilized to determine the Moxifloxacin in tablets and can be extended for the routine analysis in bulk drugs.

Background: No comprehensive short-form health literacy (HL) survey tool has been available for general use across Asia. Objective: This study aimed to develop and validate a short-form HL instrument derived from the 47-item European Health Literacy Questionnaire (HLS-EU-Q47). Methods: A population survey ( N = 10,024) was conducted from 2013 to 2015 using the HLS-EU-Q47 in 1,029 participants from Indonesia, 1,845 from Kazakhstan, 462 from Malaysia, 1,600 from Myanmar, 3,015 from Taiwan, and 2,073 from Vietnam. Validation of the short form was evaluated by principle component analysis, internal consistency, Pearson correlation, and regression analysis. Key Results: Based on responses from six countries, a 12-item short-form HL questionnaire (HLS-SF12) was developed, retaining the conceptual framework of the HLS-EU-Q47 and accounting for the high variance of the full-form (i.e., 90% in Indonesia, 91% in Myanmar, 93% in Malaysia, 94% in Taiwan, and 95% in both Kazakhstan and Vietnam)....

Iron plays a pivotal role in human physiology, while its deficiency may prove fatal in severe cases. Analytical methods for the quantitative determination of iron are thus very important. Herein, we report the estimation of iron in iron Polysaccharide complex (IPSC) using raw material and formulations, through a spectrophotometric analytical method. IPSC capsules were formulated and their stability was studied by developing a simple and validated analytical method. The process is based on the acid hydrolysis of IPSC and development of chromogen by the reaction of ammonium thiocyanate with IPSC. The chromogen was detected at 474nm. Beer's Lambert law (linearity response) was observed in the range of 10-20 μg/ml with excellent correlation coefficient of determination (R = 0.998). The quantification and detection limits were established to be 0.45 mcg/ml and 0.14 mcg/ml correspondingly. The recovery of IPSC analysis was 99.25 to 102.28 %. Percentage assay of IPSC capsules showed results around 102.34 %. The formulated IPSC capsule was stable under accelerated conditions for 6 months (% assay > 91.69). The dissolution profile over 60 minutes showed a better dissolution (94%) compared with the internationally marketed IPSC capsule (92%).