Pigment Analysis Research Articles

Microscopic analysis of overglaze green pigment on Chinese famille rose porcelain from the Imperial Palace.

Famille rose porcelain made in the imperial workshop was one of the symbols of the highest manufacturing techniques in ancient China. It was very precious and only owned by the royals for a long time. Because of its complex making technology, overglaze green pigment painted on famille rose porcelain shows many variations. In this work, selected porcelain shards that were excavated at the Forbidden City were analyzed by using microscopy observation, micro-Raman spectroscopy, scanning electron microscopy, and energy dispersive X-ray spectrometer to obtain a deep understanding of overglaze green pigment technique on famille rose porcelain. The results revealed that four types of green pigment are used in the selected samples. With different recipes and combinative ways on green pigment and opaque white pigment, craftsmen can choose the using method of pigment to get the effect they want.

Understanding the photophysics of stercobilin-Zn(II) and urobilin-Zn(II) complexes towards faecal pigment analysis

A detailed photophysical study of two faecal pigments (FPs), Urobilin (UB) and Stercobilin (SB), and their zinc complexes [FP-Zn(II)] was carried out. The enhancement of UB and SB fluorescence resulting from the formation of their Zn(II) complexes was attributed to the complexation-induced rigidity of the chromophoric units, and the corresponding decrease of nonradiative decay rate constants of the excited singlet states (knr). The effect of various physicochemical environments was also studied in detail in order to understand the fluorescence behaviour of the Zn(II) complexes. FP-Zn(II) complexes have a lower solubility in water that results in the formation of molecular aggregates. The aggregation-induced loss of fluorescence of FP-Zn(II) complexes could be overcome by using the appropriate mixture of ethanol and water (70:30). Molecular orbital calculations on the FP-Zn(II) complexes provided a good idea of the geometry of the complexes and helped rationalise the enhancement of fluorescence after complexation. This study could pave the way towards developing a convenient non-extraction aqueous phase analytical procedure for detection of FPs using Zn(II) complexation method.

Chlamydomonas reinhardtii LHCSR1 and LHCSR3 proteins involved in photoprotective non-photochemical quenching have different quenching efficiency and different carotenoid affinity

Microalgae are unicellular photosynthetic organisms considered as potential alternative sources for biomass, biofuels or high value products. However, their limited biomass productivity represents a bottleneck that needs to be overcome to meet the applicative potential of these organisms. One of the domestication targets for improving their productivity is the proper balance between photoprotection and light conversion for carbon fixation. In the model organism for green algae, Chlamydomonas reinhardtii, a photoprotective mechanism inducing thermal dissipation of absorbed light energy, called Non-photochemical quenching (NPQ), is activated even at relatively low irradiances, resulting in reduced photosynthetic efficiency. Two pigment binding proteins, LHCSR1 and LHCSR3, were previously reported as the main actors during NPQ induction in C. reinhardtii. While previous work characterized in detail the functional properties of LHCSR3, few information is available for the LHCSR1 subunit. Here, we investigated in vitro the functional properties of LHCSR1 and LHCSR3 subunits: despite high sequence identity, the latter resulted as a stronger quencher compared to the former, explaining its predominant role observed in vivo. Pigment analysis, deconvolution of absorption spectra and structural models of LHCSR1 and LHCR3 suggest that different quenching efficiency is related to a different occupancy of L2 carotenoid binding site.

Genetic Analysis of Anthocyanin Pigmentation in Sterile Lemma and Apiculus in Rice

