Magnetic Nanomaterials for Sample Preparation of Disease-related Biomarkers.

Measuring clinically relevant biomarkers is critical for disease screening, diagnosis, and therapeutic monitoring. However, analyzing trace biomarkers in complex biological fluids remains challenging. Magnetic solid phase extraction (MSPE) has recently emerged as a promising sample preparation approach due to its simplicity, efficiency, and ability to selectively isolate biomarkers. Databases, including PubMed, Web of Science, and Scopus, were systematically searched for studies on MSPE for clinical biomarkers. Key findings on nanomaterial synthesis strategies, surface modifications, and applications in biomarker isolation were synthesized. Recent research demonstrates magnetic nanoparticles with tailored surface chemistry can selectively extract biomarkers like cancer antigens, neurotransmitters, and pharmaceuticals from matrices such as plasma, urine, and serum. MSPE enables clinically relevant limits of detection, high recovery, and purification in a rapid and simple workflow. This technique shows significant potential to improve clinical diagnostics. Further research on novel magnetic materials and surface functionalization is warranted. This review provides insights for researchers aiming to develop MSPE methods for sensitive and selective analysis of clinical biomarkers.

Dispersive solid phase extraction of tacrolimus from biological samples using curcumin and iron-based metal organic frameworks nanocomposite followed by LC-MS/MS determination.

Tacrolimus is a potent immunosuppressive drug used in the prevention of tissue rejection. It has a narrow therapeutic index. Therefore, the determination of its concentration in biological fluids like plasma and urine is a very crucial issue. In this research, tacrolimus concentrations in plasma and urine samples were determined with a dispersive solid phase extraction procedure coupled to high-performance liquid chromatography-tandem mass spectrometry. For this purpose, a curcumin modified metal-organic framework was synthesized and used in extraction procedure. Tacrolimus was adsorbed onto the sorbent surface with aid of vortexing. Then, the adsorbed tacrolimus was eluted by a suitable solvent. Important parameters in extraction procedure were optimized by "one-variable-at-a-time" approach and reported as below: sorbent amount, 10 mg; sample solution pH, 2; agitation mode, vortexing; adsorption and desorption times, 1 min, and eluent (volume), methanol (200 µL). Under the optimized conditions and according to the International Council for Harmonization guidelines, the validation of the method was performed, and the results showed acceptable accuracy and precision (relative standard deviations ≤14 %), good linearity in a wide range (4-200 ng mL-1), and low limits of detection (1.2 ng mL-1 in plasma and 0.34 ng mL-1 in urine) and quantification (4.7 ng mL-1 in plasma and 1.12 ng mL-1 in urine). Finally, the validated method was successfully applied for the determination of tacrolimus in the plasma samples of the patients.

The anti-biofilm activity of hydrogen peroxide against Escherichia coli strain FL-Tbz isolated from a pharmaceutical water system.

Biofilms are considered a significant reason for the failure of disinfection strategies in industrial water systems due to their resistance to antimicrobial agents. This study is designed to investigate the anti-biofilm activity of hydrogen peroxide (H2O2) at combinations of temperatures and contact times. For this purpose, an in vitro microtiter plate (MTP)-based model system was used for biofilm formation using Escherichia coli (E. coli) strain FL-Tbz isolated from the water system of a pharmaceutical plant. To investigate the anti-biofilm activity of H2O2, it was added at different concentrations (2-7% v/v) to biofilms and incubated at different temperatures (20-60 °C) for 10-40 min to find effective conditions to eradicate biofilms. Maximum biofilms were formed when bacterial suspensions were incubated at 37 °C for 96 h. The rate of biofilm formation using an environmental isolate was higher than that of standard strain. H2O2 at concentrations of ≥6.25% (v/v) at temperatures of ≥40 °C incubated for ≥25 min significantly eradicated the biofilms.

Comparative Stability of Two Anti-hyperpigmentation Agents: Kojic Acid as a Natural Metabolite and Its Di-Palmitate Ester, Under Oxidative Stress; Application to Pharmaceutical Formulation Design.

Purpose: Kojic acid (KA) a natural metabolite and its dipalmitate ester, kojic acid dipalmitate(Kadp) are both prescribed to treat skin hyperpigmentation. Stress test reveals the intrinsicstability of active ingredients and leads to selection of the suitable formulations. This researchevaluates the comparative stability of KA and its di-palmitate ester under liquid oxidative stress. Methods: The HPLC-UV/PDA method with a C18 column was utilized. Liquid oxidative stresswas induced using hydrogen peroxide (H2O2). Degradation was separately induced for eachdrug, and they were compared to each other. Results: Kadp degraded more rapidly in similar liquid oxidative stress conditions than KA did.The superior degradation model was the first order for both drugs based on the mean percentage error (MPE) values, indicating the dependency of the reaction rate on the initial concentrationof the reactive substance. Ring opening was proposed as the most possible theory for KA andKadp oxidative degradation. Conclusion: It is suggested to use KA instead of Kadp in less stable formulations, such asextemporaneous preparations. The incorporation of antioxidant excipients in Kadp formulationsis recommended for yielding better stability results. Formulating Kadp in the internal phase ofo/w emulsion formulations may protect this susceptive molecule from oxidative degradationduring the shelf life of the pharmaceutical preparation. Further studies are required to study theexact mechanism of the degradation process.

