5-Fluorouracil (5-FU) is an effective anticancer drug widely used in the world. To improve therapy efficiency and reduce side effects, it is very important to frequently detect the concentration of 5-FU in blood samples of patients. In this work, a new type of lateral flow immunochromatographic assay (LFIA) based on surface-enhanced Raman scattering (SERS) for ultrasensitive and specific detection of 5-FU in blood samples was developed. Au@Ag/Au nanoparticles (NPs) employing Au particles as the core and Ag/Au alloy as the shell were synthesized, characterized and used as the substrate in SERS-LFIA due to their high SERS enhancement and biocompatibility. The immunoprobe was made in the form of AuMBA@Ag/Au-Ab in which mercaptobenzoic acid (MBA, a common Raman active reporter) was embedded in the core–shell layer and the monoclonal antibody (mAb) against 5-FU was immobilized on the surface. The performance of SERS-LFIA was similar to that in colloidal gold based-LFIA, and the entire assay time was within 20 min. According to the color intensity on the testing (T) lines of LFIA strips visualized by eyes, the contents of 5-FU in the samples could be qualitatively or semi-quantitatively identified. Furthermore, by measuring the characteristic Raman intensities of MBA on T lines, quantitative detection of 5-FU in the samples were achieved. The IC50 and limit of detection (LOD) of the LFIA for 5-FU were found to be 20.9 pg mL−1 and 4.4 pg mL−1, respectively. There was no cross-reactivity (CR) of the LFIA with nine relative compounds, and the CR with cytosine, tegafur and carmofur were less than 4.5%. The recoveries of 5-FU from spiked blood samples were in the range of 78.6~86.4% with the relative standard deviation (RSD) of 2.69~4.42%. Five blood samples containing 5-FU collected from the Cancer Hospital were measured by SERS-LFIA, and the results were confirmed by LC-MS/MS. It was proven that the proposed method was able to simply and rapidly detect 5-FU in blood samples with high sensitivity, specificity, accuracy and precision. 相似文献
A sensitive and selective resonance scattering spectral (RSS) assay was proposed for the determination of sudan I (SDI), using 10 nm nanogold to label the antibody against sudan I (anti-SDI Ab) to obtain a RSS probe for SDI. The immunonanogold reaction between nanogold-labelled anti-SDI Ab and SDI took place in pH 4.92 KH2PO4–Na2HPO4 buffer solution and in the presence of polyethylene glycol (PEG)-6000, and the intensity of resonance scattering peak at 580 nm decreased greatly. The decreased intensity ΔI580 nm was proportional to the concentration of SDI in the range of 0.23–45.0 ng mL?1. The linear regression equation was calculated as ΔI580nm = 1.20c + 2.01 (R = 0.9975, n = 6), with a detection limit (3σ) of 0.13 ng mL?1. The SDI in egg samples was assayed, with satisfactory results. 相似文献
The use of olaquindox (OLA) as an additive in animal feedstuffs has been prohibited in the European Union and many other countries.
In this study, a highly sensitive and specific indirect competitive enzyme-linked immunosorbent assay (ELISA) for determination
of OLA in animal feed samples was developed. OLA was activated by N′N-carbonyldiimidazole and coupled with bovine serum albumin (BSA) and ovalbumin (OVA). It was found that the sensitivity and
specificity of the two antisera were very similar, with the IC50 values of 16 ng mL−1 and 19 ng mL−1, respectively. Cross-reactivity was less than 35% for four structurally related compounds and no recognition of five other
antibiotics was observed. The better antiserum I was selected for further experiments, for example testing stability, solvent
effect, accuracy, and precision. The IC50 value for eight standard curves was in the range 12–18 ng mL−1 and the LOD at a signal-to-noise ratio of 3 (S/N = 3) was 0.31 ± 0.11 ng mL−1. The ELISA tolerated 5% methanol without significant influence on IC50 value. The recoveries of spiked OLA in five different animal feed types including auxin, pig complex feed, fish complex feed,
broiler concentrated feed, and pig premix feed were in the range 88.3–119.0% and the intra-assay relative standard deviation
(RSD) was within 4.7–33.5% (n = 3). The ELISA for unspiked feed samples was confirmed by high-performance liquid chromatography (HPLC), with a high correlation
coefficient of 0.9862 (n = 5). The proposed ELISA could be a feasible quantitative/screening method for OLA analysis in feed samples with the properties
of high sensitivity, specificity, simplicity of sample pretreatment, high sample throughput, and low expense.
