RESUMEN
A straightforward in situ detection method for dengue infection was demonstrated through the molecular imprinting of a dengue nonstructural protein 1 (NS1) epitope into an electropolymerized molecularly imprinted polyterthiophene (E-MIP) film sensor. The key enabling step in the sensor fabrication is based on an epitope imprinting strategy, in which short peptide sequences derived from the original target molecules were employed as the main template for detection and analysis. The formation of the E-MIP sensor films was facilitated using cyclic voltammetry (CV) and monitored in situ by electrochemical quartz crystal microbalance (EC-QCM). Surface properties were analyzed using different techniques including atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and polarization modulation-infrared reflection-adsorption (PM-IRRAS). The standard calibration curve (R = 0.9830) was generated for the detection of the epitope, Ac-VHTWTEQYKFQ-NH2, with a linear range of 0.2 to 30 µg/mL and detection limit of 0.073 µg/mL. A separate calibration curve (R = 0.9786) was obtained using spiked buffered solutions of dengue NS1 protein, which resulted in a linear range of 0.2 to 10 µg/mL and a detection limit of 0.056 µg/mL. The fabricated E-MIP sensor exhibited long-term stability, high sensitivity, and good selectivity towards the targeted molecules. These results indicated that the formation of the exact and stable cavity imprints in terms of size, shape, and functionalities was successful. In our future work, we aim to use our E-MIP sensors for NS1 detection in real-life samples such as serum and blood.
Asunto(s)
Virus del Dengue/aislamiento & purificación , Dengue/diagnóstico , Polímeros Impresos Molecularmente/química , Proteínas no Estructurales Virales/análisis , Adsorción , Técnicas Electroquímicas , Humanos , Límite de Detección , Impresión Molecular , Espectroscopía de Fotoelectrones , Tecnicas de Microbalanza del Cristal de Cuarzo , Proteínas no Estructurales Virales/aislamiento & purificaciónRESUMEN
Monitoring of pollution index values in sediments is crucial in assessing the environmental impacts of toxic metals in a given location. These indices are typically acquired using elaborate and tedious calibration curve-dependent techniques such as (inductively coupled plasma - optical emission spectroscopy) ICP-OES and (atomic absorption spectroscopy) AAS. In this study, laser-induced breakdown spectroscopy (LIBS) was used as a simple and fast alternative method for estimating enrichment factor (EF) and contamination factor (CF) of the sediment samples obtained from selected mining sites. Quantitative analyses of three metal targets (Cd, Pb, and Zn) were done using a calibration-free LIBS method based on the Boltzmann population distribution. Both the EF and CF values calculated from classical ICP-OES method provided significantly high correspondence with the respective EF (R2 = 0.8862-0.9770, p < 0.01-0.05) and CF (R2 = 0.9454-0.9714, p < 0.01) obtained from the developed LIBS method. The intensity-based LIBS approach identified samples AC2 and CCC as the ones with the highest and lowest pollution index values, respectively. The same observation was seen using the concentration-based ICP-OES technique which showed good correlation between the two methods. The correlation results showed the potential of the curve-fitting LIBS analysis in evaluating the level of metal contamination in an area without the preparation of matrix-matched calibration curves.
Asunto(s)
Metales/análisis , Espectrofotometría Atómica/métodos , Calibración , Rayos Láser , MineríaRESUMEN
The significance and epidemiological effects of metals to life necessitate the development of direct, efficient, and rapid method of analysis. Taking advantage of its simple, fast, and high-throughput features, we present a novel approach to metal ion detection by matrix-functionalized magnetic nanoparticle (matrix@MNP)-assisted MALDI-MS. Utilizing 21 biologically and environmentally relevant metal ion solutions, the performance of core and matrix@MNP against conventional matrixes in MALDI-MS and laser desorption ionization (LDI) MS were systemically tested to evaluate the versatility of matrix@MNP as ionization element. The matrix@MNPs provided 20- to >100-fold enhancement on detection sensitivity of metal ions and unambiguous identification through characteristic isotope patterns and accurate mass (<5 ppm), which may be attributed to its multifunctional role as metal chelator, preconcentrator, absorber, and reservoir of energy. Together with the comparison on the ionization behaviors of various metals having different ionization potentials (IP), we formulated a metal ionization mechanism model, alluding to the role of exciton pooling in matrix@MNP-assisted MALDI-MS. Moreover, the detection of Cu in spiked tap water demonstrated the practicability of this new approach as an efficient and direct alternative tool for fast, sensitive, and accurate determination of trace metal ions in real samples.