Genetic analysis of genes that regulate the color pigmentation of sterile lemma and apiculus has been conducted. “Josaengjado” has small and round grains with purple leaf, sterile lemma and apiculus. In the F2 population from a cross between Josaengjado and Daeribbyeo 1, 246 and 182 plants exhibited purple and straw-white sterile lemma, respectively. It fitted a 9:7 segregation ratio indicating that two complementary genes control the pigmentation in sterile lemma and apiculus. Genetic analysis was performed using the F2:3 and KASP (Kompetitive Allele-Specific PCR) markers. Genes for the coloration of leaf sheath, ligule, sterile lemma, and apiculus were detected on chromosomes 1 and 6. Sequence comparison showed a single nucleotide substitution C (Josaengjado) to A (Daeribbyeo 1) in the second exon of the Rd gene on chromosome 1 leading to a premature stop in Daeribbyeo 1. In C1, a 3-bp deletion in the second exon was detected in Daeribbyeo 1. Haplotype analysis was performed in the Rd and C1 genes of the 78 rice accessions. 78 accessions were divided into 14 groups. A total of 11 and 1 mutation sites were detected in OsC1 and Rd, respectively. The haplotype analysis also confirmed that two complementary genes, Rd and OsC1 are necessary to express anthocyanin pigmentation in sterile lemma and apiculus. To our knowledge, this is the first report to identify genes for the coloration of sterile lemma in rice.

Nondestructive analysis of photosynthetic pigments in forage radish and vetch

Chlorophylls and carotenoids are the main photosynthetic pigments in plants, and photosynthetic potential can be used to determine the nitrogen fixation or cycling capacity of cover crops. Therefore, this study quantified and compared the content of photosynthetic pigments in Raphanus sativus and Vicia sativa by using two different methods, namely the extraction method and measurements with a portable chlorophyll meter, and determined whether the results of these two methods showed a significant correlation. Photosynthetic pigments were measured, using both methods, in a greenhouse and laboratory, at four developmental stages: the vegetative stages I (30 days after sowing [DAS]), II (60DAS), and III (90DAS), and the reproductive stage (150DAS). The determination of the photosynthetic pigment content in R. sativus and V. sativa, obtained from the extraction method and chlorophyll index, revealed significant differences depending on the developmental stage in both species. Furthermore, the contents of chlorophyll a, chlorophyll b, and carotenoids determined using the extraction method showed high coefficients of correlation with the total chlorophyll index determined using the portable chlorophyll meter. Therefore, the portable chlorophyll meter can be used for the accurate evaluation of the photosynthetic potential of Raphanus sativus and Vicia sativa, which also saves time and reagents.

Physiological vitality of Norway spruce (Picea abies L.) stands along an altitudinal gradient in Tatra National Park

Abstract Nowadays, a large area of Norway spruce forest stands in Europe is disturbed by windstorm and, subsequently, bark beetle outbreaks. We investigated the state of three disturbed spruce stands along an altitudinal gradient in Tatra National Park (Slovakia) through various physiological processes. Tree-growth characteristics, the mineral nutrition in the needles, and photosynthetic efficiency were assessed. Two techniques of chlorophyll a fluorescence and analyses of assimilatory pigments were used to detect the changes in photosynthesis functioning. Also, the heat sensitivity of photosystem II was tested. Our results showed that these stands are located in nutrient-poor environments. We recorded similar contents of nitrogen, phosphorus, potassium, sodium, zinc, and iron in all stands. Down the vertical transect, the contents of calcium, magnesium, and manganese significantly decreased and the non-essential aluminium increased. Based on stem circumference measurements, water deficit occurred during the vegetation season in all stands, but with the smallest magnitude highest U–stand. We found some photosynthetic constraints: slightly lower chlorophyll contents in all stands were recorded; however, seasonal dynamics with increasing chlorophyll concentration in the highest U–stand were observed. Moreover, the photochemistry of the lowest D–stand was the most negatively influenced by simulated heat, as the photosynthetic performance index, and the density of the active reactions centres significantly decreased and the values of the K–step and basal fluorescence increased. Therefore, we can conclude the different levels of physiological vitality in these naturally damaged spruce stands, with the best physiological performance of the trees in the highest stand.

Weak Response of Biological Productivity and Community Structure of Phytoplankton to Mesoscale Eddies in the Oligotrophic Philippine Sea

