Stability and Photochemistry of Vitamin D2 in Model System

Oxidation of vitamin D₂ in a model system of 12% water and 88% acetone in the presence of 15 ppm riboflavin under light and dark was studied by measuring the depleted headspace oxygen. Riboflavin accelerated the oxidation of vitamin D₂ by singlet oxygen under light, but did not affect the vitamin D₂ oxidation under dark. The effect of 0 and 15 ppm riboflavin on the stability of vitamin D₂ during storage at 25 and 60°C was studied by measuring the contents of vitamin D₂ during 48h. Results indicated that photosensitized singlet oxygen oxidation of vitamin D₂ under light was temperature-independent, and triplet oxygen oxidation of vitamin D₂ both under light and in the dark was temperature-dependent.     

Quenching Mechanism of β-Carotene on the Chlorophyll Sensitized Photooxidation of Soybean Oil

The effects of 0, 5, 10, and 20 ppm β-carotene on the oxidation of 0.033, 0.066, 0.165, 0.330, or 0.660M soybean oil in methylene chloride containing 4 ppm chlorophyll have been studied by measuring headspace oxygen depletion in gas-tight sample bottles during 24 hr of light storage. The oxygen depletion was measured by a thermal conductivity gas chromatograph. The rate of singlet oxygen formation in soybean oil by 4 ppm chlorophyll was 0.17 μmoles O₂/mL head-space during first 24 hr of storage. The quenching mechanism study of g-carotene on the chlorophyll sensitized photooxidation of soybean oil showed that g-carotene minimized the oxidation of soybean oil by quenching singlet oxygen. p-carotene did not quench chlorophy11.     

Lipid oxidation in salted-dried fish: II. The effect of photosensitisers on the rate of oxidation of a fish oil

The contribution of photosensitised oxidation to the extent of lipid oxidation of a model fish oil system has been considered using a number of methods of measurement, namely peroxide value, polyene index and the initial rate of oxygen uptake. This represents the initial stages of the development of a model system which will eventually simulate the processing and storage of salted, sun-dried fish. The effects of the photosensitisers, protoporphyrin IX, riboflavin and myoglobin on the rate of oxidation of a highly unsaturated fish oil in a model system, have been studied under dark or photosynthetic light conditions at 30°C. As has previously been established, the initial rate of oxygen uptake proved to be a more reliable method than the peroxide value or the polyene index in determining the extent of lipid oxidation of the oil in the model system. Protoporphyrin IX, riboflavin and myoglobin all increased the rate of oxygen uptake significantly in photosynthetic light at 30°C, from a rate of 0–158 μl O₂g^?1for the control to 0.335 μl O₂g^?1min^?1, 0.308 μl O₂ g^?1min^?1 and 0.461 μl O₂g^?1min^?1, respectively. However, in the dark at 30°C their addition had no effect on the initial rate of oxygen uptake, showing that these components increased the rate via a photosensitised oxidation mechanism rather than by autoxidation. The addition of β-carotene, a well-established singlet oxygen quencher, to the systems containing the photosensitisers at 30°C in the light significantly reduced the rate of oxygen uptake, but not to the same rate as in the control. Photosensitised oxidation has been classified as Type I or Type II; the reduction observed in oxygen uptake rate in the presence of β-carotene shows the Type II mechanism involving singled oxygen to be important. Studying β-carotene on its own in the model system at 30°C showed an unexpected increase in the rate of oxygen uptake both in the light and in the dark.     

Contribution of minor compounds to the singlet oxygen-related photooxidation of olive and perilla oil blend

This study evaluated contribution of natural minor compounds including carotenoids, tocopherols, polyphenols, and phosphatidylcholine (PC) to the stability of chlorophyll sensitized oxidation of olive and perilla oil blend under 2,500 lx light at 25°C for 48 h. The oxidation of the oil blend was evaluated by headspace oxygen consumption and peroxide values (POV). Minor compounds were also monitored. The headspace oxygen consumption and POV of the oil blend were increased with oxidation time; however, fatty acid composition was not significantly changed. Carotenoids, tocopherols, polyphenols, and PC were degraded during the oil oxidation, which was closely related with their singlet oxygen quenching. Correlations of the oil oxidation were higher with contents of carotenoids and PC (r ²>0.93) than with those of tocopherols or polyphenols, suggesting high contribution of carotenoids and PC to decreasing singlet oxygen-related photooxidation of olive and perilla oil blend.     

