Search

Human epidermal growth factor receptor 2 (HER2) is a subtype of breast cancer that is associated with poor prognosis and low survival rates. The discovery of novel anti-cancer agents to manage this subtype of cancer is still needed. Ziziphus spina-christi (ZSC) is a plant species that is native to Qatar. It exerts various biological activities, including cytotoxicity as it contains different essential bioactive constituents, mainly rutin and quercetin. To examine the outcome of ZSC on HER2-positive breast cancer, we standardized the ZSC methanolic leaves extracted by Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) analysis using the flavonoids rutin and quercetin as marker compounds. Here we used two HER2-positive breast cancer cell lines, ZR-75-1 and SK-BR-3, and the chorioallantoic membrane as an angiogenesis model. We found that ZSC extract significantly reduces viability, alters the normal morphological phenotype of HER2-positive breast cancer cells, and inhibits cell migration as well as colony formation; this is accompanied by deregulating different apoptotic markers such as Bax/Bcl-2 and NF-κB in both cell lines. Additionally, ZSC methanolic extract significantly represses the angiogenesis of the chorioallantoic membrane model. Moreover, the molecular pathway investigations pointed out that ZSC extract represses the activity of HER2 and p38 MAPK which could be the main pathways behind the effect of ZSC in HER2-positive cells. Collectively, our results support the potential role of ZSC in the management of HER2-positive breast cancer and form the basis for future investigations. [ABSTRACT FROM AUTHOR]

The combination of the tricyclic antidepressant amitriptyline hydrochloride (AMH) and the non-selective beta-adrenergic blocker propranolol hydrochloride (PPH) is used for migraine prophylaxis. Higher doses of AMH trigger cardiac arrhythmias, anxiety, tachycardia, convulsions, hyperglycemia and anticholinergic side effects. The combined dosage formulation of AMH and PPH leads to drug–drug interactions; causes sedation, xerostomia, dysuria, insomnia and bradycardia; and results in patient non-compliance. The quantification of AMH and PPN becomes essential, especially for combination formulations, in addition to regular quality control to avoid clinical issues. Considering these facts into account, the reverse-phase -high-performance liquid chromatography (RP-HPLC) method was developed in accordance with International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use Q2(R1) guidelines for the simultaneous determination of AMH and PPH. The HPLC separation was performed on an HPLC system (Shimadzu, Japan, Prominence I series 2030C) using a Shimadzu Shim-Pack GIST C18 column (100 mm × 4.6 mm, 5 μ), which was equipped with an ultraviolet detector at the isosbestic point 238 nm. The mixture of acetonitrile and orthophosphoric acid (pH 3.5) in a ratio of 35:65 v/v with a flow rate of 0.75 mL/min was used as the mobile phase. The regression coefficients of AMH (r 2 > 0.998) and PPH (r 2 > 0.999) show good linearity between peak areas and drug concentration ranges. The limits of detection (AMH = 0.67 μg/mL, PPH = 0.67 μg/mL) and limits of quantification (AMH = 2.04 μg/mL, PPH = 2.05 μg/mL) demonstrated the higher detection sensitivity of the proposed method. [ABSTRACT FROM AUTHOR]

Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

AbstractIn this study, the chromatographic retention behavior and dissociation constants (pKa) of H2 receptor antagonists ranitidine and famotidine were determined by the green reverse phase liquid chromatography (RPLC) and hybrid micellar chromatography methods, which are widely used today. Replicate analyses were performed on a Gemini NX C18 (250 × 3.0 mm ID, 5 μm) column at constant temperature (37 °C) and flow rate (1 mL/min). The obtained data analyses were evaluated with the linear solvation energy relationship (LSER) approach and the consistency of the data was assessed by calculating the theoretical retention time values. The pKa1ss and pKa2ss values obtained in ethanol–water binary mixtures containing 11%, 13%, 15%, and 20% (v/v) ethanol were also calculated with this approach. The pKa values of these compounds in water and pure micellar environment were calculated with pKass values and solvent. It was observed that the pKaww data obtained by three different methods were compatible. Accordingly, with the hybrid micelles method, pKa2ww and pKa3ww values for famotidine were calculated as 7.305 and 9.869, respectively, and pKa1ww and pKa2ww values for ranitidine were calculated as 2.952 and 8.589. The environmental impacts of the methods applied in the study were evaluated using green metric tools. [ABSTRACT FROM AUTHOR]

Cholinergic disruptions underlie attentional deficits following traumatic brain injury (TBI). Yet, drugs specifically targeting acetylcholinesterase (AChE) inhibition have yielded mixed outcomes. Therefore, we hypothesized that galantamine (GAL), a dual-action competitive AChE inhibitor and α7 nicotinic acetylcholine receptor (nAChR) positive allosteric modulator, provided chronically after injury, will attenuate TBI-induced deficits of sustained attention and enhance ACh efflux in the medial prefrontal cortex (mPFC), as assessed by in vivo microdialysis. In Experiment 1, adult male rats (n = 10–15/group) trained in the 3-choice serial reaction time (3-CSRT) test were randomly assigned to controlled cortical impact (CCI) or sham surgery and administered GAL (0.5, 2.0, or 5.0 mg/kg; i.p.) or saline vehicle (VEH; 1 mL/kg; i.p) beginning 24-h post-surgery and once daily thereafter for 27 days. Measures of sustained attention and distractibility were assessed on post-operative days 21–25 in the 3-CSRT, following which cortical lesion volume and basal forebrain cholinergic cells were quantified on day 27. In Experiment 2, adult male rats (n = 3–4/group) received a CCI and 24 h later administered (i.p.) one of the three doses of GAL or VEH for 21 days to quantify the dose-dependent effect of GAL on in vivo ACh efflux in the mPFC. Two weeks after the CCI, a guide cannula was implanted in the right mPFC. On post-surgery day 21, baseline and post-injection dialysate samples were collected in a temporally matched manner with the cohort undergoing behavior. ACh levels were analyzed using reverse phase high-performance liquid chromatography (HPLC) coupled to an electrochemical detector. Cortical lesion volume was quantified on day 22. The data were subjected to ANOVA, with repeated measures where appropriate, followed by Newman–Keuls post hoc analyses. All TBI groups displayed impaired sustained attention versus the pooled SHAM controls (p's < 0.05). Moreover, the highest dose of GAL (5.0 mg/kg) exacerbated attentional deficits relative to VEH and the two lower doses of GAL (p's < 0.05). TBI significantly reduced cholinergic cells in the right basal forebrain, regardless of treatment condition, versus SHAM (p < 0.05). In vivo microdialysis revealed no differences in basal ACh in the mPFC; however, GAL (5.0 mg/kg) significantly increased ACh efflux 30 min following injection compared to the VEH and the other GAL (0.5 and 2.0 mg/kg) treated groups (p's < 0.05). In both experiments, there were no differences in cortical lesion volume across treatment groups (p's > 0.05). In summary, albeit the higher dose of GAL increased ACh release, it did not improve measures of sustained attention or histopathological markers, thereby partially supporting the hypothesis and providing the impetus for further investigations into alternative cholinergic pharmacotherapies such as nAChR positive allosteric modulators. [ABSTRACT FROM AUTHOR]

Vitamin K2, also known as menaquinone, has become a significant research focus, particularly in fermented products. This study aims to investigate the content of menaquinone‐7 (MK‐7), an essential form of vitamin K, and other fat‐soluble vitamins (A, D, E) during the fermentation of various fermented milk products. The vitamin contents of six fermented milk products were analyzed: two yogurt samples (produced with commercial starter and probiotic starter), two kefir samples (produced with commercial starter and kefir grain), and milk fermented with Lactobacillus acidophilus or Bifidobacterium bifidum. Throughout the fermentation process, continuous pH monitoring was conducted, and fermentation was terminated based on pH levels. Fat‐soluble vitamins and vitamins K1, K2 were extracted from samples taken at specific fermentation periods and quantitatively determined using the reverse phase liquid chromatography (RPLC) method. The developed method was validated according to ICH guidelines. Simultaneously, the microbial content of the samples was analyzed. Among the fermented dairy products analyzed, the highest MK‐7 content (4.82 μg/100 g) was found in the kefir sample produced with kefir grain (KG). The diverse microorganisms in kefir grains necessitated detailed research to explain their role in this complex mechanism. In yogurt samples, the presence of L. acidophilus in the YB sample suggests that this bacterium may be responsible for the observed differences. This is supported by the LA sample, where production with L. acidophilus strain showed an increase from 0.97 to 1.70 at the 20th hour of fermentation. It was noted that the concentration of menaquinone‐7 increased throughout the fermentation period. Given the critical health effects of essential fat‐soluble vitamins and MKs, determining their content in commonly consumed fermented dairy products and understanding the influence of starter cultures, which are potential probiotics, on vitamin production underscores the importance of this research. It is important to highlight the potential of fermented products in nutritional recommendations. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the pharmacokinetics (PK) of intravenously (IV) and subcutaneously (SC) administered nalbuphine in domestic goats. Nalbuphine hydrochloride was administered at 0.8 mg/kg for both IV and SC routes in six goats with a minimum of 10‐day washout period between sample collection phases. Eighteen plasma samples were collected over a 36‐hour period, analyzed using reverse phase high‐performance liquid chromatography (HPLC). Plasma data were analyzed using compartmental and noncompartmental approaches. Following IV nalbuphine administration, elimination half‐life, area under the plasma concentration time curve from time 0 to infinity (AUC0 − ∞), concentration at time zero (C0), and total body clearance were 120.4 ± 39.1 (min−1 ± SD), 17311.01 ± 7227.32 (min·ng·mL−1 ± SD), 675.6 ± 337.13 (ng·mL−1 ± SD), and 44.5 ± 13.8 (mL·min−1·kg−1 ± SD), respectively. After SC nalbuphine administration, elimination half‐life, area under the plasma concentration time curve from time 0 to infinity (AUC0 − ∞), and maximum plasma drug concentration were 129 ± 52.9 (min−1 ± SD), 20826.5 ± 14376.2 (min·ng·mL−1), and 368.03 ± 503.78 (ng·mL−1). Calculated bioavailability for the SC route was 138 ± 126 (% ± SD). Nalbuphine in goats is characterized by rapid elimination and high subcutaneous bioavailability and may be a safe analgesic opioid option in goats in the future. [ABSTRACT FROM AUTHOR]

