Homogeneous electrochemical ratiometric biosensor for MircoRNA detection based on UiO-66-NH2 signal probe and waste-free entropy-driven DNA machine.

The construction of efficient methods for highly sensitive and rapid detection of disease markers is essential for the early diagnosis of serious diseases. In this paper, taking advantage of the UiO-66-NH2 signal molecule in combination with a waste-free entropy-driven DNA machine, a novel homogeneous electrochemical ratiometric platform is developed to detect MircoRNA (miRNA). Metal-organic framework materials (UiO-66-NH2 MOF) and ferrocene were utilized as electrochemical signal tags and reference probes, respectively. The target-initiated waste-free three-dimensional (3D) entropy-driven DNA nanomachine is activated in the presence of miRNA, resulting in DNA-labeled-UiO-66-NH2 falling off from the electrode, leading to a decrease in the signal of UiO-66-NH2 at 0.83V. Our strategy can mitigate false positive responses induced by the DNA probes immobilized on electrodes in traditional distance-dependent signal adjustment ratiometric strategies. The proposed ratiometric platform demonstrates superior sensitivity (a detection limit of 9.8 fM), simplified operation, high selectivity, and high repeatability. The ratiometric biosensor is also applied to detect miRNA content in spiked serum samples.

Three-dimensional DNA biomimetic networks (B-3D Net)-based ratiometric fluorescence platform for cancer-related gene biosensing.

Efficient detection of cancer-related nucleic acids is pivotal for early cancer diagnosis. This study introduces a target induced three-dimensional DNA biomimetic networks (B-3D Net)-based ratiometric fluorescence platform using manganese dioxide nanosheets (MnO2 NS)/o-phenylenediamine in combination with hybridization chain reaction to detect cancer-related genes (p53 gene). The incorporation of multiple signals within the B-3D networks can significantly enhance catalytic activity and amplify the output signals, enabling a high sensitivity. Compared with traditional ratio fluorescence platforms, there is no demand to synthesize fluorescent nanoprobes due to the in-situ formation of fluorescence species, which is simple and cost-effective. The corresponding assay demonstrated exceptional sensitivity (with a detection limit as low as 2 fM), selectivity, reproducibility, and accuracy, which mitigates disturbances caused by instrument errors, an inaccurate probe count, and the microenvironment. Furthermore, the ease and straightforwardness of discerning changes in fluorescent brightness and colour by the naked eye are evident. Using the relevant software, a linear relationship between fluorescent images using a smartphone and target concentration was obtained. Hence, the novel ratiometric sensing system will demonstrate new opportunities on determination of target DNA samples in complex biological environments.

A DNA tetrahedral nanomaterial-based dual-signal ratiometric electrochemical aptasensor for the detection of ochratoxin A in corn kernel samples.

Ochratoxin A (OTA) is a highly toxic food contaminant and is harmful to human beings. Herein, a ratiometric electrochemical aptasensor based on a DNA tetrahedral nanomaterial (NTH) was developed in combination with the signal tag of a zirconium metal-organic framework (UiO-66) for the detection of OTA. In the sensor, UiO-66 and a [Fe(CN)6]3-/4- electrolyte solution were used as the signal probe and the internal reference probe, respectively. In the presence of OTA, the OTA aptamer was released from the electrode due to the specific binding of OTA. Thus, signal probe P1 labeled-UiO-66 was captured on the electrode surface by hybridization with DNA NTH. Since signal probe P1 labeled-UiO-66 was close to the electrode, it leads to an increased signal current of UiO-66 at +0.9 V. As the conductivity of the modified electrode decreased, the current signal of [Fe(CN)6]3-/4- at +0.2 V also decreased. The proposed ratiometric electrochemical aptasensor could effectively eliminate external environmental influences and could avoid electrochemical background signals. The aptasensor demonstrated high specificity for OTA, and achieved a good linear range of 1 pg mL-1-100 ng mL-1 with a detection limit of 330 fg mL-1. The developed electrochemical aptamer biosensor effectively detected OTA in corn kernel samples, verifying its practical application for the determination of OTA in actual samples.

Amino-functionalized perylenediimide derivative with dual fluorescence emission for the detection of ascorbic acid in vivo and in vitro.

