Chemical bonding of cross-linked glutaraldehyde/chitosan on the surface of a titanium wire to prepare a robust biocompatible SPME fiber for analysis of phytohormones in plants.

A robust biocompatible solid-phase microextraction (SPME) fiber, so-called Ti/APTS/GA/CS, was prepared by chemical bonding of cross-linked glutaraldehyde-chitosan to the surface of a titanium wire using APTS. The fiber was applied for sampling of phytohormones in plant tissues, followed by HPLC-UV analysis. The structure and morphology of the fiber coating was investigated by FT-IR, SEM, EDX, XRD, and TGA techniques. A Box-Behnken design was implemented to optimize the experimental variables. The calibration graphs were linear over a wide linear range (0.5-200 µg L-1) with LODs over the range of 0.01-0.06 µg L-1. The intra-day and inter-day precisions were found to be 1.3-6.3% and 4.3-7.3%, respectively. The matrix effect values ranged from 86.5% to 111.7%, indicating that the complex sample matrices had an insignificant effect on the determination of phytohormones. The fiber was successfully employed for the direct-immersion SPME (DI-SPME-HPLC) analysis of the phytohormones in cucumber, tomato, date palm, and calendula samples.

Antibiotic residues in pasteurised and Raw Cow's milk in Dhaka, Bangladesh.

This study aimed to investigate antibiotic residues such as oxytetracycline, ciprofloxacin, enrofloxacin and levofloxacin, in both pasteurised and raw cow's milk. A method using high-performance liquid chromatography with a UV detector (HPLC-UV) was developed and validated following International Conference on Harmonization (ICH) guidelines for simultaneous detection and quantification of these residues. The technique demonstrated linearity, with r2 values ranging from 0.999 to 1.00 within the 1.3-15.0 µg ml-1 range for each antibiotic. Thirty cow's milk samples, raw and pasteurised, from Dhaka's local markets were analysed, revealing the presence of enrofloxacin and levofloxacin, while oxytetracycline was absent in all samples. Notably, pasteurised milk samples contained enrofloxacin, levofloxacin and oxytetracycline, with groups P6 and P7 exceeding the Maximum Residue Limit for enrofloxacin, levofloxacin and ciprofloxacin (121 µg l-1). This study emphasises antibiotic residues in milk, with a validated method holding promise for routine analysis in industries requiring simultaneous quantitation of multiple antibiotics.

In-tube solid phase extraction with graphitic-based polyurethane sponge as a superhydrophobic sorbent and determination of drug residues in foodstuffs using high-performance liquid chromatography.

Veterinary drugs used in animal husbandry raise public health concerns due to their residues in the bodies of animals. This study employs a simple and quick sample preparation technique, in-tube solid phase extraction, to extract drug residues from foodstuffs, including eggs, honey, and water. This technique utilizes the synergy of graphitic-based materials and polyurethane sponges (PU) combined through dip coating method to make reusable sorbents for extracting drugs, including amoxicillin, paracetamol, ciprofloxacin, and cefixime. These prepared sorbents were characterized using FTIR, SEM, and XRD. HPLC analysis assessed the extraction efficiency, considering various parameters such as analyte concentration, sample solution pH, extraction time, type of eluting solvent, and graphitic-based polyurethane sponge reusability and stability. The proposed method exhibited a linear response for all three sorbents in the range of 0.03-1000 µg mL-1, with LOD 0.03-1.60 µg mL-1 and LOQ 0.18-4.84 µg mL-1. The % RSD ranged from 1.3 to 9.3 %, with recoveries of up to 98.42 %.

The reality of the use of growth hormones in fish (Rui (Labeo rohita), Catla (Catla catla), and Monosex Tilapia (Oreochromis niloticus) production.