The importance of lipids in biology continues to grow with their recent linkages to more diseases and conditions, microbiome fluctuations, and environmental exposures. These associations have motivated researchers to evaluate lipidomic changes in numerous matrices and studies. Lipidomic analyses, however, present numerous challenges as lipid species have broad chemistries that require different extraction methods and instrumental analyses to evaluate and separate their many isomers and isobars. Increasing knowledge about different lipid characteristics is therefore crucial for improving their separation and identification. Here, we present a multidimensional database for lipids analyzed on a platform combining reversed-phase liquid chromatography, drift tube ion mobility spectrometry, collision-induced dissociation, and mass spectrometry (RPLC-DTIMS-CID-MS). This platform and the different separation characteristics it provides enables more confident lipid annotations when compared to traditional tandem mass spectrometry platforms, especially when analyzing highly isomeric molecules such as lipids. This database expands on our previous publication containing only human plasma and bronchoalveolar lavage fluid lipids and provides experimental RPLC retention times, IMS collision cross section (CCS) values, and m/z information for 877 unique lipids from additional biofluids and tissues. Specifically, the database contains 1504 precursor [M + H]+, [M + NH4]+, [M + Na]+, [M-H]−, [M-2H]2−, [M + HCOO]−, and [M + CH3COO]− ion species and their associated CID fragments which are commonly targeted in clinical and environmental studies, in addition to being present in the chloroform layer of Folch extractions. Furthermore, this multidimensional RPLC-DTIMS-CID-MS database spans 5 lipid categories (fatty acids, sterols, sphingolipids, glycerolipids, and glycerophospholipids) and 24 lipid classes. We have also created a webpage (tarheels.live/bakerlab/databases/) to enhance the accessibility of this resource which will be populated regularly with new lipids as we identify additional species and integrate novel standards. [ABSTRACT FROM AUTHOR]

Nitrosamine compounds pose a significant concern as potential carcinogens, prompting heightened scrutiny from regulatory bodies, particularly regarding their presence in pharmaceuticals. The detection of unacceptable levels of N‐nitrosodiethylamine (NDMA) in ranitidine has led to widespread recalls, driving interest in alternative medications such as nizatidine, which shares a similar pharmacological class and is used to treat various gastrointestinal conditions. Despite fewer reports on NDMA levels in nizatidine, its structural similarity to ranitidine, characterized by a tertiary amine, underscores the potential for NDMA formation. Addressing the analytical challenges associated with nitrosamine detection, this study focuses on developing and validating an ultra‐high pressure liquid chromatography triple quadrupole mass spectrometry (UHPLC–MS/MS) method for quantifying NDMA in both nizatidine active pharmaceutical ingredients and tablet formulations. Method validation adheres to International Council for Harmonisation recommendations, with a demonstrated linear range of 0.25–100 ng/mL for NDMA, exhibiting excellent linearity (regression coefficient >0.999) and efficient recovery rates ranging from 95.98% to 109.57%. The method shows high sensitivity, with limits of detection and quantification of 0.25 and 0.5 ng/mL, respectively. The developed UHPLC–MS/MS method offers a simple, precise, accurate, and selective approach for monitoring NDMA levels in nizatidine formulations available in Australia, promising enhanced sensitivity and specificity with limits of quantification in the ppb and sub‐ppb ranges. [ABSTRACT FROM AUTHOR]

On the review of various research article the metformin, Teneligliptin and pioglitazone used for the diabetes mellitus and there hasn’t been any development an RP-HPLC method. Aim: For the simultaneous stability investigation of metformin, pioglitazone, teneligliptin in tablet dosage forms, a new (RP-HPLC) approach is presented in this work. We developed an easy, precise and accurate method for estimating metformin, teneligliptin, pioglitazone solid dose form simultaneously. A Kromasil C18 150x4.6mm, 5m was used to run the chromatogram. A 50:50 v/v. The column was purged with a mobile phase that included water and methanol at a rate one milliliter per minute and it was kept at 30ºC. The optimum wavelength for metformin, teneligliptin, and pioglitazone was 220.0nm. Metformin, teneligliptin and pioglitazone were shown to have retention times of 2.220min, 2.931 min and 3.409 min, respectively. It is found the mean RSD of the system precision for pioglitazone, metformin and teneligliptin is 0.7, 0.5, and 0.8, respectively. It was found that the RSD of the method precision for Pioglitazone, Teneligliptin, Metformin (.3, .2 and .3), respectively. Pioglitazone, teneligliptin and metformin recovery % was found to be 100.11%, 99.90% and 99.71%, respectively. The Metformin, Teneligliptin, and Pioglitazone regression equations obtained LOD and LOQ values (1.08, 0.05, 0.04, 3.28, 0.15 and 0.11µg/ml), respectively. Metformin Regression equation y = 4445.4x + 3095.4. that of teneligliptin y = 12258x + 936.75 and that of pioglitazone y = 77511x + 1414.9, respectively. In conclusion, when validated (RP-HPLC) model was validated in compliance with acceptance criteria. It was found to be appropriate for the quantitative analysis of metformin, teneligliptin and pioglitazone after Q2 (R1) validation of analytical techniques in pharmaceutical dosage form. [ABSTRACT FROM AUTHOR]

The quality by design enabled the development of cost‐effective, simple, precise, and rapid Reversed phase‐ high‐performance liquid chromatography techniques for determining ritonavir (RIT) and darunavir (DAR) in the tablet dosage form. Systematic method optimization was performed through central composite design by altering the flow rate and composition of the organic phase in the mobile phase as the critical method parameters for evaluating the necessary analytical attributes, namely, the tailing factor, theoretical plate count, and resolution. The optimal separation was achieved on a column with C18 in nature and dimensions of (250 mm length × 4.6 mm id × 5 µm particle size) using a mobile phase consisting of acetonitrile and water (75:25 v/v) at a flow rate of 0.9 mL/min. Detection was carried out at 247 nm. The retention time of DAR and RIT was 2.939 and 3.537 min, respectively, as per International Council for Harmonization (ICH) guidelines. The developed method was useful for the determination of bulk drug and formulations. [ABSTRACT FROM AUTHOR]

Synthesis of novel amide and ester prodrugs of Repaglinide with amino acid Leucine and Polyethylene Glycol were performed. After synthesis, prodrugcharacterization was done using IR and ¹H NMR spectroscopy. Parameters Melting point, Differential Scanning Calorimetry (DSC), and Reverse phase high-performance liquid chromatography (RP-HPLC) chromatograms were performed for synthesized prodrugs to confirm the synthesized and differentiate synthesized Repaglinide prodrugs from pure Repaglinide. [ABSTRACT FROM AUTHOR]

The current investigation was pointed at developing and progressively validating novel, simple, responsive and stable RP-UPLC method for the measurement of active pharmaceutical ingredient of viloxazine. Attention deficit hyperactivity disorder (ADHD) is children and adults are treated by viloxazine, a selective norepinephrine reuptake inhibitor and viloxazine, selective nor-epinephrine reuptake inhibitor is used for treatment of ADHD in children six years or above of age. An easy, sensitive, specific and precise method for the pharmaceutical dose estimation of viloxazine in tablet dosage form was developed using RP-UPLC method. Separation was achieved by using BEH C18 100 × 2.1 mm, 2m column. Mobile phase containing OPA: Acetonitrile taken in the ratio 60:40 was pumped through column at a flow rate of 0.3 ml/min, and temperature was maintained at 300C. Optimized wavelength selected was 218.0 nm, and retention time of viloxazine was found to be 1.678 min. % RSD was found to be 0.9, % recovery and % assay was obtained as 100.44% and 99.90%, respectively. LOD and LOQ values were obtained as 0.30 µg/ml, 0.90 µg/ml. Recovery, specificity, linearity, accuracy, robustness, ruggedness was determined as a part of method validation and the results were found to be within the acceptable range. The proposed method to be fast, simple, accurate, feasible and affordable in assay condition. [ABSTRACT FROM AUTHOR]

