Trennung und Charakterisierung von β-Carotin-Isomeren

Separation and Characterization of the cis-Isomers of β,β-Carotene A stable HPLC. system is described allowing the excellent separation of 11 different cis-isomers of β,β-carotene from the all-trans compound. The system is applied to the analysis of cis/trans mixtures obtained from plant extracts and by photoisomerization of the all-trans isomer. Al₂O₃ is used as the stationary phase while hexane with controlled H₂O content is utilized as the mobile phase. With the aid of the optimum conditions 8 sufficiently stable cis isomers were isolated and their structures shown to be the 9-, 13- and 15-cis, the 9,9′-, 9, 13-, 9, 13′- and 13,13′-di-cis and, tentatively, the 9,13,13′-tri-cis β,β-carotenes by application of 270-MHz-FT.-¹H-NMR. spectroscopy.     

Determination of carotenoids in two algae species from the saline water of Kapulukaya reservoir by HPLC

The local algae species, Chlorella vulgaris and Scenedesmus regularis, from a highly saline water body of Kapulukaya Reservoir were isolated to analyze their carotenoid composition and content using HPLC method. The gradient solvent system of methanol–acetonitrile–water (84:14:2, v/v/v) and methylene chloride (100%), used to resolve a range of carotenoids from the saponified cells, proved an acceptable separation as inferred from the retention factor (k) ranging between 0.75 and 7.76 and the separation factor (α) values greater than 1. Resolution peaks assigned to carotenoids, 21 for C. vulgaris extracts and 22 for S. regularis extracts, were reached within the duration time of 45?min. Main carotenoids identified either tentatively or positively were all-trans-lutein, 9- or 9′-cis-lutein, 13- or 13′-cis-lutein, cis-lutein, All-trans-α-carotene, 9- or 9′-cis-α-carotene, All-trans-β-carotene, 9- or 9′-cis-β-carotene in the species except for all-trans-β-cryptoxanthin found only in S. regularis. Auroxanthin, neochrome, neoxanthin, and cis-neoxanthin were identified as epoxy-containing compounds. Quantitatively, C. vulgaris was distinguished to have greater amount of lutein and cis-isomers (2.74?mg/g), 77.89% while S. regularis was predominated by β-carotene and cis isomers as major component, being 80.72% (5.76?mg/g) in total carotenoids (TC). In terms of total carotenoids, the species were considered to be efficient sources for further practical applications.     

RESONANCE RAMAN EVIDENCE FOR 15-cis TO MA,-trans PHOTOISOMERIZATION OF SPIRILLOXANTHIN BOUND TO A REDUCED FORM OF THE REACTION CENTER OF Rhodospirillum rubrum SI

The reaction center (RC) of Rhodospirillum rubrum SI, which was prepared by ultrafiltration, showed one peak in molecular-sieve HPLC, but it showed two peaks in diethylaminoethyl (DEAE) ion-exchange HPLC; they were named as RC-α and RC-β in the order of elution, Nonequilibrated isoelectric electrophoresis, together with DEAE ion-exchange HPLC, showed that RC-β is electronically more negative than RC-α. Oxidation of RC-β by addition of ferricyanide caused its transformation into RC-α, while reduction of RC-α by adding ascorbate and subsequent illumination caused its transformation into RC-β. Resonance Raman spectroscopy of the RC at liquid nitrogen temperature detected the all-trans and the 15-cis isomers in a ratio of 1:1, but HPLC analyses of the carotenoid extracted from the RC before and after the Raman measurements detected the pair of isomers in a ratio of 1:6. Thus, the 15-cis to all-trans isomerization takes place during irradiation at liquid nitrogen temperature, while the reverse isomerization takes place in the dark. The isolated RC-α and RC-β exhibited the bleaching of the 868 nm band, and contained the H, M and L subunits and 1.2-1.4 molecules of ubiquinone-10 per RC. Each RC slowly equilibrated in the dark toward a mixture of RC-α and RC-β. Generation of the all-trans isomer in the light was found not in RC-α but in RC-β.     

A 1H NMR study of tautomerism in some 1-arylamino-3-aryliminopropenes

¹H n.m.r. spectra at ambient temperatures reveal that an equilibrium exists between the ‘all-trans’ and ‘all-cis’ isomers of some of the 1-arylamino-3-aryliminopropenes. The ‘all-cis’ isomer predominates in nonpolar solvents, whereas the ‘all-trans’ isomer is favoured in hydrogen bonding solvents. From a consideration of the magnitudes of the ³J coupling constants, it is reported that the ‘cis-trans’ isomer is the most stable form of the 4-nitrophenyl derivative in dimethyl sulphoxide.     

