And the molecular formula of this chemical is C12H8N2. It is a kind of off-white powder, and belongs to the following product categories:
Derivatives of the above acids reactive toward proteins can be readily manufactured by relatively simple synthetic routes, and are useful in fluorescent immunoassay.
Description This invention relates to reagents for fluorescent assay and more especially to novel markers which can be attached to protein for purposes of fluorescent labelling such as used in immunoassay methods, and to immunoassay methods using the novel markers. Radio-immunoassay RIA methods have been used for some years, but there are hazards involved in handling, storage, and disposal of radioactive materials and there has been growing opposition against the use of radioisotopes in some countries.
Furthermore, there are disadvantages associated with radioimmunoassays methods, such as the short shelf life of I-labelled antibodies and antigens weeks arising from the short half-life of I 60 days and this necessitates frequent radioiodinations and often requires disposal of unused outdated reagents.
This increases the cost per test associated with RIAs for the end user. The various drawbacks and disadvantages have led to the current trend to replace radioimmunoassays RIAs with nonisotopic methods, the most important of which are enzyme immunoassay EIA and fluorescent immunoassay 1 10 phenanthroline.
Although EIAs have found wide applicability recently, there is great concern about their sensitivity and reproducibility. In some cases, there is uncertainty about the choice of end point which should be chosen for the enzymatic reaction after the addition of substrate.
The measurement of the enzyme activity rate constant is a more reliable method but this prolongs 1 10 phenanthroline assay and adds to its cost by requiring more sophisticated computational hardware. The high sensitivity of enzymatic activity to temperature difference is also a source of imprecision.
Another major source of error is the presence in serum of enzymes which have effects similar to the one employed for labeling or of enzyme inhibitors which would-have the opposite effect. In current fluorescent immunoassays, immunoreactive proteins, i. Generally, the sensitivity of FIAs which employ these labels is lower than that of RIAs because of Rayleigh and Raman scattering and interference due to fluorescent substances such as proteins and bilirubin in serum.
The scattering problem is partly overcome with the use of dedicated filter systems.
The underlying problem with fluorescent labels of current general use is their small Stokes shift the difference between the wavelengths of the absorbed, stimulating radiation and of the emitted, fluorescent radiation, respectively and their short fluorescent lifetimes which are comparable with those of interfering substances in serum.
For example, fluorescein has absorption and emission maxima at nm and nm, respectively, and a fluorescence lifetime of only 5 nanoseconds. Recently, the use of lanthanide chelates, especially those of europium, has been suggested as a way of eliminating background fluorescence to increase immunoassay sensitivity, for example in United States patent 4, issued February 15, in the name Soini et al.
The wavelengths of the absorption and emission maxima for these systems depend on the chelating ligands but the Stokes shifts are typically at least nm and the fluorescence lifetimes are in the order of microseconds.
Background fluorescence can be eliminated completely with the use of time-resolved fluoresence immunoassay, for example by using of a gated fluorometer wherein a time delay between a pulsed excitation and detection of emission removes short-lived fluorescence due to interfering fluorochromes.
Lanthanide chelates have the unique characteristic of absorbing light to give a high yield of linelike emission with long fluorescence lifetimes.
The emission results from intramolecular energy transfer from the lowest ligand triplet states to the metal ion, as reported by M. Time-resolved spectroscopy of europium chelates using a stroboscopic technique was performed by Bhaumik and co-workers inM.
Although the fluorescent chelates of lanthanides, as described for example in the above-mentioned United States patent 4,, provide acceptable results in fluorescence spectroscopy analysis, the chelates of this patent are difficult to manufacture as their preparation requires a multi-step process which is difficult and complex to commercialize.
We have now found that certain novel 1,phenanthroline derivatives form lanthanide chelates which fluoresce in aqueous solution, and which may be employed in fluorescent immuno- and other assays. These derivatives can be readily manufactured by relatively simple procedures.
We have found that the known compound 1,phenanthroline-2,9-dicarboxylic acid of the formula is a ligand which forms chelates with lanthanide salts which are highly fluorescent in aqueous solution.
Further, as explained in more detail below, we have found various novel derivatives of the above acid form lanthanide chelates of like fluorescent capabilities.
In some cases, the molecular structure of these derivatives enhances and advantageously modifies their radiation-absorbing and fluorescent radiation- emitting characteristics.
These derivatives include compounds containing one or more functional groups that are capable of coupling covalently with proteins, or one or more groups readily convertible to such functional groups, and which are useful as markers for conducting fluorescent immuonassay.
In the above formula R1 to R6 represent groups which may be substituted in the compound of formula I without deleteriously affecting the fluorescent activity of lanthanide chelates formed therefrom.Reaction of Ferrous and Ferric Iron with 1,Phenanthroline.
III. The Ferrous Monophenanthroline Complex and the Colorimetric Determination of Phenanthroline. The coordinating ability of 1, phenanthroline is possessed by a number of its simple derivatives.
 The quaternary mixed anion of rare earth with 2,3-dimethoxylbenzoic acid and 1,phenanthroline has been synthesized from the water-ethanol solution. Elemental analysis shows that the complexes general formula is RE(2,3-DMOBA)2NO3Phen (RE. Buy 1,Phenanthroline (CAS ), a chelating agent and metalloprotease inhibitor.
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Send questions or comments to doi. 1,Phenanthroline: a versatile ligand. Peter G. Sammes and Gokhan Yahioglu Abstract. The first page of this article is displayed as the abstract. About. Cited by. Related.
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