The novel antimalarial ganaplacide combined with lumefantrine solid dispersion formulation (LUM‐SDF) was effective and well tolerated in the treatment of uncomplicated falciparum malaria in adults, adolescents, and children in a multinational, prospective, randomized, active‐controlled Phase II study conducted between August 2017 and June 2021 (EudraCT 2020‐003284‐25, Clinicaltrials.gov NCT03167242). Pharmacokinetic data from that study are reported here.The trial comprised three parts: a run‐in part in 12 adult/adolescent patients treated with a single dose of ganaplacide 200 mg plus LUM‐SDF 960 mg assessed potential pharmacokinetic (PK) interactions between ganaplacide and lumefantrine; in Part A, adult/adolescent patients received one of the six ganaplacide‐LUM‐SDF regimens or artemether‐lumefantrine; and in Part B, three dose regimens identified in Part A, and artemether‐lumefantrine, were assessed in children aged 2 to <12 years, with body weight ≥10 kg. A rich blood sampling schedule was used for all 12 patients in the PK run‐in part and a subset of patients (N = 32) in Part A, with sparse sampling for remaining patients in Parts A (N = 275) and B (N = 159). Drug concentrations were determined by a validated protein precipitation and reverse phase liquid chromatography with tandem mass spectrometry detection method. Parameters including AUCinf, AUClast, AUC0–t, Cmax, and tmax were reported where possible, using non‐compartmental analysis.In the PK run‐in part, there was no notable increase in ganaplacide or lumefantrine exposure when co‐administered. In Parts A and B, ganaplacide exposures increased with dose, but lumefantrine exposure was numerically under dose‐proportional. Lumefantrine exposure was higher with ganaplacide‐LUM‐SDF than with artemether‐lumefantrine, although high variability was observed. Ganaplacide and lumefantrine exposures (Cmax and AUC0–24 h) were comparable across age and body weight groups. Drug exposures needed for efficacy were achieved using the dose regimen 400 mg ganaplacide plus lumefantrine 960 mg once daily for 3 days under fasted conditions. [ABSTRACT FROM AUTHOR]

The objective of the present effort is developing a fully green-assessed stability-indicating reverse phase liquid chromatography for the assessable measurement regarding Remogliflozin etabonate (RGE) & Teneligliptin (TEN) in drug substance and tablets and fully validated as per ICH criteria. Waters Atlantis T3 (150mm x 4.6, 3.5µm) column, 30 volumes acetonitrile along with 70 volumes ammonium formate (pH-3.0) mobile phase pumped at a flow rate of 1.0ml/min were used for the chromatographic separation using PDA exposure at 236 nm wavelength. Remogliflozin etabonate, Teneligliptin were separated at 2.730, 4.468 minutes of retention times respectively. ICH guidelines Q2 R1 was employed to validate the current procedure. Accuracy measurements were acceptable to intra-day and inter-day measurements. For RGE & TGN, respectively, the recommended technique was linear in the concentration ranges of 25-150µg/ml and 2.5-15 µg/ml. y=29777x+67797 and y= 30689 x +22270 respectively were the observed regression equations for RGE and TEN. The limits of detection (LOD) and quantification (LOQ) for Remogliflozin etabonate & Teneligliptin were respectively 0.3 (µg/ml), 1 (µg/ml) and 0.03(µg/ml), 0.1(µg/ml). The recovery percentage of the method was found to remain in between 98% and 100%. Q1A R2 and Q1B guidelines were followed for conducting stability-indicating studies. The technique can also be utilized for quality control and repetitive laboratories evaluation aimed at the simultaneous assessment of RGE & TEN in the drug substance and pharmaceutical dosage form. [ABSTRACT FROM AUTHOR]

A simultaneous ultra-performance liquid chromatography photodiode array detection method was developed and validated for Elvitegravir, Tenofovir and Emtricitabine and Cobicistat drugs in tablet dosage forms. Effective liquid chromatographic quantification was achieved using a Luna C18 (100 × 2.6 mm, 1.6 μ) column with an acetonitrile: 0.1% v/v trifluoro acetic acid (80:20% v/v) mobile phase and a photo diode array detector at 260 nm for a shorter run time of 3 min. The respective retention times for Elvitegravir, Tenofovir, Emtricitabine and Cobicistat were 0.311, 0.792, 1.379 and 2.045 min. The range of linearity was 37.50–225.00, 2.50–15.00, 50.00–300.00 and 37.50–225.00 μg/mL for Elvitegravir, Tenofovir, Emtricitabine and Cobicistat, respectively. A mass spectrometer was employed for quantitative analysis using electron spray ionization multiple reaction monitoring in positive mode. The developed method was validated for different parameters to meet the acceptance criteria as per ICH Q2 (R1) guidelines. Using UPLC–ESI–MS, 12 degradants were identified and their mechanisms were studied. [ABSTRACT FROM AUTHOR]

There is currently limited pharmacokinetic data for the use of famotidine in goats for treatment and prevention of abomasal ulceration. The objective of this study was to determine the pharmacokinetic parameters after a single intravenous administration of famotidine (0.6 mg/kg). Famotidine was administered to six healthy goats and plasma samples were collected over a 24‐h period. The famotidine concentration was measured using reverse phase high‐performance liquid chromatography (HPLC). Non‐compartmental analysis was then used to determine the pharmacokinetic parameters. The maximum plasma concentration was estimated at 5476.68 ± 1530.51 ng/mL and elimination half‐life was estimated at 18.455 ± 13.26 min. The mean residence time was determined to be 19.85 ± 12.14 min with the apparent volume of distribution being estimated at 321.924 ± 221.667. The area under the curve was determined to be 54230.08 ± 24947.6 min*ng/mL. Total exposure and elimination half‐life were less than what has been reported in cattle and horses. Future research evaluating the pharmacokinetics of subcutaneous administration and looking at the pharmacodynamics of famotidine in goats is needed to determine the effectiveness of famotidine on raising pH levels of the abomasum. [ABSTRACT FROM AUTHOR]

This work presents the development and validation of a solid Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method for concurrently quantifying Cinnarizine and Piracetam in pharmaceutical formulations. The technique used an Agilent 1200 series high-performance liquid chromatography (HPLC) machine equipped with a C18 column. The mobile phase composition consisted of Methanol and Orthophosphoric Acid. Correlation coefficients of 0.9992 for Cinnarizine and 0.9998 for Piracetam indicate the calibration curves exhibit a robust linear relationship. The present investigation determined the Limits of Detection (LOD) for Cinnarizine to be 0.1575 µg/ml, other than for Piracetam it was found to be 1.9926 µg/ml. The Limits of Quantification (LOQ) for both compounds were found to be 0.4774 µg/ml and 6.0384 µg/ml, respectively. In interday precision tests, the approach exhibited a high level of accuracy, as evidenced by % Relative Standard Deviation (RSD) values of 0.3601 for Cinnarizine and 0.7080 for Piracetam. Assay results for Phescetam capsules revealed 100.53% purity for Cinnarizine and 100.19% for Piracetam, with % RSD values within acceptable limits, indicating high accuracy and precision. The method proved robust and rugged, showing consistent results under varying analytical conditions and across different analysts. Stability studies confirmed the method's suitability for routine quality control, with both drugs demonstrating stability under acidic, basic, oxidative, and photolytic conditions. [ABSTRACT FROM AUTHOR]

Background: Hypertension is a prevalent health condition with severe implications for cardiovascular health. Combination therapy involving Candesartan and Nifedipine has emerged as a promising approach for managing hypertension due to the synergistic effects of these medications. Methods: This study aimed to develop a Design of Experiment (DoE) based Green High-Performance Liquid Chromatography (HPLC) method for the simultaneous estimation of Candesartan and Nifedipine in a combined dosage form. Reversed-phase HPLC was utilized with UV-Vis spectrophotometry for detection. DoE principles were applied to optimize chromatographic conditions, including pH, ethanol percentage in the mobile phase, and flow rate. Results: Optimal chromatographic separation was achieved using a composite mobile phase comprising ethanol and potassium dihydrogen phosphate buffer (45:55, v/v) at pH 3.7, with a flow rate of 1 mL/min. Statistical analysis confirmed the significance of these parameters in achieving efficient separation. The developed method exhibited high precision, accuracy, and reliability across various validation parameters, including specificity, linearity, repeatability, and robustness. Conclusion: The proposed DoE-based Green HPLC method offers a sustainable and efficient approach for the simultaneous estimation of Candesartan and Nifedipine in hypertension management. By integrating green chemistry principles with systematic method development, this method provides a reliable analytical tool for routine quality control purposes in pharmaceutical formulations containing these medications. [ABSTRACT FROM AUTHOR]