The unobservable use of hormones in fish production is becoming an alarming issue worldwide. To reveal the fact in Bangladesh, 144 fish samples (rui (Labeo rohita), catla (Catla catla), and monosex tilapia (Oreochromis niloticus)) were collected from different fish farms and markets of Mymensingh district. The market samples had two sources (Mymensingh and Rajshahi district). The steroid hormonal (testosterone, estrogen, and progesterone) residue was analyzed by HPLC-UV detection. A standard questionnaire survey was conducted where most farmers (80%) denied using the hormone in fish production. Among the analyzed samples of all three fishes, hormonal residues were detected in approximately 98% of samples, and around 92% contained residues above the ADI. Among the contaminated samples, 70% of samples had a single residue and 30% had multiple residues. The testosterone and progesterone hormonal residue was detected in all three fishes in both farm and market samples and ranged (above ADI) from 2.1 to 16.96 µg/kg and 31.47-731.57 µg/kg (p < 0.05) respectively. The estrogen hormone residue was only detected in market samples (Rajshahi district) of rui and catla and no residue was detected in tilapia fish and the hormone level (above ADI) ranged from 8.23 to 40.13 µg/kg. This study revealed that the use of hormones varies on the attitude of farmers based on the local culture pattern as estrogen hormone residue was only detected in market samples. The consumption of contaminated fish at such concentrations may cause many health hazards in humans, especially in children. Thus, this study reveals a new alarming fact to focus on, and an effective monitoring system should be implemented as soon as possible for public health concerns.

Electrospun of functionalized mesoporous UiO-66 as the selective coating of solid phase microextraction Arrow for the determination of nine alkylphenols.

A solid-phase microextraction (SPME) Arrow and high-performance liquid chromatography-UV detector (HPLC-UV, detection at 225 nm) based method was developed for the selective determination of nine alkylphenols (APs) in milk. The functionalized mesoporous UiO-66 (4-meso-UiO-66) was utilized as the new coating material, which was synthesized by post-modification of pore-expanded UiO-66-NH2 by an esterification reaction with 4-pentylbenzoic acid. It was fully characterized by X-ray photoelectron spectroscopy (XPS), fourier transformation infrared spectrometry, nitrogen sorption-desorption test, scanning electron microscopy, transmission electron microscopy, and X-ray diffractometer. The characterization results showed the ester groups and benzene rings were introduced into the 4-meso-UiO-66, and the mesoporous structure was predominant in the 4-meso-UiO-66. The extraction mechanism of 4-meso-UiO-66 to APs is the synergistic effect of Zr-O electrostatic interaction and the size exclusion effect resulting from XPS, selectivity test, and nitrogen sorption-desorption test. The electrospinning technique was utilized to fabricate the 4-meso-UiO-66 coated SPME Arrow and polyacrylonitrile (PAN) was used as the adhesive. The mass rate of 4-meso-UiO-66 to PAN and the electrospinning time were evaluated. The extraction and desorption parameters were also studied. The linear range of this method was 0.2-1000 µg L-1 with a coefficient of determination greater than 0.9989 under the optimal conditions. The detection limits were 0.05-1 µg L-1, the inter-day and intra-day precision (RSD) were 2.8-11.5%, and the recovery was 83.6%-112%. The reusability study showed that the extraction performance of this new SPME Arrow could be maintained after 80 adsorption-desorption cycles. This method showed excellent applicability for the selective determination of APs in milk.

Quick synthesis of CoFe-PBA@GO with electrochemical method as a novel, sensitive, and degradable nanocomposite applied in nanofibers for triazole extraction before HPLC-UV analysis.

Today, the wide use of triazole fungicides due to environmental damage and its side effects has raised global concern. Hence, in this research, poly-vinyl alcohol/polyacrylic-acid/CoFe-PBA@GO electrospun nanofiber was synthesized and applied as effective, degradable, and novel adsorbent at pipette-tip microextraction (PT-µSPE) method for the rapid and concurrent extraction of five of triazole fungicides in fruit and vegetable samples prior to quantitative analysis by high-performance liquid chromatography-ultraviolet. The incorporation of CoFe-PBA@GO with superporous structure and abundant functional groups in a polymer medium improves the extraction efficiency of nanofibers due to hydrogen bonding and π-π interactions formed between analytes and synthesized nano-adsorbent. Various important elements that affect the extraction yield of the target analytes were optimized utilizing a time-variable approach. Under the optimum conditions, dynamic range was attained in the range of 0.3-900.0 ng/mL with correlation coefficients ≥ 0.999. The identification limit of the PT-µSPE-HPLC-UV method ranged from 0.1 to 0.3 ng/mL.

