Drugs In Biofluids Research Articles

H‐Bonded Supramolecular Crystals‐Coated Solid‐Phase Microextraction enables the Matrix Interference‐Free Detection of the COVID‐19 Antiviral Drugs

AbstractSince the outbreak of COVID‐19, many antiviral drugs have been used to treat infected patients. Excessive drug concentrations may not only damage patients' organs, but also lead to increased mortality. The lack of data makes it difficult for them to be treated with these drugs under proper and safe dosage. However, due to the complexity of fluid matrixand structures of the antiviral drugs, it is still difficult to detect the drugs in biofluids. Herein, solid‐phase microextraction (SPME) is applied for the detection of biological matrix, which has the advantages of simplicity, efficiency and environmental protection. With abundant hydrogen bond sites and appropriate molecular volume, biocompatible graded‐pore HOF10x series materials are selected as the fiber coating for exploration. Consequently, HOF101 could achieve high enrichment effect of drugs due to the commensurate mesopores and hydrogen bond non‐covalent interaction. Moreover, the excellent extraction capacity of the HOF101 fiber is verified to be consistent in various interfering conditions. The obtained limits of detections (LODs) are 0.24–0.66 ng L−1, and the recoveries of the biofluids in three concentration levels ranged from 80.30% to 120.1%. It shows the low LODs and satisfactory recoveries of this analytical method, which is expected to be applied in medical systems for drug monitoring.

Direct coupling in-tube solid-phase microextraction with mass spectrometry using polymer coated open-tubular column for rapid analysis of antiepileptic drugs in biofluids

Development of high-throughput and rapid screening analytical method is in high demand for anti-doping and clinical point-of-care (POC) analysis. Solid-phase microextraction and mass spectrometry direct coupling (SPME-MS) has been proved as a rapid and effective way for target analysis in complex sample matrixes. An online direct coupling of in-tube SPME (IT-SPME) with MS using polymer coated open-tubular column has been developed in this work. A sharp stainless-steel needle was attached at the end of the SPME column, which enables the direct ionization of the analytes after elution from the IT-SPME column. Itaconic acid-benzene co-polymer was in-situ grown on the inner surface of the fused silica capillary and used as extraction phase. This column has low backpressure and provides both hydrophobic and weak cationic exchange interaction with the target analytes due to the chemical properties. The developed online IT-SPME-MS method showed good extraction performance towards various target analytes and good reusability at least for 60 times. As a proof-of-concept application, the above method was applied for the analysis of antiepileptic drugs (AEDs) in both plasma and urine samples with linear range (1 ng/mL-200 ng/mL), good linearity (R2 ≥ 0.99), and good reproducibility (intra-day RSDs less than 4.36%, inter-day RSDs less than 6.55%). The method exhibited high enrichment factors between 187 and 204 for the two AEDs and high sensitivity for the analysis of human plasma samples and urine samples.

Simultaneous in-situ extraction and electrochemical detection of antidepressant drug imipramine and its active metabolite in human biofluid samples

The rapidly advancing modern healthcare system necessitates the development of cutting-edge technologies for therapeutic drug monitoring. Protein-bound drugs challenge the rapid quantification of analytical methods that require additional separation and cleanup processes, which delay the development of point-of-care testing platform. Here, a human serum albumin (HSA)/iron molybdate (FeM;Fe2(MoO4)3)/reduced graphene oxide (rGO) is reported for the in-situ extraction and determination of imipramine (IMP) drug and its active metabolite desipramine (DMP) in human serum and plasma samples. HSA was used for the extraction of drugs from the complex plasma proteins, and the extracted drugs were directly detected by FeM/rGO without additional sample cleanup. The developed sensor, which was optimized for the drug therapeutic window under normal conditions, exhibited a linear range of 10–756 ng/mL as well as a remarkable limit of detection and sensitivity of approximately 4 ± 2 ng/mL and 0.0124 ± 0.0003 µA cm−2 ng−1 mL, respectively, for IMP-DMP. The spike-and-recovery method was used to validate the developed sensor in real sample analysis. Based on the results, the sensing mechanism was elucidated: protein-protein interaction facilitates drug transportation from the plasma protein to electrode surface. Thereby, the HSA-FeM/rGO modified electrode recovered more drugs in biofluids compared with HSA unmodified electrode.

