Human intake assessment of triclosan associated with the daily use of polypropylene-made antimicrobial food packaging.

In this work, we aimed to evaluate human intake of triclosan (TCS) associated with real-life use of different brands of Microban™ microwave-safe food packaging. Calculations were based on: TCS migration data (under the worst-case foreseeable conditions), MPs abundance and TCS bioaccessibility from microplastics (MPs), leached from containers under microwave heating. Bioaccessibility studies were performed with in vitro digestion of MPs, followed by liquid-liquid extraction of TCS from digestive fluids and LC-QqQ-MS analysis yielding values of 46 ± 9%. The estimated weekly intake (EWI) of TCS ranged between 11 and 42 µg/kg body weight/week, with migration being the largest contribution (0.6-2.3 mg/week), compared to leaching of MPs (75-300 µg/week). These values represent a significant source of human exposure to TCS, emphasizing the need to harmonize the ban of TCS in food contact materials worldwide and improve compliance testing of food contact articles, particularly those marketed through online sales platforms.

Assessment of the spoilage microbiota in minced free-range chicken meat during storage at 4 C in retail modified atmosphere packages.

This study assessed the evolution of spoilage microbiota in association with the changes in pH and concentrations of lactic and acetic acids in retail oxygen-free modified atmosphere (30:70 CO2/N2) packages (MAP) of minced free-range chicken meat during storage at 4 °C for 10 days. MAP retarded growth of spoilage lactic acid bacteria (LAB) below 6.5 log cfu/g and fully suppressed growth of pseudomonads, enterobacteria, enterococci, staphylococci and yeasts. Two distinct Latilactobacillus sakei strain biotypes were predominant and Leuconostoc carnosum, Carnobacterium divergens, Latilactobacillus fuchuensis and Weissella koreensis were subdominant at spoilage. The chicken meat pH ranged from 5.8 to 6.1. l-lactate (832 mg/100 g on day-0) decreased slightly on day-7. d-lactate remained constantly below 20 mg/100 g, whereas acetate (0-59 mg/100 g) increased 5-fold on day-7. All MAP samples developed off-odors on day-7 and a strong 'blown-pack' sulfur-type of spoilage on day-10. However, neither the predominant Lb. sakei nor other LAB or gram-negative isolates formed H2S in vitro, except for C. divergens.

Effects of vacuum packaging storage of minimally processed cassava roots at various temperatures on microflora, tissue structure, starch extraction by wet milling and granule quality.

BACKGROUND: Vacuum package storage is commonly applied to reduce postharvest deterioration in minimally processed cassava roots. However, the influence of vacuum packaging conditions on root end-use quality is poorly understood. Hence, the effects of vacuum packaged storage at ambient, refrigerated and freezing temperatures on microflora, cassava tissue structure and starch extraction by wet milling were studied. RESULTS: Vacuum packaged storage temperature strongly affected cassava root quality. Minimal adverse effects were obtained with frozen storage. With refrigerated storage, there was negligible microbial growth but some disruption of the parenchyma cell wall structure suggestive of chilling injury. With ambient temperature storage, there was considerable Lactobacilli dominated fermentation. This caused substantial cell degradation, probably due to the production of extracellular cellulolytic and other cell wall degrading enzymes. A benefit of this cell wall breakdown was that it substantially improved starch extraction with wet milling from the stored cassava pieces; by 18% with pieces that had been ambient vacuum packaged and wet milled using a 2000 µm opening screen. However, ambient temperature storage resulted in some starch granule pitting due to the action of extracellular amylases from the fermenting microorganisms. CONCLUSION: The best vacuum packaging storage conditions for minimally processed cassava depends on application and cost. For short-term storage, refrigeration would be best for vegetable-type products. For several months storage, freezing is best. For wet milling applications, this could be combined with subsequent short-term ambient temperature storage as it improves starch extraction efficiency and could reduce distribution energy costs. © 2021 Society of Chemical Industry.

Packaging of beef fillet with active chitosan film incorporated with ɛ-polylysine: An assessment of quality indices and shelf life.