Utilization of a pH-switchable hydrophilicity solvent for the microextraction of clomipramine from human urine samples.

Clomipramine (CLP) is a tricyclic antidepressant drug, and its determination in biological samples is of high importance in clinical and forensic evaluations to assure appropriate drug concentrations. In the present study, benzoic acid was employed as a pH-switchable hydrophilicity solvent (SHS) for the microextraction of CLP from authentic human urine samples prior to its determination by high performance liquid chromatography-ultraviolet detection (HPLC-UV). The microextraction protocol was based on the phase transition of the SHS through pH alteration that resulted in its rapid dispersion and simultaneous phase separation. The obtained solid was collected in a syringe filter, dissolved in methanol, and analyzed. The main parameters that affected the efficiency of the microextraction procedure were studied and optimized to ensure high extraction efficiency for CLP and the analytical method was validated. Under optimum conditions, good linearity was observed between 0.05 and 5.0 µg mL-1. The limit of detection and limit of quantification were found to be 0.015 and 0.05 µg mL-1, respectively. The RSD values for intra-day repeatability and inter-day precision were 2.4-8.9 % and 1.7-9.1 %, respectively. The relative recovery values were within 90.0 and 110.0 % in all cases, demonstrating good method accuracy. The proposed SHS microextraction showed cost-efficiency, handling simplicity, and rapidity resulting in enhanced sample throughput. Moreover, the proposed method exhibited a green character and good applicability based on its evaluation by Green Analytical Procedure Index and Blue Applicability Grade Index.

Quantification of quercetin from red onion (Allium cepa L.) powder via high-performance liquid chromatography-ultraviolet (HPLC-UV) and its effect on hyperuricemia in male healthy Wistar albino rats.

Onions (Allium cepa L.) contain various flavonols, including quercetin, kaempferol, anthocyanin, luteolin, and myricetin. Quercetin in onions is considered the primary bioactive component. To assess the impact of quercetin on hyperuricemia in healthy Wistar albino rats, this study used high-performance liquid chromatography with ultraviolet (HPLC-UV) to identify and measure quercetin in onion powder. Twenty-four 160 ± 10 g, six wistar albino male rats in each group were kept: NC (control sample, no onion powder), OT1, OT2, and OT3, which contained 11.13, 14.84, and 18.61 g/100 g onion powder, respectively. The treatment lasted 28 days, during which the last 7 days were for urine, feces, and blood collection. The results showed a trend of decreasing levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, total bilirubin, total cholesterol, and low-density lipoprotein in rats fed OT1, OT2, and OT3 diets. Improvements were observed in feed, water, and nutrient intake, feed conversion ratio, feed efficiency ratio, nutrient digestibility, nitrogen balance, body weight, blood urea nitrogen, creatinine, and uric acid levels (p ≤ .05). In contrast, high-density lipoprotein, triglycerides, serum total protein, neutrophils, and lymphocytes did not change (p ≥ .05). White blood cells, red blood cell count, platelet count, hemoglobin, and monocytes showed an upward trend. Based on our calculations, we determined the optimal human dosage from the most effective amount of onion powder. By taking into account the ratio of human-to-rat surface area, we estimate that the equivalent human dose of onion is 181.04 grams with 204 mg of quercetin. Additionally, when factoring in the dry matter content, the recommended dose of onion is 29.19 grams with 220 mg of quercetin.

Quantification of Arbortristoside-A isolated from Nyctanthes arbor-tristis using HPLC: Method development and pharmaceutical applications.

