Complicated Sample Pretreatment Research Articles

Analysis of rebaudioside A in probiotic-fermented milk via quantitative nuclear magnetic resonance spectroscopy

Rebaudioside A is used in the food and beverage industries owing to its sweetness, even in extremely small amounts, as well as its natural origin. Liquid chromatography is commonly used to quantify rebaudioside A; however, it suffers from complicated sample pretreatment and time consumption. External-calibration quantitative nuclear magnetic resonance (qNMR) analysis of probiotic-fermented milk has thus far not been reported. Moreover, it involves more factors contributing to errors than internal standard approaches used in the official methods, such as those recommended by the International Organization for Standardization. In this study, we developed a method for quantifying rebaudioside A in probiotic-fermented milk using internal-calibration qNMR. This method entails a simple pretreatment step with dilution and solid-phase extraction, enabling rebaudioside A determination in five types of beverages including probiotic-fermented milk. The results revealed a recovery rate and relative standard deviation of 85.4–104 % and 1.57–4.09 %, respectively, satisfying the international guidelines. The working range of qNMR was 0.80–26.7 mg/100 g of product, and the analysis time was 10–20 min. Therefore, the proposed internal-calibration qNMR method facilitates easy, rapid, and accurate quantification of rebaudioside A in probiotic-fermented milk and is applicable for analyzing various other beverages and foods.

Determination of bioequivalence between generic and reference drugs using laser-induced breakdown spectroscopy

BackgroundIn vitro bioequivalence studies are strictly limited to the comparison of dissolution performance to a reference drug. These studies are performed without considering the chemical similarity between the generic and reference drug formulations. This work has focused on developing a groundbreaking method based on the laser-induced breakdown spectroscopy (LIBS) technique for the in vitro bioequivalence determination of immediate-release solid oral dosage form generic drugs and as an alternative method for establishing the biowaiver of in vivo generic drug studies. ResultsThe novel LIBS-based methodology to determine in vitro bioequivalence is fast, easy to perform, and can be carried out without the requirement of tedious and complicated sample pre-treatment, nor expensive instrumentals and reagents, almost directly on the drug samples. Furthermore, the proposed methodology demonstrated that it is enough to identify the spectrochemical similarity of the formulation between generic drugs to a reference drug through the chemometric study of their LIBS spectra, based on the determination of the differentiation and similarity factors, f1 and f2, respectively, used in the pharmaceutical industry in this purpose. After analysing their LIBS spectra, the generic drugs selected for this work have all been shown to be in vitro bioequivalent, given their f1 values of less than 15 and f2 values greater than 50, according to the technical regulations on which the American and European medicines agencies are based for the approval of registration for generic immediate-release solid oral dosage form drugs. This has been evidenced even for drugs from Class III and Class IV of the biopharmaceutical classification system, whose active principle nominal concentration is very low as 0.1 and 0.25 mg/tablet, respectively. Significancefor the first time the LIBS technique has been successfully used in an advanced application for the pharmaceutical industry. The proposed method constitutes a reliable and specialized methodology for the establishment of formulation similarity between two drugs, without the requirement of separate identification of each of their components, which is a new and potential tool to determine the in vitro bioequivalence for generic immediate-release solid oral dosage form drugs.

Nanoengineered Nonenzymatic Paper-Based Fluorescent Platform for Pre-Dilution-Free and Visual Real-Time Home Monitoring of Urea for Early Warning of Abnormal Nitrogen-Based Unhealthy Issues.

Distorted urea levels indicate several liver, kidney, or metabolic diseases; however, traditional clinical urea detection relies on urease-based methods enslaved to well-known limitations of high-price, unstable properties, complicated sample pretreatment and analysis procedures, and difficult visual real-time monitoring. Herein, nonenzymatic paper-based fluorescent materials (UFP-BP) are strategically integrated with an on-demand fluorescent-sensor (UFP) self-aggregated nanoparticle on commercial filter paper for pre-dilution-free and visual real-time urea monitoring. The UFP is synthesized and self-aggregated into the fluorescent nanoparticles for selective urea recognition. Then, the nanoparticles are interstitially loaded on filter paper to nanoengineer the UFP-BP, achieving selective quantitative urea detection in the normal concentration range (10-1000mm). UFP and UFP-BP can successfully monitor urea levels in real rat urine, artificial simulants, and milk. The proposed sensing platform, integrated with smartphones, offers accurate, quantitative, nonenzymatic, noninvasive, pre-dilution-free, on-site, rapid, low-cost, easy-to-operate, real-time visual urea detection in food samples and human body fluids. The designed sensing system can provide early warnings of abnormal nitrogen-based health issues.

