Dual-signal output fluorescent aptasensor based on DNA programmability and gold nanoflowers for multiple mycotoxins detection.

Herein, a dual-signal output fluorescent aptamer sensor was constructed for the simultaneous detection of aflatoxin B1 (AFB1) and ochratoxin A (OTA) using the specific recognition ability of aptamers and the programmability of DNA. A functional capture probe (cDNA) was designed with the black hole quenching motif BHQ1 labeled at the 5' end and biotin (bio) labeled at the 3' end. The fluorescent dye Cy3-labeled aflatoxin B1 aptamer (AFB1-Apt) and the carboxyfluorescein FAM-labeled ochratoxin A aptamer (OTA-Apt) were used as two fluorescent probes. The cDNA is anchored to the quenching material gold nanoflowers (AuNFs) by the action of streptavidin (SA) and biotin. Its ends can be complementarily paired with two fluorescent probe bases to form a double-stranded structure. The fluorescence of Cy3 was quenched by AuNFs, and the fluorescence of FAM was quenched by BHQ1 through the fluorescence energy resonance transfer (FRET) effect, forming a fluorescence quenching system. Due to the high affinity of the target and the aptamer, the structure of the aptamer probe changes and detaches from the sensor when AFB1 and OTA are present, resulting in enhanced fluorescence. Under optimal conditions, the linear range of AFB1 was 0.1-100 ng/mL (R2 = 0.996), the limit of detection (LOD) was as low as 0.014 ng/mL, and the limit of quantification (LOQ) was 0.046 ng/mL. The linear range of OTA was 0.1-100 ng/mL (R2 = 0.995), the limit of detection (LOD) was as low as 0.027 ng/mL, and the limit of quantification (LOQ) was 0.089 ng/mL. The sensor had high accuracy in detecting both AFB1 and OTA in real sample analysis. The results of the t test show that there is no significant difference between the results of this study and the high-performance liquid phase (HPLC) method, indicating that the prepared sensor can be used as a potential platform for multiple mycotoxins detection.

A facile "turn-on" fluorescent aptasensor for simultaneous detection of dual mycotoxins in traditional Chinese medicine based on graphene oxide and FRET.

Mycotoxin is a common sort of harmful contaminant in traditional Chinese medicine (TCM), which is in a great demand of controlling. On this account, a facile "turn-on" fluorescent aptasensor based on fluorescence resonance energy transfer (FRET) for simultaneous detection of patulin (PAT) and zearalenone (ZEN) was developed. In this study, the aptamers of PAT and ZEN were labeled by FAM and Cy3, respectively, serving as fluorescence probes. Both aptamers could adsorb on the surface of graphene oxide (GO) via π-π stacking, which will consequently result in the occurrence of FRET between the fluorophores and GO. In the absence of the targets, the fluorescence would be quenched "off". In the presence of any of the dual mycotoxins, the corresponding aptamers would interact with the targets and release from GO due to the conformational variation, leading to a fluorescence "turn-on" effect. The limit of detection of this difunctional aptasensor was 2.29 nM for PAT and 0.037 nM for ZEN, respectively. This aptasensing platform exhibited satisfactory selectivity against interferents and reliability in real TCM sample detection. To our knowledge, it is the first aptasensor based on GO and FRET that realizes simultaneous detection of dual mycotoxin in TCM. Moreover, the measurement takes merely ∼60 min, does not need complicated pretreatment, and uses only inexpensive aptamer and GO as consuming materials. To sum up, this aptasensor exhibits great potential in fast, cost-effective and reliable simultaneous detection of multiple targets, and is expected to contribute to the quality and safety control of TCM.

Simultaneous electrochemical aptasensing of patulin and ochratoxin A in apple juice based on gold nanoparticles decorated black phosphorus nanomaterial.

Simultaneous detection of patulin (PAT) and ochratoxin A (OTA) in food products is in great demand, which can prevent toxins from being exposed to human and animal bodies. However, simultaneous detection of multiple targets still faces a challenge. Herein, we developed a novel electrochemical aptasensor for the simultaneous detection of PAT and OTA in apple juice based on gold nanoparticles decorated black phosphorus (AuNPs-BP) nanomaterial. AuNPs-BP function?/work? as a sensing platform for loading much different electrochemical signal molecules functionalized aptamers. In this context, methylene blue functionalized PAT aptamers (Mb-PAT-aptamers) and ferrocene functionalized OTA aptamers (Fc-OTA-aptamers) have been introduced here to fabricate the aptasensor. Fc close to electrode surface showed a strong signal, whereas Mb was far away from electrode surface so exhibited a weak signal in the absence of OTA and PAT. Two kinds of electrochemical signal changes have been recorded dependent on target of OTA and PAT concentrations. So, simultaneous detection of OTA and PAT is achieved. Under the optimum conditions, using this developed biosensor, PAT and OTA can be quantified at a linearity range of 0.01 × 10-7 µg·mL-1 ~ 0.10 µg·mL-1. In addition, it also has good selectivity, stability and repeatability. For the practical application, it shows promising performance for the simultaneous detection of PAT and OTA in apple juice.

A carbon paste electrode modified with Al2O3-supported palladium nanoparticles for simultaneous voltammetric determination of melatonin, dopamine, and acetaminophen.

