Qualitative and quantitative variations in carotenoid pigments in the ovary and hepatopancreas of Penaeus schmitti during ovarian maturation

Carotenoid pigments of Penaeus schmitti were investigated and identified in the ovaries and hepatopancreas. Their individual variations were measured in these two organs. The relative concentrations of zeaxanthin in hepatopancreas and astaxanthin in ovaries increased during the sexual development. The role of zeaxanthin monoester in carotenoids transfer from hepatopancreas to ovaries during this sexual development is discussed.     

Chromatographic analysis of carotenol fatty acid esters in Physalis alkekengi and Hippophae rhamnoides

The carotenol fatty acid esters of two potentially valuable sources of plant carotenoids, sepals of Physalis alkekengi (Chinese lantern) and fruits of Hippophae rhamnoides (sea buckthorn), were separated by column chromatography and identified by HPLC-DAD and HPLC-MS. A chemical and an enzymatic hydrolysis were employed to identify the parent carotenoids and to remove the lipid components. Zeaxanthin and beta-cryptoxanthin esters represented the main fraction in P. alkekengi sepals and an important one in H. rhamnoides fruits. Beta-Cryptoxanthin palmitate and zeaxanthin dipalmitate were identified as major compounds in both plants. In P. alkekengi, the carotenoids were mainly (> 90%) esterified with palmitic acid, and a high proportion (> 80%) of saturated medium chain fatty acids was found (by GC-MS) in the total lipid extract. Although the total lipid extract of H. rhamnoides contained significant amounts of unsaturated fatty acids, especially oleic and palmitoleic acids, the xanthophylls were mainly esterified with saturated fatty acids. The oleoresins of both species represent potential sources of carotenoid esters and can be used as food additives, cosmetic ingredients or nutraceuticals.     

Identification of carotenoid pigments and their fatty acid esters in an avian integument combining HPLC-DAD and LC-MS analyses

Yellow-orange-red ornaments present in the integuments (feathers, bare parts) of birds are often produced by carotenoid pigments and may serve to signal the quality of the bearer. Although carotenoid esterification in tissues is a common phenomenon, most of the work on avian carotenoids has been focused on the identification of free forms or have been done after sample saponification. Here we determined free and esterified carotenoid composition in a bird species with red ornaments: the red-legged partridge (Alectoris rufa). Carotenoids from leg integument were extracted and processed by TLC to separate three major carotenoid groups (free form, mono- and diesters with fatty acids), whereas saponified extracts gave only free forms of carotenoids. TLC fractions were then analyzed by HPLC-DAD with C18 phase column for a preliminary identification of carotenoid groups. The final characterization of free carotenoids and its esters with fatty acids was performed with direct extracts analyzed by LC-MS and LC-MS/MS with a C30 phase, always with a system coupled to DAD. The main carotenoid (λ(max) 478 nm and [M+H](+) at m/z 597.2) was identified as astaxanthin by comparison with standards. A second carotenoid (λ(max) between 440 and 480 nm and [M+H](+) at m/z 581.3) was not identified among any of the commercially available carotenoid standards, although it could correspond to pectenolone according to its fragmentation pattern. Both the unidentified carotenoid and astaxanthin formed monoesters with fatty acids, but only astaxanthin was in its diesterified form. Monoesters were mainly formed with palmitic, stearic, oleic and linoleic acids. Complementary analyses of fatty acid composition in partridge integument by GC-MS revealed high amounts of these and other fatty acids, such as myristic, arachidic and docosanoic acids. The combination of HPLC-DAD and LC-MS/MS spectra was especially useful to identify the carotenoids present in the esterified forms and the probable masses of the fatty acids included in them, respectively.     