Spinel structured MgAl2O4 nanoparticles as a low-cost and stable SERS substrate for rapid simultaneous detection of neurological drugs in biofluids

There is an urgent need for exploring reliable and low-cost techniques for simultaneous detection of neurological drugs to facilitate timely diagnosis and treatment of various neurological disorders. In this work, we demonstrate a low cost, one-step synthesis of spinel magnesium aluminate (MgAl2O4) nanoparticles as a chemically stable substrate for rapid, simultaneous detection of neurological drugs like nicotine, gabapentin, caffeine and carbamazepine in the biofluids via surface-enhanced Raman scattering (SERS) technique. The substrate exhibits a limit of detection (LOD) of 0.18 ng/mL, 0.78 μg/mL, 0.1 μg/mL and 0.81 μg/mL with an enhancement factor (EF) of 1.61 × 107, 9.70 × 107, 7.72 × 107 and 1.52 × 107 towards detection of nicotine, gabapentin, caffeine and carbamazepine, respectively with an accumulation time of 20 s. The substrate exhibits an excellent selectivity and a wide dynamic range of detection towards the drug molecules in presence of 0.1 mM of ascorbic acid (AA), NaCl, KCl, CaCl2, urea, and glucose. This excellent performance can be ascribed to the interaction between the metallic sties of the MgAl2O4 cubic spinel structure and the drug molecules which leads to the resonance Raman scattering-like effect through the reconstruction of the energy bandgap closer to the photon energy of the excitation laser. Simultaneous recognition of drug molecules in the blood serum samples with reliable recovery percentages is also achieved using this platform thus, proving its efficacy for developing perovskite materials-based SERS detection useful in bioanalytical applications.

Deep Learning-Assisted Three-Dimensional Fluorescence Difference Spectroscopy for Identification and Semiquantification of Illicit Drugs in Biofluids.

The fast identification and quantification of illicit drugs in biofluids are of great significance in clinical detection. However, existing drug detection strategies cannot fully meet clinical needs, and the on-site identification and quantification of various illicit drugs in biofluids remain a great challenge. Here, we report the development of a deep learning-assisted three-dimensional (3D) fluorescence difference spectroscopy for rapid identification and semiquantification of illicit drugs in biofluids. This strategy introduces highly fluorescent silver nanoclusters into the biofluids with illicit drugs as signal sources. The interaction between silver nanoclusters and drug molecules changed the fluorescence performance of the mixture. Deep learning methods were applied to grasp the subtle fingerprint information from the 3D fluorescence difference spectra to identify and semiquantify various illicit drugs in biofluids, including codeine, 4,5-methylene-dioxy amphetamine, 3,4-methylene dioxy methamphetamine, meperidine, and methcathinone. This approach can achieve a high prediction accuracy rate of 88.07% and a broad detection range from 2 μg/mL to 100 mg/mL. It opens up a new way for the detection of small molecules with or without fluorescence in complicated matrixes.

Picoanalysis of Drugs in Biofluids with Quantitative Label-Free Surface-Enhanced Raman Spectroscopy.

The enormous increase of Raman signal in the vicinity of metal nanoparticles allows surface-enhanced Raman spectroscopy (SERS) to be employed for label-free detection of substances at extremely low concentrations. However, the ultimate potential of label-free SERS to identify pharmaceutical compounds at low concentrations, especially in relation to biofluid sensing, is far from being fully realized. Opioids are a particular challenge for rapid clinical identification because their molecular structural similarities prevent their differentiation with immunolabeling approaches. In this paper, a new method called quantitative label-free SERS (QLF-SERS) which involves the formation of halide-conjugated gold nanoclusters trapping the analyte of interest near the SERS hot spots is reported, and it is demonstrated that it yields a 105 fold improvement in the detection limit over previously reported results for the entire class of clinically relevant opioids and their metabolites. Measurements of opioid concentrations in multicomponent mixtures are also demonstrated. QLF-SERS has comparable detection limits as currently existing laboratory urine drug testing techniques but is significantly faster and inexpensive and, therefore, can be easily adapted as part of a rapid clinical laboratory routine.

High-Throughput Screening and Quantitation of Target Compounds in Biofluids by Coated Blade Spray-Mass Spectrometry.

Most contemporary methods of screening and quantitating controlled substances and therapeutic drugs in biofluids typically require laborious, time-consuming, and expensive analytical workflows. In recent years, our group has worked toward developing microextraction (μe)-mass spectrometry (MS) technologies that merge all of the tedious steps of the classical methods into a simple, efficient, and low-cost methodology. Unquestionably, the automation of these technologies allows for faster sample throughput, greater reproducibility, and radically reduced analysis times. Coated blade spray (CBS) is a μe technology engineered for extracting/enriching analytes of interest in complex matrices, and it can be directly coupled with MS instruments to achieve efficient screening and quantitative analysis. In this study, we introduced CBS as a technology that can be arranged to perform either rapid diagnostics (single vial) or the high-throughput (96-well plate) analysis of biofluids. Furthermore, we demonstrate that performing 96-CBS extractions at the same time allows the total analysis time to be reduced to less than 55 s per sample. Aiming to validate the versatility of CBS, substances comprising a broad range of molecular weights, moieties, protein binding, and polarities were selected. Thus, the high-throughput (HT)-CBS technology was used for the concomitant quantitation of 18 compounds (mixture of anabolics, β-2 agonists, diuretics, stimulants, narcotics, and β-blockers) spiked in human urine and plasma samples. Excellent precision (∼2.5%), accuracy (≥90%), and linearity (R2 ≥ 0.99) were attained for all the studied compounds, and the limits of quantitation (LOQs) were within the range of 0.1 to 10 ng·mL-1 for plasma and 0.25 to 10 ng·mL-1 for urine. The results reported in this paper confirm CBS's great potential for achieving subsixty-second analyses of target compounds in a broad range of fields such as those related to clinical diagnosis, food, the environment, and forensics.