Background: Trifarotene is effective for treating acne and other skin issues. To ensure its quality and meet regulatory standards, a reverse phase liquid chromatography (RP-LC) stability-indicating method was developed and validated for its quantification in pure and topical dosage forms. An isocratic elution chromatographic method was employed, using an octadecylsilyl silica gel-packed column (150 mm × 4.6 mm, 3 µm particle size). The mobile phase was a mixture of phosphate buffer and acetonitrile (40:60 v/v). Chromatographic conditions included a flow rate of 0.5 mL/min, column temperature of 40 °C, detection at 265 nm, and injection volume of 20 µL. Results: The developed analytical method reports the retention time of trifarotene 11.2 min, higher theoretical plate count, asymmetric peak, and good resolution between the peaks of trifarotene, phenoxyethanol, and environmentally generated impurities. Conclusion: The analytical method has been found to be linear, accurate, robust, specific, and selective for impurities produced during forced degradation studies. The proposed analytical method can be utilized for routine pharmaceutical analysis of trifarotene to judge its quality and safety. [ABSTRACT FROM AUTHOR]

An accurate and precise reverse‐phase high‐performance liquid chromatographic technique for apremilast (APR) quantification in bulk, commercial tablets, and formulated APR‐loaded nanoformulations was developed and validated by the design of the experiment approach in accordance with the International Conference of Harmonization guidelines. APR was separated by using formic acid buffer (0.1%; pH 3.2):methanol (40:60, v/v) as a solvent system with a flow rate of 1 mL/min and was detected at a wavelength of 230 nm with a retention time of 7.1 min. The central composite design was employed to optimize the developed analytical method. The calibration curve for APR was linear (r2 = 0.999), with a limit of detection of 0.5 µg/mL and a limit of quantification of 1.52 µg/mL, respectively. A recovery of APR in tablet and nanoformulation was observed in the 98.56%–99.32% range. Forced degradation studies revealed the stability of the proposed analytical method, as the APR peak remained unchanged under stress‐induced degradation experiments. Hence, the proposed chromatographic technique with a cost‐effective solvent system could be employed in the pharmaceutical industry for estimating APR in various formulations. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to investigate the anticancer taxane profiles of edible and non-edible parts of seven Turkish hazelnut (Corylus avellana L.) genotypes. Hazelnut is one of the healthy foods rich in nutrients and antioxidants. Its regular consumption is associated with a reduced risk of coronary heart disease and cancer. Hazelnut has been described as a plant source that produces taxanes which are widely used in many cancers. Türkiye is a homeland of hazelnut culture and has its own cultivars. Investigation of anticancer taxane profiles in different parts of Turkish hazelnut genotypes is important to show the potential and value of this plant from the perspective of the pharmaceutical and food industries. Materials and Methods: In this study, green leafy covers (GLCs) and hard shells (HSs) (non-edible parts), skinless kernels (SKs), brown-skins (BSs), and brown-skinned kernels (BSKs) (edible parts) of Çakıldak, Sivri, Tombul, Palaz, and Kalınkara as standard and Ham and Sivri Yağlı as local genotypes were used. The five parts of each genotype were ground to powder and eliminated to a size of less than 80 mesh. Each part was extracted using hexane and methanol for 10-deacetylbaccatin III (10-DAB III), baccatin III (BAC III), cephalomannine, and paclitaxel analyses in three replicates. Samples and standards were analyzed by acetonitrile: water gradient method on NOVA Spher 100 Phenyl-Hexyl C18 column inhigh-performance liquid chromatography reverse phase system with 228 nm ultraviolet detector and 1.0 mL/min flow rate. Microsoft Office Excel, 2016, and analysis of variance Jamovi Version 2.3 were used for statistical and data analysis, consecutively. Results: Hazelnut parts differed to a very high degree from each other in terms of the highest amount of 10- DAB III (Ham HSs, 9,15 μg/g), BAC III (Kalınkara BSs, 7.24 μg/g), cephalomannine (Sivri Yağlı BSs, 6.37 μg/g), and paclitaxel (Ham BSKs, 4.36 μg/g) they contained. While HSs, BSKs, and BSs were rich in taxanes in all of the analyzed genotypes, SKs, and GLCs remain limited for anticancer taxanes. Conclusion: This is the first report that revealed the differences in taxane contents of Turkish hazelnuts including previously untested standard and local genotypes and their parts. Significant differences between genotype and hazelnut parts are expected to highlight the health benefits of consuming raw Turkish hazelnut with BSs and their possible use as a functional food. These results add more information to elucidate the bioactive potential of Turkish hazelnuts and their by-products and provide a promising resource for the food and pharmaceutical industry with an anticancer perspective. [ABSTRACT FROM AUTHOR]

Rationale: Huahong tablet, a commonly used clinical Chinese patent medicine, shows good efficacy in treating pelvic inflammation and other gynaecological infectious diseases. However, the specific composition of Huahong tablets, which are complex herbal formulations, remains unclear. Therefore, this study aims to identify the active compounds and targets of Huahong tablets and investigate their mechanism of action in pelvic inflammatory diseases. Methods: We utilised ultrahigh‐performance liquid chromatography Q‐Exactive‐Orbitrap mass spectrometry and the relevant literature to identify the chemical components of Huahong tablets. The GNPS database was employed to further analyse and speculate on the components. Potential molecular targets of the active ingredients were predicted using the SwissTargetPrediction website. Protein–protein interaction analysis was conducted using the STRING database, with visualisation in Cytoscape 3.9.1. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using the DAVID database. Additionally, a traditional Chinese medicine–ingredient–target–pathway network was constructed using Cytoscape 3.10.1. Molecular docking validation was carried out to investigate the interaction between core target and specific active ingredient. Results: A total of 66 chemical components were identified, and 41 compounds were selected as potential active components based on the literature and the TCMSP database. Moreover, 38 core targets were identified as key targets in the treatment of pelvic inflammatory diseases with Huahong tablets. GO and KEGG enrichment analysis revealed 986 different biological functions and 167 signalling pathways. Conclusion: The active ingredients in Huahong tablets exert therapeutic effects on pelvic inflammatory diseases by acting on multiple targets and utilising different pathways. Molecular docking confirmed the high affinity between the specific active ingredients and disease targets. [ABSTRACT FROM AUTHOR]

AbstractMass spectrometry (MS) enables precise identification and quantification of molecules, particularly when combined with chromatography. The advent of atmospheric pressure ionization (API) techniques allowed the efficient coupling of liquid chromatography with MS (LC-MS), extending analyses to nonvolatile and thermolabile compounds. API techniques present limitations such as low informative capacity and reproducibility of mass spectra, increasing instrument complexity and costs. Other challenges include analyzing poorly polar molecules and matrix effects (ME), which negatively impact quantitative analyses, necessitating extensive sample purification or using expensive labeled standards. These limitations prompted the exploration of alternative solutions, leading to the development of the Liquid Electron Ionization (LEI) interface. The system has demonstrated excellent robustness and reproducibility. LEI has been employed to analyze various compounds, including pesticides, drugs of abuse, phenols, polycyclic aromatic hydrocarbons (PAHs), phthalates, and many others. Its versatility has been validated with single quadrupole, triple quadrupole, and QToF detectors, operating in electron ionization (EI) or chemical ionization (CI) modes and with both reverse phase liquid chromatography (RPLC) and normal phase liquid chromatography (NPLC). LEI has also been successfully integrated with the Microfluidic Open Interface (MOI), Membrane Introduction Mass Spectrometry (MIMS), and Microfluidic Water-Assisted Trap Focusing (M-WATF), broadening its application scope and consistently demonstrating promising results in terms of sensitivity and identification power. The most recent advancement is the development of Extractive-Liquid Sampling Electron Ionization-Mass Spectrometry (E-LEI-MS), a surface sampling and real-time analysis technique based on the LEI concept. This review article offers a comprehensive and up-to-date picture of the potential of LEI. [ABSTRACT FROM AUTHOR]

A reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for simultaneous quantification of Azelnidipine (AZN) and Telmisartan (TMN) in combined tablet dosage forms. Chromatographic separation was achieved using a C18 column (100 × 4.6 mm, 2.5 µm particle size) with a mobile phase consisting of Acetonitrile: pH 3 phosphate buffer (75:25, v/v) at a flow rate of 0.9 mL/min, with detection occurring at 237 nm. The method was validated according to International Conference on Harmonisation (ICH) guidelines, demonstrating linearity over the concentration ranges of 6.4-32 µg/mL for AZN and 16-80 µg/mL for TMN. The limits of detection (LOD) and quantitation (LOQ) were determined to be 0.4399 and 1.3332 for AZN, and 0.882 and 2.6729 for TMN, respectively. The method also exhibited precision, repeatability, and accuracy, with relative standard deviations (RSD) = 2% and accuracy ranging from 99-101%. To assess the method's stability-indicating capability, forced degradation studies were conducted under four stress conditions: acidic hydrolysis, alkaline hydrolysis, oxidative degradation, and thermal degradation. The results confirmed the method's ability to accurately quantify AZN and TMN in the presence of degradation products. [ABSTRACT FROM AUTHOR]