Astaxanthin from the red crab langostilla (Pleuroncodes planipes): optical R/S isomers and fatty acid moieties of astaxanthin esters

The composition of the fatty acids of astaxanthin esters and the distribution of astaxanthin optical RS isomers in the esterified and unesterified astaxanthin fractions extracted from the meal of the pelagic red crab langostilla (Pleuroncodes planipes; Decapoda, Anomura) were determined. Astaxanthin diesters comprised approximately 70%, monoesterified astaxanthin approximately 12%, and unesterified astaxanthin approximately 10% of total carotenoids, respectively. Unidentified carotenes and minor yellow xanthophylls represented approximately 8% of the total carotenoids. Three astaxanthin diester fractions (ratio 5:4:1) and one monoester fraction were clearly distinguished by thin-layer chromatography, and fatty acid moieties were determined in all of them. Saturated fatty acids accumulated in astaxanthin diesters, but were reduced in the monoester fraction when compared to langostilla crude oil extract (CE). Astaxanthin diesters, but not monoesters were enriched in C16:0 and C18:1n-9, when compared to the CE. Astaxanthin monoesters were rich in polyunsaturated fatty acids (approximately 70% of total fatty acids), in particular C20:5n-3 and C22:6n-3. Acylation of astaxanthin in langostilla seems to be selective rather than specific. The three diesterified astaxanthin fractions of langostilla had a ratio of approximately 3:1:3 between the (3R,3'R)-, (3R,3'S)-, and (3S,3'S)-astaxanthin isomers, whereas in the monoesterified and unesterified fractions the ratio was approximately 4:1:4. The astaxanthin optical RS isomer composition indicates that langostilla is unable to racemize astaxanthin.     

Variation in lipid composition of Chinese mitten-handed crab, Eriocheir sinensis during ovarian maturation

This experiment was conducted to investigate the variation in lipid composition during the ovarian maturation of the crab Eriocheir sinensis. The Chinese mitten-handed crab broodstock was divided into six different maturation periods according to the size and color of ovary. Ovary, hepatopancreas, muscle, and hemolymph of broodstock in different maturation periods were analyzed for total lipid and fatty acids using gas chromatography, and lipid classes by thin-layer chromatography. The ovarian lipid concentration (expressed as percent wet ovarian weight) increased steadily from stage II (5.4%) to stage IV (19.1%), and decreased to the lowest levels after spawning (stage V, 6.6%). The hepatopancreatic lipid concentration (expressed as percent wet hepatopancreatic weight) increased with maturity of the ovaries, reached a maximum at stage III(2) (29.9%), and decreased during the subsequent period to spawning (16.7%). The muscular and hemolymph lipid concentration did not change markedly during the ovarian development. These results suggest the possible movement of hepatopancreatic lipids to the ovaries during the ovarian maturation. Both triacylglycerol and phosphatidylcholine were responsible for the increase in ovarian lipid concentration during sexual maturation. The fatty acids of total lipid, triacylglycerol, and phosphatidylcholine of the ovaries did not vary systematically during the ovarian maturation, but the ratio between n-3PUFA (polyunsaturated fatty acid) and n-6PUFA did change regularly with the ovarian lipid. These suggest that enough PUFA, especially n-3PUFA, should be supplied to the crab during ovarian maturation.     

Development and Quality Evaluation of Hypoallergenic Bakery Products Suitable for Patients with Wheat-associated Allergy

The influence of growth temperature (21–41°C) and light intensity on compositions of four carotenoids, and of fatty acids from carotenoid glucoside ester (carotenoid K-G-FA) and from the cellular lipids in Rhodococcus rhodochrous RNMS1 were quantitatively investigated. Lowering the temperature increased the total carotenoid content and the proportion of carotenoid K-G-FA. It increased the proportion of unsaturated fatty acids in both carotenoid K-G-FA and the cellular lipids, decreased that of saturated ones, and slightly decreased that of branched-chain ones. This bacterium adapted itself to low temperature by desaturating its fatty acids. The light intensity affected neither the content and the composition of carotenoids nor the composition of the fatty acids in carotenoid K-G-FA and the cellular lipids. This bacterium was a scotochromogenic strain.     