Nanophytosomes for enhancement of rutin efficacy in oral administration for diabetes treatment in streptozotocin-induced diabetic rats.

Rutin is a natural antioxidant compound with several therapeutic benefits. However, the application of this bioactive compound is limited due to its low stability and bioavailability. To overcome these limitations, this study aimed to encapsulate rutin into nanophytosomes (NPs) and evaluate the therapeutic potency of this nanocarrier in streptozotocin (STZ)-induced diabetic rats. The particle size, zeta potential, and encapsulation efficiency of the prepared NPs were 72.72 nm, -22 mV, and 93.7%, respectively. The in vivo study showed that the oral administration of rutin-loaded NPs (containing 25 mg rutin/kg per day) for 4 weeks was more effective than free rutin in the control of hyperglycemia and hyperlipidemia in the STZ-induced diabetic rats. Additionally, the administration of rutin-loaded NPs regulated the activities of liver marker enzymes and the levels of total hemoglobin and glycated hemoglobin in the diabetic rats. The antioxidant defenses in the diabetic rats were increased by the administration of rutin-loaded NPs more than free rutin. Moreover, the histopathological study showed that the administration of rutin-loaded NPs restored the diabetes-induced damages in kidney, liver, and pancreas. In conclusion, encapsulation of rutin with phytosomes is an effective technique to benefit from its therapeutic potential, especially to attenuate diabetic complications.

Sensing and bioimaging of lead ions in intracellular cancer cells and biomedical media using amine-functionalized silicon quantum dots fluorescent probe.

A novel amine-functionalized silica quantum dots (SiQDs) fluorescent nanoprobe was developed for sensing of lead concentration in water, plasma and cell lysate. In addition, the developed probe was utilized for bioimaging of intracellular lead ions in HT 29 cancer cells. The amine-functionalized nanoprobe exhibited fluorescence emission at 445 nm under excitation at 355 nm. Upon addition of lead ions, the fluorescence of SiQDs linearly enhanced from 50 ng/mL to 5 µg/mL and 50 ng/mL to 25 µg/mL for plasma and standard media, respectively. The synthesis and fabrication of this probe are simple and serves high sensitivity with a limit of detection down to around 20 ng/mL. In the presence of various molecular and ion interfering, reliable results are obtained, confirming the specificity of the nanoprobe for lead ion detection. Meanwhile, amine-functionalized SiQD-based nanoprobe exhibits excellent cell membrane-permeability and biocompatibility. Thus, this probe is utilized for lead tracing in HT 29 cancer live cells. Fluorescent microscopy results confirmed the attachment of the produced nanomaterials to the HT 29 cancer cells.

Evaluation of the Physicochemical and Biological Stability of Cetuximab under Various Stress Condition.

Cetuximab is a chimeric monoclonal antibody against epidermal growth factor receptor (EGFR) and it is approved for treatment of human colorectal cancer and squamous cell carcinoma of head and neck. The aim of this research was to study the stability of cetuximab finish product (5 mg/mL) under various stress conditions including mechanical, thermal, light stress, and various freeze-thaw cycles. To determine the effects of environmental stresses on the physicochemical properties and bioactivity of cetuximab, a combination of physicochemical and cell-based biological methods including size exclusion chromatography (SEC), cation exchange chromatography (CEX), flow cytometry-based binding assay, and MTS cell viability/proliferation assay was used. The results obtained by the SEC and CEX methods revealed that incubation of cetuximab at 25 and 30 °C, shaking, and various freeze-thaw cycles caused no physicochemical instability. However, functional analysis of the samples exposed to the above-mentioned conditions revealed a significant decrease in the bioactivity of cetuximab indicated by a significant reduction in the cell binding and growth inhibitory effects of cetuximab in EGFR overexpressing cancer cell line (A431). Incubation of cetuximab at 40 and 50 °C led to polymerization and fragmentation of the mAb and resulted in a significant decrease in the bioactivity of the mAb. Our findings show that the light exposure had the most destructive effects on physicochemical and biological characteristics of cetuximab. In conclusion, we found that all mentioned stress conditions significantly affect the bioactivity of cetuximab. Our finding highlights the importance of bioactivity evaluation of biopharmaceuticals in their quality control assessment.

Piroxicam cocrystals with phenolic coformers: preparation, characterization, and dissolution properties.