Kinetics for Singlet Oxygen Formation by Riboflavin Photosensitization and the Reaction between Riboflavin and Singlet Oxygen

ABSTRACT: The formation of singlet oxygen by riboflavin and the kinetics and mechanisms of riboflavin degradation in aqueous solution under light were determined. The singlet oxygen formation rate by riboflavin was 2.31 μmole oxygen/mL headspace/h of serum bottle. The degradations of riboflavin were 66% in D₂O and 40% in H₂O, respectively, under light after 24 h. The results indicate that singlet oxygen is involved in riboflavin destruction under light. The riboflavin destructions were 94.0% and 15.7% with 0 mM or 160 mM ascorbic acid, respectively, under light after 96 h. The reaction rate between riboflavin and singlet oxygen was 1.01 × 10¹⁰/M/s, which is a diffusion-controlled reaction rate. This explains the extremely fast degradation of riboflavin in foods under light. Ascorbic acid and sodium azide reduce the degradation of riboflavin under light with different quenching mechanisms. Ascorbic acid quenched both singlet oxygen and excited triplet riboflavin. Sodium azide quenched only the singlet oxygen in riboflavin solution with a quenching rate of 1.547 × 10⁷/M/s. With the involvement of both the Type-I and Type-II mechanisms in the riboflavin degradation under light, singlet oxygen quencher alone could not protect the riboflavin from degradation completely. Addition of ascorbic acid can protect riboflavin oxidation in foods exposed to light.     

Kinetic study on the singlet oxygen quenching activity of nordihydroguaiaretic acid (NDGA) using methylene blue sensitized photooxidation of α-terpinene

Singlet oxygen is highly reactive and can therefore induce rapid oxidation of a range of biological molecules, causing cell damages. The effects of nordihydroguaiaretic acid (NDGA) on the photochemical singlet oxygen oxidation of α-terpinene in methanol were studied. NDGA showed strong protective activity on the singlet oxygen oxidation of α-terpinene in methanol in a dose dependent manner. The protective activity of NDGA was considerably higher than that of butylated hydroxytoluene and 1,4-diazabicyclo[2.2.2]octane. Total singlet oxygen quenching rate constant (k _r +k _q ) of NDGA was determined by a steady state kinetic equation. The total singlet oxygen quenching rate constant of NDGA was 9.81×10⁷ M^?1 sec^?1. The result showed that NDGA possessed strong singlet oxygen quenching activity, indicating its potential for the protection of molecules, cells and nutrients from the highly reactive singlet oxygen. To the best of our knowledge, this is the first report on the singlet oxygen quenching rate constant of NDGA.     

α-, γ- and δ-Tocopherol Effects on Chlorophyll Photosensitized Oxidation of Soybean Oil

The effects of 0, 1.0, 2.0 and 4.0 (x 10^?3 M) α-, γ- or δ-tocopherol on chlorophyll b photosensitized oxidation of soybean oil in methylene chloride were studied by peroxide values and headspace oxygen. As concentrations of tocopherols increased, peroxide values decreased and headspace oxygen increased (P < 0.05). At 1.0 × 10^?3 M, α-tocopherol showed highest antioxidant effect, γ-tocopherol second and then δ-tocopherol. α- and -γ-Tocopherols had similar effects and δ-tocopherol had lower effect at 2.0 × 10^?3 M (P < 0.05). However, the three tocopherols were not different (P > 0.05) at 4.0 × 10^?3 M. α-Tocopherol quenched singlet oxygen to reduce the photosensitized oxidation of oil. The quenching rate constants of α-tocopherol were 2.7 × 10⁷M^?1sec^?1 by peroxide value and 2.6 × 10⁷ M^?1sec^?1 by headspace oxygen.     