Phospholipids from the hepatopancreas of Indian horseshoe crab Carcinoscropius rotundicauda

Total phospholipids were extracted from the heart, hepatopancreas, and hemolymph of the Indian horseshoe crab Carcinoscorpius rotundicauda by the conventional method. Characteristic group reaction and 2-dimensional thin-layer chromatography on silica gel were used for identification of different phospholipids. The phospholipid profile obtained from hemolymph and 2 major organs are comparable and show phosphatidyl choline (PC) and phosphatidyl ethanolamine to be the major phospholipids. A phospholipid has been consistently detected migrating immediately below the PC in the thin-layer chromatogram of lipids extracted from the hepatopancreas. When mixed methyl esters of this slower moving PC are resolved on a silica gel plate ran in hexane ether:acetic acid 80:20:1, with appropriate controls, an additional spot is seen just below the normal methyl ester, indicating a difference between the fatty acid compositions of 2 PC (e.g., regular and slower). The slower mixed methyl esters were found to comprise mainly the 4 saturated fatty acids: lauric, myristic, palmitic, and stearic. The slow moving PC seems to consist mainly of molecular species with the above-mentioned saturated fatty acids at both Sn 1 and Sn 2 positions.     

Time- and media-dependent secondary carotenoid accumulation in Haematococcus pluvialis

The green microalgae Haematococcus pluvialis synthesizes secondary carotenoids after exposure to environmental stress, a process that is used for the biotechnological production of astaxanthin (Ax). This study reports, for the first time, the medium-dependent changes in the carotenoid pattern throughout the cultivation process as well as the exact composition of carotenoids and their fatty acid mono- and diesters using LC-MS. Secondary carotenoid formation started immediately upon exposure to nutrient depletion and high light conditions. Ax and its corresponding mono- and diesters were detected simultaneously. After 15 days of cultivation, no significant changes were detected in carotenoid composition; however, the ratio between carotenoid mono- and diesters still varied. Main carotenoids were identified as Ax linolenate and Ax oleate, but also five adonirubin and one lutein monoester were detected. The influence of three different autotroph media was studied on carotenoid content, which reached a maximum 16.1 mg/g dry weight. The results indicate that media composition has an influence on the ratio of Ax mono- to diester but not on the qualitative composition of secondary carotenoids in H. pluvialis. Beside the pathway via echinenone, canthaxanthin and adonirubin the results indicate that Ax biosynthesis takes place via another route: from beta-carotene via beta-cryptoxanthin, zeaxanthin and adonixanthin.     

Changes of dolichol and dolichyl fatty acyl esters during the germination and development of soybeans.

The amounts of dolichol and dolichyl fatty acyl esters and their composition in various parts of soybean seedlings were determined during germination and development. The dolichol content of cotyledons decreased during germination. Dolichyl fatty acyl esters were identified in cotyledons and the amount was estimated by high performance liquid chromatography. The relative amounts of short-chain dolichols of 15, 16, and 17 isoprene units increased during development of the seedlings. The homologue distribution of free dolichol was different from that of dolichyl fatty acyl esters. The relative amounts of dolichols with 16, 17, and 18 isoprene units were greater in free dolichol than in dolichyl fatty acyl esters. The percentages of long-chain saturated fatty acids in dolichyl fatty acyl esters, specifically 21:0, 22:0, 23:0, 24:0, and 25:0, increased during development. These fatty acids represented more than 40% of the fatty acids in dolichyl fatty acyl esters in stems. These results suggest that dolichyl fatty acyl esters are not a storage form of dolichol. The large accumulation of dolichol and dolichyl fatty acyl esters in the leaves, where photosynthesis takes place, suggests some other function.     

Changes in the lipid and fatty acid composition of hemolymph and ovaries during the reproductive cycle of Labidura riparia

Lipid metabolism was investigated during the reproductive cycle of Labidura riparia (Pallas). The lipid classes and their constitutive fatty acids present in hemolymph and ovaries were measured using thin‐layer chromatography and gas‐liquid chromatography. In the hemolymph, total lipids increase steadily from the previtellogenic period to vitellogenic arrest. These lipids are predominantly diacylglycerols and phospholipids. In the ovaries, total lipids increase during vitellogenesis then decrease during the vitellogenesis arrest period. The major lipids are triacylglycerols, followed by phospholipids. In both hemolymph and ovaries, all lipid classes contained variable proportions of seven main fatty acids: the saturated fatty acids myristic acid (14:0), palmetic acid (16:0), and stearic acid (18:0); the monounsaturated fatty acids palmitoleic acid (16:1) and oleic acid (18:1); and the polyunsaturated fatty acids linoleic acid (18:2) and linolenic acid (18:3). Unsaturated fatty acids predominate throughout the reproductive cycle. The percentage compositions of total and triacylglycerol fatty acids do not change markedly during the reproductive cycle in hemolymph nor in ovaries, with 18:2, 18:1 and 16:0 fatty acids being the major components. However, for diacylglycerols and phospholipids, the proportions of fatty acids vary systematically. For phospholipids during the vitellogenesis period, 18:2 increases considerably whereas other fatty acids decrease; for diacylglycerols, these fatty acids vary in the reverse way.     

