The photoyellowing of stilbene-derived fluorescent whitening agents--mass spectrometric characterization of yellow photoproducts.

The application of fluorescent whitening agents (FWAs) significantly accelerates the photoyellowing of wool and silk under exposure to the ultraviolet and visible components of sunlight <500 nm. The photochemistry involved in this process is poorly understood, particularly the role of photoproducts derived directly from the FWA itself. Hydroxylation was identified as the key initial mechanism of photodegradation leading to coloration of the solution in the irradiation of the stilbene-derived FWA 4,4'-bis(2-sulfostyryl)biphenyl (DSBP) in the presence of hydrogen peroxide (H2O2). Polyhydroxylated DSBP derivatives were implicated as critical intermediates in the formation of yellow photoproducts under these conditions. The formation of trace quantities of DSBP quinone derivatives subsequent to hydroxylation was identified as the key cause of DSBP photoyellowing. These results are the first successful characterization of yellow photoproducts resulting directly from irradiation of a stilbene-based FWA. Formation of these yellow stilbene-based FWA-derived photoproducts may occur on the surface of FWA-treated wool exposed to simulated sunlight, as previous work has shown that H2O2 is photogenerated when wet FWA-treated wool is exposed to light. These results therefore suggest that yellow FWA-derived photoproducts contribute to the accelerated photoyellowing of FWA-treated wool.     

The Photoyellowing of Stilbene-derived Fluorescent Whitening Agents—Mass Spectrometric Characterization of Yellow Photoproducts

The application of fluorescent whitening agents (FWAs) significantly accelerates the photoyellowing of wool and silk under exposure to the ultraviolet and visible components of sunlight <500 nm. The photochemistry involved in this process is poorly understood, particularly the role of photoproducts derived directly from the FWA itself. Hydroxylation was identified as the key initial mechanism of photodegradation leading to coloration of the solution in the irradiation of the stilbene-derived FWA 4,4'-bis(2-sulfostyryl)biphenyl (DSBP) in the presence of hydrogen peroxide (H₂O₂). Polyhydroxylated DSBP derivatives were implicated as critical intermediates in the formation of yellow photoproducts under these conditions. The formation of trace quantities of DSBP quinone derivatives subsequent to hydroxylation was identified as the key cause of DSBP photoyellowing. These results are the first successful characterization of yellow photoproducts resulting directly from irradiation of a stilbene-based FWA. Formation of these yellow stilbene-based FWA-derived photoproducts may occur on the surface of FWA-treated wool exposed to simulated sunlight, as previous work has shown that H₂O₂ is photogenerated when wet FWA-treated wool is exposed to light. These results therefore suggest that yellow FWA-derived photoproducts contribute to the accelerated photoyellowing of FWA-treated wool.     

Determination of photo-oxidation products within photoyellowed bleached wool proteins

Photo-oxidative processes occurring in wool can lead to significant photoyellowing of the fiber. In particular, wool that has been chemically bleached photoyellows more rapidly and to a greater degree than untreated wool. Direct identification of the chromophores responsible for such yellow discoloration in irradiated wool has proven to be elusive for many years. This article describes the characterization and location of yellow photo-oxidation products within the proteins of photoyellowed bleached wool fabric, using advanced protein chemistry techniques. The discolored fabric was enzymatically digested and chromatographed by high-pressure liquid chromatography, with monitoring at 400 nm, to select out fractions containing yellow chromophoric species. Thorough tandem mass spectrometric analysis was then used to sequence peptides and, in turn, to characterize modifications to key amino acid residues that had resulted in yellow chromophore formation. In total, 11 separate yellow chromophoric species were identified, ten derived from tryptophan residues and one from tyrosine. The tryptophan-derived modifications characterized included hydroxytryptophan, N-formylkynurenine, hydroxyformylkynurenine, kynurenine, hydroxykynurenine, carbolines, tryptophandiones and nitrotryptophan. The tyrosine-derived modification of tyrosine to dopa was also identified. The range of photomodifications we observed provides insight into the photo-oxidation pathways occurring within irradiated fibrous proteins leading to the formation of yellow chromophores.     