In the current study, the effect on packaged beef fillets (1 × 5 × 8 cm) of using active chitosan film (1%) was investigated. The fillets were stored at 4 °C for 12 days, and the film contained ɛ-polylysine (ɛ-PL) (0.3, 0.6, and 0.9% w/w). Chemical, microbiological, sensory properties, and quality indices of the fillets were investigated. Added to these factors was an assessment of the influence of ɛ-polylysine incorporation on the optical, structural, barrier, and mechanical specifications (elongation at break and tensile strength) of chitosan films. Based on the findings, a significant difference among the corresponding values to thickness, color, water vapor permeability (WVP), and mechanical specifications between the treated films by ɛ-PL and untreated films were noted. In addition, higher values of thickness and tensile strength were correlated with ɛ-PL added active chitosan films while compared with control samples. Additionally, no significant differences regarding the proximate composition (including protein, moisture, and fat) among beef fillet samples were observed. In this regard, due to significantly lower levels of pH, TVB-N, and TBARS ɛ-PL in enriched films, this technique demonstrated some protective effects on beef fillets. Another observation was that lower levels of the total viable count, coliform, mold, yeasts, and higher sensory properties were significantly associated with samples with added ɛ-PL (0.9%). Therefore, adding ɛ-PL into chitosan films could be introduced as an effective technique to extend the shelf life of beef fillets and maintain their quality indices during refrigerated storage.

Assessment of silver release and biocidal capacity from silver nanocomposite food packaging materials.

In this study, silver release from commercially available food-contact materials in food simulants (water, acetic acid, ethanol-water and olive oil) and meats (tuna, ham, and turkey) was assessed. Additionally, the antimicrobial capacity of migrated silver was examined in meats. Largest silver release was observed in simulants from food touch papers (25 ± 11 mg/kg) as compared to bag, cutting board and containers. Silver ion and silver nanoparticles were released from food touch paper in food simulants. Food touch paper released the highest amount of silver in tuna (0.5 ± 0.02 mg/kg) than ham (0.2 ± 0.08 mg/kg) or turkey (0.3 ± 0.08 mg/kg) in the same conditions. Tuna exhibited the lowest pH and higher number of bacterial populations on day 0 compared with other foods. Nonetheless, a significant antibacterial capacity of released silver was noticed predominantly in turkey for Gram-negative bacteria. Our study suggests silver released in food simulants indicate an overestimation of silver migration; thus, precaution should be maintained when extrapolating such findings to "real" food. Moreover, further investigations are needed to determine if the amount of silver released from food touch paper in certain foods (for example turkey in the present study) possess any risk to human health.

The role of smart packaging system in food supply chain.

Food supply chain is a rapidly growing integrated sector and covers all the aspects from farm to fork, including manufacturing, packaging, distribution, storing, as well as further processing or cooking for consumption. Along this chain, smart packaging could impact the quality, safety, and sustainability of food. Packaging systems have evolved to be smarter with integration of emerging electronics and wireless communication and cloud data solutions. Although there are many factors causing the loss and waste issues for foods throughout the whole supply chain of food and there have been several articles showing the recent advances and breakthroughs in developing smart packaging systems, this review integrates these conceptual frameworks and technological applications and focuses on how innovative smart packaging solutions are beneficial to the overall quality and safety of food supply by enhancing product traceability and reducing the amount of food loss and waste. We start by introducing the concept of the management for the integrated food supply chain, which is critical in tactical and operational components that can enhance product traceability within the entire chain. Then we highlight the impact of smart packaging in reducing food loss and waste. We summarize the basic information of the common printing techniques for smart packaging system (sensor and indicator). Then, we discuss the potential challenges in the manufacturing and deployment of smart packaging systems, as well as their cost-related drawbacks and further steps in food supply chain.

Genotoxicological safety assessment of puree-only edible films from onion bulb (Allium cepa L.) for use in food packaging-related applications.

