Lourdes Gallardo-Guerrero

Pigments present in virgin olive oil

The qualitative and quantitative control of pigments in ripe olives and in extracted virgin olive oil has increased our knowledge of the influence on these compounds in the areas of ripening of the fruit, storage time in the factory and the oil extraction process. As the harvesting time of the fruits increases, pigment content decreases. During storage, the presence of lipoxygenase has been detected, as well as a considerable decrease in chlorophylls and a small decrease in carotenoids. During the extraction process, the chlorophyllic fraction is destroyed in the greater part, and although the carotenoid fraction is also affected, its concentration increases in the oil with respect to that in the fresh fruit. In the pigment degradation, in addition to the acid-catalyzed reaction, the presence of lipoxygenase suggests a role for this enzyme.

Action of chlorophylls on the stability of virgin olive oil

Virgin olive oil was used as substrate to study the influence of chlorophylls on its oxidative stability in light and in darkness. Chlorophylls a and b were added to this substrate, after which oils were stored at 36 ± 2°C for three months under artificial light (1340 lux) or in darkness. The effect of light was greater than that of the additives. The prooxidant action of chlorophylls in the presence of other pigments of the oil was not observed in this assay. During early storage, the rate of peroxide formation was lower in the samples with added chlorophylls, but later it equalled that of the control. In darkness, stability was greater in the samples containing chlorophylls, indicating a slight antioxidant effect, which was more marked for chlorophyll a.

Role of chlorophyllase in chlorophyll metabolism in olives cv. Gordal

Abstract During the development of the olive two stages in chlorophyll evolution can be distinguished: one of synthesis when the fruit is in the growth phase, and a degradative stage which begins when the fruit is completely developed. Although the enzyme chlorophyllase is present in olives throughout their development cycle, its activity reaches a maximum at both the beginning and end of the vegetative growth phase. During these two periods chlorophyll synthesis prevails, so the two maxima in enzyme activity are associated with the involvement of chlorophyllase in chlorophyll synthesis. Small amounts of chlorophyllides a and b have been detected during the initial growth period of the fruit, which coincides with a phase of great chlorophyll synthesis. Later, synthetic and degradative mechanisms may overlap, making the detection of dephytylated compounds impossible. At the same time allomerized chlorophyllic derivatives—13 2 -hydroxychlorophyll a , 13 2 -methoxychlorophyll a , 13 2 -methoxychlorophyll b , 15 1 -methoxylactone-chlorophyll a and 15 1 -methoxylactone-chlorophyll b —can be detected, suggesting the presence of chlorophyll oxidase.

Carotenoid profiling in tubers of different potato (Solanum sp) cultivars: accumulation of carotenoids mediated by xanthophyll esterification.

The carotenoid profile of sixty potato cultivars (commercial, bred, old and native cultivars) has been characterised in order to provide information to be used in selective breeding programs directed to improve the nutritional value of this important staple food. Cultivars were segregated into three groups according to the major pigment in the carotenoid profile: violaxanthin (37 cultivars; especially those with higher carotenoid content), lutein (16 cultivars), and neoxanthin (7 cultivars). Other minor carotenoids were antheraxanthin, β-cryptoxanthin and β-carotene, while zeaxanthin was absent in all sample. The total carotenoid content ranged from 50.0 to 1552.0 μg/100 g dry wt, with an average value of about 435.3 μg/100 g dry wt. Sipancachi, Poluya and Chaucha native cultivars showed the highest carotenoid content (1020.0, 1478.2 and 1551.2 μg/100 g dry wt, respectively). Xanthophyll esters were present in most cultivars, mainly as diesterified forms, being observed a direct correlation between the carotenoid content and the esterified fraction, suggesting that the esterification process facilitates the accumulation of these lipophilic compounds within the plastids. Therefore, the presence of xanthophyll esters should be a phenotypic character to be included in the breeding studies, and more efforts should be dedicated to the understanding of the biochemical process leading to this structural modification of carotenoids in plants.

Control of olive oil adulteration with copper-chlorophyll derivatives

The present work proposes an analytical method able to detect in an adulterated olive oil sample the addition of the copper complexes of chlorophylls (E 141i). The method consists of a pigment extraction in liquid phase and subsequent analysis by HPLC-DAD. The profile of chlorophyll pigments of an olive oil is determined essentially by its content in pheophytins (a and b), but in no case any copper derivative. Different samples of colorant E 141i have been analyzed, the natural coloring additives used to adulterate vegetable oils. The 99.59+/-0.52% of the chlorophyll pigments present in the different samples of E 141i colorant are not those of an olive oil (more than 75% are cupro-derivatives). Thus, the simple detection of one of the compounds in an olive oil indicates adulteration. The major chlorophyll derivative in all the E 141i colorants samples is Cu-pyropheophytin a and its limit of detection (LOD) defined at a signal-to-noise ratio of about 3 was 6.58 ng/g.

Involvement of copper and zinc ions in green staining of table olives of the variety gordal

The presence of metalochlorophyllic complexes of copper has been detected in table olives showing the alteration known as green staining. These compounds are absent in the healthy fruit. The possible implication of fungicidal treatment of olive trees in this alteration has been studied. No alteration was produced in table olives prepared with fruit from trees with and without fungicidal treatment and the differences found between copper levels in the fruit were not significant. The possibility that the copper involved in this alteration is of extraneous origin was, therefore, discarded. On the other hand, there were no significant differences in the levels of copper in random samples of fruits with and without green staining. Therefore, although the green-staining alteration is the result of the formation of complexes of copper with chlorophyll derivatives, it seems clear that the simple presence in the fruits of copper, by itself, does not lead to the appearance of green-staining.