Acrylamide is an important chemical in great global demand for the synthesis of polyacrylamide. A facile and benign approach of concentrating high osmotic acrylamide aqueous solution at a low temperature is needed to replace the current energy-intensive and cost-expensive thermal flash evaporation process. For the first time, a multi-stage forward osmosis (FO) process has been developed to concentrate acrylamide solution from 200 g/L to 600 g/L. Thin-film composite (TFC) membrane was fabricated and used for the multi-stage FO process. Acrylamide feed solution (FS) with various concentrations was systematically characterized in terms of viscosity and osmotic pressure (OP). Draw solutes including NaCl and MgCl2 were tested, and their reverse salt fluxes were measured with the quantification of their accumulative contents in the resultant concentrated acrylamide solution. Different operation modes including AL-FS (active layer facing FS) and AL-DS (active layer facing DS) were explored to optimize the system efficiency of the FO concentration process. Both single- and multi-stage FO operations were investigated, and their performances were quantified to assess the efficiency of the concentration of acrylamide solution. The results demonstrate that the multi-stage FO operation could dramatically improve the system efficiency for the concentration of acrylamide solution better than the single-stage FO process, and the draw concentration renewal at Stages II, III, and IV led to a water flux increase of 18.56%, 17.52%, and 18.43%, respectively. Moreover, the accumulated MgCl2 in the final 600 g/L acrylamide solution was below 3.7 g/L, less than 0.62 wt% impurity in the product of acrylamide. Our work provides a practical insight into the viability and optimization of a multi-stage FO process for concentrating high osmotic chemicals. [ABSTRACT FROM AUTHOR]

Background The topical veterinary drug product containing fipronil and permethrin provides an effective repellent protection and high insecticidal efficacy for dogs. Objective The objective of this study was to develop a stability-indicating high-performance liquid chromatography (HPLC) method for simultaneous detection and quantification of fipronil, permethrin, their key degradation products, and butylated hydroxytoluene (BHT) in a topical drug product. Method The two active ingredients, their degradation products, and the antioxidant (BHT) were separated by a gradient elution on a Phenomenex Kinetex C18 column (150 × 3 mm, 2.6 µm particle size) maintained at 37°C with H2O “acetonitrile “isopropyl alcohol “85% H3PO4 (65.5 + 32.5 + 4/0.0053, v/v/v/v) as mobile phase A and acetonitrile (100%) as mobile phase B. The flow rate was 0.9 mL/min, and analytes were detected and quantified at 235 nm. Results The specificity of the method was demonstrated by adequate separation of fipronil, permethrin, their degradation products, and BHT in the forced degraded finished product. The linearity of the method was demonstrated in the range of 0.2% to 150% of target analytical concentration of both active ingredients and 50% to 150% for BHT. Excellent recoveries of fipronil, permethrin, and BHT in placebo spiked active ingredient solutions in the linearity range showed sufficient accuracy of the method. The LOQ and LOD of the method were determined to be 0.2% and 0.07% of the analytical concentration. A robustness study did not identify any critical parameter that adversely affected the separation and quantification. Conclusions Here, we report the development and validation of a robust, stability-indicating HPLC method for identification and assay of fipronil, permethrin, and BHT, including estimation of fipronil's and permethrin's degradation products in a topical drug product for dogs. Highlights The new HPLC method permits the acquisition of data for all analytes of interest for a topical finished drug product containing fipronil, permethrin, and BHT. [ABSTRACT FROM AUTHOR]

Background Estimation of the drug and development of the method is a critical aspect of formulation development and a critical factor for analytical scientists. Gefitinib is a poorly soluble anticancer drug. Objective The present research focuses on the topic of the development of innovative quality by design methods for the estimation of gefitinib (GF) from bulk, pharmaceutical tablet formulation, and complex nanoformulations. Methods To simplify the estimation of poorly soluble drugs such as GF, response surface methodology (RSM) was adopted with effective leverages to obtain precise computation design space using the Box-Behnken design (BBD) model. The major three mixed-effect independent factors (percentage of buffer, pH of buffer, and flow rate) were screened with three prominent dependent responses (viz. theoretical plate, retention time, and tailing factor) selected for optimal analysis. Furthermore, co-processed steps were employed for the estimation of the analyte from the complex formulation. Results The RP-HPLC method uses the quality by design (QbD) approach can effectively estimate the analyte concentration of less than 4.5 min. The developed method was economically robust and sensitive and shows a relative standard deviation (RSD, %) of less than 2% for all the selected validation parameters. The estimated design space suggests the highest desirability (R2—0.998) at 60% of buffer in the mobile phase, pH 4.25, and flow rate of 0.7 mL/min. Conclusions The QbD approach was used to design and develop the method by understanding the interaction between dependent and independent variables to get the optimum values. The developed method was validated successfully and can be useful for formulation scientists to estimate drug concentration and drug release profiles from complex nanoformulations. Highlights The analytical approach was designed and quantified using a quality-by-design approach to make the RP-HPLC method more robust and efficient for the estimation of analytes from complex nanoformulations. The method is also useful to eliminate the interfering molecule during estimation by employing co-processing steps. The developed method saves time and cost of solvent and employs QbD as a requirement of recent regulatory concern. [ABSTRACT FROM AUTHOR]

Vigna trilobata, belonging to the Fabaceae family, has been extensively used in traditional medicine in different societies due to its diverse medicinal attributes and potential therapeutic applications. The current study aimed to assess the antioxidant and antibacterial efficacy of protein extract derived from Vigna trilobata seeds. The extracted seed protein sample was precipitated using the ammonium sulphate method, at 80% concentration. Subsequent analysis of the crude sample was conducted using RP-HPLC (reverse phase high-performance liquid chromatography) and LC–MS (liquid chromatography–mass spectrometry). The antioxidant assays demonstrated that at a concentration of 10 mg/ml, the maximum percentage of inhibition of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical was found to be 87.88 ± 0.01%. The ABTS (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) assay revealed the most significant scavenging activity (22.69 ± 0.22%) at a concentration of 10 mg/ml of crude protein. Furthermore, the FRAP (ferric reducing antioxidant power) assay identified the highest scavenging activity in the crude protein at a concentration of 25 mg/ml, amounting to 2.344 mol Fe2+/mg. The anti-inflammatory efficacy was assessed by the protein denaturation and proteinase inhibition assays. The antibacterial activity of protein extract was also evaluated against Gram-positive and Gram-negative bacteria. In conclusion, the evaluation of the protein extract of Vigna trilobata shows its potential as a natural health-promoting agent due to its antioxidant, anti-inflammatory, and antibacterial properties. The protein can help in reducing oxidative stress related to chronic diseases. It also helps in alleviating inflammation-linked ailments. Additionally, its antibacterial activity shows that it helps in combating microbial infections. Further analysis and purification of the protein extract could impact the food industry, where the purified proteins could potentially be used as a natural preservative, to improve the longevity and safety of food products. [ABSTRACT FROM AUTHOR]

For the first time a new QbD-assisted green stability indicating ultra-high-performance liquid chromatography (UHPLC) method was developed and validated for quantifying Tolvaptan. The method is simple, quick, cost-effective, and stable, and it was used to formulate a quality target product profile (QTPP) with strategically defined critical analytical attributes (CAAs) to meet specific criteria. Chromatographic separation was undertaken using a 10 cm long column of ACE excel super C18 with an interior diameter of 2.1 mm and particle size of 1.7 µm. The analysis was performed under controlled conditions at 25 ℃ with the mobile phase flowing at a rate of 0.2 mL/min and detection occurring at 220 nm. Injected 3 µL of standard by using an isocratic mobile phase system consisting of acetonitrile and water in a 95:5 v/v ratio. The diluents, prepared by mixing acetonitrile with water at a 90:10 volumetric ratio, were utilized. The analyte's retention time was determined to be 1.63 min. The developed method provided reliable results with accuracy exceeding 99% and a correlation coefficient exceeding 0.999 ranged between 10 and 150 µg/mL across the range for LOQ—150% levels. Notably, during forced degradation testing, Tolvaptan exhibited susceptibility to acidic hydrolysis. The method effectively separated degradation products during stress testing, demonstrating its stability-indicating status. Environmental sustainability assessment of the developed method was conducted through the investigation of various indicators of Complex GAPI, Analytical Eco scale and Analytical GREEness and it was concluded the optimized method aligns with environmentally friendly practices. [ABSTRACT FROM AUTHOR]