长江刀鲚生殖洄游期间脂肪酸组成及含量变化分析

为探究长江刀鲚生殖洄游过程中脂肪酸组成及其含量变化规律,研究选择洄游距离、卵巢发育和规格大小3个影响因子设置梯度,对长江刀鲚肝胰腺、肌肉和卵巢的脂肪酸组成及含量进行实验分析。实验结果显示, 62尾雌性刀鲚3个组织均检测出28种脂肪酸,以单不饱和脂肪酸(MUFA)含量最高,大于各组织总脂肪酸含量的56.23%,各类脂肪酸中的C18:1、C16:0、C16:1、DHA和EPA含量较高,为主要脂肪酸。在生殖洄游过程中,刀鲚肝胰腺总脂肪含量随洄游距离的延长呈上升趋势,从崇明江段的(526.61±38.50) mg/g增加至安庆江段的(587.21±124.72) mg/g,而肌肉和卵巢总脂肪酸含量呈显著下降趋势,分别下降了33.03%和57.09%(P<0.05)。在各体长组中,肌肉总脂肪酸、SFA、MUFA和PUFA含量与体长呈正相关(P<0.05),而肝胰腺和卵巢总脂肪酸及各类脂肪酸含量与体长无显著相关性(P>0.05)。在卵巢发育过程中,刀鲚肝胰腺和肌肉总脂肪酸、MUFA和PUFA含量随卵巢由Ⅱ期发育至Ⅳ期均呈下降趋势,总脂肪酸含量分别减少了47.56%和22.40%,...     

Phospholipids from the hepatopancreas of Indian horseshoe crab Carcinoscorpius rotundicauda

Total phospholipids were extracted from the heart, hepatopancreas, and hemolymph of the Indian horseshoe crab Carcinoscorpius rotundicauda by the conventional method. Characteristic group reaction and 2-dimensional thin-layer chromatography on silica gel were used for identification of different phospholipids. The phospholipid profile obtained from hemolymph and 2 major organs are comparable and show phosphatidyl choline (PC) and phosphatidyl ethanolamine to be the major phospholipids. A phospholipid has been consistently detected migrating immediately below the PC in the thin-layer chromatogram of lipids extracted from the hepatopancreas. When mixed methyl esters of this slower moving PC are resolved on a silica gel plate ran in hexane ether:acetic acid 80:20:1, with appropriate controls, an additional spot is seen just below the normal methyl ester, indicating a difference between the fatty acid compositions of 2 PC (e.g., regular and slower). The slower mixed methyl esters were found to comprise mainly the 4 saturated fatty acids: lauric, myristic, palmitic, and stearic. The slow moving PC seems to consist mainly of molecular species with the above-mentioned saturated fatty acids at both Sn 1 and Sn 2 positions.     

Fatty acids of astaxanthin esters in krill determined by mild mass spectrometry

Krill is a major source of astaxanthin, which has strong antioxidant activity. Fractions with astaxanthin monoesters and diesters of Antarctic krill Euphausia superba were isolated. Astaxanthin esters were separated by C18-HPLC depending on the number of carbons and double bonds of esterified fatty acid(s). Small amounts of other lipids remained in the samples, but relative molecular masses of carotenoid esters could be measured by field desorption mass spectrometry without fragmentation and interference from contaminant lipids. The fatty acids were determined by calculation of difference between astaxanthin and astaxanthin esters. Only five kinds of fatty acids, dodecanoate, tetradecanoate, hexadecanoate, hexadecenoate and octadecenoate, were detected. Fast atom bombardment mass spectrometry and secondary ion mass spectrometry showed similar spectra. The fatty acid composition in astaxanthin esters was different from those in krill lipids. Therefore, determination of fatty acids in carotenoid esters by a combination of HPLC elution profile and mild mass spectrometry is found to be a useful tool.     