The production of films and coatings from onion (Allium cepa L.) to be applied as packaging is attractive, due to its high nutritional and therapeutic value. Also, it can collaborate to minimize environmental impacts caused by the improper disposal of products made from plastics. However, despite it being an innovative and novel proposal, onion films for the development of edible packaging should be evaluated before being considered nontoxic and safe for human consumption. Thus, the objective of the present study was to elucidate the cytotoxic and mutagenic profile of eluates of polymer films of Allium cepa L. obtained by the casting process and to verify their safety for commercial purposes. The analysis of cellular viability demonstrated greater cytotoxicity for unwashed hydrothermally treated pulp (HTP) than for films of washed hydrothermally treated pulp (W-HTP). Regarding the mutagenic activity, the HTP and W-HTP films were not able to statistically increase the frequencies of the biomarkers for chromosome damage (micronucleus test) at the tested concentrations. However, the HTP films showed signs of mutagenicity in the Ames test (gene mutations), suggesting caution in their use. The detection of genotoxicity is highly recommended in order to avoid the risk of genotoxic exposure to mutagens and carcinogens. In conclusion, the absence of mutagenicity and cytotoxicity observed in this study is extremely relevant, because it provides support for toxicogenic properties of the Allium cepa films with promising applicability in the food industry. PRACTICAL APPLICATION: The bioplastics made from onion bulbs are multifunctional materials, which requires safety profile assessment. The results of the mutagenicity and cytotoxicity tests suggests that especially the W-HTP films are harmless, supporting at the first level of evidence, its safety potential to be used in the food industry (food films), biodegradable packaging, and biomaterials (substrates for drug delivery system).

Assessment of the Impact of Accelerated Migration Testing for Coated Food Cans Using Food Simulants.

In this study, an accelerated migration test on food can coatings into food simulants was investigated. Food simulants covering a wide range of polarity were used to conduct migration tests at 60 °C with storage times ranging from 4 h to 30 days. Epoxy-resins, acrylic-phenolic, polyester, and vinyl coatings were exposed to water, 3% acetic acid, 50% ethanol, and Miglyol 812®. Using liquid chromatography coupled to a variety of detectors (UHPLC-Q-Orbitrap-MS, UFLC-MS/MS, and HPLC-DAD), migration of several monomers and previously identified oligomers, as well as some unidentified migrants, were determined during the experiment. The data from this study was compared to our findings from previous long-term migration studies with storage times ranging from 24 h to 540 days at 40 °C using the same can coating applications. The results illustrate that performing migration experiments for short time periods at 60 °C may mimic migration results that would be obtained at 40 °C after long-term migration tests (up to 1.5 years) from food can coatings into food simulants.

Banana starch nanocomposite with cellulose nanofibers isolated from banana peel by enzymatic treatment: In vitro cytotoxicity assessment.

The potential use of cellulose nanofibers (CNFs) as a reinforcing agent in banana starch-based nanocomposite films was investigated. CNFs were isolated from banana peel (Musa paradisiaca) by enzymatic hydrolysis. Banana starch-based nanocomposite films were prepared with CNFs using the casting method. CNFs effect on cell viability and on nanocomposite films properties' was investigated. The cytotoxicity of CNFs was assessed on Caco-2 cell line. CNFs were not cytotoxic at 50-2000 µg/mL. However, CNFs above 2000 µg/mL significantly decreased cell viability. Topography analysis showed that the incorporation of CNFs modified the film structure. The nanocomposites exhibited a complex structure due to strong interactions between CNFs and starch matrix, promoting a remarkable improvement on mechanical and water barrier properties, opacity and UV light barrier compared to the control film. CNFs can offer a great potential as reinforcing material for starch-based nanocomposite films, producing a value-added food packaging from a waste material.

Citrus essential oils: Extraction, authentication and application in food preservation.

Citrus EOs is an economic, eco-friendly and natural alternatives to chemical preservatives and other synthetic antioxidants, such as sodium nitrites, nitrates or benzoates, commonly utilized in food preservation. Citrus based EOs is obtained mainly from the peels of citrus fruits which are largely discarded as wastes and cause environmental problems. The extraction of citrus oils from the waste peels not only saves environment but can be used in various applications including food preservation. The present article presents elaborated viewpoints on the nature and chemical composition of different EOs present in main citrus varieties widely grown across the globe; extraction, characterization and authentication techniques/methods of the citrus EOs; and reviews the recent advances in the application of citrus EOs for the preservation of fruits, vegetables, meat, fish and processed food stuffs. The probable reaction mechanism of the EOs based thin films formation with biodegradable polymers is presented. Other formulation, viz., EOs microencapsulation incorporating biodegradable polymers, nanoemulsion coatings, spray applications and antibacterial action mechanism of the active compounds present in the EOs have been elaborated. Extensive research is required on overcoming the challenges regarding allergies and obtaining safer dosage limits. Shift towards greener technologies indicate optimistic future towards safer utilization of citrus based EOs in food preservation.

