New SERS feature of β‐carotene: consequences for quantitative SERS analysis

While recording SERS spectra of pure β‐carotene at sub‐micromole concentrations for reference purpose, we discovered an unusual spectral response never reported before. In pre‐resonance conditions with the 532‐nm line, SERS of β‐carotene with AgNPs exhibits among the strong υ(CC) mode at 1512 cm⁻¹ unshifted from normal Raman spectrum, additional strong bands at 1649, 1575 and 1387 cm⁻¹ as well as other medium bands not observed in the Raman spectrum of the crystalline powder. Such behavior is explained in terms of selection rules relaxation upon cyclohexene terminal rings of the β‐carotene interaction with the NP surface. AFM images of the SERS system suggested dimers and trimers clustering of the nanoparticles with adsorbed β‐carotene. In light of the new SERS feature the consequences in correct interpretation of the SERS imaging from complex biosystems containing carotenoids are discussed. Relative intensity ratio of the β‐carotene band at 1512 cm⁻¹ and water against concentration allowed a reliable SERS calibration curve for 50 to 500 nmol l⁻¹ concentration range and provided quantitative SERS assessment of the carotenoid content in the sea urchin (Paracentrotus lividus) gonads extracts. Copyright © 2015 John Wiley & Sons, Ltd.     

Solvatochromism of 2‐(N,N‐dimethylamino)‐7‐nitrofluorene and the natural dye β‐carotene: application for the determination of solvent dipolarity and polarizability

The effects of solvents on chemical phenomena (rate and equilibrium constants, spectroscopic transitions, etc.) are conveniently described by solvation free‐energy relationships that take into account solvent acidity, basicity and dipolarity/polarizability. The latter can be separated into its components by manipulating the UV–vis spectra of two solvatochromic probes, 2‐(N,N‐dimethylamino)‐7‐nitrofluorene (DMANF) and a di‐(tert‐butyl)‐tetramethyl docosanonaen probe (ttbP9) whose synthesis is laborious and expensive. Recently, we have shown that the natural dye β‐carotene can be conveniently employed instead of ttbP9 for the determination of solvent polarizability (SP) of 76 molecular solvents and four ionic liquids. In the present work, we report the polarizabilities of further 24 solvents. Based on the solvatochromism of β‐carotene and DMANF, we have calculated solvent dipolarity (SD) for 103 protic and aprotic molecular solvents, and ionic liquids. The dependence of SD and SP on the number of carbon atoms in the acyl‐ or alkyl group of several homologous series (alcohols; 2‐alkoxyethanols; carboxylic acid‐ anhydrides, and esters, ionic liquids) is calculated and briefly discussed. Copyright © 2013 John Wiley & Sons, Ltd.     

Vitamin A and Bone Health: A Review on Current Evidence

Vitamin A is a fat-soluble micronutrient essential for growth, immunity, and good vision. The preformed retinol is commonly found in food of animal origin whereas provitamin A is derived from food of plant origin. This review summarises the current evidence from animal, human and cell-culture studies on the effects of vitamin A towards bone health. Animal studies showed that the negative effects of retinol on the skeleton were observed at higher concentrations, especially on the cortical bone. In humans, the direct relationship between vitamin A and poor bone health was more pronounced in individuals with obesity or vitamin D deficiency. Mechanistically, vitamin A differentially influenced the stages of osteogenesis by enhancing early osteoblastic differentiation and inhibiting bone mineralisation via retinoic acid receptor (RAR) signalling and modulation of osteocyte/osteoblast-related bone peptides. However, adequate vitamin A intake through food or supplements was shown to maintain healthy bones. Meanwhile, provitamin A (carotene and β-cryptoxanthin) may also protect bone. In vitro evidence showed that carotene and β-cryptoxanthin may serve as precursors for retinoids, specifically all-trans-retinoic acid, which serve as ligand for RARs to promote osteogenesis and suppressed nuclear factor-kappa B activation to inhibit the differentiation and maturation of osteoclasts. In conclusion, we suggest that both vitamin A and provitamin A may be potential bone-protecting agents, and more studies are warranted to support this hypothesis.     

