Eosin derivatives

Several derivatives of Eosin have been prepared and employed to study biological systems. Their main applications are as singlet energy acceptors and as triplet probes [93-97] to measure the rotational mobility of virus particles [98] and proteins in membranes and in solution. Examples of proteins studied using Eosin derivatives include myosin [99,100], band 3 protein [101,102], pyruvate dehydrogenase [103,104], and Sarcoplasmic... 

Reticulum ATPase [105,106], Owing to the long-lived nature of the triplet state, Eosin derivatives are suitable to study protein dynamics in the microsecond-millisecond range. Rotational correlation times are obtained by monitoring the time-dependent anisotropy of the probe s phosphorescence [107-112] and/or the recovery of the ground state absorption [113— 118] or fluorescence [119-122], The decay of the anisotropy allows determination of the mobility of the protein chain that cover the binding site and the rotational diffusion of the protein, the latter being a function of the size and shape of the protein, the viscosity of the medium, and the temperature. 

Determination of translational diffusion rates of proteins requires measurements at longer timescales, one-tenth of a second to several minutes. Eosin derivatives are also commonly used to measure translational diffusion coefficients using the Fluorescence Recovery After Photobleaching technique [138-141],... 

Fluorescein is obtained by condensing phthahc anhydride (1 mol) with resorcinol (2 mols) in the presence of anhydrous zinc chloride. The tetra-bromo derivative, readily prepared by the addition of the calculated quantity of bromine, is eosin. 

Eosin (Tetrabromofluorescein). Place 16 5 g. of powdered fluorescein and 80 ml. of rectified (or methylated) spirit in a 250 ml. flask. Support a small dropping funnel, containing 36 g. (12 ml.) of bromine, above the flask make sure that the stopcock of the funnel is well lubricated before charging the latter with bromine. Add the bromine diopwise during about 20 minutes. When half the bromine has been introduced, and the fluorescein has been converted into dibromofluor-escein, all the solid material disappears temporarily since the dibromo derivative is soluble in alcohol with further addition of bromine the tetrabromofluorescein (sparingly soluble in alcohol) separates out. Allow the reaction mixture to stand for 2 hours, filter ofiF the eosin at the pump, wash it with alcohol, and dry at 100°. The yield of eosin (orange-coloured powder) is 25 g. 

Eosin Flavonoids Morin Flavonol, fisetin, robinetin Quercetin Rutin condensation products of urea, formaldehyde and methanol [126], pesticide derivatives [127] sweetening agents [128, 129] anion-active and nonionogenic surface-active agents [130] steroids, pesticides [29,132, 133] pesticides [134—137] vanadium in various oxidation states [138] uracil derivatives [139]... 

The basis of this method is that when normal seawater is chlorinated at the usual levels of 1 to 10mg/l of chloride, the bromine in seawater (8.1 x 10 4 M, 65 mg/1 at salinity = 35%o) is rapidly and quantitatively oxidised to Br() and HBrO. If 50 mg/1 of bromide is added to distilled or fresh waters containing HCIO plus C1CT, then HBrO plus BrO" are both formed. The HBrO plus BrO" will in turn rapidly brominate fluorecein (9-[o-carboxyphenyl]-6-hydroxy-3-isoxanthenone) to give the pink tetrabromo derivative eosin yellow (2,4,5,7-tetrabromo-9-[o-carboxyphenyl]-6-hydroxy-3-isoxanthenone), provided the molar ratio of bromide to fluorescein is 4 1. The resultant increase in eosin can be measured visually or spectrophotometrically, and the decrease in fluo-roscein measured fluorometrically. If the molar ratio of bromide to fluoroscein is < 4 1, then the mono-, di-, and tri-bromo derivatives are formed repro-ducibly. These derivatives have extinction coefficients close to eosin and are accounted for in the standardisation. 

In 1888, Walter studied the quenching of fluorescence, by the concentration effect, of fluorescein solutions. Nicols and Merrit observed in 1907, in solutions of eosine and resoruflne, the symmetry existing between their absorption and fluorescence spectra. In 1910, Ley and Engelhardt determined the fluorescence quantum yield of various benzene derivatives, values that were still referred to until recent years [18], The works by Lehmann and Wood, around 1910, marked the beginning of analysis based on fluorescence [4],... 

Brominating fluorescein in the presence of sodium chlorate affords the red tetrabromo derivative, whose sodium salt is known as eosine (Caro 1871). Sodium chlorate reoxidizes, evolving hydrogen bromide, to bromine which can react again ... 

The bromination product is actually contaminated with a certain percentage of mono to tribromo derivative. Although impurities, these side products apparently furnish the good tinctorial strength of P.R.90. Eosine itself is only a tinctorially weak pigment. 

Like quinone itself, its derivatives are also converted by reducing agents into colourless benzenoid hydrogenated products (in the case of the dyes they are called leuco-compounds ). The following scheme expresses this process for eosin as well as for the other dye ... 