Mechanisms involved in the intestinal absorption of dietary vitamin A and provitamin A carotenoids

Vitamin A is an essential nutrient for humans and is converted to the visual chromophore, 11-cis-retinal, and to the hormone, retinoic acid. Vitamin A in animal-derived foods is found as long chain acyl esters of retinol and these are digested to free fatty acids and retinol before uptake by the intestinal mucosal cell. The retinol is then reesterified to retinyl esters for incorporation into chlylomicrons and absorbed via the lymphatics or effluxed into the portal circulation facilitated by the lipid transporter, ABCA1. Provitamin A carotenoids such as β-carotene are found in plant-derived foods. These and other carotenoids are transported into the mucosal cell by scavenger receptor class B type I (SR-BI). Provitamin A carotenoids are partly converted to retinol by oxygenase and reductase enzymes and the retinol so produced is available for absorption via the two pathways described above. The efficiency of vitamin A and carotenoid intestinal absorption is determined by the regulation of a number of proteins involved in the process. Polymorphisms in genes for these proteins lead to individual variability in the metabolism and transport of vitamin A and carotenoids. This article is part of a Special Issue entitled Retinoid and Lipid Metabolism.     

Carotenoids in ripe green and in autumn senescing leaves of apple tree: I - Qualitative composition of free carotenoids,xanthophyll esters and fatty acids of esters

Abstract

By different chemical, spectral and chromatographic procedures seven carotenoids, two carotenes (α- and β-carotene) and five free xanthophylls (violaxanthin, lutein, antheraxanthin, zeaxanthin and neoxanthin) have been isolated and identified in green apple leaves. In autumn senescing leaves, besides the above seven carotenoids, the occurrence of at least seven xanthophyll esters has also been proved. Four esters, violaxanthin mono- and diester and lutein mono- and diester, (≥90% of total esters) and a minor component, neoxanthin triester, have been identified. Another minor component is most probably a monoester of antheraxanthin; the seventh, present in traces, has not been identified.

The fatty acids combined with the above xanthophyll esters have been analysed by gas-chromatography. The unusual presence of low molecular weight fatty acids, C7-C11, has been registered (1,5-8% of the total); caprilic and capric acids have been identified. Lauric acid with three minor homologous (2-7% and myristic acid (9-15%) are also present. The major component, palmitic acid, with palmitoleic and two other minor components, reaches 36-44%. In addition a remarkable presence of oleic acid (12-26%) has been observed together with stearic (8-10%), linolenic (5-6%), arachidic (0,7-2,5%) and linolenic-gadoleic acids (2,5-9,3%).     

The qualitative composition of carotenoids and their seasonal dynamics in tissues of the bivalve <Emphasis Type="Italic">Anadara kagoshimensis</Emphasis> (Tokunaga, 1906)

A total of six carotenoids, viz., β-carotene, pectenol A, pectenolone (trans- and cis-isomers), zeaxanthin, diatoxanthin, and alloxanthin, as well as esters of alloand diatoxanthin, have been detected in total carotenoid extracts from the tissues of the bivalve Anadara kagoshimensis (Tokunaga, 1906) using the methods of thin-layer chromatography, high-performance liquid chromatography, mass spectrometry, UV-VIS spectroscopy, and characteristic reactions for the identification of chemical groups. The major group (over 90% of the total carotenoids) is comprised of alloxanthin, pectenolone, and allo- and diatoxanthin esters. Tissues of A. kagoshimensis are typically characterized by cyclic variations in the level of carotenoids over the period from winter to summer, with the maxima in February and June and the minimum in April. The largest contribution to the seasonal carotenoid dynamics is made by the major group of pigments (R ² = 0.75–0.99), which depends on the pattern of succession of diatomic microalgae during the annual cycle. The pathways of metabolic transformation of the carotenoids in tissues of this bivalve are discussed.     