Growth profile of stimulated Raman anti‐Stokes scattering influenced by fluorescence seeding in liquid‐core optical fiber

We demonstrated the growth profile of stimulated Raman anti‐Stokes scattering (SRS) of carbon disulfide (CS₂) influenced by fluorescence seeding of all‐trans‐β‐carotene and rhodamine‐B (RhB) in liquid‐core optical fiber (LCOF). The pump energy which was needed to build up the first‐order anti‐Stokes radiation of CS₂ solutions with the fluorescence seeding was lower than that of CS₂ solutions without fluorescence seeding because of the fluorescence enhancement effect on the intensity of the first‐order Stokes radiation. The first‐order anti‐Stokes radiation of the RhB solution (10⁻⁸ M ) was built up at a lower pump energy than that of the all‐trans‐β‐carotene solution (10⁻⁶ M ), and the intensity of the first‐order anti‐Stokes radiation of the RhB solution was higher than that of the all‐trans‐β‐carotene solution. Simultaneously, the coupled wave differential equations were obtained by the theoretical derivation, and the growth profile of the first‐order anti‐Stokes radiation was theoretically calculated with and without the fluorescence seeding by these equations. Copyright © 2009 John Wiley & Sons, Ltd.     

1H HR-MAS NMR of carotenoids in aqueous samples and raw vegetables

Carotenoids are linear C40 tetraterpenoid hydrocarbons and represent a wide category of natural pigments. They are components of the pigment system of chloroplasts and are involved in the primary light absorption and the photon canalization of photosynthesis. Moreover, they also behave as quenchers of singlet oxygen, protecting cells and organisms against lipid peroxidation.Carotenoids have a strong lipophilic character and are usually analyzed in organic solvents. However, because of their biological activity, the characterization of these compounds in an aqueous environment or in the natural matrix is very important.One of the most important dietary carotenoids is beta-carotene, which has been extensively studied both in vivo and in model systems, but because of the low concentration and strong interaction with the biological matrix, beta-carotene has never been observed by NMR in solid aqueous samples.In the present work, a model system has been developed for the detection and identification of beta-carotene in solid aqueous samples by 1H HR-MAS NMR. The efficiency of the model has led to the identification of beta-carotene in a raw vegetable matrix.     

A stable ‘sandwich’ system of Surface‐Enhanced Resonance Raman Scattering for the analysis of β‐carotenes in a photosynthetic pigment‐protein complex

In plants, Photosystem I (PSI) is composed of a core complex and a membrane‐associated antenna complex light‐harvesting complex I that captures light and funnels its energy to the core complex. To obtain Raman structural information on β‐carotenes embedded in the PSI core complex, a ‘sandwich’ system of roughened silver slice: target protein complexes: single silver nanoparticles was fabricated for Surface‐Enhanced Resonance Raman Scattering (SERRS) measurements. This study provided a method to overcome spectral irreproducibility, which is the main drawback of Surface‐Enhanced Raman Scattering/SERRS‐based studies. The Raman spectra of β‐carotenes embedded in the PSI core complex can be obtained at very low sample concentrations (1–5 µg Chl/ml) and high signal/noise ratios. The β‐carotenes in the spinach PSI core complex were predominantly all‐trans configuration. The membrane protein‐mediated adsorption of silver nanoparticles induced the uniform distribution of a large number of single nanoparticles, which contributed to achieving highly reproducible SERRS spectra. This study is the first to apply single silver nanoparticle‐based SERRS analysis in membrane proteins. Copyright © 2013 John Wiley & Sons, Ltd.     