Preparation of sterically hindered geometric isomers of 7-cis-β-ionyl and β-ionylidene derivatives in the vitamin A series

The preparation of the relatively unknown 7-cis isomers of β-ionyl and β-ionylidene derivatives via one way sensitized geometric isomerization is described. These highly sterically crowded isomers, once prepared do not readily thermally revert back to the trans. But the trienes, at temperatures ≥ 100° undergo irreversible cyclization to cyclohexadienes. 7-cis Isomers of β-ionylideneacetone and acetaldehyde as well as higher tetraenes and pentaenes in this series could not be prepared by sensitized isomerization. Partial reduction of 7-cis-β-ionylideneacetonitrile isomers led to the preparation of 7-cis- and 7,9-dicis-β-ionylideneacetaldehyde; and methyl Grignard reaction with the same nitrile gave the triene-methyl ketones.     

Photoisomerization of zeta-carotene stereoisomers in cells of Euglena gracilis mutant W3BUL and in solution

Abstract— Dark-grown cells of Euglena gracilis var. bacillaris bleached mutant W₃BUL are shown by a novel alumina HPLC method to accumulate a cis isomer of ζ-carotene (cis I; δ_max at 286,296,377,398,422 nm; ε_mM= 97 at 398 nm). Illumination of cells with saturating blue light converts nearly all of this to the trans isomer (δ_max at 379,400,425 nm) and a small amount of a second cis isomer (cis II; δ_max at 285.5,296, 374, 394.5, 419 nm; ε_mM= 111 at 394.5 nm) with no significant changes in any other carotenoids. Photoisomerization of the purified isomers in hexane yields the same mixture of stereoisomers in all three cases, and this mixture is similar to that produced in the cells. Photoisomerization of the purified cis isomers in hexane occurs readily with first order kinetics indicating that no additional photosensitizer or catalyst is necessary for the reaction in vivo. Wild-type cells grown in darkness in the presence of 72 μM J₃₃₄ accumulate ζ-carotene almost exclusively with approximately equal amounts of the cis I and trans isomers, thus cis I is not unique to the mutant. Cis I is identified as 15-cis-ζ-carotene by UV, visible, infrared and mass spectra; cis II may be the 13-eis isomer. Since W₃BUL also accumulates cis isomers of phytoene and phytofluene while the other carotenoids are trans, it is suggested that, in Euglena, ζ-carotene is the point of isomerization from cis to trans in the biosynthetic pathway.     

Molecular iodine: An efficient and environment-friendly catalyst for the synthesis of calix[4]resorcinarenes

Iodine catalyzes the cyclocondensation of various aldehydes with resorcinol to give tetrameric cyclic products, resorcinarenes. Through the reaction of resorcinol with aromatic aldehydes, the product is obtained as a mixture of two isomers, the all-cis isomer (rccc) and the cis-trans-trans isomer (rctt), whereas a single diastereomer, the all-cis, is formed with aliphatic aldehydes. Besides excellent isolated yields, the use of iodine makes this procedure simple, convenient, cost-effective and practical.     

The photoisomerization of retinal

Abstract— –Quantum efficiencies have been measured for the photoisomerization of four stereoisomers of retinal (all-trans, 13-cis, 11 cis, and 9-cis) in two solvents at different wavelengths of irradiation and at various temperatures. In heane at 25°C the quantum efficiencies for isomerization at 365 nm are: 9-cis to trans, 0.5; 13-cis to trans, 0.4; 11-cis to trans, 0.2; all-trans to monocis isomers, 0.2-0.06, depending upon assumptions made regarding the stereo-isomeric composition of the product. These values vary somewhat with the wavelength of the irradiating light. The quantum efficiency for the photoisomerization of all-trans retinal in hexane decreases by a factor of 30 when the temperature is lowered from 25° to – 65°C; the activation energy for this photoisomerization is about 5 kcal/mole. The quantum efficiencies for the isomerization of the monocis isomers to all-trans retinal in hexane are virtually independent of temperature. In ethanol the rates of photoisomerization from trans to cis or cis to trans depend only slightly on the temperature between 25° and – 65°C. The photosensitivities of the stereoisomers of retinal are of the same order of magnitude as those of the retinylidene chromophores of rhodopsin (11 -cis), metarhodopsin I (all-trans), and isorhodopsin (9-cis); but it is not yet possible to derive the photochemistry of rhodopsin uniquely and quantitatively from that of retinal.     