Imprinted membrane-covalent organic framework platform for efficient label-free visual detection of Listeria monocytogenes and Salmonella typhimurium in food samples

BackgroundRapid and sensitive detection of foodborne pathogens in food plays a crucial role in controlling outbreaks of foodborne diseases, of which Listeria monocytogenes and Salmonella typhimurium are representative and notable pathogens. Thus, it's of great importance to achieve the effective detection of these pathogens. However, the most common detection methods (culture-based technique, Polymerase Chain Reaction and immunological methods) have disadvantages that cannot be ignored, such as time-consuming, laborious, complex sample preparation process, and the possibility of cross-reaction. Hence, it is essential to develop a facile detection method for the pathogens with high sensitivity and specificity to avoid the above-mentioned disadvantages. ResultsWe report a label-free visual platform for the simultaneous capture and detection of Listeria monocytogenes and Salmonella typhimurium. For the first time, we have prepared polydimethylsiloxane-Chromotrope 2R membrane which serves as the substrate for bacterial capture and enrichment through the formation of specific recognition sites. The positively charged Pt-covalent organic framework combines with the pathogens through surface charge interaction, thereby the label-free sandwich platform is formed. Remarkable peroxidase activity of Pt-covalent organic framework converts the conversion of bacterial quantity into amplified color signal by catalyzing 3,3′,5,5′-Tetramethylbenzidine to oxidized 3,3′,5,5′-Tetramethylbenzidine. The platform demonstrates the capability to identify two representative food-borne pathogens within a time frame of 100 min, exhibiting high sensitivity and excellent specificity without the interference from non-target bacteria. The limit of detection of the visual platform toward Listeria monocytogenes and Salmonella typhimurium was 1.61 CFU mL−1 and 1.31 CFU mL−1, respectively. And the limit of quantification toward Listeria monocytogenes and Salmonella typhimurium was 4.94 CFU mL−1 and 2.47 CFU mL−1, respectively. The relative standard derivations of the visual platform for both bacteria were lower than 4.9 %. Furthermore, our proposed platform has obtained reliable and satisfactory results on analyzing diverse food samples. SignificanceThis research expands the application of a label-free platform combined with unlabeled nanocomponents in the rapid isolation and detection of diverse of food-borne pathogens. The platform possesses the advantages of simple operation and real-time monitoring, without complicated sample pretreatment process. The whole detection process can realize the simultaneous monitoring of Listeria monocytogenes and Salmonella typhimurium within 100 min. Furthermore, it is also of reference significance for the detection of other common pathogens.

CeO2 nanocages with tetra-enzyme mimetic activities for dual-channel ratiometric colorimetric detection of microcystins-LR

BackgroundMicrocystin-leucine-arginine (MC-LR) produced by various cyanobacteria during harmful algal bloom poses serious threats to drinking water safety and human health. Conventional chromatography-based detection methods require expensive instruments and complicated sample pretreatment, limiting their application for on-site detection. Colorimetric aptasensors are simple and rapid, and are amenable to fast detection. However, they provide only one output signal, resulting in poor sensitivity and accuracy. Dual-channel ratiometric colorimetric method based on the peroxidase-like activity of nanozyme can achieve self-calibration by recording two reverse signals, providing significantly enhanced sensitivity and accuracy. ResultsCeO2 nanocages (CeO2 NCs) with tetra-enzyme mimetic activities (oxidase-, peroxidase-, catalase- and superoxide dismutase-like activities) were facilely synthesized using zeolitic imidazolate framework-67 (ZIF-67) as sacrificial template. The peroxidase-like activity of CeO2 NCs can be regulated by DNA, and it showed opposite response to two chromogenic substrates (2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 3,3′,5,5′-tetramethylbenzidine (TMB)), which was mainly attributed to the changed affinity. On the basis of MC-LR aptamer-tunable peroxidase-like activity of CeO2 NCs in TMB and ABTS channel, a dual-channel ratiometric colorimetric aptasensor was constructed for detection of MC-LR. Compared with conventional single-signal colorimetric assays, the proposed method showed lower limit of detection (0.66 pg mL−1) and significantly enhanced sensitivity. Moreover, the practicability of the ratiometric colorimetric assay was demonstrated by detecting MC-LR in real water samples, and satisfactory recoveries (94.9–101.9 %) and low relative standard deviations (1.6–6.3 %) were obtained. SignificanceThis work presents a nanozyme-based ratiometric colorimetric aptasensor for MC-LR detection by recording the reverse responses of two chromogenic reactions. Benefiting from the self-calibration function, the method can achieve higher sensitivity and accuracy. The short detection time and practical application in real water samples show great potential for environmental monitoring.

