Jian-Lian Chen

Comparative syntheses of tetracycline-imprinted polymeric silicate and acrylate on CdTe quantum dots as fluorescent sensors.

The amphoteric drug molecule tetracycline, which contains groups with pKa 3.4-9.9, was used as a template for conjugating molecularly imprinted polymers (MIPs) and as a quencher for CdTe quantum dot (QD) fluorescence. Two MIP-QD composites were synthesized by a sol-gel method using a silicon-based monomer and a monomer linker between the MIP and QD, i.e., tetraethoxylsilane/3-mercaptopropyltriethoxysilane (MPS) and tetraethoxylsilane/3-aminopropyltriethoxysilane (APS). Another MIP-QD composite was synthesized by the chain-growth polymerization of methacrylic acid (MAA) and an allyl mercaptan linker. The prepared MIP-QDs were characterized by FTIR and SEM and utilized at 0.33 mg/mL to determine the tetracycline content in phosphate buffers (pH 7.4, 50mM) through the Perrin and Stern-Volmer models of quenching fluorometry. The Perrin model was applied to tetracycline concentrations of 7.4 μM-0.37 mM for MIP-MPS-QD, 7.4 μM-0.12 mM for MIP-APS-QD, and 7.4 μM-0.10mM for MIP-MAA-QD (R(2)=0.9988, 0.9978, and 0.9931, respectively). The Stern-Volmer model was applied to tetracycline concentrations of 0.12-0.37 mM for MIP-APS-QD (R(2)=0.9983) and 0.10-0.37 mM for MIP-MAA-QD (R(2)=0.9970). The detection limits were 0.45 μM, 0.54 μM, and 0.50 μM for MIP-MPS-QD, MIP-APS-QD, and MIP-MAA-QD, respectively. Equilibrium times, differences between imprinted and nonimprinted polymers, and MIP-QD quenching mechanisms were discussed. Finally, specificity studies demonstrated that MIP-MAA-QD exhibited optimal recoveries of 96% from bovine serum albumin (n=5, RSD=3.6%) and 91% from fetal bovine serum (n=5, RSD=4.8%).

Determination of urinary malondialdehyde by isotope dilution LC-MS/MS with automated solid-phase extraction A cautionary note on derivatization optimization

A highly sensitive quantitative LC-MS/MS method was developed for measuring urinary malondialdehyde (MDA). With the use of an isotope internal standard and online solid-phase extraction, urine samples can be directly analyzed within 10 min after 2,4-dinitrophenylhydrazine (DNPH) derivatization. The detection limit was estimated as 0.08 pmol. This method was further applied to assess the optimal addition of DNPH for derivatization and to measure urinary MDA in 80 coke oven emission (COE)-exposed and 67 nonexposed workers. Derivatization optimization revealed that to achieve complete derivatization reaction, an excess of DNPH is required (DNPH/MDA molar ratio: 893-8929) for urine samples that is about 100 times higher than that of MDA standard solutions (molar ratio: 10-80). Meanwhile, the mean urinary concentrations of MDA in COE-exposed workers were significantly higher than those in nonexposed workers (0.23±0.17 vs 0.14±0.05 μmol/mmol creatinine, P<0.005). Urinary MDA concentrations were also significantly associated with the COE (P<0.005) and smoking exposure (P<0.05). Taken together, this method is capable of routine high-throughput analysis and accurate quantification of MDA and would be useful for assessing the whole-body burden of oxidative stress. Our findings, however, raise the issue that derivatization optimization should be performed before it is put into routine biological analysis.

Hydrophilic ionic liquid-passivated CdTe quantum dots for mercury iondetection

A hydrophilic ionic liquid, 1-ethyl-3-methylimidazolium dicyanamide (EMIDCA), was used as a medium for the synthesis of highly luminescent CdTe nanocrystals (NCs) capped with thioglycolic acid (TGA). The synthesis was performed for 8 h at 130 °C, was similar to nanocrystal preparation in an aqueous medium, and used safe, low-cost inorganic salts as precursors. After the reaction, the photoluminescence quantum yield of the CdTe NCs (NC(IL-130)) prepared in EMIDCA was significantly higher than that of the nanocrystals prepared in water (NC(w)) at 100 °C (86% vs. 35%). Moreover, the emission wavelength and particle size of NC(IL-130) were smaller than NC(w) (450 nm vs. 540 nm and 4.0 nm vs. 5.2 nm, respectively). The activation of NC(IL-130) was successful due to the coordinated action of two ligands, EMIDCA and TGA, in the primary steps of the NC formation pathway. An increase or decrease in the synthesis temperature, to 160 °C or 100 °C, respectively, was detrimental to the luminescence quality. However, the quenching effect of Hg²⁺ on the fluorescence signals of the NC(IL-130) was distinctively unique, whereas certain interfering ions, such as Pb²⁺, Fe³⁺, Co²⁺, Ni²⁺, Ag⁺, and Cu²⁺, could also quench the emission of the NC(w). Based on the Perrin model, the quenching signals of NC(w) and NC(IL-130) were well correlated with the Hg²⁺ concentrations in the phosphate buffer (pH 7.5, 50 mM). In comparison with the NC(w), the NC(IL-130) had a high tolerance of the interfering ions coexisting with the Hg²⁺ analyte, high recovery of Hg²⁺ spiked in the BSA- or FBS-containing medium, and high stability of fluorescence quenching signals between trials and days. The NC(IL-130) nanocrystals can potentially be used to develop a probe system for the determination of Hg²⁺ in physiological samples.