AbstractIn the Philippine Sea, mesoscale eddies have been frequently observed, but little is known about their contributions to the biological productivity and community structure of phytoplankton in the region. In situ carbon and nitrogen uptake rates and phytoplankton community structure were investigated in the Philippine Sea based on the 13C‐15N dual‐tracer technique and photosynthetic pigment analysis from late August to September 2018. Based on the comparison of the phytoplankton community structure among the cold eddy, warm eddy, and outer area, we found that the phytoplankton composition was different in the warm eddy with relatively lower contributions of large phytoplankton. However, a distinct enhancement in phytoplankton biomass over the euphotic zone was not detected even in eddy occurrence. In terms of community, the picoplankton Prochlorococcus was persistently dominant, which was attributed to a low supply and chemical composition of nutrients. The average integrated primary productions (IPP) were 80.4, 75.9, and 76.3 mg C m−2d−1 for the cold eddy, warm eddy, and outer area, respectively. The IPP under the eddy condition was not largely different from that in reference sites, even though the vertical distribution of productivity maximum was found to be different depending on eddy condition. These weak biological responses to mesoscale eddies might be attributed to the sustaining picoplankton‐dominant community structure and no substantial upward inputs of nutrients to the euphotic zone. Our findings represent an important contribution to understanding the biological response of mesoscale eddies in the oligotrophic Philippine Sea.

Chemical composition analysis and antioxidant activity of black rice pigment.

The purpose of this research was conducted to determine anthocyanin constituent from the grains of Purple black rice No. 6. Moreover, the in vitro antioxidant activity of black rice pigment (BRP) was evaluated. The crude extract of black rice was isolated and purified by silica gel thin layer chromatography. Anthocyanins were elucidated using ultraviolet visible spectroscopy, infrared spectroscopy, nuclear magnetic resonance spectroscopy, and high-performance liquid chromatography. Meanwhile, salicylic acid Fenton system, superoxide radical system, H2 O2 scavenging system, DPPH-free radical scavenging system, and Prussian blue method were used to determine the free radical scavenging ability and total reducing ability of BRP. Overall results suggest that the main component of BRP is cyanidin-3-glucoside, which is a kind of anthocyanins. It also had significant scavenging capacity to OH, O2- , H2 O2 , DPPH, and its scavenging rate and reducing ability increased with the increase in pigment concentration.

Chemical Composition Analysis, Antioxidant, and Antibacterial Activities of Eggplant Leaves.

Agricultural by-products represent one of the biggest pollutants for the environment, therefore, investigating new sources for the extraction of bioactive compounds is a topic of great interest for the entire scientific community. This work is aimed at studying the chemical composition (pigments, phenolics, volatile compounds) and biological activities (antioxidant and antimicrobial properties) of young and mature leaves of five eggplant cultivars (Solanum melongena L.) grown in Tunisia (Tizona, A1014, Nawel, A336, and Mirval). Pigment analysis showed no statistical variations between the two types of leaves, whereas higher levels of anthocyanins were recorded in young leaves of three out of five cultivars (24.7-52.8 mg 100 g-1 ). Total phenolic compounds (966.5-2072.6 mg 100 g-1 ) and o-diphenols (343.9-741.2 mg 100 g-1 ) contents varied significantly according to leaf age and cultivar. However, no statistically significant variation between young and mature leaves was found in flavonols and tannins content. As for volatile profiles, 62 constituents were characterized with non-terpene derivatives (52.0-61.6 %) and apocarotenes (19.9-35.6 %) as major chemical classes. Regarding the antioxidant activity, methanol extracts of Tizona cultivar mature leaves displayed important antiradical properties using DPPH, ABTS, and phosphomolybdenum assays (IC50 0.1, 0.2 and 0.01 mg mL-1 , respectively). With regards to reducing power determination, the most active extract was obtained from mature A1014 leaves. Further, young Mirval leaves proved to be significantly active from a bactericidal point of view against Candida albicans and Staphylococcus aureus of 3.1 mg mL-1 and 0.2 mg mL-1 MBC, respectively. In conclusion, eggplant leaves are a noteworthy source of antioxidant and antimicrobial compounds with potential use in the pharmaceutical, the cosmetics and the food industries.

