Antioxidants and protein oxidation

Proteins are susceptible to oxidation by reactive oxygen species, where the type of damage induced is characteristic of the denaturing species. The induction of protein carbonyls is a widely applied biomarker, arising from primary oxidative insult. However, when applied to complex biological and pathological conditions it can be subject to interference from lipid, carbohydrate and DNA oxidation products. More recently, interest has focused on the analysis of specific protein bound oxidised amino acids. Of the 22 amino acids, aromatic and sulphydryl containing residues have been regarded as being particularly susceptible to oxidative modification, with L-DOPA from tyrosine, ortho-tyrosine from phenylalanine; sulphoxides and disulphides from methionine and cysteine respectively; and kynurenines from tryptophan. Latterly, the identification of valine and leucine hydroxides, reduced from hydroperoxide intermediates, has been described and applied. In order to examine the nature of oxidative damage and protective efficacy of antioxidants the markers must be thoroughly evaluated for dosimetry in vitro following damage by specific radical species. Antioxidant protection against formation of the biomarker should be demonstrated in vitro. Quantification of biomarkers in proteins from normal subjects should be within the limits of detection of any analytical procedure. Further to this, the techniques for isolation and hydrolysis of specific proteins should demonstrate that in vitro oxidation is minimised. There is a need for the development of standards for quality assurance material to standardise procedures between laboratories. At present, antioxidant effects on protein oxidation in vivo are limited to animal studies, where dietary antioxidants have been reported to reduce dityrosine formation during rat exercise training. Two studies on humans have been reported last year. The further application of these methods to human studies is indicated, where the quality of the determinations will be enhanced through inter-laboratory validation.

Synthesis and evaluation of novel polymer modifiers for biodegradable polymers

The effects of ester plasticizers and copolymers on the mechanical properties of the natural biodegradable polymers, poly(3-hydroxybutyrate) [PHB] and poly(lactic acid) [PLA] have been studied after subjecting to melt processing conditions. Ester plasticizers were synthesized from citric, tartaric and maleic acids using various alcohols. A variety of PLA copolymers have also been prepared from poly(ethylene glycol) derivatives using stannous octanoate catalysed ring opening polymerisations of DL-lactide. A novel PLA star copolymer was also prepared from an ethoxylated pentaerythritol. The structures of these copolymers were determined by NMR spectroscopy. The plasticizing effect of the synthesised additives at various concentrations was determined. While certain additives were capable of improving the mechanical properties of PLA, none were effective in PHB. Moreover, it was found that certain combinations of additives exhibited synergistic effects. Possible mechanisms are discussed. Biotic and abiotic degradation studies showed that the plasticizers (esters and copolymers) did not inhibit the biodegradability of PHB or PLA in compost at 60°C. Simple toxicity tests carried out on compost extract and its ability to support the growth of cress seeds was established. PLA was found to be susceptible to limited thermal degradation under melt processing conditions. Conventional phenolic antioxidants showed no significant effect on this process, suggesting that degradation was not predominantly a free radical process. PLA also underwent photo-oxidative degradation with UV light and the process could be accelerated in the presence of a photoactivator such as iron (III) diisononyl dithiocarbamate. The mechanisms for the above processes are discussed. Finally, selected compounds were prepared on a pilot plant scale. Extruded and blown films were prepared containing these additives with conventional polymer processing equipment. The mechanical properties were similar to those obtained with laboratory produced compression moulded films.