Figure Polyclonal antibody based ELISA for detection of olaquindox 相似文献
A selective molecularly imprinted solid-phase extraction (MISPE) for indomethacin (IDM) from water samples was developed.
Using IDM as template molecule, acrylamide (AM) or methacrylic acid (MAA) as functional monomer, ethylene dimethacrylate (EDMA)
as crosslinker, and bulk or suspension polymerization as the synthetic method, three molecularly imprinted polymers (MIPs)
were synthesized and characterized with a rebinding experiment. It was found that the MIP of AM-EDMA produced by bulk polymerization
showed the highest binding capacity for IDM, and so it was chosen for subsequent experiments, such as those testing the selectivity
and recognition binding sites. Scatchard analysis revealed that at least two kinds of binding sites formed in the MIP, with
the dissociation constants of 7.8 μmol L−1 and 127.2 μmol L−1, respectively. Besides IDM, three structurally related compounds — acemetacin, oxaprozin and ibuprofen — were employed for
selectivity tests. It was observed that the MIP exhibited the highest selective rebinding to IDM. Accordingly, the MIP was
used as a solid-phase extraction sorbent for the extraction and enrichment of IDM in water samples. The extraction conditions
of the MISPE column for IDM were optimized to be: chloroform or water as loading solvent, chloroform with 20% acetonitrile
as washing solution, and methanol as eluting solvent. Water samples with or without spiking were extracted by the MISPE column
and analyzed by HPLC. No detectable IDM was observed in tap water and the content of IDM in a river water sample was found
to be 1.8 ng mL−1. The extraction efficiencies of the MISPE column for IDM in spiked tap and river water were acceptable (87.2% and 83.5%,
respectively), demonstrating the feasibility of the prepared MIP for IDM extraction.
Figure Molecularly imprinted polymer-based solid-phase extraction for indomethacin 相似文献
We present a new approach for simulating the motions of flexible polyelectrolyte chains based on the continuous kink-jump Monte Carlo technique coupled to a lattice field theory based calculation of the Poisson-Boltzmann (PB) electrostatic free energy "on the fly." This approach is compared to the configurational-bias Monte Carlo technique, in which the chains are grown on a lattice and the PB equation is solved for each configuration with a linear scaling multigrid method to obtain the many-body free energy. The two approaches are used to calculate end-to-end distances of charged polymer chains in solutions with varying ionic strengths and give similar numerical results. The configurational-bias Monte Carlo/multigrid PB method is found to be more efficient, while the kink-jump Monte Carlo method shows potential utility for simulating nonequilibrium polyelectrolyte dynamics. 相似文献
We describe a method for the preparation of water-soluble gold nanoclusters (Au-NCs) from chloroauric acid using denatured-casein as both a reducing and stabilizing agent. The resulting Au-NCs were characterized by photoluminescence, UV–vis absorption, and X-ray photoelectron spectroscopies, and by transmission electron microscopy. The Au-NCs have an average diameter of 1.7 ± 0.2 nm and exhibit orange-red fluorescence emission peaking at 600 nm (with a Stokes’ shift as large as 237 nm), a quantum yield of 4.3 %, and good stability over the physiologically relevant range of pH values and ionic strength. Cytotoxicity studies showed the Au-NCs to display negligible effects in terms of altering cell proliferation or triggering apoptosis. Fluorescence imaging of HeLa cancer cells was accomplished by loading such cells with the Au-NCs. The fluorescence of the Au-NCs is found to be strongly quenched by Hg(II) ions, and thus the Au-NCs can be used for detecting and, possibly, imaging of Hg(II). An assay was worked out for the determination of Hg(II), and its limit of detection is 1.83 nM, which is 5.5 times lower than the maximum allowed concentration of Hg(II) in drinking water as defined by the US EPA.