Detection Of Methotrexate Research Articles

Folate-functionalized CoFe2O4 nanozyme inspired by folate antagonist effect for methotrexate detection with high anti-interference ability and recyclability

Methotrexate (MTX) is a widely used chemotherapeutic drug to treat cancer and autoimmune diseases, which has a narrow therapeutic window and can cause systemic toxicity. The monitoring of MTX is therefore of crucial importance in clinical practice and precision medication. However, current detection techniques suffer from limitations in cost and time effectiveness, making them unsuitable for out-of-lab and point-of-care testing. Inspired by the antimetabolite mechanism of MTX, a signal-on colorimetric sensing platform based on CoFe2O4 nanozyme functionalized with folic acid (FA@CoFe2O4) was elaborately developed to monitor MTX. Due to the ability of the immobilized folate to be reduced by dihydrofolate reductase (DHFR) to tetrahydrofolate, it poses competitive effect on the chromogenic substrate by its strong reducibility when FA@CoFe2O4 exerts its peroxidase-like activity, thereby inhibiting the discoloration of the substrate. Once MTX is introduced, the active sites of DHFR will preferentially bind to it, leading to suppression in the generation of tetrahydrofolate and further restoring the substrate’s color rendering. Based on this principle, the developed sensing strategy exhibited a broad range from 0.1 μM to 40 μM towards MTX detection. In addition, the magnetic property endowed the FA@CoFe2O4 nanozyme excellent recyclability and allowed the MTX detection with high anti-interference ability from other drugs and coexisting antioxidants. The feasibility of the method was validated by monitoring MTX level in human serums. This work paves a new way towards therapeutic drug monitoring that serves personalized administration.

Methotrexate Detection in Serum at Clinically Relevant Levels with Electrochemically Assisted SERS on a Benchtop, Custom Built Raman Spectrometer.

Therapeutic drug monitoring (TDM) is an essential clinical practice for optimizing drug dosing, thereby preventing adverse effects of drugs with a narrow therapeutic window, slow clearance, or high interperson pharmacokinetic variability. Monitoring methotrexate (MTX) during high-dose MTX (HD-MTX) therapy is necessary to avoid potentially fatal side effects caused by delayed elimination. Despite the efficacy of HD-MTX treatment, its clinical application in resource-limited settings is constrained due to the relatively high cost and time of analysis with conventional analysis methods. In this work, we developed (i) an electrochemically assisted surface-enhanced Raman spectroscopy (SERS) method for detecting MTX in human serum at a clinically relevant concentration range and (ii) a benchtop, Raman detection system with an integrated potentiostat, software, and data analysis unit that enables mapping of small areas of SERS substrates and quantitative SERS-based analysis. In the assay, by promoting electrostatic attraction between gold-coated nanopillar SERS substrates and MTX molecules in aqueous samples, a detection limit of 0.13 μM with a linear range of 0.43-2 μM was achieved in PBS. The implemented sample cleanup through gel filtration proved to be highly effective, resulting in a similar detection limit (0.55 μM) and linear range (1.81-5 μM) for both PBS and serum. The developed and optimized assay could also be used on the in-house built, Raman device. We showed that MTX detection can be carried out in less than 30 min with the Raman device, paving the way toward the TDM of MTX at the point-of-need and in resource-limited environments.

Relationship of color change to permeation of target compound in polydiacetylene vesicle system

AbstractPolydiacetylene (PDA) systems, which showed color changes due to the permeation of target molecules, were reported previously by our research group. To use the systems for controlled drug delivery systems, the relationship of the color change to the permeation time of the PDA vesicle must be characterized precisely. Therefore, this relationship was investigated and characterized in this study. Methotrexate (MTX) was used as the target compound and the detection of MTX is an important process in anticancer study. N‐(2‐aminoethyl)pentacosa‐10,12‐diynamide (AEPCDA) was used as the diacetylene surfactant. Dimethyldioctadecylammonium bromide (DOAB) was used as an assistant surfactant. The color change occurred when the mixed surfactant vesicles, with DOAB, were used. When a lower ratio of DOAB was added to the AEPCDA vesicle, the color change rate was slower than that when using the higher ratio of DOAB. The color change rate constants were estimated for several different ratios (AEPCDA:DOAB). Surfactants with different carbon numbers to DOAB were used as the assistant surfactants in this study. The results showed that the color change rate constant decreased with the increasing carbon number of the assistant surfactant. This means that the color change rate can be controlled by changing the assistant surfactant.