Arbortristoside-A (Arbor-A) is a naturally occurring iridoid glycoside and herbal-based lead molecule with proven medicinal potential. Aiming at the development of an efficient analytical tool for the quantification of Arbor-A in pharmaceutical dosage forms, in the presented work, we developed an economical, fast, and sensitive RP-HPLC-UV method and validated the procedure as per the ICH guidelines, Q2(R1). The chromatographic separation was accomplished under the optimised experimental conditions using an HPLC system with an LC-2010 autosampler, a PDA detector, and a Phenomenex C18 column with the mobile phase composed of a 70:30 (v/v) water-acetonitrile mixture eluting isocratically at a flow rate of 1 mL/min at ambient temperature, and UV detection at 310 nm. Arbor-A showed a sharp peak at the retention time of 5.60 min and exhibited linearity (R2 = 0.9988) with LOD and LOQ of 0.50 µg/mL and 1.50 µg/mL, respectively. The accuracy of the method was 98.33-101.36 % with acceptable intra-day and inter-day precisions as well as robustness (<2% RSD). To ratify the applicability of the presented approach in emerging pharmaceuticals, a nanoformulation loaded with Arbor-A was designed and analysed utilising the provided methodology. The method has also enabled to determine the degradation kinetics of Arbor-A under stress conditions, etcetera, employing forced degradation and short term stability studies.

Mesoporous organosilicas with highly-content tyrosine framework as extractive phases for non-steroidal anti-inflammatory drugs in aquatic media.

BACKGROUND: Attentions regarding ordered mesoporous silica materials (OMSs), with large specific surface areas and narrow pore size distribution, which are prepared via self-assembly techniques, have been raised in sorption, separation, and sample preparation. However, in order to extend and improve their applications, a functionalization step is required. Organic units can be anchored on the inner or outer surface as well as in the silica wall framework by co-condensation-, grafting-, and periodic mesoporous organosilica (PMO) preparation approaches. Apparently, by synthesizing PMO with extensive and flexible organic bridging groups within the mesoporous wall, an efficient extractive phase can be achieved. RESULTS: We employed tyrosine amino acid to synthesize a PMO-based extractive phase. The FT-IR, 1H NMR, HR-ESI-MS, Low angle-XRD, TEM, FESEM, BET, and EDX-MAP analyses confirmed the successful synthesis of PMO within the salt-assisted templating method. A comprehensive study on sorption behavior of PMO was performed and its efficiency was evaluated against the grafting and co-condensation methods. Then, it was implemented to the pipette tip-micro solid phase extraction (PT-µ-SPE) of widely used non-steroidal anti-inflammatory drugs (NSAIDs) in water/wastewaters. Limits of detection and quantification were obtained in the range of 0.1-1.5 and 0.3-5 µg L-1, respectively. The calibration plots are linear in the 1-1000, 3-1000, 10-750, and 3-750 µg L-1, respectively. The intra-and inter-day precision at 50 and 200 µg L-1 levels are 2.9-7.1 % and 3.5-8%, while recoveries are between 84 and 111 %. SIGNIFICANCE: High-capacity tyrosine functionalized PMO with 2D hexagonal symmetry silica mesoporous structures found to be highly efficient extractive media. Despite the bulkiness and flexibility of the bridging group within the mesoporous wall, the synthesis condition was optimized in order to load more organic content in the PMO structure. The PMO performance was superior over organically modified ordered mesoporous silica materials prepared by the grafting and co-condensation methods.

Screening of natural inhibitors against peptidyl arginine deiminase 4 from herbal extracts by a high-performance liquid chromatography ultraviolet-visible based method.