The rapid, sensitive, and selective detection of ascorbic acid (AA) is of significance in medical assays and diagnostics. In this work, a new aminoperylenediimide (APDI) derived ratiometric fluorescent probe based on the specific redox reaction of cobalt oxyhydroxide (CoOOH) and AA was constructed. APDI exhibited dual fluorescence emission peaks at 549 and 596 nm with an excitation wavelength of 494 nm. In the presence of CoOOH, the dual fluorescence could be quenched. The dominant fluorescence quenching mechanism was caused by the inner filter effect. Using the red emission as a reference, the fluorescence intensity ratio (F549 /F596 ) was linearly correlated with the concentration of AA over a range of 0.05 to 1 µM. The limit of detection for AA was found to be 17 nM. Importantly, the probe was successfully used to detect AA in living cells. Therefore, this high sensitivity and selectivity strategy could directly survey the AA levels in real samples.

Modulation of inner filter effect between persistent luminescent particles and 2, 3-diaminophenazine for ratiometric fluorescent assay of ascorbic acid and ascorbate oxidase activity.

Ascorbate oxidase (AAO) and ascorbic acid (AA) play an important role in delaying lives senescence and metabolism. In this study, a sensitive ratiometric fluorescence sensing system based on the inner filter effect (IFE) between persistent luminescent particles (PLPs) and 2, 3-diaminophenazine (DAP), was designed for the detection of AA and AAO activity. Wherein, PLPs emit blue fluorescence at 475 nm with an excitation wavelength of 370 nm. CoOOH nanosheets with oxidase-like activity can oxidize o-phenylenediamine (OPD) to produce 2, 3-diaminophenazine (DAP) with orange fluorescence at 558 nm. The generated DAP quenched the fluorescence of PLPs by an inner filter effect (IFE). When AA was introduced to the system, CoOOH nanosheets were destroyed and reduced to Co2+, thereby inhibiting the oxidization of OPD and effectively preserving the blue fluorescence of PLPs at 475 nm. Besides, AAO can catalyse AA to produce the oxided dehydroascorbic acid (DHA). The dissipative AA can recover orange fluorescence of DAP with weakening the blue fluorescence of PLPs. Therefore, a sensitive ratio fluorescence sensing strategy was established by using PLPs as the reference signal and DAP as a reported signal for the detection of AA and AAO activity. Under optimal conditions, the obtained linear ranges were 1-45 µM and 1-20 mU/mL, and detection limits were 0.2 µM and 0.25 mU/mL, respectively. Finally, this proposed ratiometric fluorescent analytical strategy was used to detect AA in real samples (lemon, orange, tomato), which exhibited satisfactory results comparing with commercial kit.

[Determination of lambda-cyhalothrin residue tea and soil using gas chromatography].

A gas chromatographic (GC) method was established for the determination of lambda-cyhalothrin residue in tea and soil. Tea and soil samples were extracted with hexane, separated by capillary column and determined by gas chromatography-electron capture detector (GC-ECD). The average recoveries of lambda-cyhalothrin in tea and soil were 89.0% - 94.1% and 89.8% - 94.7%, respectively at the spiking levels of 0.02 to 2.00 mg/kg. The corresponding relative standard deviations (RSDs, n = 5) were 3.0% -4.9% and 2.5% -4.2%, respectively. The limit of detection (S/N = 3) was 0.002 mg/kg for lambda-cyhalothrin. The degradations of 2.5% lambda-cyhalothrin microemulsion in tea and soil in Changsha, Hunan were investigated and the degradation equations were Y = 3.199 6e(-0.339 4x) and Y = 0.122 4e(-0.103 6x) with the correlation coefficients of 0.995 6 and 0.924 7, respectively. The half-lives of lambda-cyhalothrin in tea and soil were 2.04 days and 6.69 days, respectively.

[Determination of nitenpyram residue in cabbage and soil using gas chromatography].

An analytical method for the determination of nitenpyram residue in cabbage and soil using gas chromatography was established. The nitenpyram residue was extracted from cabbage and soil with acetone-water (4 : 1, v/v), cleaned up by a Florisil column, and then detected by gas chromatography-electron capture detection (GC-ECD). At the spiked level range from 0.02 to 2.00 mg/kg, the average recoveries of nitenpyram were 88.73%-94.3% and 90.82%-96.27% with the relative standard deviations (RSDs) of 3.09%-7.39% and 2.01%-4.92% in cabbage and soil, respectively. The limit of detection of nitenpyram was 0.02 mg/kg. The method is fast, sensitive, simple, reproducible and practical for the determination of nitenpyram residue in environmental systems.