Active packaging films with natural antioxidants to be used in meat industry: A review.

Spoilage of meat products during processing, distribution and exposure in the markets have an important negative impact on meat industry from an economic point of view. Two of the main problems of meat and products during processing and subsequent storage are lipid oxidation and deterioration due to microorganism growth. In this context, several packaging alternatives have been developed by meat industry in order to limit these losses and to extend the meat products´ shelf life. Over the last years, the use of active packaging has been proposed as an alternative to traditional packaging. The principle of active packaging, particularly antioxidant active packaging, consists of including active agents in the packaging which interact with meat and/or its environment, either by trapping pro-oxidant compounds or by releasing antioxidant compounds in order to delay degradation due to lipid oxidation. Therefore, the use of active packaging is presented as a future option to solve the problems derived from oxidative deterioration of meat and meat products. However, its use will depend on the costs involved in the development of this active packaging. Therefore, this review will give an overview about the use of active packaging and natural antioxidants, the active film development techniques, as well as the use of biopolymers as substitutes for synthetic polymers and their direct application in the meat industry.

Effect of organic agronomic techniques and packaging on the quality of lamb's lettuce.

BACKGROUND: This research focused on the effect of organic production systems on initial quality and postharvest performance of lamb's lettuce leaves stored in air or under modified atmosphere at refrigerated temperature. Different strategies of organic soil fertility management were compared under the same environmental conditions: (i) a simplified organic production system based on organic commercial fertilizers to recover crop uptake (SB); (ii) an organic production system based on organic matter amendment mainly supplied by animal manure (AM); and (iii) an organic production system based on organic matter amendment supplied by green waste compost (AC). Fully developed lamb's lettuce leaves were harvested and then packed into perforated bags (control in AIR) or in modified atmosphere packaging (MAP) and stored at 4 °C. RESULTS: At harvest, the yield of lamb's lettuce in the AM and AC systems was higher than that in SB. Phenol and dehydroascorbic acid accumulation was observed in the system with the lowest initial supply of organic amendment (SB). Regarding the effect of packaging, AIR conditions maintained the initial quality attributes for a longer period than MAP, which developed off-odours above the threshold of acceptability at 11 days, irrespective of the production system used. CONCLUSION: In general, the initial differences among the production systems were minimal during the postharvest storage. As for the tested packaging systems, AIR successfully maintained the initial quality attributes for a longer period than the MAP. © 2018 Society of Chemical Industry.

Integrating bioassays and analytical chemistry as an improved approach to support safety assessment of food contact materials.

Food contact materials (FCM) contain chemicals which can migrate into food and result in human exposure. Although it is mandatory to ensure that migration does not endanger human health, there is still no consensus on how to pragmatically assess the safety of FCM since traditional approaches would require extensive toxicological and analytical testing which are expensive and time consuming. Recently, the combination of bioassays, analytical chemistry and risk assessment has been promoted as a new paradigm to identify toxicologically relevant molecules and address safety issues. However, there has been debate on the actual value of bioassays in that framework. In the present work, a FCM anticipated to release the endocrine active chemical 4-nonyphenol (4NP) was used as a model. In a migration study, the leaching of 4NP was confirmed by LC-MS/MS and GC-MS. This was correlated with an increase in both estrogenic and anti-androgenic activities as measured with bioassays. A standard risk assessment indicated that according to the food intake scenario applied, the level of 4NP measured was lower, close or slightly above the acceptable daily intake. Altogether these results show that bioassays could reveal the presence of an endocrine active chemical in a real-case FCM migration study. The levels reported were relevant for safety assessment. In addition, this work also highlighted that bioactivity measured in migrate does not necessarily represent a safety issue. In conclusion, together with analytics, bioassays contribute to identify toxicologically relevant molecules leaching from FCM and enable improved safety assessment.

Nanotechnology for Food Packaging and Food Quality Assessment.

Nanotechnology has paved the way to innovative food packaging materials and analytical methods to provide the consumers with healthier food and to reduce the ecological footprint of the whole food chain. Combining antimicrobial and antifouling properties, thermal and mechanical protection, oxygen and moisture barrier, as well as to verify the actual quality of food, e.g., sensors to detect spoilage, bacterial growth, and to monitor incorrect storage conditions, or anticounterfeiting devices in food packages may extend the products shelf life and ensure higher quality of foods. Also the ecological footprint of food chain can be reduced by developing new completely recyclable and/or biodegradable packages from natural and eco-friendly resources. The contribution of nanotechnologies to these goals is reviewed in this chapter, together with a description of portable devices ("lab-on-chip," sensors, nanobalances, etc.) which can be used to assess the quality of food and an overview of regulations in force on food contact materials.

