PubMed · 5403352
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P Bonnin. 1969. [Fluorography and fluoroscopy].. https://pubmed.ncbi.nlm.nih.gov/5403352/
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A new method for the determination of trace mercury by solid substrate-room temperature phosphorimetry (SS-RTP) quenching method has been established. In glycine-HCl buffer solution, xylenol orange (XO) can react with Sn4+ to form the complex [Sn(XO)6]4+. [Sn(XO)6]4+ can interact with Fin- (fluorescein anion) to form the ion associate [Sn(XO)6]4+.[(Fin)4]-, which can emit strong and stable room temperature phosphorescence (RTP) on polyamide membrane (PAM). Hg2+ can catalyze H2O2 oxidizing the ion association complex [Sn(XO)6]4+.[(Fin)4]-, which causes the RTP to quench. The DeltaIp value is directly proportional to the concentration of Hg2+ in the range of 0.016-1.6 fg spot(-1) (corresponding concentration: 0.040-4.0 pg ml(-1), 0.40 microl spot(-1)), and the regression equation of working cure is DeltaIp=10.03+83.15 m Hg2+ (fg spot(-1)), (r=0.9987, n=6) and the detection limit (LD) is 3.6 ag spot(-1)(corresponding concentration: 9.0 x 10(-15) g ml(-1), the sample volume: 0.4 microl). This simple, rapid, accurate method is of high selectivity and good repeatability, and it has been successfully applied to the determination of trace mercury in real samples. The reaction mechanism for catalyzing H2O2 oxidizing the ion association complex ([Sn(XO)6]4+.[(Fin)4]-) SS-RTP quenching method to determine trace mercury is also discussed.
Three compounds structurally related to the fluorescent zinc sensor Zinpyr-1 (ZP1) have been synthesized and characterized. In each of these ZinAlkylPyr (ZAP) analogues, an alkyl group (methyl, benzyl) replaces one of the metal-binding picolyl moieties in ZP1. The methyl-for-picolyl substitutions in ZAP1 and ZAP2 have a negligible effect on the optical spectrum of the fluorophore but elevate the quantum yields (Phi = 0.82 (ZAP1), 0.74 (ZAP2)) to values near that of Zn2+-saturated ZP1 (Phi = 0.92). The benzyl-for-picolyl substitution in ZAP3 similarly enhances the quantum yield (Phi = 0.52) relative to that of metal-free ZP1 (Phi = 0.38). As previously observed for methylated ZP1 sensors, methylation of the 6-position of the pyridyl ring diminishes the emission by lowering both the molar extinction coefficient and the quantum yield. Although these new ZAP compounds cannot detect Zn2+ fluorimetrically at neutral pH, complexation of Zn2+ does occur, as evidenced by sizable changes in the optical spectra. The ZAP1-3 probes can detect Zn2+ fluorimetrically at pH 9, indicating that proton-induced background emission obscures any Zn2+-induced fluorescence at pH 7. The tertiary amine groups in ZAP1-3 are less basic than those in ZP1, which implies that the additional pyridine rings are responsible for the emissive response to Zn2+ at pH 7.0.
Reduced "leuco" dyes such as dichlorodihydrofluorescein (DCFH(2)) are widely used as profluorescent probes for oxidative stress, although they require a catalyst to be oxidized by hydrogen peroxide and react indiscriminately with oxidizing radicals and the fluorescent product (DCF) is a potential photosensitizer of superoxide generation. In this study, key properties of the radical intermediate in oxidation ("semiquinone," DCFH(.-)/DCF(.)(-)) were measured, to help understand the reactions that can occur in biological systems. The intermediate was generated by oxidizing DCFH(2) or reducing DCF by radiolytically generated radicals and monitoring the reactions using kinetic spectrophotometry. The semiquinone showed pH-sensitive absorption spectral changes, decay kinetics (both in the absence and in the presence of oxygen), and reduction potential, all corresponding to prototropic dissociations with pK(a)'s of approximately 7.1 and 9.0. DCFH(2) has pK(a)'s in a similar region (8-9) and hence pH variations are potentially important in the use of this probe. The rate constant for reaction of the semiquinone with oxygen at pH 7.4 is 5.3 x 10(8) M(-1) s(-1): this reaction, rather than disproportionation of DCFH(.-)/DCF(.)(-), generates DCF in biological systems, concomitantly forming superoxide and hence H(2)O(2) to cycle the catalyst. The midpoint reduction potential of the couple DCF,H(+)/DCFH() is approximately -0.75 V vs. NHE at pH 7.4; DCF is unlikely to be reduced rapidly by common flavoprotein reductases.