In this study, protonation constant values and liquid chromatographic behaviors of hydrophobic cyclizine, chlorcyclizine, hydroxyzine, cinnarizine, cetirizine, meclizine, and buclizine in some water‐organic solvent binary mixtures were examined for the first time using classical and green reverse phase liquid chromatography methods. In the isocratic study, the relationship of the retention time and mobile phase pH in water‐organic solvent binary mixtures containing acetonitrile (45, 50, 55, 60, 65%, v/v), methanol (60, 65, 70, 75%, v/v) and ethanol (45, 50, 55, 56, 59, 60, 62, 65%, v/v) were determined at 37 °C. In the study, XBridge C18 and Gemini NX C18 columns suitable for the chemical properties of basic compounds were used. The obtained liquid chromatographic data were analyzed using the linear solvation energy relationship methodology and the SOLVER program. The aqueous protonation constant values of the investigated compounds were calculated using the linear relationship between the protonation constant data calculated in studied binary mixtures and some macroscopic constant values of the solvents used. The greenness of methods developed using three different solvents was evaluated with the Analytical Greenness Metric Approach, the Green Analytical Procedures Index, and the Green Solvent Selection Tool approaches. [ABSTRACT FROM AUTHOR]

The development of analytical procedures, in line with the recent regulatory requirements ICH Q2 (R2) and ICH Q14, is progressing, and it must be able to manage the entire life cycle of the methodology. This is also applicable to and especially challenging for combinations of drug substances and dosage form. A reliable and efficient, stability-indicating, MS-compatible, reverse-phase ultra-performance liquid chromatographic (UPLC®) method was developed for the determination of carvedilol and felodipine in a combination oral dosage form. The development of the method, performed using analytical quality by design (AQbD) principles, was in line with the future regulatory requirements. Furthermore, the fixed-dose combination dosage forms are a clear solution to the polypharmacy phenomenon in the elderly population. The main factors evaluated were the mobile phase buffer, organic modifier, column, flow, and column temperature. The optimum conditions were achieved with a Waters Acquity HSS T3 (100 × 2.1 mm i.d., 1.8 µm) column at 38 °C, using ammonium acetate buffer (5 mM, pH 4.5) (Solution A) and MeOH (Solution B) as mobile phases in gradient elution (t = 0 min, 10% B; t = 1.5 min, 10% B; t = 12.0 min, 90% B; t = 13.0 min, 10% B; t = 15.5 min, 10% B) at a flow rate of 0.2 mL/min and UV Detection of 240 and 362 nm for carvedilol (CAV) and felodipine (FLP), respectively. The linearity was demonstrated over concentration ranges of 30–650 µg/mL (R2 = 0.9984) (CAV) and 32–260 µg/mL (R2 = 0.9996) (FLP). Forced degradation studies were performed by subjecting the samples to hydrolytic (acid and base), oxidative, and thermal stress conditions. Standard solution stability was also performed. The proposed validated method was successfully used for the quantitative analysis of bulk, stability, and fixed-dose combination dosage form samples of the desired drug product. Using the AQbD principles, it is possible to generate methodologies with improved knowledge, leading to high-quality data, lower operation costs, and minimum regulatory risk. Furthermore, this work paves the way for providing a platform of robust analytical methods for the simultaneous quantification of innovative on-demand new dose combinations. [ABSTRACT FROM AUTHOR]

• Extracted root protein from resistant and susceptible tobacco genotype at 60 DAI of nematode infection. • 2DE and in-solution digested proteins analyzed by RPLC-MS/MS to identify the unknown proteins. • Comparative proteomics revealed RFK1 and RPP 13 like candidate genes exclusively present in resistant genotype. • Out of 306 proteins, 79 proteins were up-regulated and 51 proteins were down-regulated. • These candidate genes can be exploited for the development to resistant crop against nematode disease. Root knot nematode (RKN) is an obligate endoparasitic pest of many vegetable, fruits, non-food cash crops etc. It causes more economic losses in terms of production and productivity worldwide. A range of management strategies including synthetic nematicides are available which can be harmful to human health, environment, wildlife, beneficial organisms and may induce resistance in plant populations. Therefore, identification of resistant gene is an alternative strategy to improve the crop either through breeding or by transgenic technology. In present investigation, an attempt was made to identify the candidate genes resistant to root knot nematode through proteomics approach. Comparative proteome was executed using RKN resistant ABT-10 and susceptible A-119 genotype. Proteomics was performed on nematode-challenged root tissue using 2D gel electrophoresis, in-solution trypsin digestion, and reverse phase liquid chromatography mass spectrometry. A total of 306 proteins were identified of which 79 proteins were up-regulated, 51 were down-regulated and 176 proteins showed no significant expression in resistant genotype. Out of 51 up-regulated proteins, two genes namely LRR Receptor-Like Serine/Threonine-Protein Kinase RFK1 and Putative Disease Resistance RPP13-Like Protein were identified in RKN resistant genotype. In addition to these two genes, RT-PCR analysis showed that three additional genes—Putative Wall-Associated Receptor Kinase-Like 11, Alpha-mannosidase, and TMV Resistance Protein N-Like gene—had significantly higher gene expression levels in the resistant root sample, exhibiting 14.89, 9.95, and 6.81 fold expression, respectively. The identified candidate genes can be exploited further for development of nematode resistant genotype in vegetables as well as other field crops through transgenic approaches. [ABSTRACT FROM AUTHOR]

Enantioseparation of α -hydroxy acids is essential since specific enantiomers of these compounds can be used as disease biomarkers for diagnosis and prognosis of cancer, brain diseases, kidney diseases, diabetes, etc., as well as in the food industry to ensure quality. HPLC methods were developed for the enantioselective separation of 11 α -hydroxy acids using a superficially porous particle–based teicoplanin (TeicoShell) chiral stationary phase. The retention behaviors observed for the hydroxy acids were HILIC, reversed phase, and ion-exclusion. While both mass spectrometry and UV spectroscopy detection methods could be used, specific mobile phases containing ammonium formate and potassium dihydrogen phosphate, respectively, were necessary with each approach. The LC–MS mode was approximately two orders of magnitude more sensitive than UV detection. Mobile phase acidity and ionic strength significantly affected enantioresolution and enantioselectivity. Interestingly, higher ionic strength resulted in increased retention and enantioresolution. It was noticed that for formate-containing mobile phases, using acetonitrile as the organic modifier usually resulted in greater enantioresolution compared to methanol. However, sometimes using acetonitrile with high ammonium formate concentrations led to lengthy retention times which could be avoided by using methanol as the organic modifier. Additionally, the enantiomeric purities of single enantiomer standards were determined and it was shown that almost all standards contained some levels of enantiomeric impurities. [ABSTRACT FROM AUTHOR]

The current study highlights the systematic quality by design-assisted creation of an efficient analytical technique for the estimation of embelin and vilangin in Vidanga (Embelia ribes) tablets. Response surface methodology in the design of experiments was used to identify key material attributes and critical process parameters that influence the designated critical analytical attributes. Separation was achieved on the X-Bridge (100 × 2.1 mm, 2 μm). The effects of acetonitrile content (v/v), flow rate, and column temperature on the retention times of the two drugs and their resolution, number of theoretical plates, and tailing factor were investigated and optimized. The optimal chromatographic conditions within the design space were found to be an isocratic mobile phase consisting of buffer and methanol (57.77:42.33, v/v) with a flow rate of 0.2 mL/min and a run time of 3 min. The retention times of embelin and vilangin were found to be 1.31- and 1.7-min. Different validation parameters were established, and the approach was validated in agreement with ICH and FDA requirements. Data analysis using statistical methods has revealed that the method is reliable, accurate, and robust. [ABSTRACT FROM AUTHOR]

Pyrimethamine (PYM) is an antiparasitic drug used in the treatment of congenital toxoplasmosis, available only as tablets. Therefore, preparing extemporaneous formulations and their quality control is essential for safe pediatric treatment. This study aimed to develop and validate a stability-indicating method by ultra-performance liquid chromatography (UPLC) to quantify PYM and methylparaben (MP) in extemporaneous oral suspension and evaluate its applicability to the tablets assay. The analytical conditions included RP-C18 column, mobile phase composed of 0.05 M acetate buffer pH 4.5 and organic mixture (acetonitrile:methanol), at 60:40 (v/v), flow rate of 0.2 mL/min, injection volume of 2 µL and detection at 272 nm. PYM was detected at 3.9 min and MP at 4.7 min, without interference of the degradation products or other excipients. Linearity was observed in the range of 0.5 to 25 μg/mL. The method showed accuracy (mean recovery between 98.0 and 102.0%) and precision (RSD < 2%) for suspension and tablets; the method was robust regarding the variations in the analytical conditions. The developed method met the requirements of current guidelines, was indicative of stability, and was suitable for PYM determination in extemporaneous preparations and tablets. [ABSTRACT FROM AUTHOR]