Migration assessment and the 'threshold of toxicological concern' applied to the safe design of an acrylic adhesive for food-contact laminates.

The suitability of an acrylic adhesive used on food packaging was studied. Six potential migrants were identified using GC-MS and UPLC-QTOF. Five compounds were intentionally added (2-butoxyethanol and 2,4,7,9-tetramethyl-5-decyne-4,7-diol 10 (TMDD) and TMDD ethoxylates). One of the compounds identified as 2-(12-(methacryloyloxy) dodecyl)malonic acid was a non -intentionally added substance (NIAS), which could be a methyl metacrylate derivative. A migration study from multilayers containing paper-adhesive-film was carried out. The films used were polyethylene (PE), polypropylene, polyethylene terephthalate, polylactic acid (PLA) and Ecovio F2223®, which is a mixture of biodegradable polyester with PLA. All the non-volatile compounds, including the identified NIAS, migrated into the dry food simulant Tenax ®. Five surfactants based on TMDD were found to migrate from all laminates into Tenax at levels from 0.05 to 0.6 mg kg-1. The results showed that the lowest migration (0.01 mg kg-1 for 2-(12-(methacryloyloxy)dodecyl)malonic acid to 0.07 for TMDD mg kg-1) occurred when the compounds passed through PLA, demonstrating its functional barrier properties to these compounds. In contrast, PE showed the worst barrier properties to these compounds. To evaluate the migration results, the threshold of toxicological concern strategy was applied. The migration values of the surfactant identified were above 0.09 mg kg-1. Thus, it was decided to remove this surfactant from the formulation.

Optimizing and developing a continuous separation system for the wet process separation of aluminum and polyethylene in aseptic composite packaging waste.

The consumption of milk in China is increasing as living standards rapidly improve, and huge amounts of aseptic composite milk packaging waste are being generated. Aseptic composite packaging is composed of paper, polyethylene, and aluminum. It is difficult to separate the polyethylene and aluminum, so most of the waste is currently sent to landfill or incinerated with other municipal solid waste, meaning that enormous amounts of resources are wasted. A wet process technique for separating the aluminum and polyethylene from the composite materials after the paper had been removed from the original packaging waste was studied. The separation efficiency achieved using different separation reagents was compared, different separation mechanisms were explored, and the impacts of a range of parameters, such as the reagent concentration, temperature, and liquid-solid ratio, on the separation time and aluminum loss ratio were studied. Methanoic acid was found to be the optimal separation reagent, and the suitable conditions were a reagent concentration of 2-4 mol/L, a temperature of 60-80°C, and a liquid-solid ratio of 30 L/kg. These conditions allowed aluminum and polyethylene to be separated in less than 30 min, with an aluminum loss ratio of less than 3%. A mass balance was produced for the aluminum-polyethylene separation system, and control technique was developed to keep the ion concentrations in the reaction system stable. This allowed a continuous industrial-scale process for separating aluminum and polyethylene to be developed, and a demonstration facility with a capacity of 50t/d was built. The demonstration facility gave polyethylene and aluminum recovery rates of more than 98% and more than 72%, respectively. Separating 1t of aluminum-polyethylene composite packaging material gave a profit of 1769 Yuan, meaning that an effective method for recycling aseptic composite packaging waste was achieved.

Evaluation and simulation of silver and copper nanoparticle migration from polyethylene nanocomposites to food and an associated exposure assessment.