FRET-Based Host-Guest Supramolecular Probe for On-Site and Broad-Spectrum Detection of Pyrethroids in the Environment.

The massive use of pyrethroid pesticides in agriculture has brought growing concerns about food safety due to their several harmful effects on human health, especially through the accumulation of the food chain. To date, most of the available analytical methods for pyrethroids still suffer from insufficient detection universality, complicated sample pretreatment, and detection processes, which severely limit their practical applications. Herein, a novel Förster resonance energy transfer (FRET)-assisted host-guest supramolecular nanoassembly is reported, for the first time, successfully realizing ratiometric fluorescent detection of pyrethroids in real samples through the indicator displacement assay (IDA) mechanism. This method is capable of detecting a broad spectrum of pyrethroids, including bifenthrin, cyfluthrin, cypermethrin, deltamethrin, etofenprox, fenvalerate, and permethrin, with ultrahigh detection sensitivity, great selectivity, high anti-interference ability, and, in particular, distinct emission color response from red to green. Such a large chromatic response makes this method available for fast and on-site detection of pyrethroids in real samples with the aid of several simple portable analytical apparatuses.

Real-time optical detection of mercury contamination in drinking water using an amphiphilic recognition probe at liquid crystal/aqueous interfaces.

Mercury contamination is a global environmental issue due to its toxicity and persistence in ecosystems. It poses a particular risk in aquatic systems, where it bioaccumulates and biomagnifies, leading to serious health impacts on humans. Therefore, effective detection technologies for mercuric ions in natural water resources are highly desirable. However, most existing detection methods are time-consuming, require complicated sample pre-treatment, and rely on expensive equipment, which hinders their widespread use in real-time detection. Here, we present a convenient, rapid, portable, user-friendly, and cost-effective sensing system for detecting Hg2+ ion contamination in water. This system utilizes a highly selective, amphiphilic, and structurally simple molecular probe, N-dodecylamine-di-thiocarbamate (DDC). DDC molecules align at the interface between the liquid crystal (LC) and water, inducing a homeotropic LC orientation. In water samples contaminated with Hg2+, a bright optical texture is observed, indicating the alignment of the 5CB LC in a planar manner at the LC/aqueous boundary. The minimum detectable concentration (LOD) for Hg2+ ions is 5.0 μM in distilled water, with a broad detection range from 5.0 μM to 2 mM. The sensor selectively detects Hg2+ ions over other common interfering metal ions, including Pb2+, Co2+, Ni2+, Cu2+, Cd2+, Zn2+, Cr2+, Mg2+, Na+, K+, and Ca2+. Boolean logic gates, bar graphs, and truth tables are employed to explain the selectivity of this liquid crystal-based sensor. This work demonstrates the significant potential of the sensor for monitoring mercuric ions in natural water resources, offering a promising strategy for controlling mercury pollution.