Role of Plants in Biomonitoring Vehicular Emissions

Introduction: Air pollution caused by the pollutants present in the atmosphere which is harmful to theenvironment and the living population. The plants showed stunted growth, animal’s causes various formof diseases and the pollutants harmfully affect the human beings by causing respiratory problems and thewhole environment is totally affected causing depletion of ozone layer and that directly or indirectly affectsthe living system present in the environment.Materials and Method: The leaf samples were collected from the Sholinganallur area and authenticated.Later the samples were characterized for morphological characters, pigment analysis, Biochemicalparameters and the score of tolerance index of Air pollution was calculated were tabled and graphed as perthe standard protocol.Results: The samples were collected from sholinganallur traffic zonal area and totally 14 leaf samples weregathered from the experimental site and was authenticated. The morphological characters of length andwidth of leaf samples were calculated and in Biochemical parameters, pH of leaf, Relative water contentand ascorbic acid content showed good activity in evergreen shrub. In pigment analysis of chlorophyll,the maximum and minimum amount was found in evergreen shrub Bougainvillea glabra with 68.72mg/gand Tecoma capensis in 14.72mg/g. The APTI value was maximum and minimum in evergreen shrubTabernaemontana divaricata with 114.66 and Nerium oleander with 48.05.Conclusion: The plants found in and around the experimental area possess good activity against the airpollution and these plants can be kept at the roadside to prevent and withstand the air pollution in theenvironment.

The decline of Fraxinus angustifolia Vahl in a Mediterranean salt meadow: Chlorophyll fluorescence measurements in long-term field experiment

The ecological transition zones also known as ecotones are specific ecosystems but how global change will affect their biodiversity and functioning is poorly understood. In the case of the areas at the interface between riparian forest and salt meadows the biodiversity conservation remains a major issue. The aim of this work was to evaluate the effects of a salty environment on Fraxinus angustifolia a non-halophyte species. For this, the photochemistry of PSII was in situ explored during the summer between 2007 and 2016. The predawn maximum quantum efficiency of PSII (Fv/Fm) was always greater than 0.8 which indicates an absence of chronic photoinhibition induced by salinity. During these years, the light kinetic of the actual quantum yield of PSII (ΦPSII) was studied. It appears that this yield is generally higher in salty environments than in the riparian forest. Multivariate analysis, showed that the dissipation regulated light energy faction (ΦNPQ), the photochemical quenching (qP) and the Fd/Fs ratio constitute the main explanatory variables of ΦPSII changes. The subsequent two-way-ANCOVA carried out demonstrated that ΦNPQ was weaker in saline environments, suggesting a better efficiency of photosystem collecting antennas. Meanwhile, the analysis of photosynthetic pigments did not exhibited a salt effect except the significant accumulation of chlorophyll a in 2016. In order to address the question of the salt-related toxicity, sodium ion contents in the leaves were found to be identical in the two sites, suggesting the establishment of an energy-dependent mechanism of salt exclusion/retention in roots. The growth rings of F. angustifolia in saline areas highly follows the annual water deficit changes. Between 2007 and 2016, the biomass in the salt meadow was higher with a peak in 2011 representing 60% of the biomass of control individuals. But, since 2011 a continuous decline of growth was recorded. Water conditions seem to constitute a limiting factor even if the photosynthetic apparatus generates the excess metabolic energy necessary to cope with the saline toxicity. This study thus highlighted the plasticity of the photochemistry of the PSII depending to the environmental conditions of the salt meadows. It also shows the vulnerability of an ecotone through the decline in the productivity of a glycophyte with climate change. This augurs for changes in the composition of plant communities which is in favor of strict halophytes.

Spatial distribution of planktonic ciliates in the western Pacific Ocean: along the transect from Shenzhen (China) to Pohnpei (Micronesia).

Planktonic ciliates have been recognized as major consumers of nano- and picoplankton in pelagic ecosystems, playing pivotal roles in the transfer of matter and energy in the microbial loop. However, due to the difficulties in identification, the species composition of ciliate assemblages, especially for the small, fragile, and naked species that usually dominate the ciliate communities in the oceanic waters, remains largely unknown. In the present study, 22 stations along the transect from Shenzhen (China) to Pohnpei (Micronesia) were sampled for the enumeration of picoplankton and nanoflagellates. In addition, pigment analysis of major phytoplankton groups along with the measurements of environmental variables including temperature, salinity, and nutrients were also carried out. Ciliates were identified at species level using quantitative protargol stain to reveal the species composition and their distribution patterns from off-shore to open ocean. Ciliate abundance was positively correlated with phosphate, silicate, and pico-sized pigmented eukaryotes (PPEs), whereas the biomass was closely related with PPEs, heterotrophic nanoflagellates, and chlorophytes. The combination of silicate and pigmented nanoflagellates was identified as the major factor driving the ciliate community composition. The close relationship between silicate and ciliate abundance and community structure needs further validation based on more data collected from oceanic waters. Our study showed the necessity of using techniques that can reveal the community composition at higher taxonomic resolutions in future studies on ciliates.