Peptidyl arginine deiminase 4 (PAD4) is an important biocatalytic enzymes involved in the conversion of protein arginine to citrulline, its dysregulation has a great impact on many physiological processes. Recently, PAD4 has emerged as a potential therapeutic target for the treatment of various diseases including rheumatoid arthritis (RA). Traditional Chinese Medicines (TCMs), also known as herbal plants, have gained great attention by the scientific community due to their good therapeutic performance and far fewer side effects observed in the clinical treatment. However, limited researches have been reported to screen natural PAD4 inhibitors from herbal plants. The color developing reagent (COLDER) or fluorescence based methods have been widely used in PAD4 activity assay and inhibitor screening. However, both methods measure the overall absorbance or fluorescence in the reaction solution, which are easy to be affected by the background interference due to colorful extracts from herbal plants. In this study, a simple, and robust high-performance liquid chromatography ultraviolet-visible (HPLC-UV) based method was developed to determine PAD4 activity. The proposed strategy was established based on COLDER principle, while used hydrophilic l-arginine instead of hydrophobic N-benzoyl-l-arginine ethyl ester (BAEE) as a new substrate to determine PAD4 inhibition activity of herbal extracts. The herbal extracts and PAD4 generated hydrophobic l-citrulline were successfully separated by the HPLC, and the developed method was optimized and validated with a known PAD4 inhibitor (GSK484) in comparison with COLDER assay. The IC50 value of GSK484 measured by HPLC-UV method was 153 nM, and the detection limit of the citrulline was 0.5 nmol, respectively, with a linear range of 0.5 nmol to 20 nmol. The IC50 value of the HPLC-UV method was improved by nearly three times compared with COLDER assay (527 nM), and the results indicated the reliability of PAD4 inhibition via HPLC-UV method. The inhibitory effect against PAD4 were fast and accurately screened for the twenty-four extracts from eight herbs. Among them, Ephedra Herba extracts showed significant inhibitory activity against the PAD4 with the IC50 values of three extracts (ethanol, ethyl acetate and water) ranging from 29.11 µg/mL to 41.36 µg/mL, which may help researchers to discover novel natural compounds holding high PAD4 inhibition activity.

Comparative study on the enzymatic degradation of phenolic esters: The HPLC-UV quantification of tyrosol and gallic acid liberated from tyrosol acyl esters and alkyl gallates by hydrolytic enzymes.

HPLC-UV analysis was used to evaluate the enzymatic degradation characteristics of tyrosol acyl esters (TYr-Es) and alkyl gallates (A-GAs). Among various hydrolytic enzymes, TYr-Es can be hydrolyzed by pancrelipase, while A-GAs cannot be hydrolyzed by pancrelipase. Interestingly, carboxylesterase-1b (CES-1b), carboxylesterase-1c (CES-1c) and carboxylesterase-2 (CES-2) are able to hydrolyze TYr-Es and A-GAs, and thus to liberate tyrosol (TYr) and gallic acid (GA). By contrast, the degrees of hydrolysis (DHs) of TYr-Es and A-GAs by CES-1b and CES-1c were significantly higher than those by CES-2. Meanwhile, the DHs of TYr-Es were much higher than those of A-GAs. Especially, the DHs firstly increased and then decreased with the increasing alkyl chain length. Besides, DHs positively correlated with the unsaturation degree at the same chain length. Through regulating carbon length, unsaturation degree and the ester bond structure, controlled-release of phenolic compounds and fatty acids (or fatty alcohols) from phenolic esters will be easily achieved.

Magnetic dendritic mesoporous silica nanoparticles based integrated platform for rapid and efficient analysis of saccharides.

BACKGROUND: As an essential compound in living organism, saccharides have attracted enormous attentions from scientists in various fields. Understanding the distribution of saccharides in various samples is of great scientific importance. However, the low signal response and lack of specific recognition technology of saccharides and the complex matrix of samples make the analysis of saccharides a very challenge task. Thus, the development of a simple and straightforward strategy for the analysis of saccharides would represent a great contribution to the field. RESULTS: In this study, by employing the sulfonyl functionalized magnetic dendritic mesoporous silica nanoparticles as the substrate, we develop an integrated platform for analysis of saccharides. The construction of the platform mainly relied on multi-functional boronic acid, which serves as separation and derivation ligands at the same time. In the general procedure, the boronic acid is first immobilized onto the surface of substrate, then the selective enrichment of saccharides can be realized via boronate affinity separation. Finally, by the rational choice of the solution, we are able to elute the labelled complex (boronic acid-saccharide) from the substrate, which can be direct subjected to HPLC-UV analysis. The reliable precision (<15 %), accuracy (80-100 %), reproducibility (<10 %), improved sensitivity (20x) and limited time-consuming (down to minutes) of the proposed platform are experimentally demonstrated. SIGNIFICANCE AND NOVELTY: The successful quantification of different saccharides (alditols, glucose) in real samples is achieved. The proposed strategy is not only straightforward and fast, but also avoid the requirement of special equipment. With these attractive features, we believe that this strategy will greatly prompt the analysis of saccharides in various samples (eg. food, pharmaceutics and biosamples).