Silver nanoparticles (nanosilver) and copper nanoparticles (nanocopper) exhibit antimicrobial activity and have been incorporated into polymers to create antimicrobial packaging materials. Their use in conjunction with food has caused concerns regarding the potential risk of particle migration, resulting in human exposure to nanoparticles. A migration experiment was carried out to investigate the effect of time and temperature on the migration of nanosilver and nanocopper particles from polyethylene (PE) nanocomposites to boneless chicken breasts. Migration of silver ranged from 0.003 to 0.005 mg/dm², while migration of copper ranged from 0.024 to 0.049 mg/dm², for a set of four different scenarios representing typical storage conditions. Effects of time and temperature were not significant (p > 0.1). A migration and exposure model was developed on the basis of mathematical relationships defining migratability and subsequent migratables using the Williams-Landel-Ferry equation for time-temperature superposition. The results of the model accurately predicted the nanosilver levels detected in the laboratory migration tests (R values ranging from 0.43 to 0.99); however, the model was less accurate in predicting nanocopper levels (R values ranging from 0.65 to 0.99), probably because of the highly variable background levels of copper observed in the real food matrix. The 95th percentile of the simulated human exposure to nanosilver based on laboratory experimental results of four scenarios ranged from 5.89 × 10⁻5 to 8.9 × 10⁻5 mg kg(bw)⁻¹ day⁻¹. For the measured migration of copper under the same storage conditions, the exposure ranged from 2.26 × 10⁻5 to 1.17 × 10⁻4 mg kg(bw)⁻¹ day⁻¹. This study highlights the potential migration of nanoparticles from PE composite packaging to a food material and the potential for simulation models to accurately capture this migration potential; however, variable background levels of copper in the food matrix can make prediction more difficult for trace migration of nanocopper.

Determination of bisphenol a migrating from canned food and beverages in markets.

The determination of bisphenol A (BPA) in foods and beverages sold in Turkish markets was carried out using high performance liquid chromatography. In this research, foods packed in packages with an inner surface covered with plastic film, such as milk, fruit juice, cream, pudding and tuna samples were used. Furthermore, foods in glass jar and metal cans such as green peas, garniture, corn, tomato paste, pepper paste, pickles, mushroom and bean samples were also used. BPA concentrations were 21.86±0.80-1858.71±8.24µg kg(-1) for canned foodstuffs, 36.48±0.95-554.69±3.18µgkg(-1) for foods in paper box, "not detected" - 399.21±3.26µgkg(-1)for foods in glass jar. The change in the amount of bisphenol A in all of these food, based on expiration date, the amount of glucose and sodium chloride in it has been determined. We see that in these kind of food the amount of bisphenol A increases with an increase in the amount of glucose, NaCl and expiration date.

Migration and exposure assessment of silver from a PVC nanocomposite.

Nanotechnology is the manipulation of matter at the nanoscale, generally between 1 and 100 nm. The discovery of unique nanomaterial properties has lead to novel applications in the food industry, one of which is antimicrobial food packaging materials. The objective of this study was to evaluate the migration of silver from plasticised polyvinyl chloride (PVC) nanocomposites to chicken meat following varying storage time and temperature conditions. The silver content of the chicken was quantified using inductively coupled plasma mass spectroscopy (ICPMS) and migration was found to occur within a range of 0.03-8.4 mg/kg. An exposure assessment revealed that human exposure to silver (assuming a worst case scenario that all silver is in its most harmful nanoform), is likely to be below current migration limits for conventional migrants and a provisional toxicity limit; however it is acknowledged there is still considerable uncertainty about the potential harmful effects of particles at the nanoscale. A sensitivity analysis revealed that silver migration from the nanocomposite to the food surface was influenced most by the percentage fill (p<0.01), followed by storage time (p<0.01) and storage temperature (p<0.05). This study represents an initial and much needed attempt to quantify human risks from the use of nanomaterials in the food industry.

Novel water-resistant UV-activated oxygen indicator for intelligent food packaging.

For the first time, alginate polymer has been applied to prevent dyes from leaching out of colorimetric oxygen indicator films, which enable people to notice the presence of oxygen in the package in an economic and simple manner. The dye-based oxygen indicator film suffers from dye leaching upon contact with water. In this work, UV-activated visual oxygen indicator films were fabricated using thionine, glycerol, P25 TiO2, and zein as a redox dye, a sacrificial electron donor, UV-absorbing semiconducting photocatalyst, and an encapsulation polymer, respectively. When this zein-coated film was immersed in water for 24h, the dye leakage was as high as 80.80±0.45%. However, introduction of alginate (1.25%) as the coating polymer considerably diminished the dye leaching to only 5.80±0.06%. This is because the ion-binding ability of alginate could prevent the cation dye from leaching into water. This novel water-resistant UV-activated oxygen indicator was also successfully photo-bleached and regained colour fast in the presence of oxygen.