Cultivation of juvenile Colossoma macropomum in different colored tanks in recirculating aquaculture system (RAS): Effects on performance, metabolism and skin pigmentation

This study assessed the effects of different tank colors (white, blue, green and black) on the performance, metabolism and skin pigmentation of Colossoma macropomum cultivated in recirculating aquaculture system (RAS). In Phase 1 of the experiment, 224 juvenile C. macropomum (4.04 ± 0.32 g) were cultivated for a period of 30 days at a density of 0.5 juveniles/L. In Phase 2 of the experiment, the juveniles (34.32 ± 12.14 g) were cultivated within each treatment for another 20 days at a density of 0.25 juveniles/L. Biometrics were performed for all juveniles after 30 and 50 days of cultivation. Blood was also collected after 50 days of cultivation and skin pigmentation analysis and determination of the hepatosomatic index (HIS) and lipid concentration in the liver (HL) were performed. Juveniles grown in white tanks had the highest final weight (FW), total length (TL) and weight gain (WG) among treatments at the end of the both phases. At the end of Phase 2, juveniles grown in white tanks had higher levels of triglycerides (p < 0.05) than juveniles of the other treatments. Juveniles grown in black tanks had darker pigmentation and higher hemoglobin and total plasma protein (p < 0.05) than the other treatments. Tank color was found to affect the performance, pigmentation and metabolism of juvenile C. macropomum cultivated in RAS, with white tanks providing the best environment for cultivation and welfare.

A Novel Spectral Matching Approach for Pigment: Spectral Subsection Identification Considering Ion Absorption Characteristics

Background: Hyperspectral technology has made it possible to perform completely non-invasive investigations on pigment analysis, in particular, on pigment identification. The most commonly used method of pigment identification is to compare the spectral similarity between ones of unknown target and ones in spectral library, which requires a comprehensive and complete spectral library and is based on overall shape of the spectrum. To a certain extent, it may ignore some of the key absorption characteristics of the spectrum. Methods: A novel spectral matching method was proposed based on the spectrum divided into subsections for identification according to the main ion absorption characteristics. Main works: (1) establishing a spectral library suitable for typical pigment identification of painting; (2) discussing the main components, as well as the absorption positions of the ions and functional groups contained in pigments frequently used by artists; (3) presenting a novel spectral matching algorithm carried on spectral subsections for pigment identification; (4) verifying the feasibility and applicability of proposed method by a Chinese painting and a fresco. Conclusions: The proposed method can correctly identify the main pigments or components contained in the mixed area, which is better than the traditional method and more convenient than the unmixing method, except for some limitations in detecting white and black pigments.

Transcriptomic analysis of eyelid pigmentation in Hereford cattle.

The study of skin color in cattle holds both economic and scientific interest. Several ocular diseases of cattle have been associated with low pigmentation of the eyelids, including ocular squamous cell carcinoma and infectious keratoconjunctivitis, the two most common ocular diseases affecting cattle production. Although low eyelid pigmentation is a well-known risk factor for various ocular diseases, the genetic and biological basis of this relationship is largely unknown. We investigated the transcriptome of eyelid skin in Hereford cattle using RNA-sequencing technology. Two contrasting groups were evaluated: steers that were completely pigmented and steers with no pigmentation in both eyelids. Most of the up-regulated genes in pigmented samples are directly implicated in melanogenesis and melanosome development, whereas up-regulated genes in non-pigmented samples are implicated in cancer development and the immune system, among other functions. Interestingly, network analysis comparing pigmented vs. non-pigmented samples revealed significant differences in the co-expression patterns of genes related to melanosome, pigmentation and defense response to bacteria, showing higher gene activity, greater co-expression patterns and tighter co-regulation mechanisms in pigmented samples. Overall, our findings indicate that bovine eyelid pigmentation depends on the expression of many genes involved not only in pigmentation and melanosome function but also related to inflammatory response, infection and tumoral pathways.