Effects of Riboflavin Photosensitized Oxidation on the Volatile Compounds of Soymilk

ABSTRACT: Soymilks with or without added riboflavin in serum bottles were stored under light or in dark at 20 °C. The headspace oxygen and volatile compounds were determined by gas chromatography. Riboflavin had significant effects on the headspace oxygen depletion and volatile compounds formation in soymilk under light ( P < 0.05). Riboflavin did not have significant effects on the formation of volatile compounds and the depletion of headspace oxygen in dark ( P > 0.05). The volatile compounds increased under light, but not in dark as the added riboflavin increased. Storage temperature at 4 °C or 20 °C did not have significant difference in the effect of riboflavin on the headspace oxygen depletion in soymilk under light. Hexanal, an important beany flavor compound, was identified as the major volatile compound in the riboflavin photosensitized soymilk. Singlet oxygen oxidation was involved in the formation of volatile compounds in soymilk under light. Hexanal could be formed by singlet oxygen oxidation. Ascorbic acid, a quencher for singlet oxygen and the excited triplet sensitizer, significantly inhibited the formation of hexanal and total volatiles in soymilk under light.     

Direct Spectroscopic Observation of Singlet Oxygen Quenching and Kinetic Studies of Physical and Chemical Singlet Oxygen Quenching Rate Constants of Synthetic Antioxidants (BHA,BHT, and TBHQ) in Methanol

Abstract: Singlet oxygen quenching by synthetic antioxidants (BHA, BHT, and TBHQ) was directly observed by spectroscopic monitoring of luminescence at 1268 nm. The luminescence data showed unambiguous evidence of singlet oxygen quenching by synthetic phenolic antioxidants with the highest activity for TBHQ, followed by BHA and BHT. The protective activities of these synthetic antioxidants on α-terpinene oxidation with chemically-induced singlet oxygen under dark further confirmed their singlet oxygen quenching abilities. Total singlet oxygen quenching rate constants (k_r + k_q) of BHA, BHT, and TBHQ were determined in a system containing α-terpinene (as a singlet oxygen trap) and methylene blue (as a sensitizer) during light irradiation, and the values were 5.14 × 10⁷, 3.41 × 10⁶, and 1.99 × 10⁸ M⁻¹s⁻¹, respectively. After the k_r value of α-terpinene was first determined, the k_r values of the synthetic antioxidants were calculated by measuring their relative reaction rates with singlet oxygen to that of α-terpinene under the identical conditions. The k_r values of the BHA, BHT, and TBHQ were 3.90 × 10⁵, 1.23 × 10⁵, and 2.93 × 10⁶, M⁻¹s⁻¹. The percent partition of chemical quenching over total singlet oxygen quenching (k_r × 100)/(k_r + k_q) for BHA, BHT, and TBHQ were 0.76%, 3.61%, and 1.47%, respectively. The results showed that the synthetic antioxidants quench singlet oxygen almost exclusively through the mechanism of physical quenching. This represents the first report on the singlet oxygen quenching mechanism of these synthetic antioxidants. Practical Application: The synthetic antioxidants, especially TBHQ, have been found to have a strong singlet oxygen quenching ability. This article also clearly showed that singlet oxygen quenching by synthetic antioxidants was mainly by the physical quenching mechanism. The results suggested that these synthetic antioxidants, especially TBHQ, could be used practically for the protection of the food components such as edible oils and vitamins against singlet oxygen induced oxidations without significant losses of antioxidant activity during storage under light.     

Effects of riboflavin-photosensitization on the formation of volatiles in linoleic acid model systems with sodium azide or D2O

Effects of riboflavin-photosensitization on volatile formations were studied in linoleic acid model systems with sodium azide or D₂O. Linoleic acid with riboflavin solution were stored under light or in the dark and volatiles were analyzed by a combination of solid phase microextraction (SPME), gas chromatography (GC), and mass spectrometry (MS). Riboflavin-photosensitization produced 27.9% more total volatiles than samples stored in the dark for 16 h. Addition of sodium azide decreased the total volatiles by 28.4% compared to samples without sodium azide. Linoleic acid in D₂O had higher increases in total volatiles than samples in H₂O system. Decrease of total volatiles in samples with sodium azide and increase of total volatiles in D₂O sample indirectly showed the presence of singlet oxygen in riboflavin-photosensitization. Hexanal, 2-heptenal, 1-octen-3-ol, 2-octenal, and 2,4-decadienal were greatly influenced by the addition of sodium azide and in D₂O, which indicates that formation of these volatiles was related to singlet oxygen oxidation. Especially, 2-heptenal showed the highest peak area among volatiles under riboflavin-photosensitized linoleic acid for 16 h.     