The authors have modified a carbon paste electrode with Al2O3-supported palladium nanoparticles (PdNP@Al2O3) to obtain a sensor for simultaneous voltammetric determination of melatonin (MT), dopamine (DA) and acetaminophen (AC). The PdNP@Al2O3 was characterized by scanning electron microscopy and energy-dispersive X-ray spectra. The sensor can detect DA, AC, MT and their mixtures by giving distinct signals at working voltages of typically 236, 480 and 650 mV (vs. Ag/AgCl), respectively. Differential pulse voltammetric peak currents of DA, AC and MT increase linearly in the 50 nmol L-1 - 1.45 mmol L-1, 40 nmol L-1 -1.4 mmol L-1, and 6.0 nmol L-1 - 1.4 mmol L-1 concentration ranges. The limits of detection are 36.5 nmol L-1 for DA, 36.5 nmol L-1 for AC, and 21.6 nmol L-1 for MT. The sensor was successfully used to detect the analytes in (spiked) human serum and drug samples. Graphical abstract Schematic presentation of Al2O3-supported palladium nanoparticles (PdNP@Al2O3) for modification of a carbon paste electrode (CPE) to develop a voltammetric sensor for the simultaneous determination of dopamine (DA), acetaminophen (AC) and melatonin (MT).

Magneto-controlled aptasensor for simultaneous electrochemical detection of dual mycotoxins in maize using metal sulfide quantum dots coated silica as labels.

Currently there is an urgent need for multi-mycotoxin detection methods due to the co-occurrence of multiple mycotoxins in food raw materials and their augmented toxicity. Herein, a magneto-controlled aptasensor has been developed for simultaneous electrochemical detection of ochratoxin A (OTA) and fumonisin B1 (FB1), two typical mycotoxins found in food crops world-wide. This aptasensor was designed using the high specificity between the target and aptamer with heavy CdTe or PbS quantum dots (QDs) coated silica as labels and the complementary DNA functionalized magnetic beads as capture probes. In presence of targets, the aptamer preferred to form the target-aptamer binding which forced the partial release of the preloaded labels from the magnetic beads. After a one-step incubation and a simple magnetic separation, the electrochemical signals of Cd2+ and Pb2+ dissolved from the reserved labels which had negative correlation with targets contents, was measured based on the difference of peak potentials. This aptasensor provided a wide detection range of 10pgmL-1 to 10ngmL-1 for OTA and 50pgmL-1 to 50ngmL-1 for FB1, and succeeded in real maize samples. This method provides a new avenue for high throughput screen of mycotoxins due to the advantages of simple instrument, low sample consumption, short assay times, and lower detection costs per assay. Moreover, it could be readily expanded for the simultaneous detection of a large panel of mycotoxins by using different metal sulfide QDs when their specific aptamers are available.

A novel "dual-potential" electrochemiluminescence aptasensor array using CdS quantum dots and luminol-gold nanoparticles as labels for simultaneous detection of malachite green and chloramphenicol.

A novel type of "dual-potential" electrochemiluminescence (ECL) aptasensor array was fabricated on a homemade screen-printed carbon electrode (SPCE) for simultaneous detection of malachite green (MG) and chloramphenicol (CAP) in one single assay. The SPCE substrate consisted of a common Ag/AgCl reference electrode, carbon counter electrode and two carbon working electrodes (WE1 and WE2). In the system, CdS quantum dots (QDs) were modified on WE1 as cathode ECL emitters and luminol-gold nanoparticles (L-Au NPs) were modified on WE2 as anode ECL emitters. Then the MG aptamer complementary strand (MG cDNA) and CAP aptamer complementary strand (CAP cDNA) were attached on CdS QDs and L-Au NPs, respectively. The cDNA would hybridize with corresponding aptamer that was respectively tagged with cyanine dye (Cy5) (as quenchers of CdS QDs) and chlorogenic acid (CA) (as quenchers of l-Au NPs) using poly(ethylenimine) (PEI) as a bridging agent. PEI could lead to a large number of quenchers on the aptamer, which increased the quenching efficiency. Upon MG and CAP adding, the targets could induce strand release due to the highly affinity of analytes toward aptamers. Meanwhile, it could release the Cy5 and CA, which recovered cathode ECL of CdS QDs and anode ECL of L-Au NPs simultaneously. This "dual-potential" ECL strategy could be used to detect MG and CAP with the linear ranges of 0.1-100 nM and 0.2-150 nM, with detection limits of 0.03 nM and 0.07 nM (at 3sB), respectively. More importantly, this designed method was successfully applied to determine MG and CAP in real fish samples and held great potential in the food analysis.

Highly-sensitive and rapid detection of ponceau 4R and tartrazine in drinks using alumina microfibers-based electrochemical sensor.

Alumina microfibers were prepared and used to construct an electrochemical sensor for simultaneous detection of ponceau 4R and tartrazine. In pH 3.6 acetate buffer, two oxidation waves at 0.67 and 1.01 V were observed. Due to porous structures and large surface area, alumina microfibers exhibited high accumulation efficiency to ponceau 4R and tartrazine, and increased their oxidation signals remarkably. The oxidation mechanisms were studied, and their oxidation reaction involved one electron and one proton. The influences of pH value, amount of alumina microfibers and accumulation time were examined. As a result, a highly-sensitive, rapid and simple electrochemical method was newly developed for simultaneous detection of ponceau 4R and tartrazine. The detection limits were 0.8 and 2.0 nM for ponceau 4R and tartrazine. This new sensor was used in different drink samples, and the results consisted with the values that obtained by high-performance liquid chromatography.