Electrochemical detection of methotrexate in serum sample based on the modified acetylene black sensor

A novel acetylene black (AB) sensor modified with stearyltrimethylammoniumbromide (STAB) was fabricated and applied to the determination of methotrexate (MTX) in serum. For the strong adsorption of STAB and the high catalytic capacity of AB, the prepared sensor performed better adsorption and electrochemical catalysis capacity compared with acetylene black paste electrode (ABPE). Several factors affecting the performance of the sensor, such as the construction of the sensor and the parameters affect the electrochemical catalysis were optimized carefully on sequences in this study. The prepared sensor showed favorable performance in the determination of MTX with the linear range of 0.005 μM to 7.0 μM and the detection limit of 3.07 nM. Furthermore, the fabricated sensor exhibited good stability and repeatability, which was of importance in the electrochemical detection. Finally, the established electrode was employed in the detection of MTX in real sample of serum, which demonstrated the reliable and accurate of the prepared method. It is obvious that the prepared sensor existed great potential in the determination of MTX, which could provide a new approach for the fast and low-coat detection for MTX.

Development and validation of stability indicating reversed-phase liquid chromatographic method for simultaneous quantification of methotrexate and teriflunomide in nanoparticles and marketed formulation.

Methotrexate (MTX) and teriflunomide (TEF) are the two most effective disease-modifying antirheumatic drugs used as combination therapy for rheumatoid arthritis and no robust high-performance liquid chromatography (HPLC) method is available for their simultaneous estimation to date. Therefore, we have developed and validated an isocratic reversed-phase HPLC method for simultaneous analysis of MTX and TEF spiked in the form of active pharmaceutical ingredients, tablets and nanoformulations. The best separation was achieved on a BDS, C18 , 4.6 × 250 mm, 5 μm analytical column (Thermo Hypersil) with methanol-ethylammonium formate-potassium dihydrogen phosphate buffer (55 mm, pH 3.5; 65:5:30, v/v) as mobile phase at a flow rate of 0.8 mL/min. All the samples were subjected to force degradation studies. Responses of MTX and TEF were found to be a linear function of concentration over the range 1-50 μg/mL (r2 = 0.9976 and 0.9982). The limits of detection and limit of quantification were 7.74 and 25.82 ng/mL and 10.74 and 35.80 ng/mL, respectively. Degradation products produced under the stress studies did not interfere with the detection of MTX and TEF and therefore the developed method can be regarded as stability indicating.

Novel tungsten phosphide embedded nitrogen-doped carbon nanotubes: A portable and renewable monitoring platform for anticancer drug in whole blood

Biosensors based on converting the concentration of analytes in complex samples into single electrochemical signals are attractive candidates as low cost, high-throughput, portable and renewable sensor platforms. Here, we describe a simple but practical analytical device for sensing an anticancer drug in whole blood, using the detection of methotrexate (MTX) as a model system. In this biosensor, a novel carbon-based composite, tungsten phosphide embedded nitrogen-doped carbon nanotubes (WP/N-CNT), was fixed to the electrode surface that supported redox cycling. The electronic transmission channel in nitrogen doped carbon nanotubes (N-CNT) and the synergistic effect of uniform distribution tungsten phosphide (WP) ensured that the electrode materials have outstanding electrical conductivity and catalytic performance. Meanwhile, the surface electronic structure also endows its surprisingly reproducible performance. To demonstrate portable operation for MTX sensing, screen printing electrodes (SPE) was modified with WP/N-CNT. The sensor exhibited low detection limits (45 nM), wide detection range (0.01–540 μM), good selectivity and long-term stability for the determination of MTX. In addition, the technique was successfully applied for the determination of MTX in whole blood.

Determination of Trace Methotrexate and 7-OH-Methotrexate in Plasma by High-Performance Liquid Chromatog-Raphy with Fluorimetric Detection

Abstract A sensitive and reliable HPLC method consisting of a solid-phase extraction, post-column photoreaction of the analytes in a polyethylene tubing by UV lamp, and fluorimetric detection for the simultaneous determination of methotrexate (MTX) and 7-OH-methotrexate (7-OH-MTX) in plasma was developed. The linear relationships between the peak area and MTX (0.1 ng/ml-1000 ng/ml, r=0.9997) or 7-OH-MTX (6.25 ng/ml-400 ng/ml, r=0.9995) concentrations were obtained. The average coefficients of variation for the intraday and interday replications were less than 11% and the absolute recoveries were 96.8%-101.2% for MTX and 76.4%-86.7% for 7-OH-MTX for the calibration ranges used. The limit of detection for MTX in plasma was 0.05 ng/ml. This most sensitive HPLC method reported so far was used for the trace assay of plasma MTX concentrations in dogs following a topical application of MTX in gel.

Utility of serum bile acid levels as measured by bioluminescence for detection of methotrexate induced liver injury

Utility of serum bile acid levels as measured by bioluminescence for detection of methotrexate induced liver injury