An NMR study of some N-arylaminopropenylylidenearylammonium salts

The preparation, characterisation and ¹H NMR spectra of some N-arylaminopropenylylidenearylammonium salts are reported. The NMR data show that in DMSO the cations exist as conjugated amino–imines, generally as the ‘all-trans’ geometrical isomer. An exception is found in the case of the 4-nitrophenyl derivative which produces a mixture of the ‘all-trans’ and ‘cis-trans’ isomers with slow exchange between them at room temperature. The relative stability of the ‘all-trans’ isomer increases as the salt becomes more ionic. By varying the temperature, pH and nature of the anion it is found that exchange of the N? H protons on the cation controls the appearance of spin-spin coupling between the N? H and 1,3-propene protons.     

PHOTOISOMERIZATION OF SIXTEEN ISOMERS OF RETINAL. INITIAL PRODUCT DISTRIBUTION IN DIRECT AND SENSITIZED IRRADIATION. PHOTOCHEMISTRY OF POLYENES 31*

Abstract— Initial product distributions of all 16 geometric isomers of retinal in hexane solutions have been determined. With direct irradiation, the product ratios are characterized by a preference for isomerization at the 13,14-bond. In particular, all isomers containing the 13-cis geometry give the corresponding 13-trans isomer as the major product. Preference for one-photon-one-bond isomerization was also noted, although a substantial amount of the all-trans isomer was detected for all poly-cis, 13-trans isomers. In sensitized irradiation, the initial mixture shows extensive one-photon-two(or multiple)-bond isomerization to the corresponding unhindered isomers. In cases of hindered isomers, multiple-bond isomerized products are dominant. The different results are accountable by the different shapes of the excited state potential curves for singlet and triplet states.     

Solvolysis and Isomerization of cis- and trans-1-Bromo-1-(p-anisyl)-propene (α-Bromoanethole) Mesomeric Vinyl Cations,Part V

The influence of a β-methyl group on the reactivity of two stereoisomeric vinyl bromides has been studied. In 80% ethanol cis-( 8 ) and trans-α-bromoanethole ( 9 ) undergo first order reactions leading to p-methoxypropiophenone ( 15 ), 1-ethoxy-1-(p-anisyl)-propene ( 16 ) and p-anisylpropyne ( 12 ). Solvolysis of the cis isomer 8 is accompanied by isomerization to the more stable trans isomer 9 which is approx. eight times less reactive than 8 . Cis-trans isomerization is also observed in nitrobenzene at 150°. These results are in agreement with the unimolecular substitution-elimination (S_N1?E1) mechanism which competes with cis-trans isomerization at the ion pair stage. The solvolysis rate of 9 is slightly lower and that of 8 somewhat higher than the rate of α-bromo-p-methoxystyrene ( 3c ). In the absence of other effects a β-methyl group therefore slightly depresses the ionization rate, presumably by steric hindrance of solvation. These results confirm the negligible polar influence of a β-methyl substituent on the stability of vinyl cations.     

Temperature and Concentration Dependent Circular Dichroism of Mono- and Di-cis Isomers of (3S,3′S)-Astaxanthin Diacetate

The circular dichroism (CD.) spectra of all-trans-(3S, 3′S) astaxanthin diacetate and its 9-cis, 13-cis, 9,9′-di-cis, 9,13′-di-cis, and 9,13-di-cis isomers conform to the rules previously formulated for optically active carotenoids with a 4-oxo-β-end ring containing an asymmetric C-atom [1]. Thus the CD. bands of the all-trans and the di-cis isomers show the same signs whereas those of the mono-cis isomers have opposite signs. The CD. spectra of all the astaxanthin diacetate isomers invert sign upon cooling to ?180°. The CD. spectra of the 9-mono-cis and 9,9′-di-cis isomers and to a lesser extent also those of the 9, 13′-di-cis and 9, 13-di-cis isomers are concentration dependent at ?180°, with the longest wavelength band giving at the higher concentration a bisignate CD. curve under the main absorption characteristic of aggregation. This phenomenon has been observed only in isomers with a 9-cis linkage. It is suggested that steric hindrance prevents such aggregation taking place in the other isomers.     

9-CIS, 11-CIS-RETINAL* FROM DIRECT IRRADIATION OF ALL-TRANS-RETINAL. NEW GEOMETRIC ISOMERS OF VITAMIN A AND CAROTENOIDS 9

Abstract The irradiation mixture of all-trans-retinal in acetonitrile was shown to contain four cis,cis isomers as well as the four cis isomers reported earlier. The relatively new 9-cis, 11-cis-retinal was isolated and characterized.     