三疣梭子蟹卵巢发育过程中主要类胡萝卜素组成变化及其与抗氧化性能的关系

实验研究了三疣梭子蟹(Portunus trituberculatus)卵巢发育过程中(Ⅰ—V期)组织色泽、类胡萝卜素组成与含量、抗氧化和非特异性免疫指标的变化规律,进一步探讨了肝胰腺中类胡萝卜素含量变化与抗氧化指标的关系。结果表明:(1)卵巢发育期间,卵巢指数(GSI)显著增加(P<0.05),肝胰腺指数(HSI)整体呈先升后降的趋势;卵巢的红度(a*)值和黄度(b*)值、肝胰腺的亮度(L*)值和b*值均呈显著上升趋势,但卵巢中L*值呈下降趋势。(2)在卵巢发育过程中,卵巢中的总类胡萝卜素、虾青素、叶黄素和β-胡萝卜素含量均为先升后降的趋势,虾青素含量在Ⅳ期最高;肝胰腺中的虾青素含量呈上升趋势,而β-胡萝卜素含量为显著下降趋势;内表皮中的总类胡萝卜素、虾青素、叶黄素、海胆烯酮和β-胡萝卜素含量均呈现下降趋势;就不同组织中类胡萝卜素含量而言,内表皮中虾青素含量最高。卵巢和肝胰腺的L*值与总类胡萝卜素含量呈显著负相关,卵巢的a*值和b*值均与总类胡萝卜素、虾青素、叶黄素、海胆烯酮和β-胡萝卜素含量呈显著正相关。肝胰腺的a*值与各类胡萝卜素含量无显著相关性, b*值仅与虾青素含量呈显...     

Incorporation of carotenoid esters into liposomes

Carotenoid esters are investigated for their interaction with liposomal membranes and compared with their corresponding free (non-esterified) carotenoids. A monoester (beta-cryptoxanthin) and two diesters (zeaxanthin and lutein) were chosen. Egg yolk phosphatidylcholine liposomes served as the membrane model. We measured the sizes of the liposomes by photon correlation spectroscopy. The incorporation yields were determined spectrophotometrically. From liposomes simultaneously doped with the fluorescent dye Laurdan, fluidity changes of the liposomes were obtained. In summary, the results indicate that the carotenoid esters: (i) get incorporated, but at a lower yield than their corresponding free carotenoids, (ii) also increase the membrane rigidity as do the free carotenoids, and (iii) increase the liposome sizes significantly, but after extrusion through an 0.1 mum filter the sizes resemble with the exception of the liposomes incorporated with lutein diesters, they remain bigger indicating an elastic property due to two different accessible locations in the membrane.     

Conserved Fatty Acid Composition of Proteolipid Protein During Brain Development and in Myelin Subfractions

Myelin proteolipid protein (PLP) is modified after translation by the attachment of long-chain fatty acids to several cysteine residues. In this study, the amount and pattern of fatty acids covalently bound to rat PLP were determined during brain development and in myelin subfractions. For this purpose, PLP was isolated by gel-filtration chromatography in organic solvents, subjected to alkaline methanolysis, and the released fatty acid methyl esters were analyzed by gas-liquid chromatography. At all ages examined, PLP had the same amount of covalently-bound fatty acids (3–4% w/w) and palmitate, oleate and stearate were always the major acyl chains. In contrast to myelin lipids, the fatty acid composition of PLP showed only minor changes between 15-days and 90-days of age. The amount and pattern of fatty acids bound to PLP prepared from three myelin subfractions were also indistinguishable. The conservation of a characteristic PLP-fatty acid make-up during brain development and in various myelin compartments suggests that this post-translational modification is essential for the normal functioning of the protein.     

Carotenoids and fatty acids in red yeasts Sporobolomyces roseus and Rhodotorula glutinis

Rhodotorula glutinis and Sporobolomyces roseus, grown under different aeration regimes, showed differential responses in their carotenoid content. At higher aeration, the concentration of total carotenoids increased relative to biomass and total fatty acids in R. glutinis, but the composition of carotenoids (torulene > beta-carotene > gamma-carotene > torularhodin) remained unaltered. In contrast, S. roseus responded to enhanced aeration by a shift from the predominant beta-carotene to torulene and torularhodin, indicating a biosynthetic switch at the gamma-carotene branch point of carotenoid biosynthesis. The overall levels of total carotenoids in highly aerated flasks were 0.55 mol-percent and 0.50 mol-percent relative to total fatty acids in R. glutinis and S. roseus (respectively), and 206 and 412 microg g(-1) dry weight (respectively).