The new drug phenomenon

The new drug phenomenon

Restricted-access media development for direct analysis of drugs in biofluids using capillary liquid chromatography

In analytical sciences the design of novel materials and stationary phases for the sample preparation and separation of analytes from biological fluids is needed. In this work we present different strategies for modification of stationary phases to produce tailored solutions for the analytical problem. In this context a novel shielded polymeric reversed-phase monolithic material was prepared in the presence of different numbers of reactive groups and concentrations of the coating polymer. Chromatographic experiments were performed using benzoic acid propyl ester in order to characterize the hydrophobicity and efficiency of the different restricted-access continuous beds prepared. Inverse size-exclusion chromatography was used for investigation of the pore structure properties of the beds. Capillary columns were applied for nanochromatography of biological fluids containing a mixture of nitrazepamum and medazepamum.

THERMAL BEHAVIOR OF ANTI-HYPERTENSIVES DRUGS IN SOLUTION AND HUMAN PLASMA USING DSC-COOLING

The present study proposes the use of DSC-cooling as a fast tool to evaluate the drug behavior in biofluids submitted to freeze-thaw studies. Captopril, captopril+ascorbic acid and propranolol in plasma were submitted to 3 cycles of freeze-thaw using DSC-cooling up to 243 K. Captopril-plasma has suffered calorimetric alteration and revealed a considerable interaction front to freezing and heating processes. The presence of additive can stabilize the analyte in plasma. Propranolol-plasma presented reducing in phase transition enthalpy comparable to the plasma denatured by perchloric acid in the last cycles of freezing. DSC-cooling data demonstrated great potentiality for the freeze-thaw studies of drugs in biofluids.

Solid-phase micro-extraction in bioanalysis, exemplified by lidocaine determination

Solid-phase micro-extraction (SPME) is a never sample preparation technique that can be used for gaseous, liquid or solid samples in conjunction with GC, HPLC or CE (e.g. [1]). The use of SPME for the analysis of drugs in biofluids is also becoming popular (e.g. [2]). The principle is that a fused silica fibre coated with a polymer such as polydimethylsiloxane (PDMS) is put directly into the sample or placed in the headspace above it (e.g. [3]), analyte desorption is done thermally (for GC) or with liquid. As a model compound to study conditions we have investigated lidocaine in human urine using GC or LC [4] and, with attention to protein binding, in plasma mainly using GC [5].

Determination of phase II drug metabolites in equine urine by micellar electrokinetic capillary chromatography

Micellar electrokinetic capillary chromatography (MECC) using diode array detection has been investigated for the determination of phase I and phase II metabolites of drugs in biofluids. Methods were optimised for the determination of morphine, morphine-3-glucuronide, morphine-6-glucuronide, normorphine, meclofenamic acid and its metabolites in equine urine. Solid-phase extraction procedures were developed to concentrate and purify the analytes from spiked and post administration urines for MECC analysis. A simple on-line procedure for monitoring the kinetics of hydrolysis of morphine-glucuronide conjugates by β-glucuronidase was demonstrated.

Rapid determination of drugs in biofluids by capillary electrophoresis Measurement of antipyrine in saliva for pharmacokinetic studies

A micellar electrokinetic capillary chromatography method was developed that permitted the resolution of antipyrine from endogenous compounds and its quantitation in neat saliva in as little as 1 min. Final conditions were: SpectraPhoresis 1000, 30(23) cm × 50 μm silica capillary, 50 m M sodium phosphate pH 9.6, 50 m M SDS, 10 s hydrodynamic load, detection scanning 200–300 nm or 260 nm, run 25 kV. To overcome the effects of Joule heating the capillary was cooled to 15°C. Sensitivity was <10 μM and linearity extended to 350 μM. Comparison with an HPLC assay demonstrated that hydrodynamic injection gave a loading bias unless samples and standards were of equal viscosity. For 75 samples from five subjects the correlation of CE vs. HPLC was then r = 0.99.

Polymeric reagents for derivatizations in micellar electrokinetic chromatography.

A polymer immobilized o-nitrobenzophenone reagent was prepared for analysis of amine drugs in micellar electrokinetic chromatography (MEKC). A model compound, propylamine, was used to characterize the reagent's performance in MEKC. Derivatizations were performed on the CE instrument with reagent in the sample vial. The yielded derivative was directly sampled from the reaction mixture, and directly injected onto the MEKC system. The derivatization reagent was also applied to the derivatization of n-alkyl amine mixtures and amino acids. The method was validated for adamantanamine in urine and in plasma by single-blind spike analysis. Precisions and accuracies for all samples were less than 6.0% for urine samples and 10% for plasma samples. The procedure was a direct injection technique requiring minimal sample preparation for the analysis of drugs in biofluids.