Chemistry and Reaction of Singlet Oxygen in Foods

Singlet oxygen is a highly reactive, electrophilic, and nonradical molecule. It is different from diradical triplet oxygen in its electron arrangement. Photosensitizers can form singlet oxygen from triplet oxygen in the presence of light. The reaction rate of singlet oxygen with foods is much greater than that of triplet oxygen due to the low activation energy. Singlet oxygen oxidation produces undesirable compounds in foods during processing and storage. However, carotenoids and tocopherols in foods can minimize singlet oxygen oxidation. The in‐depth scientific knowledge on the formation, reactions, quenching mechanisms, and kinetics of singlet oxygen can greatly improve the quality of foods by minimizing the oxidation during processing and storage. The single oxygen oxidation of foods has contributed to the explanation of several important chemical reactions in the reversion flavor in soybean oil, sunlight flavor in milk products, and the rapid losses of vitamin D, riboflavin, and ascorbic acid in milk under light storage.     

Mechanisms and Factors for Edible Oil Oxidation

ABSTRACT: Edible oil is oxidized during processing and storage via autoxidation and photosensitized oxidation, in which triplet oxygen (³O₂) and singlet oxygen (¹O₂) react with the oil, respectively. Autoxidation of oils requires radical forms of acylglycerols, whereas photosensitized oxidation does not require lipid radicals since ¹O₂ reacts directly with double bonds. Lipid hydroperoxides formed by ³O₂ are conjugated dienes, whereas ¹O₂ produces both conjugated and nonconjugated dienes. The hydroperoxides are decomposed to produce off‐flavor compounds and the oil quality decreases. Autoxidation of oil is accelerated by the presence of free fatty acids, mono‐ and diacylglycerols, metals such as iron, and thermally oxidized compounds. Chlorophylls and phenolic compounds decrease the autoxidation of oil in the dark, and carotenoids, tocopherols, and phospholipids demonstrate both antioxidant and prooxidant activity depending on the oil system. In photosensitized oxidation chlorophyll acts as a photosensitizer for the formation of ¹O₂; however, carotenoids and tocopherols decrease the oxidation through ¹O₂ quenching. Temperature, light, oxygen concentration, oil processing, and fatty acid composition also affect the oxidative stability of edible oil.     

Kinetic Study of the Quenching Reaction of Singlet Oxygen by Common Synthetic Antioxidants (tert-Butylhydroxyanisol, tert-di-Butylhydroxytoluene, and tert-Butylhydroquinone) as Compared with α-Tocopherol

ABSTRACT: Effects of synthetic phenolic antioxidants (BHA, BHT, and TBHQ) on the methylene blue (MB) sensitized photooxidation of linoleic acid as compared with that of α‐tocopherol have been studied. Their antioxidative mechanism was studied by both ESR spectroscopy in a 2,2,6,6‐tetramethylpiperidone (TMPD)‐methylene blue (MB) system and spectroscopic analysis of rubrene oxidation induced by a chemical source of singlet oxygen. Total singlet oxygen quenching rate constants (k_ox?Q+k_q) were determined using a steady state kinetic equation. TBHQ showed the strongest protective activity against the MB sensitized photooxidation of linoleic acid, followed by BHA and BHT. TBHQ (1 × 10^?3 M) exhibited 86.5% and 71.4% inhibition of peroxide and conjugated diene formations, respectively, in linoleic acid photooxidation after 60‐min light illumination. The protective activity of TBHQ against the photosensitized oxidation of linoleic acid was almost comparable to that of α‐tocopherol. The data obtained from ESR and rubrene oxidation studies clearly showed the strong singlet oxygen quenching ability of TBHQ. The k_ox?Q+k_q of BHA, BHT, and TBHQ were determined to be 3.37 × 10⁷, 4.26 × 10⁶, and 1.67 × 10⁸ M^?1 s^?1, respectively. The k_ox?Q+k_q of TBHQ was within the same order of magnitude of that of α‐tocopherol, a known efficient singlet oxygen quencher. There was a high negative correlation (r² = ?0.991) between log (k_ox?Q+k_q) and reported oxidation potentials for the synthetic antioxidants, indicating their charge‐transfer mechanism for singlet oxygen quenching. This is the 1st report on the kinetic study on k_ox?Q+k_q of TBHQ in methanol as compared with other commonly used commercial synthetic antioxidants and α‐tocopherol.     