A novel sorbent based on electrospun for electrically-assisted solid phase microextraction of six non-steroid anti-inflammatory drugs, followed by quantitation with HPLC-UV in human plasma samples.

BACKGROUND: Although NSAIDs possess notable therapeutic and pharmaceutical qualities, it's essential to acknowledge that excessive doses can result in toxicity within the human body. Moreover, the importance lies in identifying and measuring their trace amounts. Due to their existence within intricate matrices, the creation of novel electrospun nanofibers as sorbents for electrically-assisted solidphase microextraction (EA-SPME) becomes vital. This innovation caters to the requirement for the effective pre-treatment of NSAID samples, providing a strategic approach to managing the complexities associated with trace quantities found in various matrices. RESULTS: First, polyvinylalcohol/casein/tannic acid/polyaniline/titanium dioxide nanoparticles (PVA/CAS/TA/PANI/TiO2 NPs) electrospun nanofibers were prepared for EA-SPME on pewter rode and then, trace amounts of six NSAIDs (Acetaminophen, Caffeine, Naproxen, Celecoxib, Ibuprofen and mefenamic acid) were adsorbed chemically on these nanofibers. In the next step, the desorption of six NSAIDs was electrochemically done from prepared electrospun nanofibers on a pewter rod which was as working electrode at three electrodes system. Finally, these drugs were quantified from different human plasma samples with HPLC-UV. The synthesis of electrospun nanofibers was confirmed through a series of analytical techniques including field emission-scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy with elemental mapping analysis (EDX-Mapping), X-ray diffraction (XRD), and Fourier transform-infrared (FT-IR). The optimal percentage of additive compounds to PVA/CAS for electrospinning, as well as the factors influencing adsorption and desorption processes, were determined through both of Design Expert software and MATLAB programming language. SIGNIFICANCE: Under optimum conditions, the wide linear range was 27-8000 ng mL-1 with R2≥ 0.9897, low detection limits were ranged from 8 to 27.3 ng mL-1 based on S/N = 3 and significant enrichment factors were acquired. The intra-day and inter-day RSDs% were obtained within the 4.51% - 5.68% and 4.28%-5.45%, respectively. Finally, The effectiveness of the EA-SPME-HPLC-UV method was assessed for determining NSAIDs in plasma samples, demonstrating good recoveries ranging from 90.2% to 105.2%.

Development of a simple high-performance liquid chromatography-ultraviolet detection method for olaparib in patients with ovarian cancer.

Olaparib is a small-molecule inhibitor of poly(ADP)-ribose polymerase (PARP) used as maintenance therapy for recurrent ovarian cancer and newly diagnosed advanced ovarian cancer after initial chemotherapy. An exposure-toxicity correlation has been reported between the probability of anemia, a common adverse event associated with olaparib, and the steady-state minimum plasma concentration (Cmin) as well as the predicted maximum plasma concentration (Cmax). On the other hand, olaparib exhibits high interpatient variability with regard to the area under the concentration-time curve, Cmax, and Cmin. Therefore, we developed a simple and sensitive assay based on high-performance liquid chromatography with ultraviolet light (HPLC-UV) for the therapeutic drug monitoring of olaparib. The analysis was performed on an octadecylsilyl column with a mobile phase consisting of 0.5% KH2PO4 (pH 4.5) and acetonitrile (71:29, v/v), at a flow rate of 0.8 mL/min. Olaparib and an internal standard (imatinib) were well separated from the co-extracted material, with retention times of 13.6 and 11.5 min, respectively. The calibration curve for olaparib showed linearity over the concentration range of 0.10-10.0 µg/mL (r2 = 0.9998). The intra- and inter- day validation coefficients ranged from 1.79 to 4.13% and 1.37 to 3.55%, respectively. Measurement accuracy ranged from - 6.07 to 3.26%, with a recovery rate of more than 91.06%. The developed method was then applied to evaluate the plasma olaparib concentrations in patients with ovarian cancer. Our findings demonstrate that HPLC-UV is an economical, simple, and sensitive method for clinical application and holds promise for the effective drug monitoring of olaparib during ovarian cancer treatment.