4-Aminophenylboronic acid dimer sensitized carbon nanotubes for the construction of portable and disposable sulfide electrochemical sensors

Abnormal sulfide levels in serum can cause kidney failure, Alzheimer's, Parkinson's, stroke, diabetes, and even cancer. Therefore, maintaining proper sulfide balance is crucial for managing and preventing these conditions. Electrochemical approaches have emerged as potential alternatives for sulfide detection. However, the adsorption of elemental sulfur on the working electrode leads to surface fouling and passivation, posing a significant challenge in developing reliable electrochemical methods for sulfide detection. In this study, we present a portable and surface fouling-free electrochemical system that utilizes functionalized multi-walled carbon nanotubes and 4-aminophenylboronic acid dimers for accurate sulfide measurement. The sensor showed excellent electrocatalytic activity to sulfide and antifouling activity to possible interferences. The method had a linear range from 10 µM to 2 mM and a detection limit of 2.3 µM. The effectiveness and applicability of this method were evaluated by analyzing sulfide levels in different serum samples and comparing the results with their glucose levels. As both blood glucose and sulfide levels can serve as indicators of certain health conditions, such as diabetes or sulfur metabolism disorders, this comparison provides valuable insights. Importantly, no complicated sample pretreatment was involved, thus this method can be easily adopted as a point-of-care diagnostic device. Additionally, the study also revealed a higher concentration of sulfide in cancer cells than in normal cells, offering a new development direction for differentiating between cancer cells and normal cells.

Ag-MXene as peroxidase-mimicking nanozyme for enhanced bacteriocide and cholesterol sensing

Reactive oxygen species (ROS) are ideal alternative antibacterial reagents for rapid and effective sterilization. Although a variety of ROS-based antimicrobial strategies have been developed, many are still limited by their inefficiency. Herein, we report the synthesis of the Ag-MXene nanozyme, which have superior peroxidase-like activity for antibacterial applications. As a result, Ag-MXene nanozyme can efficiently increase the level of intracellular ROS, converting H2O2 into hydroxyl radicals that effectively kill both Gram-negative and Gram-positive bacteria and disrupting the bacterial biofilm formation. Moreover, a sensitive and selective colorimetric biosensor was constructed for assaying cholesterol based on the Ag-MXene’s prominent peroxidase-mimicking activity and the cholesterol oxidase cascade reaction. The biosensor exhibits high performance with a linear cholesterol detection range of 2–800 μM, and a detection limit of 0.6 μM. Ag-MXene nanozyme can be used for the rapid detection of cholesterol in serum without complicated sample pretreatment. Collectively, it is conceivable that the proposed Ag-MXene nanozyme could be used as a biocide and as a cholesterol sensor. This study provides a broad prospect for the rapid detection and sterilization of MXene nanozymes in the biomedical field.

Diclofenac-Impregnated Mesoporous Carbon-Based Electrode Material for the Analysis of the Arsenic Drug Roxarsone.

This paper describes a novel electrode material, diclofenac-impregnated mesoporous carbon modified with a cationic surfactant, cetyltrimethylammonium bromide (DF-CMK-3/CTAB), for ultratrace analysis of the arsenic drug roxarsone (ROX). DF-CMK-3 amorphous carbon is a material with a high specific surface area and well-defined, hexagonally ordered, thin mesopores. The functional groups attached to the carbonaceous surface, such as chromene and pyron-like oxygen groups, lactam, and aromatic carbon rings, have the basic character and they can donate electrons. Modification of DF-CMK-3 with a CTAB layer significantly increases the analytical signal due to electrostatic interactions between the cationic surfactant and the anion form of ROX in the acidic medium. The voltammetric procedure at the glassy carbon sensor modified with DF-CMK-3/CTAB exhibited excellent sensitivity (limit of detection of 9.6 × 10-11 M) with a wide range of linearity from 5.0 × 10-10 to 1.0 × 10-4 M. Analysis of real samples (treated municipal wastewater and river water) showed recoveries from 96 to 102% without applying the complicated sample pretreatment step. The sensor demonstrated excellent sensitivity in the analysis of the arsenic drug ROX in the presence of interferences in environmental water samples.

Polydopamine- and MOFs-based molecularly imprinted microfiltration membranes in syringe filter for selective capture of thiamphenicol, florfenicol and chloramphenicol in milk samples