Combined pigment and metatranscriptomic analysis reveals highly synchronized diel patterns of phenotypic light response across domains in the open oligotrophic ocean.

Sunlight is the most important environmental control on diel fluctuations in phytoplankton activity, and understanding diel microbial processes is essential to the study of oceanic biogeochemical cycles. Yet, little is known about the in situ temporal dynamics of phytoplankton metabolic activities and their coordination across different populations. We investigated diel orchestration of phytoplankton activity in photosynthesis, photoacclimation, and photoprotection by analyzing pigment and quinone distributions in combination with metatranscriptomes in surface waters of the North Pacific Subtropical Gyre (NPSG). We found diel cycles in pigment abundances resulting from the balance of their synthesis and consumption. These dynamics suggest that night represents a metabolic recovery phase, refilling cellular pigment stores, while photosystems are remodeled towards photoprotection during daytime. Transcript levels of genes involved in photosynthesis and pigment metabolism had synchronized diel expression patterns among all taxa, reflecting the driving force light imparts upon photosynthetic organisms in the ocean, while other environmental factors drive niche differentiation. For instance, observed decoupling of diel oscillations in transcripts and related pigments indicates that pigment abundances are modulated by environmental factors extending beyond gene expression/regulation reinforcing the need to combine metatranscriptomics with proteomics and metabolomics to fully understand the timing of these critical processes in situ.

Bioactive metabolites produced from Streptomyces enissocaesilis SSASC10 against fish pathogens

Abstract In this study Marine pigment producing actinobacteria were screened for bioactive metabolite against fish pathogens. Totally ten morphologically different actinobacteria was isolated from sediment sample of Mudasalodai, Chidambaram, India. The isolates were preliminary tested for antibacterial activity by agar plug method against Pseudomonas sp, Vibrio anguillarum, Aeromonas hydrophilia, Vibrio paraheamolyticus and Vibrio harveyi. The brown pigment producing isolate SSASC found to inhibit all the tested fish pathogens and hence selected for further study. The bioactive metabolite was extracted using different solvents such as methanol, chloroform, ethyl acetate and n-hexane in which ethyl extract showed promising antibacterial activity. The minimum inhibitory concentration (MIC) of the ethyl acetate extract ranged from 15.6 μg/ml to 125 μg/ml and minimum bactericidal concentration (MBC) found to be in the range of 62.5 μg/ml to 250 μg/ml. The ethyl acetate extract was purified using TLC and a single spot with Rf value of 0.45 was observed in TLC. The FT-IR spectrum exhibits characteristic signals at 16 peaks corresponding to functional groups indicating phosphenes, amino acids, and presence of alkyl ether. GC-MS analysis of brown pigment from SSASC showed 25 peaks. The potent strain SSASC10 was identified as Streptomyces enissocaesilis based on morphological, biochemical and molecular characteristics. In this present finding, the isolated Streptomyces enissocaesilis SSASC10 strain found to have excellent antibacterial activity against fish pathogens hence can be used for treating infectious fish disease.

Spatiotemporal Variation in Phytoplankton Community Driven by Environmental Factors in the Northern East China Sea