AGGREGATION OF COLLAGEN EXPOSED TO UVA IN THE PRESENCE OF RIBOFLAVIN: A PLAUSIBLE ROLE OF TYROSINE MODIFICATION

Riboflavin-sensitized photodynamic modification of collagen led to significant formation of cross-linked molecules. Sodium azide or l,4-diazabicyclo(2,2,2)octane, which are known to be singlet oxygen quenchers, and catalase could not inhibit the modification. Surprisingly, the collagen modification was accelerated in the presence of superoxide dismutase. The aggregation was accompanied by the loss of tyrosine and histidine residues in the collagen. An inhibitory effect of dissolved oxygen on the modification of collagen was observed. Similarly, the loss of tyrosine residues in the irradiated collagen was inhibited in the presence of dissolved oxygen. Dityrosine formation was also observed with the loss of tyrosine. These results indicate that photodynamic modification of tyrosine probably contributes to the riboflavin-sensitized cross-linking of collagen through the formation of dityrosine.     

PHOTOCHEMICAL REACTIONS OF THYMINE WITH CARBOXYLIC ACIDS. A MODEL FOR NUCLEIC ACID-PROTEIN PHOTOCROSSLINKING

When nucleic acid bases are UV-irradiated in the presence of carboxylic acids or carboxylate anions new photoproducts are formed as compared to the bases irradiated in the absence of carboxylic acids. The behavior of thymine and thymidine has been examined in detail. At least four photoproducts are formed in the presence of propionic acid and three in the presence of butyric acid. None of them appears to be a cyclobutyl dimer. From the concentration dependence of the rate of photoproduct formation it is concluded that the reactive excited species is the first excited singlet state of thymine. When ¹⁴C-labelled thymine is irradiated in the presence of polyglutamic acid an important part of the radioactive material is covalently linked to the polymer. Photochemical reaction of thymine with glutamic (or aspartic) acid could thus induce crosslinks between proteins and nucleic acids. It is also shown that these photoproducts are stable under usual conditions of acidic hydrolysis of UV-irradiated DNA.     

PHOTOCHEMICAL REACTIONS OF THYMINE, URACIL, URIDINE, CYTOSINE AND BROMOURACIL IN FROZEN SOLUTION AND IN DRIED FILMS*

Abstract— The photochemistry of uracil, uridine, cytosine, thymine and broinouracil has been investigated in frozen aqueous solution and in dried films. Essentially the same photoproducts were obtained in the two conditions; however, the yield of photoproducts was considerably greater in frozen solution. Uracil forms a dimer which can exist in two forms. Some kinetic data are presented for the interconversion of these two forms, The mixed dimer of thymine and uracil can also exist in two forms. Uridine forms only one acid stable photoproduct and does not appear to form mixed photoproducts under the conditions used. Two new photoproducts of thymine other than the dimer are described. Cytosine was at first considered to be completely inert hut using more sensitive detecting equipment it has recently been found to form uracil dinier as a result of dinierization and deamination. The most remarkable response was shown by bromouracil. When irradiated by itself it formed no photoproducts but when irradiated in the presence of uracil, uridine, cytosine or NaOH it formed many photoproducts. Most of these products were devoid of bromide, but two still contained bromine. One of these has been identified as the mixed dimer of uracil and bromouracil while the other has been tentatively identified as the dimer of bromouracil. Dimers of thymine or bromouracil were not formed by X-rays.     

Determination of fluorescent whitening agents in laundry detergents and surface waters by solid-phase extraction and ion-pair high-performance liquid chromatography

A simple method was developed to detect four stilbene-type disulfonate and one distyrylbiphenyl-type fluorescent whitening agents (FWAs) in household laundry detergents and surface waters by ion-pair high-performance liquid chromatography. The FWA concentrations in detergents were measured directly. The contents of FWAs in water samples were extracted by solid-phase extraction (C18-SPE) with ion-pairing reagent, and were then determined by an isocratic ion-pair chromatography (IPC) using a C18 column, applying tetrabutylammonium hydrogensulfate (TBA) as the ion-pairing reagent in mobile phase, and equipped with fluorescence detection. Water samples at various pH conditions for SPE were evaluated. Experimental results indicate that the proposed method is precise and sensitive in analyzing FWAs, and enables quantitation of 0.01-0.1 microg/l in 100 ml water samples. The recovery rates of FWAs in spiked water samples were between 73 and 89%, and the precision (RSD) ranged from 2.6 to 8.9%. Over 7200 microg/g of 4,4'-bis(2-sulfostryl)-biphenyl (DSBP) and 2320 microg/g of 4,4'-bis[(4-anilino-6-morpholino-1,3,5-triazine-2-yl)-amino]stilbene-2,2'-disulfonate (DAS1) were detected in household laundry detergents. Trace amounts of DSBP were detected in surface water samples ranging from 0.2 to 3.7 microg/l.     