In this study, molecularly imprinted microfiltration membranes (MIMFMs) based on polydopamine and metal–organic framework were innovatively proposed, and successfully prepared for capturing thiamphenicol, florfenicol and chloramphenicol from milk samples through syringe filter device. The provision of PDA and coupling of MOF particles enabled the filter membranes to have great permeability and binding capacity. Cleverly, hydrophilicity and heightened rebinding selectivity (IF = 2.42) of filter membranes was provided using two functional monomers (methacrylate and acrylamide), and microfiltration-grade composite membranes were generated by adjusting the imprinting polymerization time. The combination of the selective filter membranes and the syringe filter devices can effectively simplify the operation process, minimize the extraction time and reduce the tested sample volume. The method was validated by coupling with ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) and obtained a good correlation (r2 > 0.99). The limit of detection (LOD) of three analytes was 0.0800–0.6583 µg kg−1 and the mean recovery ranged from 85.5% to 91.2% within relative standard deviations (RSDs) less than 8.2%. Finally, an effective MIMFMs-UHPLC-MS/MS method was established for the recognition and detection of three amphenicol antibiotics in milk samples. The MIMFMs filtration technique offered in this work provides valuable science references and potential application prospects for complicated sample pretreatment.

Coconut shell biochar application in liquid-solid microextraction of triazine herbicides from multi-media environmental samples

A rapid, fast, widely applicable liquid-solid microextraction and purification method of triazine herbicides (TRZHs) in muti-media samples using salting-out assisted liquid-liquid extraction (SALLE) combined with self-assembled monolithic spin columns-solid phase micro extraction (MSC-SPME) was developed. Environmentally friendly coconut shell biochar (CSB) was used as the adsorbents of MSC-SPME. Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was the separation and determination method. The adsorption kinetics and isotherms were investigated to indicate the interaction between CSB and TRZHs. Several parameters influencing the liquid-solid microextraction efficiency, such as sample pH, salting-out solution volume and pH, sample loading speed, elution speed, elution ratio and volume of eluent were systematically investigated with the aid of orthogonal design. The whole extraction process was operated within 10 min. Under the optimum extraction and determination conditions, good linearities for three TRZHs were obtained in a range of 0.10–200.00 ng mL−1, with linear coefficients (R2) greater than 0.999. The limits of detection (LODs) and limits of quantification (LOQs) were in the range of 6.99–11.00 ng L−1 and 23.33–36.68 ng L−1, respectively. The recoveries of the three TRZHs in multi-media environmental samples were ranged from 69.00% to 124.72%, with relative standard deviations (RSDs) lower than 0.43%. This SALLE-MSC-SPME-UPLC-MS/MS method was successfully applied to the determination of TRZHs in environmental and food samples and exhibited the advantages of high efficiency and sensitivity, low cost, and environmental friendliness. Compared with the methods published before, CSB-MSC was green, rapid, easy-operated, and reduced the whole cost of the experiment; SALLE combined MSC-SPME eliminated the matrix references effectively; what's more, the SALLE-MSC-SPME-UPLC-MS/MS method could be applied to various sample without complicated sample pretreatment procedure.

All-solid-state potentiometric salicylic acid sensor for in-situ measurement of plant.

Using PEDOT as the conductive polymer, an innovative small-scale sensor for directly measuring salicylate ions in plants was developed, which avoided the complicated sample pretreatment of traditional analytical methods and realized the rapid detection of salicylic acid. The results demonstrate that this all-solid-state potentiometric salicylic acid sensor is easy to miniaturize, has a longer lifetime (≥1month), is more robust, and can be directly used for the detection of salicylate ions in real samples without any additional pretreatment. The developed sensor has a good Nernst slope (63.6 ± 0.7mV/decade), the linear range is 10-2 ~ 10-6 M, and the detection limit can reach (2.8 × 10-7M). The selectivity, reproducibility, and stability of the sensor were evaluated. The sensor can perform stable, sensitive, and accurate in situ measurement of salicylic acid in plants, and it is an excellent tool for determining salicylic acid ions in plants in vivo.

Raman spectroscopy: Principles and recent applications in food safety.

Food contaminant is a significant issue because of the adverse effects on human health and economy. Traditional detection methods such as liquid chromatography-mass spectroscopy for detecting food contaminants are expensive and time-consuming, and require highly-trained personnel and complicated sample pretreatment. Raman spectroscopy is an advanced analytical technique in a manner of non-destructive, rapid, cost-effective, and ultrasensitive sensing various hazards in agri-foods. In this chapter, we summarized the principle of Raman spectroscopy and surface enhanced Raman spectroscopy, the methods to process Raman spectra, the recent applications of Raman/SERS (surface-enhanced Raman spectroscopy) in detecting chemical contaminants (e.g., pesticides, antibiotics, mycotoxins, heavy metals, and food adulterants) and microbiological hazards (e.g., Salmonella, Campylobacter, Shiga toxigenic E. coli, Listeria, and Staphylococcus aureus) in foods.