In this work, an eco-friendly simple, precise reverse phase high-performance liquid chromatography (HPLC) method has been developed and validated for Favipiravir in bulk and tablet dosage form followed by its force degradation study. The proposed method was validated to obtain official requirements including stability, accuracy, precision, linearity, robustness and selectivity as per International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) Guidelines. The estimation was developed on C (18) column reversed-phase using the mobile phase composition as methanol:water (10:90 v/v). The flow rate was set as 1 ml/min, and the maximum absorption was observed at 323 nm using Shimadzu Photo Diode Array detector. The Favipiravir, drug showed a precise and good linearity at the concentration ranges of 10–50 μg/mL. The Revearse Phase High Perforance Liquid Chromatography assay showed the highest purity ranging from 99.90 to 100.02% for Favipiravir, tablet dosage form, and 100.15% was the mean percentage purity. The percent recovery was found within the acceptance limit of (98.6–100.0%). Intra- and inter-day precision studies of the method were less than the maximum allowable limit percentage of relative standard deviation ≤ 2.0. The Favipiravir retention time was found to be 5.00 min. To examine the stability of the drug, various forced degradation studies were conducted on Favipiravir Active Pharmaceutical Ingredient. The developed method was validated according to the ICH guidelines. A very quick, cost-effective, precise and accurate HPLC method for the determination of Favipiravir has been developed and validated in compliance with ICH guidance Q2. [ABSTRACT FROM AUTHOR]

Ageratum conyzoides L., recognized globally as a noxious weed, holds significant value in traditional medicine, particularly in treating cuts, wounds, and tick infestations. This study focuses on identifying specific germplasms of this plant, rich in therapeutically valuable metabolites, across various phytogeographical zones in India. Using the Reverse Phase High-Performance Liquid Chromatography with Photodiode Array Detection (RP-HPLC-PDA) method, we analyzed 110 samples to quantify five key bioactive markers: Precocene-I (PRC-I), Coumarin (CM), Caryophyllene (CAR), Caryophyllene Oxide (CAO), and Humulene (HUM). Our findings revealed distinct geographical variations in these compounds. The highest concentrations of PRC-I (7.86±1.36 %) was found in sample NAC-12, CM (1.50±0.46 %) and HUM (0.0099±0.0004 %) in NAC-104, while CAR (6.33±0.87 %) and CAO (6.23±0.18 %) were predominant in NAC-01 and NAC-02, respectively. Ecological Niche Modelling (ENM) was used to predict the climatic suitability of regions for high yield of these metabolites. The ENM results showed that annual precipitation (Bio 12), precipitation of the coldest quarter (Bio 19) and minimum temperature of coldest month (Bio 6) were the principal climatic factors influencing metabolite content. Geographically, the Indo-Gangetic plain, Indus plain, and Central India were identified as suitable for higher content of PRC-I, CAO, CAR, whereas the North Eastern region favoured high content of CM and HUM. This study underscores the importance of bioactive marker-based assessment in understanding chemotypic diversity and demonstrates the effective use of ENM in predicting optimal habitats for harvesting specific chemotypes. These insights pave the way for location-specific cultivation strategies and targeted procurement of metabolite-rich germplasms, contributing to sustainable sourcing and conservation efforts, as well as to the advancement of herbal product standardization and efficacy. [Display omitted] • Five bioactive markers were quantified in 110 Ageratum conyzoides samples. • Precocene I was in maximum concentration followed by caryophyllene. • Central India region was favorable for precocene and caryophyllenes. • North Eastern region of India favoured high content of coumarin and humulene. [ABSTRACT FROM AUTHOR]

Background: Despite young children's widespread use of mobile devices, little research exists on this use and its association with children's language development. The aim of this study was to examine the associations between mobile device screen time and language comprehension and expressive language skills. An additional aim was to examine whether three factors related to the domestic learning environment modify the associations. Methods: The study uses data from the Danish large-scale survey TRACES among two- and three-year-old children (n = 31,125). Mobile device screen time was measured as time spent on mobile devices on a normal day. Measurement of language comprehension and expressive language skills was based on subscales from the Five to Fifteen Toddlers questionnaire. Multivariable linear regression was used to examine the association between child mobile device screen time and language development and logistic regression to examine the risk of experiencing significant language difficulties. Joint exposure analyses were used to examine the association between child mobile device screen time and language development difficulties in combination with three other factors related to the domestic learning environment: parental education, reading to the child and child TV/PC screen time. Results: High mobile device screen time of one hour or more per day was significantly associated with poorer language development scores and higher odds for both language comprehension difficulties (1–2 h: AOR = 1.30; ≥ 2 h: AOR = 1.42) and expressive language skills difficulties (1–2 h: AOR = 1.19; ≥ 2 h: AOR = 1.46). The results suggest that reading frequently to the child partly buffers the negative effect of high mobile device screen time on language comprehension difficulties but not on expressive language skills difficulties. No modifying effect of parental education and time spent by the child on TV/PC was found. Conclusions: Mobile device screen time of one hour or more per day is associated with poorer language development among toddlers. Reading frequently to the child may have a buffering effect on language comprehension difficulties but not on expressive language skills difficulties. [ABSTRACT FROM AUTHOR]

A sensitive ultra‐high‐performance liquid chromatography (UHPLC) method was developed for the quantification of trace levels of nilotinib and its related substances (RSs) in the nilotinib capsule dosage form. Critical method parameters were optimized using the design of experiments, employing the ACQUITY UHPLC BEH Phenyl column (2.1 × 100 mm, 1.7 μm) at a constant flow rate of 0.6 mL/min. The isocratic mobile phase, consisting of aqueous and organic phases, achieved efficient chromatographic separation within 8 min at 261 nm. The mean retention time for nilotinib was 6.1339, and for RSs, it ranged from 2.10 to 6.91 min, with a resolution exceeding 2 for all peaks. The method demonstrated robustness, separating known impurities and degradation products. The linearity was assessed over a concentration range of 0.02–80 ppm for nilotinib and 0.015–0.12 ppm for impurities. The limit of detection and limit of quantitation (LOQ) for nilotinib were 0.01 and 0.02 μg/mL, respectively, and 0.01 and 0.015 μg/mL for individual impurities. Recovery studies at LOQ, 100%, and 150% of the specification limit yielded percent recoveries ranging from 92.27% to 102.45%. Precision results showed low relative standard deviations, below 2% for nilotinib and below 8% for impurities. This method is deemed suitable for pharmaceutical formulation quantification. [ABSTRACT FROM AUTHOR]

This paper described a short and precise reverse‐phase high‐performance liquid chromatography (HPLC) method for quantifying mafenide in a film‐forming spray and preliminary in‐vitro release study. Following the International Council for Harmonization guidelines Q2 (R1), the method was validated. The chromatographic separation was achieved using a 10 mM potassium dihydrogen phosphate solution (mobile phase A) mixed with HPLC‐grade methanol (mobile phase B) in an 85:15 v/v ratio. Inert Sustain C8 (4.6 × 250 mm), 5 μm column was selected as the stationary phase. The flow rate was set as 0.8 mL/min, and detection was carried out at a wavelength of 222 nm. The developed HPLC method showed an accuracy between 98% and 102%. The primary diffusion study of film‐forming spray was performed using the Franz vertical diffusion cell apparatus. It was observed that an average of 26.94% of drug releases at 24 h. This indicates the drug has a slower release rate and shows local pharmacological action. The Weibull dissolution model was more fitting with regression square 0.9953. Furthermore, the drug excipient compatibility study revealed no interactions as the only drug peak was found in the chromatographic method, indicating that the chosen excipients were appropriate for a stable formulation. [ABSTRACT FROM AUTHOR]

Objectives: The aim of this study was to develop a simple, accurate, and precise method for the estimation of bupropion and dextromethorphan in a fixed-dose combination of tablets and robust high-performance liquid chromatography for assay analysis of such a fixed combination. Materials and Methods: Chromatographic analysis was performed and separations were achieved on a Denali C18 150 × 4.6 mm, 5 micron using a mobile phase composition of ortho-phosphoric acid and acetonitrile in the ratio of 600:400 (v/v), flow rate of 1.0 mL/min, injection volume is 10 µL and run time of 6 min in isocratic elution. Ultraviolet (UV) detection was performed at a wavelength of 221 nm. The temperature was maintained at 30 °C. Well-resolved peaks were observed with a high number of theoretical plates, lower tailing factor, and reproducible relative retention time. The method was validated, and all validation parameters were found to be within the acceptance limits. Results: A simple, accurate, and precise method has been developed for estimating bupropion and dextromethorphan in a fixed dose combination of tablets. The optimized method included the following parameters: column temperature of 30 °C, 40% acetonitrile as the mobile phase, and flow rate of 1.0 mL/min. Retention times were 2.25 min and 3.12 min for bupropion and dextromethorphan, respectively. The method was found to be linear in the range of 17.5-105 µg/mL [for R2 < 0.999) and 7.5-45 µg/mL (for R2 > 0.999] for bupropion and dextromethorphan, respectively. Both active pharmaceutical ingredients dissolved more than 90% within 5 min. Conclusion: The current study describes a new, simple, reliable, and economical elution reversed-phase high performance liquid chromatography method for estimating bupropion and dextromethorphan in a fixed combination tablet dosage form. The forced degradation studies were conducted using several degradation conditions such as acidic, alkali, oxidation, thermal, UV, and neutral conditions; the proposed method was effectively employed from the resolution of sample peaks. To the best of our knowledge, no such detailed and stability-indicating method has been reported for a fixed tablet dosage form. [ABSTRACT FROM AUTHOR]