Photooxidation of cellulose treated with amino compounds

The effect of short wavelength UV radiation on celluloses treated with different amino compounds was studied. Samples of cotton cellulose were treated in aqueous solutions of different pH with polyethylenimine (PEI), hexamethylenetetramine (HMTA) and ethylenediamine (EDA), and irradiated in air with a low pressure mercury vapour lamp for different times. The photoyellowing was measured using standard colorimetry and the degradation processes and products were characterized using diffuse reflectance (FTIR and UV-visible) spectroscopy. The degradation processes were compared with those observed in the same materials during thermal treatments. Celluloses treated with EDA and PEI show an intense photoyellowing which depends on the pH of the treating solution. Colour changes in these materials correlate well with the appearance of imine groups and the disappearance of amino groups, determined by FTIR measurements. However, HMTA-treated cellulose, which suffers an intense thermal yellowing, shows only insignificant photoyellowing. These results were explained considering that the photoyellowing is due to the coloured Schiff bases produced in the condensation between amino and carbonyl groups.     

Photochemical studies on a phenolic phenylcoumarone lignin model molecule in relation to the photodegradation of lignocellulosic materials. Part 1. Structure of the photoproducts

The contribution of phenols, absorbing above 300 nm, to the photodegradation of lignin was approached by studying a phenolic phenylcoumarone lignin model molecule PCO. The compound was irradiated in solution and adsorbed on filter paper. The isolation and analysis of the photoproducts formed during the irradiation indicate the presence of a catechol structure (involving demethylation of the starting material) and two dimeric stilbene compounds: a ketone and a conjugated quinone methide giving high colouration to the irradiated solution. Those compounds have been detected as well in the solid state. The structure elucidation of the photoproducts was mainly based on mass spectrometry and 2D NMR experiments at 500 MHz. The major role played by the phenol group was supported by studying the O-methylated analogue PCOMe.     

Changes induced by ultraviolet light in fluorescence of collagen in the presence of β-carotene

The influence of UV radiation (253.7 nm) on collagen fluorescence in the absence, and presence, of β-carotene was investigated. It was found that UV radiation of 253.7 nm causes irreversible destruction of tyrosyl and phenylalanyl residues. The fluorescence of collagen (excitation at 275 nm, emission at 305 nm) decreased rapidly during irradiation and a new fluorescence large band at 400–500 nm formed under UV radiation. Smaller changes in the fluorescence of collagen in the presence of β-carotene suggest that it makes collagen less sensitive to the action of UV radiation.     

Effect of UV irradiation on the physico-chemical properties of iron crosslinked collagen

Collagen is the main component of connective tissue and finds immense applications as a biomaterial. In this study, effect of UV irradiation on collagen crosslinked with iron has been carried out. The physical and optical properties of crosslinked collagen affected by UV irradiation were analyzed using electrospectral and fluorescence studies. The electronic spectral studies showed that the photoproducts formed on UV radiation decrease in the presence of iron. Circular dichroic studies revealed that the conformational changes brought about in the protein due to UV irradiation have been reduced owing to the crosslinking with iron. However, prolonged irradiation does bring about conformational changes to the protein.     

UVA- and UVB-induced changes in collagen and fibronectin biosynthesis in the skin of hairless mice.

The modifications induced in hairless mouse skin by chronic UV irradiation were investigated. Skin explant cultures were used to study UVA- and UVB-induced changes occurring in interstitial collagen (type I and type III) and fibronectin biosynthesis. To study the long-term effects, albino hairless mice were irradiated with UVA radiation alone from two sources with different spectral qualities or with UVB. UVA and UVB radiation produced a significant increase in the ratio of type III to type I collagen (more than 100% for UVA-irradiated skin and about 60% for UVB-irradiated skin) accompanied by a significantly increased fibronectin biosynthesis (50% or more in all irradiated groups). Irradiation with either UVA or UVB alone had no significant effect on the total collagen synthesis and resulted in only a slight decrease in the total collagen content of the skin determined as hydroxyproline. This decrease was significant only in the case of the group irradiated with UVA (xenon) (decrease of 25%, expressed as micrograms of hydroxyproline per milligram wet weight). A significant decrease in collagen hydroxylation (expressed as radioactive hydroxyproline/radioactive hydroxyproline plus proline in neosynthesized collagen) was observed of about 50% in skin irradiated with UVA (xenon) but not in UVB-treated skin. Several of the above modifications (increased fibronectin biosynthesis, increased collagen type III to type I ratio) correspond to the modifications observed during the aging of non-irradiated hairless mice. Therefore it appears that UV irradiation accelerates the modifications of extracellular matrix biosynthesis observed during aging.     