Suppressed DNA base pair stacking assembly of gold nanoparticles in an alcoholic solvent for enhanced ochratoxin A detection in Baijiu.

The currently established DNA nanoprobes for the detection of mycotoxin from beverages have been limited by complicated sample pretreatment and uncontrollable nanoparticle flocculation in complex systems. We develop a rapid colorimetric approach for ochratoxin A (OTA) detection in Baijiu in a sample-in/"yes" or "no" answer-out fashion through target-modulated base pair stacking assembly of DNA-functionalized gold nanoparticles (DNA-AuNPs). The colorimetric signification of OTA relies on the competition of OTA with the AuNP surface-grafted DNA in binding with an OTA-targeted aptamer. The specific recognition of OTA by the aptamer prevents DNA duplex formation on the AuNP surface, thereby inhibiting the base pair stacking assembly of the DNA-AuNPs and giving rise to a "turn-on" color. By further suppressing DNA hybridization using a bulged loop design and an alcohol solution, the DNA-AuNPs exhibit an improved reproducibility for OTA sensing while maintaining excellent susceptivity to OTA. A detection limit of 88 nM was achieved along with high specificity towards OTA, which is lower than the maximum tolerated level of OTA in foodstuffs defined by countries worldwide. The entire reaction time, avoiding sample pretreatment, is less than 17 min. The DNA-AuNPs with anti-interference features and sensitive "turn-on" performance promise convenient on-site detection of mycotoxin from daily beverages.

A simple fluorescent sensor for highly sensitive detection of UO22+

Extensive application of nuclear energy has caused widespread environmental uranium contamination. New detection approaches without complicated sample pretreatment and precision instruments are in demand for on-site and in-time determination of uranyl ions in environmental monitoring, especially in an emergency situation. In this work, a simple and effective fluorescent sensor (Z)-N'-hydroxy-4-(1,2,2-triphenylvinyl)benzimidamide (TPE-A) with aggregation-induced emission (AIE) character was established and studied. It could realize to detect UO22+via quenching the fluorescence of its aggregation-induced emission, with good selectivity and sensitivity. Such strategy shows a wide linear range from 5.0 × 10−8 mol/L to 4.5 × 10−7 mol/L (R2 = 0.9988) with exceptional sensitivity reaching 4.7 × 10−9 mol/L, which is far below the limit for uranium in drinking water (30 µg/L, ca. 1.1 × 10−7 mol/L) stipulated by the WHO. A response time less than four minutes make it rapid for uranyl ion measurement. It was applied for detection of uranyl ion in spiked river water samples with recoveries in the range of 98.7%-104.0%, comparable to those obtained by ICP-MS. With the advantages of portable apparatus, rapid detection process and high sensitivity, TPE-A can serve as a promising fluorescent sensor for the detection of UO22+ in environmental water samples.

Electrochemical Sensor Based on Au NPs@NiPc-Cu MOFs Modified Electrode for the Rapid Detection of Luteolin

In this study, a novel electrochemical sensor was designed to detect luteolin (Lu) with the composite of gold nanoparticles and nickel phthalocyanine-based 2D conductive metal-organic frameworks (Au NPs@NiPc-Cu MOFs) for the first time. The NiPc-Cu MOFs exhibit excellent conductivity, large specific surface area, and porous structure, which can accelerate the mass transfer process of target molecules. To further improve the sensitivity of the sensing platform, Au NPs with outstanding conductivity were introduced to the surface of NiPc-Cu MOFs to prepare Au NPs@NiPc-Cu MOFs. The synergistic effect of NiPc-Cu MOFs and Au NPs endows the sensor with excellent electrocatalytic performance and outstanding sensitivity. Under optimal conditions, the electrochemical sensor has a wide linear range (0.1–40 μM). Moreover, the prepared sensor possesses good stability and anti-interference ability. This method does not require complicated sample pretreatment, simple operation, and short detection time, which can provide a new method for the rapid detection of Lu.