Reactive processing of polyolefins using antioxidant systems

Various monoacrylic compounds containing a hindered phenol function (e.g.3,5-di-tert.-butyl-4-hydroxy benzyl alcohol, DBBA and vinyl-3-[3',5'-di-tert.-butyl-4-hydroxy phenyl] propionate, VDBP), and a benzophenone function (2-hydroxy-4-[beta hydroxy ethoxy] benzophenone, HAEB) were synthesised and used as reactive antioxidants (AO's) for polypropylene (PP). These compounds were reacted with PP melt in the presence of low concentration of a free radical generator such a peroxide (reactive processing) to produce bound-antioxidant concentrates. The binding reaction of these AO's onto PP was found to be low and this was shown to be mainly due to competing reactions such as homopolymerisation of the antioxidant. At high concentrations of peroxide, higher binding efficiency resulted, but, this was accompanied by melt degradation of the polymer. In a special reactive processing procedure, a di- or a trifunctional reactant (referred to as coagent), e.g.tri-methylol propane tri-acrylate, Tris, and Divinyl benzene, DVB, were used with the antioxidant and this has led to an enhanced efficiency of the grating reaction of antioxidants on the polymer in the melt. The evidence suggests that this is due to copolymerisation of the antioxidants with the coagent as well as grafting of the copolymers onto the polymer backbone. Although the 'bound' AO's containing a UV stabilising function showed lower overall stabilisation effect than the unbound analogues before extraction, they were still much more effective when subjected to exhaustive solvent extraction. Furthermore, a very effective synergistic stabilising activity when two reactive AO's containing thermal and UV stabilising functions e.g. DBBA and HAEB, were reactively processed with PP in the presence of a coagent. The stabilising effectiveness of such a synergist was much higher than that of the unbound analogues both before and after extraction. Analysis using the GPC technique of concentrates containing bound-DBBA processed in the presence of Tris coagent showed higher molecular weight (Mn), compared to that of a polymer processed without the coagent, but was still lower than that of the control processed PP with no additives. This indicates that Tris coagent may inhibit further melt degradation of the polymer. Model reactions of DBBA in liquid hydrocarbon (decalin) and analysis of the products using FTIR and NMR spectroscopy showed the formation of grafted DBBA onto decalin molecules as well as homopolymerisation of the AO. In the presence of Tris coagent, copolymerisation of DBBA with the Tris inevitably occured; which was followed by grafting of the copolymer onto the decalin, FTIR and NMR results of the polymer concentrates containing bound-DBBA processed with and without Tris, showed similar behaviour as the above model reactions. This evidence supports the effect of Tris in enhancing the efficiency of the reaction of DBBA in the polymer melt. Reactive procesing of HAEB in polymer melts exhibited crosslinking formation In the early stages of the reaction, however, in the final stage, the crosslinked structure was 'broken down' or rearranged to give an almost gel free polymer with high antioxidant binding efficiency.

Effect of processing on sulphur antioxidants in polyolefins

The effect of processing on the antioxidant activity of sulphur-containing compounds, with particular reference to nickel dialkyldithiophosphates and their corresponding di sulphides, were studied in polyolefins under melt, thermal and photo-oxidative conditions. These compounds were evaluated both at low (normal) and high (concentrates) concentrations. In general, the dithiophosphates were found to be very efficient melt stabilisers at normal concentrtion levels, and compare quite favourably with the best commercially available systems. The nickel dithiophosphates were also found to be very efficient thermal stabilisers for polyolefins, but their activity is highly dependent on the alkyl substituent in the molecule. The corresponding disulphides on the other hand showed very little activity under thermal oxidative conditions, and this was attributed to their inefficiency in scavenging alkyl peroxyl radicals since both compounds possess similar peroxidolytic activity. Furthermore, the nickel dithiophosphates were found to be excellent photo stabilisers for mildly-processed polyolefins while the corresponding disulphides only offer slight protection to the polymer. Oxidative processing of the disulphide, however, results in a dramatic improvement in their photo antioxidant activity. Thionophospho-ric acid, a major oxidation product of dithiophosphates, was also shown to have photo antioxidant activity similar to that of the disulphides. A combination of a U.V. absorber with the nickel complex and/or the disulphide resulted in a synergistic stabiliser system which was further augmented by oxidative processing. Moreover, the dilute analogues of such multicomponent stabiliser concentrates also showed excellent melt, thermal and photo-stabilising activity. The mechanistic studies carried out on the nickel complex and the corresponding disulphide clearly identified the thionophosphoric acid a a major transformation product although various triesters were formed as reaction intermediates. The mechanisms of the antioxidant action of the dithiophosphates, which is believed to involve a cyclical process similar to that shown for simple alkyl sulphides and nitroxyls, are discussed.

Rosemary : a raw material for a new genre of bio-based antioxidants for sustainable applications

The potential replacement, partially or fully, of synthetic additives by bio-based alternatives derived from indigenous renewable non-food crop resources offers a market opportunity for a green supply of raw materials for different industrial and health products, with greater involvement of the farming community in crop production while addressing the ever more stringent environmental and pollution laws that now require the use of less potentially toxic/harmful ingredients, even if they are present in relatively small quantities. The work presented here relates to developing a new genre of environmentally-sustainable bio-based antioxidants (AO) for industrial uses that are obtained from extracts of UK-grown rosemary (Rosmarinus officinalis) plant. The performance of these AOs was tested, and their efficacy compared with some common and benchmark synthetic AOs from the same chemical class, in different products including polymers especially for packaging, as well as lubricants, cosmetics and health products. One of the main active ingredients in rosemary is Rosmarinic acid which is a water-soluble compound. This was chemically transformed into a number of ester derivatives, Rosmarinates, targeted for different applications. The parent and the modified antioxidants (the rosmarinates) were characterised and their antioxidancy were examined and tested in linear low-density polyethylene (LLDPE) and in polypropylene (PP) and compared with compounds of similar structure and with other well known synthetic antioxidants used commercially in polyolefins. The results show that antioxidants sourced from rosemary have the added benefit of being highly efficient and intrinsically more active than many synthetic and bio-based alternatives.