The East China Sea (ECS) is the largest marginal sea in the northern western Pacific Ocean. In comparison to various physical studies, little information on the seasonal patterns in community structure of phytoplankton is currently available. Based on high performance liquid chromatography (HPLC) pigment analysis, spatiotemporal variations in phytoplankton community compositions were investigated in the northern ECS. Water temperature and salinity generally decreased toward the western part of the study area but warmer conditions in August led to strong vertical stratification of the water column. In general, major inorganic nutrient concentrations were considerably higher in the western part with a shallow water depth, and consistent with previous results, had no discernable vertical pattern during our observation period except in August. This study also revealed PO4-limited environmental conditions in May and August. The monthly averaged integral chlorophyll-a concentration varied seasonally, highest (35.2 ± 20.22 mg m−2) in May and lowest (5.2 ± 2.54 mg m−2) in February. No distinct vertical differences in phytoplankton community compositions were observed for all the sampling seasons except in August when cyanobacteria predominated in the nutrient-deficient surface layer and diatoms prevailed at deep layer. Canonical correlation analysis results revealed that nutrient distribution and the water temperature were the major drivers of the vertical distribution of phytoplankton communities in August. Spatially, a noticeable difference in phytoplankton community structure between the eastern and western parts was observed in November with diatom domination in the western part and cyanobacteria domination in the eastern part, which were significantly (p &lt; 0.01) correlated with water temperature, salinity, light conditions, and nutrient concentrations. Overall, the two major phytoplankton groups were diatoms (32.0%) and cyanobacteria (20.6%) in the northern ECS and the two groups were negatively correlated, which holds a significant ecological meaning under expected warming ocean conditions.

Differences in diversity and photoprotection capability between ice algae and under-ice phytoplankton in Saroma-Ko Lagoon, Japan: a comparative taxonomic diatom analysis with microscopy and DNA barcoding

Sea ice algae, comprised mainly of diatoms, are the main primary producers in polar ecosystems, and they are generally distributed with the highest biomass at the bottom of ice. The taxonomy of ice algae has been traditionally investigated using light microscopy, but molecular techniques, including pigment analysis, have recently provided new insights into the diversity and physiology of ice algae. However, no comparative taxonomic survey has been conducted for ice algae thus far. Here, we investigated differences and similarities in (1) the diversity and (2) the photosynthetic strategies of diatom communities in sea ice and the underlying seawater of Saroma-Ko Lagoon, Hokkaido, Japan, using algal pigment signatures determined by ultra-high performance liquid chromatography, light and scanning electron microscopy and Ion Torrent next-generation sequencing techniques targeting the 18S rRNA gene (i.e., DNA barcoding). Ice algae typically possessed greater biomass (> 20-fold) and chlorophyll (Chl) a breakdown products than under-ice phytoplankton, suggesting that the ice algae formed blooms, and cell senescence and disruption could be significant mitigating factors. At the genus level, the diversity of diatoms in sea ice was higher than in under-ice seawater, although the evenness was comparable or lower in ice algae than in under-ice phytoplankton. Ice algae had a larger xanthophyll pool size and a higher ratio of photoprotective to photosynthetic pigments (11-fold and 4.5-fold higher, respectively) than under-ice phytoplankton. The results indicated that ice algae were well adapted to changes in the light regime, which could partly support their survival capability and high taxonomic diversity.

From pigments to coloured napkins: comparative analyses of primary aromatic amines in cold water extracts of printed tissues by LC-HRMS and LC-MS/MS

ABSTRACT A collaborative study was conducted to understand the correlation between pigments purity profile (primary aromatic amine content of the pigments) and the behaviour of these PAAs during cold water extraction (CWE) tests according to EN 645. From a selection of organic pigments based on seven colour indexes (PR122, PR184, PO13, PY74, PY111, PY138 and PY155), the pigment purity profile was established according to European Resolution AP (89) 1, then mono-pigmented inks were prepared and napkins printed with these inks. In a second step, cold water extraction and PAA determination were performed by two independent laboratories. In one laboratory, an analytical method based on LC-MS/MS was used, whereas in the other laboratory a method based on LC-HRMS using Orbitrap technology was developed for the simultaneous analysis of 35 PAAs. Good qualitative results were obtained if we consider that at significant levels the PAAs were positively detected in both laboratories, except for 3-amino-4-methoxybenzanilide and 8-amino-2-methyl-quinoline, for which inter-laboratory differences were observed. It was also shown that no contamination from unexpected PAAs was detected. The comparison between pigment analysis and CWE results shows that if the pigment purity profile is of major importance, other parameters such as pigment surface treatment, ink grinding process or ink formulation could have an important influence on the CWE results. For such sensitive applications, for example napkins or other Food Contact Materials (FCM), it is therefore recommended not only to select a pigment with a good purity profile but also to test the pigment in the final application. Finally, this work highlights the difficulty of validating a product on a single analysis and shows the importance of a multilevel global assessment on worst case application.