Digestive stability, micellarization, and uptake by Caco-2 human intestinal cell of chlorophyll derivatives from different preparations of pea (Pisum sativum L.).

The digestive stability, efficiency of micellarization, and cellular accumulation of the chlorophyll pigments of different preparations of pea were investigated, using an in vitro digestion procedure coupled with human intestinal Caco-2 cells. Fresh pea (FP), cooked fresh pea (CFP), frozen pea (FZP), cooked frozen pea (CFZP), and canned pea (CP) were subjected to simulated digestion. Although after digestion the pigment profile was modified for all samples, except CP, allomerization reactions and greater destruction of chlorophylls were observed in only FP, which should be due to enzymes in FP that were denaturalized in the rest of the test foods. A pigment extract of CFZP was also subjected to in vitro digestion, showing a positive effect of the food matrix on the pigment digestive stability. The transfer of the chlorophyll pigments from the digesta to the micellar fraction was significantly more efficient in CFZP (57%, p < 0.0001), not significantly ( p > 0.05) different between CFP, FZP, and CP (28-35%), and lowest in FP (20%). Pheophorbide a stood out as the most-micellarized chlorophyll derivative in all of the samples, reaching levels of up to 98%. Incubation of Caco-2 cells with micellar fractions at the same concentration prepared from each test food showed that pigment absorption was considerably lower ( p < 0.006) in cells incubated with FP, whereas there were no differences among the rest of the preparations. Therefore, factors associated with the food matrix could inhibit or mediate the chlorophyll pigment absorption. These results demonstrated that the industrial preservation processes of peafreezing and canningas well as the cooking have a positive effect on the bioaccessibility and bioavailability of the chlorophyll pigments with respect to the FP sample, emphasizing CFZP with greater bioaccesibilty degree.

The chloroplast NADPH thioredoxin reductase C, NTRC, controls non-photochemical quenching of light energy and photosynthetic electron transport in Arabidopsis.

High irradiances may lead to photooxidative stress in plants, and non-photochemical quenching (NPQ) contributes to protection against excess excitation. One of the NPQ mechanisms, qE, involves thermal dissipation of the light energy captured. Importantly, plants need to tune down qE under light-limiting conditions for efficient utilization of the available quanta. Considering the possible redox control of responses to excess light implying enzymes, such as thioredoxins, we have studied the role of the NADPH thioredoxin reductase C (NTRC). Whereas Arabidopsis thaliana plants lacking NTRC tolerate high light intensities, these plants display drastically elevated qE, have larger trans-thylakoid ΔpH and have 10-fold higher zeaxanthin levels under low and medium light intensities, leading to extremely low linear electron transport rates. To test the impact of the high qE on plant growth, we generated an ntrc-psbs double-knockout mutant, which is devoid of qE. This double mutant grows faster than the ntrc mutant and has a higher chlorophyll content. The photosystem II activity is partially restored in the ntrc-psbs mutant, and linear electron transport rates under low and medium light intensities are twice as high as compared with plants lacking ntrc alone. These data uncover a new role for NTRC in the control of photosynthetic yield.

Composition of pigments and colour changes in green table olives related to processing type

Brownish colourations in Natural green table olives (non-treated with alkali) make this product less attractive to consumers than Spanish-style green table olives (treated with alkali), which develop a more appreciated bright golden-yellow colour. These colour differences were studied in relation to changes in the composition of chlorophyll and carotenoid pigments, as well as polyphenolic compounds and polyphenol oxidase enzyme (PPO) activity. Natural green olives showed a different chlorophyll profile than Spanish-style. However, all the chlorophyll pigments formed in both processing types were Mg-free derivatives (mostly pheophytins) with similar colourations, ranging from grey to green brownish. In the carotenoid fraction no appreciable differences were found between both processing types. The fruits brownish colour was mainly due to polymeric substances with a size of >1000 daltons and polyphenolic nature, resulting from an enzymatic oxidation by PPO of the o-diphenolic compounds present in the fresh fruits.

Anomalous transformation of chloroplastic pigments in gordal variety olives during processing for table olives

The results of a qualitative and quantitative study of pigments, carried out during the processing of Gordal variety olives for table use, has provided valuable information on the type, extent, and mechanism of degradation of the chlorophylls and carotenoids present in the fresh fruit. Unexpected results were obtained, since the initial treatment of the fruits with NaOH did not provoke chlorophyllase activity. However, the alkaline pH brought about the oxidation of chlorophylls, giving rise to Mg-phytyl-chlorin e6 and Mg-phytyl-rhodin g7. Subsequently, as a consequence of the acid pH generated in the fermentation medium, these compounds were transformed into the corresponding Mg-free derivatives, phytyl-ch1orin e6 and phytyl-rhodin g7. At the same time, those chlorophylls which initially escaped transformation were converted into their corresponding pheophytins. In addition, small amounts of pyropheophytin a, pheophorbides a and b, and pyropheophorbide a were detected, As far as the carotenoid fraction is concerned, β-carotene and lutein remained unaltered throughout processing, and only those components with molecular structures sensitive to the acid medium were affected. The total balance of pigment material indicated that there was a slow but progressive decrease in the concentration of the chlorophyllic and carotenoid fractions, which indicates that a certain amount of these pigments is degraded into colorless products.