Polymer optical fiber grating as water activity sensor

Controlling the water content within a product has long been required in the chemical processing, agriculture, food storage, paper manufacturing, semiconductor, pharmaceutical and fuel industries. The limitations of water content measurement as an indicator of safety and quality are attributed to differences in the strength with which water associates with other components in the product. Water activity indicates how tightly water is "bound," structurally or chemically, in products. Water absorption introduces changes in the volume and refractive index of poly(methyl methacrylate) PMMA. Therefore for a grating made in PMMA based optical fiber, its wavelength is an indicator of water absorption and PMMA thus can be used as a water activity sensor. In this work we have investigated the performance of a PMMA based optical fiber grating as a water activity sensor in sugar solution, saline solution and Jet A-1 aviation fuel. Samples of sugar solution with sugar concentration from 0 to 8%, saline solution with concentration from 0 to 22%, and dried (10ppm), ambient (39ppm) and wet (68ppm) aviation fuels were used in experiments. The corresponding water activities are measured as 1.0 to 0.99 for sugar solution, 1.0 to 0.86 for saline solution, and 0.15, 0.57 and 1.0 for the aviation fuel samples. The water content in the measured samples ranges from 100% (pure water) to 10 ppm (dried aviation fuel). The PMMA based optical fiber grating exhibits good sensitivity and consistent response, and Bragg wavelength shifts as large as 3.4 nm when the sensor is transferred from dry fuel to wet fuel. © 2014 Copyright SPIE.

New genre of antioxidants from renewable natural resources:synthesis and characterisation of rosemary plant-derived antioxidants and their performance in polyolefins

Several ester derivatives of rosmarinic acid (rosmarinates) were synthesised, characterised (1D and 2D NMR, UV and FTIR spectroscopy) and tested for their potential use as antioxidants derived from a renewable natural resource. The intrinsic free radical scavenging activity of the rosmarinates was assessed, initially using a modified DPPH (2, 2-diphenyl-1-picrylhydrazyl radical) method, and found to be higher than that of commercial synthetic hindered phenol antioxidants Irganox 1076 and Irganox 1010. The thermal stabilising performance of the rosmarinates in polyethylene (PE) and polypropylene (PP) was subsequently examined and compared to that of samples prepared similarly but in the presence of Irganox 1076 (in PE) and Irganox 1010 (in PP) which are typically used for polyolefin stabilisation in industrial practice. The melt stability and the long-term thermo-oxidative stability (LTTS) of processed polymers containing the antioxidants were assessed by measuring the melt flow index (MFI), melt viscosity, oxidation induction time (OIT) and long-term (accelerated) thermal ageing performance. The results show that both the melt and the thermo-oxidative stabilisation afforded by the rosmarinates, and in particular the stearyl derivative, in both PE and PP, are superior to those of Irganox 1076 and Irganox 1010, hence their potential as effective sustainable bio-based antioxidants for polymers. The rosmarinic acid used for the synthesis of the rosmarinates esters in this study was obtained from commercial rosemary extracts (AquaROX80). Furthermore, a large number of different strains of UK-grown rosemary plants (Rosmarinum officinalis) were also extracted and analysed in order to examine their antioxidant content. It was found that the carnosic and the rosmarinic acids, and to a much lesser extent the carnosol, constituted the main antioxidant components of the UK-plants, with the two acids being present at a ratio of 3:1, respectively.

Grafting functional antioxidants on highly crosslinked polyethylene

The problem of interference of antioxidants, such as hindered phenols, with peroxide-initiated crosslinking of polyethylene was addressed through the use of functional (reactive) graftable antioxidants (g-AO). Reactive derivatives of hindered phenol and hindered amine antioxidants were synthesised, characterised and used to investigate their grafting reactions in high density polyethylene; both non-crosslinked (PE) and highly peroxide-crosslinked (PEXa). Assessment of the extent of in-situ grafting of the antioxidants, their retention after exhaustive solvent extraction in PE and PEXa, and the stabilising performance of the grafted antioxidants (g-AO) in the polymer were examined and benchmarked against conventionally stabilised crosslinked & non-crosslinked polyethylene. It was shown that the functional antioxidants graft to a high extent in PEXa, and that the level of interference of the g-AOs with the polymer crosslinking process was minimal compared to that of conventional antioxidants which bear the same antioxidant function. The much higher level of retention of the g-AOs in PEXa after exhaustive solvent extraction, compared to that of the corresponding conventional antioxidants, accounts for their superior long-term thermal stabilising performance under severe extractive conditions.