This study explores the preparation and investigation of dissolution properties of piroxicam cocrystals. Differential scanning calorimetry (DSC) was used to determine the capability of resorcinol (RES), methylparaben (MPB), and vanillin (VAN) to form cocrystals with piroxicam (PRX). Generation of cocrystals was attempted by liquid assisted grinding and slurry methods. Cocrystals were characterized by thermal methods, powder X-ray diffraction, and Fourier-transform infrared spectroscopy. Apparent solubility, intrinsic dissolution rate (IDR), and powder dissolution profile of cocrystals were compared with anhydrous piroxicam, piroxicam monohydrate (PRXMH), and previously reported piroxicam-succinic acid cocrystal. Contact angles and particle sizes of the studied solids were also measured. Based on the DSC screening results, we prepared and characterized PRX-RES and PRX-MPB cocrystals. Interestingly, the cocrystals not only failed to improve apparent solubility and IDR of PRX but also showed lower values than PRX that were attributed to induction of phase transformation of PRX to PRXMH. In contrary, cocrystals performed better than PRX in powder dissolution studies. The higher dissolution rates of cocrystals were explained by improved wettability and reduced sizes. This study has highlighted the complexity of solid state properties of cocrystals and has provided new evidence for the in-solution stability issues of cocrystals.

Recent advances in improving oral drug bioavailability by cocrystals.

Introduction : Oral drug delivery is the most favored route of drug administration. However, poor oral bioavailability is one of the leading reasons for insufficient clinical efficacy. Improving oral absorption of drugs with low water solubility and/or low intestinal membrane permeability is an active field of research. Cocrystallization of drugs with appropriate coformers is a promising approach for enhancing oral bioavailability. Methods : In the present review, we have focused on recent advances that have been made in improving oral absorption through cocrystallization. The covered areas include supersaturation and its importance on oral absorption of cocrystals, permeability of cocrystals through membranes, drug-coformer pharmacokinetic (PK) interactions, conducting in vivo-in vitro correlations for cocrystals. Additionally, a discussion has been made on the integration of nanocrystal technology with supramolecular design. Marketed cocrystal products and PK studies in human subjects are also reported. Results : Considering supersaturation and consequent precipitation properties is necessary when evaluating dissolution and bioavailability of cocrystals. Appropriate excipients should be included to control precipitation kinetics and to capture solubility advantage of cocrystals. Beside to solubility, cocrystals may modify membrane permeability of drugs. Therefore, cocrystals can find applications in improving oral bioavailability of poorly permeable drugs. It has been shown that cocrystals may interrupt cellular integrity of cellular monolayers which can raise toxicity concerns. Some of coformers may interact with intestinal absorption of drugs through changing intestinal blood flow, metabolism and inhibiting efflux pumps. Therefore, caution should be taken into account when conducting bioavailability studies. Nanosized cocrystals have shown a high potential towards improving absorption of poorly soluble drugs. Conclusions : Cocrystals have found their way from the proof-of-principle stage to the clinic. Up to now, at least two cocrystal products have gained approval from regulatory bodies. However, there are remaining challenges on safety, predicting in vivo behavior and revealing real potential of cocrystals in the human.

Feasibility of electrospray deposition for rapid screening of the cocrystal formation and single step, continuous production of pharmaceutical nanococrystals.

OBJECTIVES: This study employed electrospray deposition (ESD) for simultaneous synthesis and particle engineering of cocrystals. SIGNIFICANCE: Exploring new methods for the efficient production of cocrystals with desired particle properties is an essential demand. METHODS: The possibility of cocrystal formation by ESD was examined for indomethacin-saccharin, indomethacin-nicotinamide, naproxen-nicotinamide, and naproxen-iso-nicotinamide cocrystals. Solutions of the drug and coformer at stoichiometric ratios were sprayed to a high electric field which caused rapid evaporation of the solvent and the formation of fine particles. The phase purity, size, and morphology of products were compared with reference cocrystals. Experiments were performed to evaluate the effects of stoichiometric ratio, concentration and solvent type on the cocrystal formation. Physical stability and dissolution properties of the electrosprayed cocrystals were also compared with reference cocrystals. RESULTS: ESD was found to be an efficient and rapid method to produce cocrystals for all studied systems other than indomethacin-nicotinamide. Pure cocrystals only formed at a specific drug:coformer ratio. The solvent type has a weak effect on the cocrystal formation and morphology. Electrosprayed cocrystals exhibited nano to micrometer sizes with distinct morphologies with comparable physical stability with reference cocrystals. Nanococrystals of indomethacin-saccharin with a mean size of 219 nm displayed a threefold higher dissolution rate than solvent evaporated cocrystal. CONCLUSION: ESD successfully was utilized to produce pure cocrystals of poorly soluble drugs with different morphologies and sizes ranging from nano to micrometer sizes in one step. This study highlighted the usefulness of ESD for simultaneous preparation and particle engineering of pharmaceutical cocrystals.