氧化镁和氧化铝柱层析法分离胡萝卜素的实验效果对比分析

通过比较氧化镁和氧化铝柱层析法分离胡萝卜素的实验效果,为实验选择最佳吸附剂。分别以粉末状氧化镁和氧化铝作为吸附剂,采用柱层析法分离菠菜叶中的胡萝卜素。采用氧化镁作吸附剂,可将菠菜叶中的色素清晰地分为3条色带,从层析柱前向后依次为胡萝卜素、叶绿素a和叶绿素b,各色带致密,边缘整齐,彼此界限清晰;采用氧化铝作吸附剂,同样分离出3条色带,但色带均弥散较宽,颜色较浅,边缘欠整齐,以胡萝卜素带最显著。结果表明,柱层析法分离胡萝卜素时,氧化镁作为吸附剂实验效果优于氧化铝。     

Reaction of carotenoids with CCl<Subscript>3</Subscript>OO<Superscript>.</Superscript> by using pulse radiolysis

The interactions of carotenoids (bixin, β-carotene and lycopene) with CCl₃OO^. in aqueous and i-propylalcohol solution saturated with air have been studied by pulse radiolysis. For bixin and β-carotene reaction products from forming process, absorbing in the region of 650 nm, is observed with concomitant carotenoid bleaching (bixin at 500 nm, β-carotene at 450 nm). Their rate constants from forming process are 1.78 ×10⁸ and 7.8 ×10⁷ mol^-1 · L · s^-1 respectively. However, in the case of lycopene, no such a forming process of reaction as bixin and β-carotene can be observed although there is the bleaching reaction (rate constant 4 ×10⁷ mol^-1 · L · s^-1). The results suggest that the carotenoid radical cation and an additional radical are produced in the case of bixin and β-carotene, whereas lycopene undergoes electron transfer with CCl₃OO^., forming cation radical.     

Comparative analysis of plant lycopene cyclases

Carotenoids are essential isoprenoid pigments produced by plants, algae, fungi and bacteria. Lycopene cyclase (LYC) commonly cyclize carotenoids, which is an important branching step in the carotenogenesis, at one or both end of the backbone. Plants have two types of LYC (β-LCY and ϵ-LCY). In this study, plant LYCs were analyzed. Based on domain analysis, all LYCs accommodate lycopene cyclase domain (Pf05834). Furthermore, motif analysis indicated that motifs were conserved among the plants. On the basis of phylogenetic analysis, β-LCYs and ϵ-LCYs were classified in β and ϵ groups. Monocot and dicot plants separated from each other in the phylogenetic tree. Subsequently, Oryza sativa Japonica Group and Zea mays of LYCs as monocot plants and Vitis vinifera and Solanum lycopersicum of LYCs as dicot plants were analyzed. According to nucleotide diversity analysis of β-LCY and ϵ-LCY genes, nucleotide diversities were found to be π: 0.30 and π: 0.25, respectively. The result highlighted β-LCY genes showed higher nucleotide diversity than ϵ-LCY genes. LYCs interacting genes and their co-expression partners were also predicted using String server. The obtained data suggested the importance of LYCs in carotenoid metabolism. 3D modeling revealed that depicted structures were similar in O. sativa, Z mays, S. lycopersicum, and V. vinifera β-LCYs and ϵ-LCYs. Likewise, the predicted binding sites were highly similar between O. sativa, Z mays, S. lycopersicum, and V. vinifera LCYs. Most importantly, analysis elucidated the V/IXGXGXXGXXXA motif for both type of LYC (β-LCY and ϵ-LCY). This motif related to Rossmann fold domain and probably provides a flat platform for binding of FAD in O. sativa, Z mays, S. lycopersicum, and V. vinifera β-LCYs and ϵ-LCYs with conserved structure. In addition to lycopene cyclase domain, the V/IXGXGXXGXXXA motif can be used for exploring LYCs proteins and to annotate the function of unknown proteins containing lycopene cyclase domain. Overall results indicated that a high degree of conserved signature were observed in plant LYCs.