Antioxidant mechanisms of Trolox and ascorbic acid on the oxidation of riboflavin in milk under light

Antioxidant activities and the mechanism of water-soluble Trolox and ascorbic acid on the oxidation of riboflavin in milk were studied. Trolox or ascorbic acid at 0, 100, 250, 500, or 1000 ppm was added to milk with or without added 50 ppm riboflavin and stored under light at 27 °C. Headspace oxygen was analysed by GC and Trolox, ascorbic acid, and riboflavin were determined by HPLC. The headspace oxygen of milk with added 50 ppm riboflavin depleted faster than that of milk without added riboflavin (p < 0.05). Trolox and ascorbic acid decreased during storage under light and riboflavin was completely destroyed within 24 h. As the concentration of Trolox or ascorbic acid increased, the riboflavin loss decreased. Riboflavin, Trolox, and ascorbic acid competed to react with singlet oxygen which was formed in the presence of riboflavin under light. Trolox and ascorbic acid protected riboflavin in milk under light by reacting with singlet oxygen.     

Riboflavin Photosensitized Singlet Oxygen Oxidation of Vitamin D

Samples containing various levels of vitamin D and riboflavin, with and without ascorbic acid or a-tocopherol were prepared in a model system. Samples were stored in the light or in the dark at 45®C for up to 8 h. Headspace oxygen was determined by gas chromatography with thermal conductivity detection. Oxidation of vitamin D was not observed in samples without riboflavin stored in the light nor in samples with riboflavin stored in the dark. Vitamin D with riboflavin was oxidized under light. α-Tocopherol quenched singlet oxygen at a rate of 2.52 × 10⁸ M^-1 sec^-1, whereas ascorbic acid quenched singlet oxygen at a rate of 2.23 × 10⁷ M^-1 sec^-1.     

Effects of Trolox and ascorbic acid on the riboflavin photosensitised oxidation of aromatic amino acids

The effects of 0, 100, 250, 500, 750 or 1000 ppm Trolox and ascorbic acid on the singlet oxygen oxidation of aromatic amino acids viz. phenylalanine, tryptophan, tyrosine in the presence of 25 ppm riboflavin were determined by measuring depleted headspace oxygen by gas chromatography and aromatic amino acid content by high performance liquid chromatography. The coefficients of variations (CVs) for headspace oxygen analysis and HPLC analysis of aromatic amino acids were 1.4% and 4.7%, respectively. Samples were stored under light (1000 lux) at 30 °C for 10 h. Both Trolox and ascorbic acid acted as antioxidants. As the concentration of Trolox and ascorbic increased from 0 to 1000 ppm, the head space oxygen depletion increased. This was due to the oxidation of Trolox and ascorbic acid along with amino acids in the presence of riboflavin. High performance liquid chromatography analysis of the samples clearly indicated that both Trolox and ascorbic acid minimised the degradation of phenylalanine, tryptophan, tyrosine significantly (p < 0.05), but did not prevent their oxidation completely. Trolox acted as a better antioxidant than ascorbic acid in protecting phenylalanine, tryptophan and tyrosine. Type-I mechanism was mainly responsible for riboflavin photosensitised degradation of aromatic amino acids.     

Lipid oxidation in salted-dried fish: The effect of temperature and light on the rate of oxidation of a fish oil

In relation to studies of lipid oxidation during the processing and storage of salted sun-dried fish, the measurement of the initial rate of oxygen uptake has been studied on a model system consisting of a highly polyunsaturated fish oil. This parameter has been used for assessing the effects of temperature and light conditions on the model system and has been compared to changes in the peroxide value and the polyene index. Assessment of oxygen uptake using a Gilson respirometer was found to give a clearer indication than other methods of assessment of the effect of different conditions on fish oil oxidation. At 30°C the initial rate of oxygen uptake increased from 0-103 μl O₂, g^?1 miir^?1 to 0-158 μl O₂, g^?1 min^?1 with a change in light conditions from dark to photosynthesis light. At 40°C, the rate of oxygen uptake was fester than at 30 °C, and the effect of light was more pronounced, giving values of 0.188 μl O₂, g^?1 min^?1 and 0.483 μ O₂, g^?1 min^?1, for dark and photosynthesis light, respectively. The observed results for the measurement of the initial rate of oxygen uptake suggest its use as a sensitive and reliable method for the investigation of the contribution of the many components in salted sun-dried fish towards the rate of oxidation and subsequent rancidity.     