P-AMINOBENZOIC ACID CAN SENSITIZE THE FORMATION OF PYRIMIDINE DIMERS IN DNA: DIRECT CHEMICAL EVIDENCE

Thymine-containing photoproducts with chromatographic properties similar to those of cyclobutyl pyrimidine dimers can be formed in [³H]-thymine-labeled DNA in solution by 313 nm ultraviolet radiation in the presence of para-aminobenzoic acid (PABA), a compound used in sunscreen preparations. In the absence of PABA, similar fluences of 313 nm radiation do not produce significant numbers of these photoproducts. The thymine-containing photoproducts can be reversed by 254 nm radiation so that the tritium label migrates with the mobility of thymine monomer, a behavior characteristic of thymine-containing cyclobutyl pyrimidine dimers. This result supports previous, but less direct, data from other laboratories indicating that PABA can sensitize dimer formation in the DNA of bacterial and mammalian cells.     

Model Systems for Evidencing the Mediator Role of Riboflavin in the UVA Cross-Linking Treatment of Keratoconus

Riboflavin under UVA radiation generates reactive oxygen species (ROS) that can induce various changes in biological systems. Under controlled conditions, these processes can be used in some treatments for ocular or dermal diseases. For instance, corneal cross-linking (CXL) treatment of keratoconus involves UVA irradiation combined with riboflavin aiming to induce the formation of new collagen fibrils in cornea. To reduce the damaging effect of ROS formed in the presence of riboflavin and UVA, the CXL treatment is performed with the addition of polysaccharides (dextran). Hyaluronic acid is a polysaccharide that can be found in the aqueous layer of the tear film. In many cases, keratoconus patients also present dry eye syndrome that can be reduced by the application of topical solutions containing hyaluronic acid. This study presents physico-chemical evidence on the effect of riboflavin on collagen fibril formation revealed by the following methods: differential scanning microcalorimetry, rheology, and STEM images. The collagen used was extracted from calf skin that contains type I collagen similar to that found in the eye. Spin trapping experiments on collagen/hyaluronic acid/riboflavin solutions evidenced the formation of ROS species by electron paramagnetic resonance measurements.     

Phototransformation of imidacloprid on isolated tomato fruit cuticles and on tomato fruits

Imidacloprid, a neonicotinoid insecticide, is widely used in plant protection to prevent crop losses. The objective of this study was to show the photochemical fate of imidacloprid on plant surfaces by irradiation experiments on isolated tomato fruit cuticles and tomato fruits (Lycopersicon esculentum Mill.). Imidacloprid spiked samples were irradiated both under sunlight simulator and natural sunlight conditions for 24 h, which resulted in recoveries of 23% and 24%, respectively, if isolated cuticles were studied. On whole tomato fruits, recoveries were 33% and 71%, respectively. Similar results were obtained on cuticles spiked with the formulation Confidor and irradiated under natural sunlight. However, on tomato fruits the application of Confidor resulted in a higher loss (56%) of imidacloprid. During all irradiation experiments both on cuticles and whole fruits, 1-[(6-chloropyridin-3-yl)methyl]imidazolidin-2-imine was generally formed at 10–14 mol%, but different other photoproducts were also detected in low amounts, whereas N-nitrosoimidacloprid was only detected under natural sunlight conditions. Rapid photodegradation of imidacloprid could be demonstrated in all experiments. The identified photoproducts, 1-[(6-chloropyridin-3-yl)methyl]imidazolidin-2-imine and N-nitrosoimidacloprid, are possible reaction partners for plant cuticle constituents to form cuticle bound residues.     

A SURVEY OF PHOTOPRODUCTS OF AN IRRADIATED OLIGODEOXYNUCLEOTIDE BY MONOCHROMATIC PHOTONS WITH THE ENERGY RANGED FROM 6.5 TO 22.5 eV