Bidirectional Hiyama–Denmark Cross-Coupling Reactions of Bissilyldeca-1,3,5,7,9-pentaenes for the Synthesis of Symmetrical and Non-Symmetrical Carotenoids

The construction of the carotenoid skeleton by Pd-catalyzed Csp²−Csp² cross-coupling reactions of symmetrical and non-symmetrical 1,10-bissilyldeca-1,3,5,7,9-pentaenes and the corresponding complementary alkenyl iodides has been developed. Reaction conditions for these bidirectional and orthogonal Hiyama–Denmark cross-coupling reactions of bisfunctionalized pentaenes are mild and the carotenoid products preserve the stereochemical information of the corresponding oligoene partners. The carotenoids synthesized in this manner include β,β-carotene and (3R,3′R)-zeaxanthin (symmetrical) as well as 9-cis-β,β-carotene, 7,8-dihydro-β,β-carotene and β-cryptoxanthin (non-symmetrical).     

Optimization of the isocratic non-aqueous reverse phase (NARP) HPLC separation of trans/cis-α-and β-carotenes

Non-aqueous reverse phase chromatography was used to optimize the separation of all-trans-α- and β-carotenes and their cis isomers extracted from dried carrots. The technique can easily be applied to routine assays. Optimization was carried out by modifying the concentrations of methylene chloride and acetonitrile, and by studying the influence of temperature on the capacity factors and selectivity. The effect of the type of C 18 stationary phase bonding (mono- and polyfunctional) on separation was also examined. The need for polymeric bonding on the stationary phase is stressed for the realization of the trans/cis isomer separation.     

Nitration reactions of astaxanthin and β-carotene by peroxynitrite

The in vitro reactivities of astaxanthin and β-carotene toward peroxynitrite were investigated and the reaction products after scavenging with peroxynitrite were analyzed. A series of carotenoids substituted with nitro group, 14′-s-cis-15′-nitroastaxanthin, 10′-s-cis-11′-cis-11′-nitroastaxanthin, 14′-s-cis-15′-nitro-β-carotene and 10′-s-cis-11′-cis-11′-nitro-β-carotene, were isolated from the reaction products of carotenoids with peroxynitrite. Carotenoids with nitro derivatives were reported for the first time. These results indicated that astaxanthin and β-carotene could catch peroxynitrite or nitrogen dioxide radical (NO₂) in their molecule to form nitrocarotenoids.     

Lanthanide chemical shift reagents as tools for determining isomer distributions in 2,4-hexadienoates and related compounds

Quantitative analysis of geometrical isomers of unsaturated esters and alcohols is facilitated with Eu(fod)₃. Commercial sorbic acid exists in the all-trans form, while sorbyl alcohol contains c. 10% of the cis-4,5 isomer.     

Kinetics of the radical scavenging activity of β-carotene-related compounds

To clarify the non-enzymatic radical-scavenging activity of β-carotene-related compounds and other polyenes, we used differential scanning calorimetry to study the kinetics of radical polymerization of methyl methacrylate (MMA) by 2,2′-azobisisobutyronitrile (AIBN) or benzoyl peroxide (BPO) in the absence or presence of polyenes under nearly anaerobic conditions at 70°C, and analyzed the results with an SAR approach. The polyenes studied were all-trans retinol, retinol palmitate, calciferol, β-carotene and lycopene. Polyenes produced a small induction period. The stoichiometric factor (n) (i.e. the number of radicals trapped by each inhibitor molecule) of polyenes was close to 0. Tetraterpenes (β-carotene, lycopene) suppressed significantly more of the initial rate of polymerization (R ^inh) than did diterpenes (retinol, retinol palmitate). The inhibition rate constants (k ^inh) for the reaction of β-carotene with AIBN-or BPO-derived radicals were determined to be 1.2–1.6?×?10⁵ l?/?mol?s, similar to published values. A linear relationship between k ^inh and the kinetic chain length (KCL) for polyenes was observed; as k ^inh increased, KCL decreased. KCL also decreased significantly as the number of conjugated double bonds in the polyenes increased. Polyenes, particularly β-carotene and lycopene, acted as interceptors of growing poly-MMA radicals.     

Structure and reactivity of α,β-unsaturated ethers. III. Cationic copolymerizations of alkenyl alkyl ethers

The relative cationic polymerizabilities of the geometrical isomers of various alkenyl alkyl ethers were studied both in copolymerizations with each other and in their respective copolymerizations with vinyl isobutyl ether as standard. Copolymerizations were carried out in methylene dichloride at ?78°C. with boron trifluoride etherate as catalyst. The cis isomers have been found to be more reactive than the corresponding trans isomers. A primary alkyl substituent on the β-cis position of vinyl ethyl ether enhances the reactivity. Yet the steric effect is noticeable when the substituents are bulky. Compounds substituted with cis-β-isobutyl and with β-dimethyl showed little tendency to homopolymerization. It was proved that the polymer ends derived from cis and from trans monomers are respectively different in character because of the restricted rotation of the end unit around the terminal carbon–carbon bond. The alternation tendency, remarkable in the copolymerization of cis monomers with vinyl ether, was explained in terms of the cis-opening mechanism.