Quenching Mechanisms and Kinetics of α-Tocopherol and β-Carotene on the Photosensitizing Effect of Synthetic Food Colorant FD&C Red No. 3

ABSTRACT: Effects of FD&C Red No. 40, Red No. 3, Yellow No. 5, Yellow No. 6, Green No. 3, Blue No. 1 and Blue No. 2 on 0.03M soybean oil oxidation in acetone at 25 °C under light were studied by measuring headspace oxygen depletion. As Red No. 3 increased from 0 to 5, 20, 100 and 200 ppm, the headspace oxygen was reduced by 2 to 70, 73, 77 and 77%, respectively, for 4 h. Only Red No. 3 acted as a photosensitizer to produce singlet oxygen in the oil. The quenching rates of α-tocopherol and β-carotene for the singlet oxygen by Red No.3 were 4.1 × 10⁷ M⁻¹s⁻¹ and 7.3 × 10⁹ M⁻¹s⁻¹, respectively. When β-carotene was below 1.86 × 10⁻⁶ M, β-carotene quenched singlet oxygen, but it quenched both singlet oxygen and Red No. 3 at or above 3.72 × 10⁻⁶ M. However, α-tocopherol quenched singlet oxygen only.     

Singlet molecular oxygen quenching by the antioxidant dimethylmethoxy chromanol in solution and in ex vivo porcine skin

Singlet‐oxygen is a non‐radical reactive oxygen species believed to play a major role in many photooxidation processes in connection with diverse photo‐biological processes such as skin ageing or photocarcinogenesis. Dimethylmethoxy chromanol (3,4‐dihydro‐6‐hydroxy‐2,2‐dimethyl‐7‐methoxy‐1(2H)‐benzopyran) is a potent antioxidant used in cosmetic and pharmaceutical formulations. We have assessed the singlet oxygen quenching ability of dimethylmethoxy chromanol, by monitoring the near‐IR phosphorescence of singlet‐oxygen in solution and in ex vivo porcine skin samples. Dimethylmethoxy chromanol quenches singlet oxygen with a rate constant of (1.3 ± 0.1) × 10⁸ M^?1 s^?1 in solution. Consistent with this, a clear reduction in the singlet oxygen lifetime and emission intensity was observed when ex vivo porcine skin samples were treated with dimethylmethoxy chromanol.     

Effect of ultraviolet‐B irradiation on antioxidative properties of aqueous extracts from shiitake (Lentinus edodes) mushrooms

Antioxidant properties of aqueous extracts from Lentinus edodes treated with UV‐B irradiation were examined in vitro systems including DPPH, ABTS and oxygen radical absorbance capacity (ORAC) assays and in riboflavin‐photosensitised oil‐in‐water (O/W) emulsions. Changes in total phenolics, total flavonoids and vitamin C were also analysed. Lentinus edodes receiving 25 kJ m^?2 UV‐B treatment showed high radical scavenging ability based on DPPH and ABTS assays compared with samples with 0, 50 and 75 kJ m^?2, while those with 50 kJ m^?2 had higher antioxidant capacities than other samples from ORAC assays. Samples with 25 kJ m^?2 UV treatment had significantly 7.1% higher total phenolic content, 12.0% higher total flavonoid content and 8.0% higher vitamin C content than UV‐B‐untreated sample (P < 0.05), respectively. In O/W emulsions under riboflavin photosensitisation, 25 and 50 kJ m^?2 UV treatment significantly increased the oxidative stability compared with other samples based on headspace oxygen content and lipid hydroperoxide analyses (P < 0.05). Aqueous extracts of UV‐B‐treated mushrooms possessed enhanced antioxidant properties compared with untreated mushrooms.