We have irradiated 2'-deoxyadenylyl-(3'-5')-2'-deoxyadenosine (dephospho(dA)₂) in solid state with a newly developed synchrotron irradiation system over 6.5 to 22.5 eV photon energy range (190 to 55 nm in wavelength). Densitometric analysis of thin-layer chromatogram of photoproducts indicated that adenine and deoxyadenosine monophosphate were commonly produced above 7.3 eV (170 nm) in approximately equal amount. Deoxyribose was not detected by the fluorescence method with scratched materials from the chromatogram at any location except deoxyadenosine monophosphate and parent oligonucleotide positions. The photoproducts above mentioned were not observed at 6.5 eV (190 nm) even after very high radiation fluence. The energy dependence of this degradation indicates that the shorter the wavelength in use the smaller the fluence it requires. Careful examination of irradiated oligonucleotide on the chromatograms shows that the major portion of deoxyadenosine monophosphate so produced was the 5'phospho derivative. This site-selective degradation, independent of photon energy, seemed to be a remarkable characteristic of vacuum-UV induced effects above 7 eV. From these results a working hypothesis was advanced that the deoxypentose moiety was first destroyed in one of the nucleoside residues due to the strong absorption in the sugar-phosphate group, liberating adenine on the one hand and leaving 5'-deoxyadenosine monophosphate on the other hand.     

Identification of phototransformation products of sildenafil (Viagra) and its N‐demethylated human metabolite under simulated sunlight

Recent publications on pharmaceutical monitoring are increasingly covering the field of illicit drugs and lately the forensic evaluation of designing illegal analogs of lifestyle drugs like the phosphodiesterase type 5 (PDE‐5) inhibitors Viagra (sildenafil), Levitra (vardenafil) and Cialis (tadalafil). Recently, the presence of all three erectile dysfunction treatment drugs has been reported in wastewaters at very low concentrations. In the environment, contaminants undergo various physical or chemical processes classified into abiotic (photolysis, hydrolysis) and biotic (biodegradation) reactions. Thus, changes in the chemical structure lead to the formation of new transformation products, which may persist in the environment or be further degraded. This study describes the photolysis of sildenafil (SDF) and its human metabolite N‐demethylsildenafil (DM‐SDF) under simulated solar radiation (Xenon lamp). Following chromatographic separation of the irradiated samples, eight photoproducts in the SDF samples and six photoproducts for DM‐SDF were detected and characterized. The combination of ultra performance liquid chromatography‐electrospray ionization‐quadrupole time‐of‐flight‐mass spectrometry (UPLC‐ESI‐QToF‐MS), liquid chromatography‐atmospheric pressure chemical ionization‐triple quadrupole mass spectrometry (LC‐APCI‐QqQ‐MS) and hydrogen/deuterium‐exchange experiments allowed to propose plausible chemical structures for the photoproducts, taking into account the characteristic fragmentation patterns and the accurate mass measurements. These mass spectral data provided sound evidence for the susceptibility of the piperazine ring toward photodegradation. A gradual breakdown of this heterocyclic structure gave rise to a series of products, which in part were identical for SDF and DM‐SDF. The sulfonic acid, as the formal product of sulfonamide hydrolysis, was identified as key intermediate in the photolysis pathway. In both drug/metabolite molecules, phototransformation processes taking place beyond the sulfonamide group were deemed to be of minor relevance. Copyright © 2012 John Wiley & Sons, Ltd.     

电子受体H2O2和O2对TiO2光催化甘油氧化反应中活性氧物种1O2和OH?的影响

在室温条件下研究了电子受体H2O2和O2对TiO2光催化甘油氧化反应中的活性氧物种、甘油转化率和产物分布的影响。当在紫外光辐射和TiO2的体系中不存在电子受体时,只产生HO?自由基。而当在此体系中有电子受体存在时,则产生了HO?自由基和1O2,但它们的浓度不同,这取决于电子受体的浓度。以H2O2为电子受体时甘油转化率的提高大于以O2为电子受体时。甘油转化生成有价值产物的类型则与体系中的活性氧物种浓度有关。     

Excision of cytosine-thymine adduct from the DNA of ultraviolet-irradiated Micrococcus radiodurans

Abstract— 6-4‘-[pyrimidin-2’-one]-thymine (PO-T), a deamination product of a cytosine–thymine adduct accounted for about 17 per cent of the total detectable thymine-derived photoproducts in u.v.-irradiated Micrococcus radiodurans. On incubation of the irradiated cells, oligonucleotides containing the photoproducts were released into the medium. After various periods of incubation the different photoproducts were isolated both from the cells and from the medium. Analysis of these different photoproducts showed that during the excision process the ratio of PO–T to total thymine-derived products remained constant both in the cells and in the medium. This shows that the precursor to PO–T is excised at the same rate as the other thymine-derived photoproducts by the dark repair mechanisms exhibited by this radiation-resistant organism.