Optimization and validation of a bioanalytical HPLC-UV technique for simultaneous determination of underivatized phenylalanine and tyrosine in the blood for phenylketonuria diagnosis and monitoring.

This study aimed to develop a fast, accurate, and precise high-performance liquid chromatography with UV detection method for simultaneous analysis of underivatized phenylalanine (Phe) and tyrosine (Tyr) in biological samples. Separation of the analytes was accomplished using a Discovery HS F5-3 column, which offered better retention and peak symmetry for the tested analytes. Chromatographic conditions were optimized using central composite experimental design, and three factors were investigated: the concentration of ammonium acetate (A), the acetonitrile proportion in the mobile phase (B) and the column oven temperature (C). The approach was verified using ß-expectation tolerance intervals for total error measurement that did not exceed 15%. Optimal settings were A = 50 mm, B = 24% and C = 28°C. The method applicability was determined using human plasma from 75 volunteers. The limits of detection and quantification of the technique were satisfactory at 9 and 29 µm for Phe and 4 and 13 µm for Tyr. The mean analytical bias in spiking levels was acceptable, ranging from -1.649 to +1.659% for both substances, with RSD <5% in all instances. The suggested approach was successfully used to analyze Phe and Tyr in human blood samples and calculate the Phe/Tyr ratio.

Exploring the optimal electropolymerization strategy for the preparation of solid-phase microextraction fibers using pyrrole-dopamine copolymers.

In-situ electropolymerization of conductive polymers on the surface of stainless-steel substrates is a well-established but promising procedure for the preparation of solid-phase microextraction (SPME) tools. Herein, different electrochemical methods including constant potential (CP), constant potential pulse (CPP), and cyclic voltammetry (CV) were utilized to fabricate SPME fibers by in-situ electropolymerization of pyrrole-dopamine copolymers (PPY/PDA) on the surface of stainless-steel fibers. The coated fibers were characterized and applied for the direct-immersion SPME (DI-SPME) sampling of ultra-trace amounts of plant hormones including abscisic acid (ABA), gibberellic acid (GA3), and indole acetic acid (IAA) in fruit juices, followed by HPLC-UV determination. The results showed that CV electropolymerization is significantly more efficient than the two other methods. The coatings created by the CV method were satisfactorily uniform, adhesive, and durable and exhibited higher extraction performance compared to the CP and CPP procedures. The important experimental variables of the proposed DI-SPME-HPLC method were evaluated and optimized using response surface methodology with a Box-Behnken design. The developed method showed wide-range linearities, spanning from 0.05 to 20µg mL-1 for GA3, and 0.02 to 20µg mL-1 for ABA and IAA. The limits of detection were obtained 0.01µg mL-1 for GA3, and 0.005µg mL-1 for ABA and IAA. The fiber was successfully employed for the simultaneous DI-SPME-HPLC analysis of plant hormones in fruit juice samples.

Poly-ε-caprolactone based nanoparticles for delivery of genistein in melanoma treatment