Wooden-Tip Electrospray Mass Spectrometry Characterization of Human Hemoglobin in Whole Blood Sample for Thalassemia Screening: A Pilot Study

Traditional analytical methods for thalassemia screening are needed to process complicated and time-consuming sample pretreatment. In recent decades, ambient mass spectrometry (MS) approaches have been proven to be an effective analytical strategy for direct sample analysis. In this work, we applied ambient MS with wooden-tip electrospray ionization (WT-ESI) for the direct analysis of raw human blood samples that were pre-identified by gene detection. A total of 319 whole blood samples were investigated in this work, including 100 α-thalassemia carriers, 67 β-thalassemia carriers, and 152 control healthy samples. Only one microliter of raw blood sample was directly loaded onto the surface of the wooden tip, and then five microliters of organic solvent and a high voltage of +3.0 kV were applied onto the wooden tip to generate spray ionization. Multiply charged ions of human hemoglobin (Hb) were directly observed by WT-ESI-MS from raw blood samples. The signal ratios of Hb chains were used to characterize two main types of thalassemia (α and β types) and healthy control blood samples. Our results suggested that the ratios of charged ions to Hb chains being at +13 would be an indicator for β-thalassemia screening.

Determination of glutamate using paper-based microfluidic devices with colorimetric detection for food samples

A paper-based device (PAD) capable of colorimetric detection was developed to determine the presence of glutamate in various food samples. The PAD employs an enzymatic reaction with glutamate followed by an oxidation reaction with N-benzoyl leucomethylene blue (BLMB) in the presence of horseradish peroxidase. The designed PAD consists of a sample introduction zone connected to a channel that transports a sample solution to three detection zones. The detection zones contain pre-deposited reagents: glutamate oxidase, horseradish peroxidase, BLMB, a phosphate buffer, and poly(acrylic acid). The PAD is perpendicularly immersed into a sample solution and bent at a right angle using a 3D-printed holder to allow the sample to simultaneously flow into three different detection zones. When the PAD is immersed into a sample containing glutamate, glutamate oxidase produces hydrogen peroxide, which changes the pale blue color of BLMB to a deep blue color in the presence of horseradish peroxidase. Under the optimum conditions, the calibration curve between the logarithm of the glutamate concentrations and the color intensity was linear within a range of from 5 × 10-6 mol L-1 to 10-2 and with a correlation coefficient of 0.994. Using this system, the PAD successfully determined glutamate in soup stocks, sauces, snacks, and tomato juice without the need of complicated sample pretreatment. These results agreed with those of a commercially available glutamate assay kit, which was employed as a certification method (tstat = 1.95, tcrit = 2.57). The developed PAD is simple, easy to fabricate, portable, and could be used outside of equipped laboratories to determine the presence of glutamate in food samples.

Reusable ring-like Fe3O4/Au nanozymes with enhanced peroxidase-like activities for colorimetric-SERS dual-mode sensing of biomolecules in human blood

Peroxidase−like nanozymes have led to important progress in biosensing, but most of nanozyme sensing systems are currently established by a single−signal output mode, which is susceptible to environmental and operational factors. Thus construction of a dual−signal output nanozyme sensing system is essential for obtaining reliable and robust performance. In this study, a novel peroxidase mimicking nanozyme was developed by decorating magnetic ring-like Fe3O4 with gold nanoparticles (R–Fe3O4/Au) for the colorimetric and surface-enhanced Raman scattering (SERS) dual-mode detection of biomolecules in human serum. The R–Fe3O4/Au nanozymes served as mimetic peroxidase which can catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine by hydrogen peroxide, and concomitantly as SERS substrates for detecting the Raman signals of oxidized products, providing an effective approach to investigate the reaction kinetics of enzymes. Based on the redox reactions, the nanozymes achieved colorimetric−SERS dual−mode sensing of glutathione (GSH) and cholesterol with detection limits as low as 0.10 μM and 0.08 μM, respectively. Furthermore, the nanozymes enabled rapid detection of GSH and cholesterol in serum without any complicated sample pretreatment. The R–Fe3O4/Au catalyst still displayed excellent peroxidase activity even after repeated use for 5 times. The proposed colorimetric-SERS dual-mode sensors exhibited good accuracy and reproducibility, which provides a new avenue for exploiting multifunctional sensors and has a great application prospect in biosensing.