Efficient stripping of photoresist on metallized wafers by a pause flow of supercritical fluid.

Utilization of supercritical fluids (SCFs) is studied here on the premises of a saving of hazardous organic solvents and of the specification for stripping the photoresist (PR) on metallization layers, which is one of the integrated circuit processing modules. By using factorial experimental designs with five factors and four level ranges, this research focuses on determining an optimized recipe with high stripping efficiency and to determine the stripping mechanism. In the case of PR on an aluminum layer, the initial use of the pulse flow mode could increase the extraction ratio remarkably when compared to the conventional continuous flow mode. Based on the limitation of a total volume of 30 mL purging SCF-CO(2) for economical considerations, the optimum conditions can be summarized as follows: 120 degrees C, oven temperature; 350 atm, CO(2) pressure; 0.2 mL of ethylacetate spiking to SCF-CO(2); 2.0 min, static equilibrium time; and five cycles of dynamic flow pausing. A recovery of 94.6% (n=3, RSD=6.5%) was obtained, while the diffusion of stripped PR from substrate matrix prevailed over the dissolution of binding PR into the SCF medium. In the case of copper, the optimum parameters in a pause flow mode were 140 degrees C, oven temperature; 500 atm, CO(2) pressure; 0.75 mL, ethylacetate spiking volume; 5.0 min, static time; and six cycles of flow pausing. These extreme parameters still did not produce an SCF environment suitable for diffusion or dissolution mass transfer, and thus a recovery of 76.2% (n=3, RSD=7.5%) was only obtained. Removing PR coated on a Cu layer was harder than that on an Al layer.

Sensitive Detection of 8-Nitroguanine in DNA by Chemical Derivatization Coupled with Online Solid-Phase Extraction LC-MS/MS

8-Nitroguanine (8-nitroG) is a major mutagenic nucleobase lesion generated by peroxynitrite during inflammation and has been used as a potential biomarker to evaluate inflammation-related carcinogenesis. Here, we present an online solid-phase extraction (SPE) LC-MS/MS method with 6-methoxy-2-naphthyl glyoxal hydrate (MTNG) derivatization for a sensitive and precise measurement of 8-nitroG in DNA. Derivatization optimization revealed that an excess of MTNG is required to achieve complete derivatization in DNA hydrolysates (MTNG: 8-nitroG molar ratio of 3740:1). The use of online SPE effectively avoided ion-source contamination from derivatization reagent by washing away all unreacted MTNG before column chromatography and the ionization process in mass spectrometry. With the use of isotope-labeled internal standard, the detection limit was as low as 0.015 nM. Inter- and intraday imprecision was <5.0%. This method was compared to a previous direct LC-MS/MS method without derivatization. The comparison showed an excellent fit and consistency, suggesting that the present method has satisfactory effectiveness and reliability for 8-nitroG analysis. This method was further applied to determine the 8-nitroG in human urine. 8-NitroG was not detectable using LC-MS/MS with derivatization, whereas a significant false-positive signal was detected without derivatization. It highlights the use of MTNG derivatization in 8-nitroG analysis for increasing the method specificity.

Comprehensive analysis of the formation and stability of peroxynitrite-derived 8-nitroguanine by LC-MS/MS: Strategy for the quantitative analysis of cellular 8-nitroguanine

Peroxynitrite is a major oxidizing and nitrating biological agent formed at sites of inflammation. Peroxynitrite can cause DNA damage and is thought to contribute to inflammation-related carcinogenesis. This study describes a sensitive and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the direct determination of peroxynitrite-derived 8-nitroguanine (8-nitroGua) in DNA hydrolysates. This method exhibited a sensitive detection limit of 3 fmol and inter- and intraday imprecision of <10% and was applied to systemically examine the formation and stability of peroxynitrite-derived 8-nitroGua in different DNA substrates under various conditions. The 8-nitroGua formation was maximal at pH 8. The formation rate of 8-nitroGua in different DNA substrates decreased in the order of monodeoxynucleoside>single-stranded DNA>double-stranded DNA. A stability test revealed that the half-life for the depurination of 8-nitroGua from DNA was short and affected by both the temperature and DNA structure. When present in monodeoxynucleoside, the half-life of 8-nitroGua was estimated to be ~6min at 25°C and 2.3h at ~0°C. In single-stranded DNA, the half-life varied from 1.6h at 37°C to 533h at -20°C, whereas the half-life increased from 2.4h at 37°C to 1115h at -20°C in double-stranded DNA. We demonstrated that the measurement of 8-nitroGua in isolated DNA is not practicable because 8-nitroGua is unstable and lost during DNA extraction from cell. Therefore, we suggest that directly detecting cellular 8-nitroGua following nuclear membrane lysis is an alternative measure of the nitrative damage of nucleic acids, accounting for both DNA and RNA lesions within cells.