The second family ofxanthene dyes is fluorescein and its derivatives. Fluorescein itself is only slightly fluorescent in alcohol solution. In contrast, the alkali salt obtained by addition of alkali exhibits the well-known yellow-green fluorescence characteristic of the fluorescein dianion (uranin). Fluorescein and its derivatives, e.g. eosin Yand erythrosin Y, are known to be very sensitive to pH and can thus be used as pH fluorescent probes (see Chapter 10). 

Photooxygenation of a-terpinene 155 in the presence of eosin (equation 87) produces ascaridole 156, a constituent of the essential oil Chenopodium ambrosioides L 1. The ewrfo-peroxide 157 derived from cyclopentadiene is a crystalline solid, stable at —100 °C78 above this temperature it rearranges to a mixture of the bis-epoxide 158 and the epoxyaldehyde 159 (equation 88)79,80. 

These classes of chemicals contain not only some of the oldest dyes, dating back to the 19th centnry, bnt also some of the most important flnorescent chromophores. The earliest of these discoveries was flnorescein (3.21), which was later brominated to give the polybromo derivative eosin (3.22) (Fignre 3.10). They are of little importance in textile coloration processes bnt are nsed in some fluorescent pigments and extensively in analysis and biological methods and also in tracing the course of water in drains and other water outlets. 

Eosin is a dye that is obtained by reaction of fluorescein with bromine (Eq. 9). This reaction has been carried out in batch and flow reactors, and both eosin and erythrosine (the tetraiodine derivative) have been obtained. The mechanism and... 

A nanocrystalline Sn02 solar cell sensitized by a perylene derivative produced 0.9% efficiency under AM 1.5 (7 = 3.26 mA/cm2 and = 0.45 V) [132]. A nanocrystalline Ti02 solar cell (1 cm2) sensitized by eosin Y, oneof the... 

Eosin Y stands for Eosin Yellowish and it is the disodium salt of tetrabromofluorescein. There are a number of other fluorescein derivatives which are also referred to as Eosins. Eosin B or bluish is 4,5-dibromo-3,7-dinitrofluorescein. The investigator is best advised to buy dyes on the basis of structure rather than name. In the case of the other dyes in the table the literature confusion is less imposing. 

Fluorescein is an energy acceptor for chromophores such as naphthalene and anthracene and acts as energy donor toward Eosin and Rhodamine, so derivatives have been used for singlet-singlet energy transfer studies. According to Forster s theory [68] the rate constant for energy transfer increases... 

The localization of the sensitizer has a significant effect on the photodynamic activity. Eosin is scarcely taken up by cells, and Pooler and Girotti [131] report that Eosin isothiocyanate is 50-100 times more efficient for inducing photohemolysis of human erythrocytes. The spectral properties and quantum yields for singlet oxygen production are identical for both compounds. Therefore, they attribute the difference in photodynamic activity to the ability of the isothiocyanate derivative to bind covalently to band 3 protein. 

The photochemistry of Eosin under both reductive and oxidative conditions has been studied by several groups [145-151], Photoreduction by amines such as tribenzylamine (R = CH2, R" = ) produces two leuco analogues, the dihydro derivative, and the cross-coupled product formed from the amine radical and the dye radical anion (2) [152], In addition, debromination of Eosin is reported during photobleaching with amines and phenols. The reader however is referred to the extensive studies of Rose Bengal dehalogenation by Paczkowski and Neckers [153]. Radiolysis of Eosin in methanol shows that debromination is a consequence of the photochemical decomposition of semireduced Eosin [154],... 

Rose Bengal differs in reactivity from fluorescein and Eosin because of the iodines at C-2, C-4, C-5, and C-7 which differentiate the reactivity of the phenoxide from the carboxylate and make substitution at C-2 possible in lieu of substitution at C-6. Essentially three different kinds of Rose Bengal derivatives have been synthesized. The most common include derivatives with gegen ions other than sodium at both C-2 and C-6. Many esters of C-2 have been synthesized and compounds with a number of different gegen ions at C-6 have been reported. The rarest derivatives of Rose Bengal are the C-6 ethers, of which only three have been reported. 

Several derivatives of xanthene are of value as colouring matters and the topic has been reviewed (B-52MI22400). For example, eosin (701 R = Br) (Acid Red 87 C.I. 45380) is used in inks, lipsticks and nail varnish in addition to its value as a dye for silk and paper. 

Dyes, parphinc derivative used for chlorophyll, phlhalocyantnes. and hemin. t Dyes, xanthene used for eosin. fluorcscein-iype phthatcins. rlttxiamines. and mse hengal. 

Arakawa et al.82) employed various 9-phenyl xanthene derivatives as the dye for DSC, and found that Eosin Y (EY Dye 18) shows relatively high conversion efficiency of 1.3% under 100 mW-cm"2 irradiance. They clarified the efficiency of 9-phenyl xanthene derivatives as follows EY > Dibromofluorescein > Fluorescein = Fluorescin > Rhodamin B > Dichlorofluorescein > Uranine >... 

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