An isocratic HPLC method has been developed and validated for estimating paracetamol and diclofenac sodium simultaneously with three skeletal muscle relaxants, namely, methocarbamol, tizanidine hydrochloride and chlorzoxazone in their pure standard mixtures and in different multi-component dosage forms in a single chromatographic run. HPLC separation was achieved on a C18 Inertsil ODS-3V 5 μm column (250 × 4.6 mm) using a mobile phase mixture containing acetonitrile and 25 mM phosphate buffer (pH 7.4 adjusted with NaOH) in the proportion of (39.7:60.3, v/v) pumped at 1.2 mL/min flow rate with UV detection at 220 nm. An experimental design was used by applying Plackett–Burman design for screening the most critical predictors affecting the chromatographic separation and Box–Behnken design for optimizing the selected predictors and creating the response surface between the selected predictors and the interested responses. ICH recommendations were applied for validating the proposed method with regard to linearity, precision, accuracy, selectivity, limits of detection and quantitation, and robustness. There are many applications for the optimized method that can be applied for routine estimation of the cited drugs in laboratories of quality control and pharmaceutical industries to save money and time and to reduce material waste and effort. [ABSTRACT FROM AUTHOR]

A reverse phase high-performance liquid chromatography (HPLC) method has been developed for the quantification of a typical drug Dasatinib (DST) and its related impurities in pharmaceuticals. Kinetex C18 (4.6 × 150 mm, 5 μm) column was used in the chromatographic separations, using buffer (1.36 g of KH2PO4 in 1000 mL of water, pH = 7.8; adjusted with diluted KOH solution) with solvent as acetonitrile and mode of elution as the gradient. The flow rate is 0.9 mL/min, column oven temperature as 45°C and the overall gradient run time as 65 min. The developed method was found to produce symmetric and good separation between the process-related and degradation impurities. Method optimization is achieved with photodiode array at 305 nm over the concentration range of 0.5 mg/mL and degradation studies were carried out under acidic, alkaline, oxidative, photolytic and thermal conditions to demonstrate the stability indicating capability of the method. Two major impurities were found in forced degradation studies in the HPLC analysis, the unknown, acid degradants were enriched and isolated by preparative HPLC, then characterized through high-resolution mass spectrometry, nuclear magnetic resonance spectroscopy and Fourier transform infrared spectroscopy. The unknown acid degradation impurity was showing Exact Mass of 521.11, molecular formula C22H25Cl2N7O2S and its chemical name as 2-(5-chloro-6-(4-(2-hydroxyethyl) piperazin-1-yl)-2-methylpyrimidin-4-ylamino)-N-(2-chloro-6-methylphenyl) thiazole-5-carboxamide. Another impurity (oxidative degradant) found as known DST N-oxide Impurity-L and its chemical name as 4-(6-((5-((2-chloro-6-methylphenyl) carbamoyl) thiazol-2-yl) amino)-2-methylpyrimidin-4-yl)-1-(2-hydroxyethyl) piperazine 1-oxide. The analytical HPLC method was further validated as per ICH guidelines. [ABSTRACT FROM AUTHOR]

This study presents the validation of a reverse phase high performance liquid chromatographic technique (RP-HPLC) for the simultaneous measurement of hydrochlorothiazide and telmisartan in tablet and bulk dose forms. An ODS Inertsil C18 (150 × 4.6 mm, 5μ particle size) column was used for chromatography in isocratic mode. The mobile phase included 0.1% trifluoroacetic acid: acetonitrile (70:30) ratio. The eluent was measured at 225 nm and the flow rate was 1.2 ml/min. It was discovered that the chosen chromatographic conditions successfully separated hydrochlorothiazide (RT-2.70) and telmisartan (RT-9.98 min). It was discovered that the suggested approach was reliable, accurate, economical and resilient. It can be applied to the simultaneous study of these medications in tablet form. [ABSTRACT FROM AUTHOR]

The present paper describes a simple method for the determination of uranium in uranium process stream solutions by reversed-phase liquid chromatographic method using C18 column, mixture of α-hydroxy isobutyric acid and methanol as eluent and flow-through spectrophotometer detector. This method has an advantage that interfering elements like rare-earth elements do not retain on the column and elute near the solvent peak thereby not interfering with uranium elution peak. Different eluents and parameters like pH, concentration of eluent are studied for the purpose. The spiking standard recovery was found to be more than 90 %. The developed method has been compared with other techniques and found to be in agreement. [ABSTRACT FROM AUTHOR]

The objective of this study is to develop simple, rapid and sensitive reversed-phase high performance liquid chromatography method for bilastine in tablet dosage form by quality by design approach. In reversed-phase high performance liquid chromatography method, chromatographic conditions such as mobile phase composition and flow rate were optimized with the help of Design-Expert software using central composite design. In this method, bilastine was estimated using octadecylsilane column (250×4.6 mm, 5 µm) and methanol:acetonitrile (90:10 % v/v) as mobile phase and flow rate 1 ml/min which are optimized using Design-Expert software. Method was developed at 280 nm detection wavelength. The retention time was found to be 3.484 min. The proposed method was successfully applied to the determination of bilastine in tablet dosage form. High linearity of developed method was confirmed over concentration range of (20-120) µg/ml and correlation co-efficient is 0.9997. The percentage relative standard deviation for precision and accuracy of the method was found to be <2 %. The recovery was in the range of 98.8 %-99.7 %; limit of detection was found to be 0.1352 µg/ml and limit of quantification was found to be 0.4098 µg/ml. Bilastine was found to degrade under oxidation condition. There was no interference of excipient and degradation product in retention time, so the method was specific. Analytical parameters such as precision, accuracy, limit of detection, limit of quantification and robustness were determined according to International Conference on Harmonization guidelines. [ABSTRACT FROM AUTHOR]

A simple, specific, accurate, precise and robust ultra-performance liquid chromatography-photodiode array detection method for quantifying lunasin peptide in pure form or in plant based vegan supplements was developed and validated. Chromatographic separation was achieved on Accucore C18 column (50 mm×4.6 mm, 2.6 µm) with 0.1 % formic acid in water (A) and 0.1 % formic acid in acetonitrile (B) delivered isocratically (50:50 v/v) at a flow rate of 0.4 ml/min for 10 min and the detection was monitored at 216 nm. Lunasin was eluted at a retention time of 4.94 min with a total runtime of less than 6 min. The chromatographic parameters were optimized using the Box-Behnken design with a sample set of 30 experimental runs. The developed ultraperformance liquid chromatography-photodiode array detection method was validated in accordance with the International Council for Harmonisation Q2(R1) guideline and evaluated the parameters such as system suitability, linearity, sensitivity, accuracy, precision, specificity and robustness. The method was found to be linear within the concentration range of 0.50-10.0 µg/ml. The developed method could be a viable candidate for routine quantification of lunasin peptide in nutraceutical formulations. [ABSTRACT FROM AUTHOR]

Qiangli Dingxuan (QLDX) tablet is a widely recognized traditional Chinese medicine formula that has been extensively used in China for decades to treat vertigo, tinnitus, and dizziness owing to its outstanding therapeutic outcomes. However, the complexity of the chemical components in this tablet makes it challenging to separate and identify these components. This study presented an effective and sensitive strategy for the rapid separation and simultaneous structural identification of QLDX tablet components using ultra‐performance liquid chromatography with quadrupole time‐of‐flight tandem mass spectrometry and the UNIFI platform. Based on retention times, accurate masses, fragment ions, related literature, and authentic standards, 119 compounds were identified or tentatively characterized; these included 9 iridoids, 12 lignans, 21 phenylpropanoids, 27 flavonoids, 7 phthalides, and 43 others. Among them, 36 were confirmed using reference standards. The representative compounds with various chemical structures were studied by analyzing their fragmentation patterns and characteristic ions. In conclusion, this study established a rapid approach for characterizing the chemical constituents in QLDX tablet. The proposed approach provides a basis for qualitative analysis and quality control in the manufacturing process and is beneficial for advancing investigations into the efficacy and mechanism of action of this tablet. [ABSTRACT FROM AUTHOR]