Trace Analysis of Biochemical Substances by Near-Infrared Semiconductor Laser Fluorimetry

1.激光荧光法灵敏度高，适用于痕量分析。然而，激光器体积大且价格昂贵，因此它尚未被广泛用作光谱分析的常规工具。在这项研究中，我开发了一种新的光谱方法，使用一个小而便宜的半导体激光器。吲哚菁绿色在近红外区有一个吸收带，OH自由基可使其猝灭。用此反应方案可灵敏地测定产生OH自由基的过氧化物酶。该方法允许在10^ U水平下检测过氧化物酶<-4>。该方法也适用于酶免疫分析。基于夹心法，在U/ml水平上测定胰岛素.近红外半导体激光器的振荡波长被限制在780-900 nm，其分析应用受到很大限制。当激光紧密聚焦到非线性晶体中时，它会产生二次谐波发射（390-450 nm）。当使用KDP晶体时， ...更多信息 效率为2.5 × 10 - 4<-6>。当<-11>激光介质本身产生二次谐波发射时，它减小到1.7 × 10 ~ 4。这种发射被用作光纤传感器的光源，并用于中和滴定终点的检测。4.在近红外区域，只有少数物质是荧光的，并且它们中的大多数在水溶液中是化学不稳定的。因此，很难将其应用于实际分析应用。最近可用的可见光半导体激光器（670 nm）用于克服这个问题。用离子对试剂罗丹明800萃取间二甲苯后测定表面活性剂。可以检测到10 μ M水平的十二烷基硫酸钠<-7>。为了将可见半导体激光荧光法应用于生物化学分子，研究了可见染料荧光标记蛋白质的各种方法。结果表明，恶嗪750最适合作为本研究的标记试剂。少

1. Laser fluorimetry is very sensitive and is useful for trace analysis. However, a laser is large and expensive, so that it has not been widely used as a conventional tool for spectrometric analysis. In this study I develop a new spectrometric method using a small and less-expensive semiconductor laser.2. Indocyanine green has an absorption band in a near-infrared region, ant it is found to be quenched by OH radical. By using this reaction scheme peroxidase generating OH radical is sensitively determined. This method allows detection of peroxidase at 10^<-4> U levels. This method is also applied to enzyme immunoassay. Based on sandwich assay, insulin is determined at U/ml levels.3. The oscillating wavelength of the near-infrared semiconductor laser is restricted to 780-900 nm, and then its analytical application is quite limited. When the laser is tightly focused into a non-linear crystal, it generates second harmonic emission (390-450 nm). When a KDP crystal is used, the conversion e … More fficiency was 2.5 x 10^<-6>. It decreases to 1.7 x 10^<-11> when second harmonic emission is generated by the laser medium itself. This emission is used as a light source for an optical fiber sensor and for detection of the end-point in neutralization titration.4. In the near-infrared region only minor species are fluorescent, and most of them are chemically unstable in an aqueous solution. Therefore, it is difficult to apply it to practical analytical applications. A visible semiconductor laser (670 nm), which becomes recently available, is used to overcome this problem. A surfactant is determined after solvent extraction into m-xylene with an ion-pair reagent of Rhodamine 800. It is possible to detect sodium lauryl sulfate at 10^<-7> M levels. For application of visible semiconductor laser fluorimetry to a biochemical molecule, a various procedures are investigated for fluorescence labeling of protein with visible dyes. It is found that oxazine 750 is most suitable as a labeling reagent in this study. Less