Abstract We developed poly-ε-caprolactone (PCL)-based nanoparticles containing D-α-tocopherol polyethylene glycol-1000 succinate (TPGS) or Poloxamer 407 as stabilizers to efficiently encapsulate genistein (GN). Two formulations, referred to as PNTPGS and PNPol, were prepared using nanoprecipitation. They were characterized by size and PDI distribution, zeta potential, nanoparticle tracking analysis (NTA), GN association (AE%), infrared spectroscopy (FT-IR), and differential scanning calorimetry (DSC). PNTPGS-GN exhibited a particle size of 141.2 nm, a PDI of 0.189, a zeta potential of -32.9 mV, and an AE% of 77.95%. PNPol-GN had a size of 146.3 nm, a better PDI than PNTPGS-GN (0.150), a less negative zeta potential (-21.0 mV), and an AE% of 68.73%. Thermal and spectrometric analyses indicated that no new compounds were formed, and there was no incompatibility detected in the formulations. Cellular studies revealed that Poloxamer 407 conferred less toxicity to PCL nanoparticles. However, the percentage of uptake decreased compared to the use of TPGS, which exhibited almost 80% cellular uptake. This study contributes to the investigation of stabilizers capable of conferring stability to PCL nanoparticles efficiently encapsulating GN. Thus, the PCL nanoparticle proposed here is an innovative nanomedicine for melanoma therapy and represents a strong candidate for specific pre-clinical and in vivo studies.

Optimization and validation of high-performance liquid chromatography using modified QuEChERS to determine anthelmintic drugs in mutton.

The aim of this study was to rapidly determine the presence of anthelmintic drugs in sheep meat using the optimized high-performance liquid chromatography-ultraviolet (HPLC-UV) method with modified QuEChERS (quick, easy, cheap, effective, rugged, safe) technology. Fifty fresh sheep meat samples from different slaughterhouses were collected. A double extraction procedure (QuEChERS/HPLC-UV technology) was used to extract the target analytes. A multilevel calibration curve from 1 to 1000 g/kg was used to establish instrument linearity for rafoxanide, albendazole, and closantel, whereas 0.1-100 µg/kg was used for ivermectin, levamisole, and oxyclozanide to find the lowest concentration, maximum residue limit (MRL), and occupied range for targeted analytes. The concentration levels were used to investigate the linearity, whereas several certified reference materials were applied to determine accuracy. The process was linear for all combinations, from the limit of quantification (LOQ) to the maximum concentration. The LOQ was established at 0.5 µg/kg for ivermectin, levamisole, and oxyclozanide and 10 µg/kg for rafoxanide, albendazole, and closantel. Recovery values were 70%-120%, and repeatability/reproducibility stated in relative standard deviation was obtained at less than 20%. QuEChERS method revealed that most meat samples contained anthelmintic drug residues, of which the majority exceeded the MRLs. Thus, the drugs should be used correctly in animals to avoid residues in food for human consumption.

Simultaneous and trace-level quantification of four benzene sulfonate potential genotoxic impurities in doxofylline active pharmaceutical ingredients and tablets using high-performance liquid chromatography with ultraviolet detection.

In the production of doxofylline, the common occurrence of toxic p-toluene sulfonate generation prompted the development and validation of a method using HPLC with ultraviolet detection (HPLC-UV). This method is designed for detecting four potential genotoxic impurities (PGIs) present in both doxofylline drug substance and tablets, with a focus on the UV-absorbing group p-toluene sulfonate. The four impurities were methyl 4-methylbenzenesulfonate (PGI-1), ethyl 4-methylbenzenesulfonate (PGI-2), 2-hydroxyethyl 4-methylbenzenesulfonate (PGI-3), and 2-(4-methylphenyl)sulfonyloxyethyl 4-methylbenzenesulfonate (PGI-4). In this method, chromatographic separation was achieved using a Waters Symmetry C18 column (250 mm × 4.6 mm, 5 µm). The mobile phases consisted of 20% acetonitrile as mobile phase A and pure acetonitrile as mobile phase B, operating in gradient elution mode at a flow rate of 1.0 mL/min. According to the guidelines of the International Conference on Harmonization, it was determined that this method could quantify four PGIs at 0.0225 µg/mL in samples containing 60 mg/mL. The validated approach demonstrated excellent linearity (R2 > 0.999) across the concentration range of 30%-200% (relative to 0.075 µg/mL doxofylline) for the four PGIs. The accuracy of this method for the four PGIs ranged from 94.8% to 100.4%. The reverse-phase-HPLC-UV analytical method developed in this study is characterized by its speed and precision, making it suitable for the sensitive analysis of benzene sulfonate PGIs in doxofylline drug substances and tablets.