Lutein identification

Dachtler, M., Kohler, K., and Albert, K. 1998. Reversed-phase high-performance liquid chromatographic identification of lutein and zeaxanthin stereoisomers in bovine retina using a C30 bonded phase. J. 

The assignment of the lutein absorbing at 495 nm as lutein 1 has helped with the identification of an excitation energy quencher in LHCII, when the complex is in aggregated form. 

Khachik, F., Bernstein, P., and Garland, D. 1997. Identification of lutein and zeaxanthin oxidation products in human and monkey retinas. Invest. Ophtalmol. 38 1802-1811. 

Fig. 2.16. HPLC profile of carotenoids in an extract of vegetable soup. An expansion of the profile from 30 to 39 is shown in the inset (A). Monitored wavelengths were 436, 440, 464, and 409 nm for peaks 9,10,11,12, and 14, respectively, in the inset (A). Peak identification 1 + 1" = all-trans-lutein and cw-lutein 2 = 5,6-dihydroxy-5,6-dihydrolycopene (lycopene-5,6-diol) 3 = j3-apo-8 -carotenal (internal standard) 4 = lycopene 1,2-epoxide 5 = lycopene 5,6-epoxide 6 = 1,2-dimethoxyproly-copene (tentative identification) 7 = 5,6-dimethoxy-5,6-dihydrolycopene 8 = lycopene 9 = pheo-phytin b 10 = neurosporene 11 = (-carotene 12 = pheophytin a 13 = (-carotene 14 = pheophytin a isomer and (-carotene 15 = a-carotene 16 and 16" = all-trans-/fcarotene, cis-/J-carotene 17 and 17" = all-trans- or cA-phytofluene 18 and 18" = all-trans- or cw-phytoene. Reprinted with permisson from L. H. Tonucci et al. [40].

Fig. 2.17. Saponified carotenoids in orange juice. Chromatographic conditions are given in text. Chromatograms from absorbance monitoring at 430, 486 and 350 nm, respectively, are shown, all at identical attenuation. Peak identification 1, 3, 5, 8, 26 and 29 = unidentified peaks 4 = valen-ciaxanthin 6 = neochrome 7 = trollichrome 9 = antherxanthin 11 = c/s-anthexanthin 12 = neoxanthin 19 = auoxanthin B 20 = c/s-violaxanthin 22 = leutoxanthin 23 = mutatoxan-thin A 24 = mutatoxanthin B 25 = lutein 27 = zeaxanthin 28 = isolutein 31 = a-cryptoxanthin 33 = /J-cryptoxanthin 34 = phytofluene 35 = a-carotene 36 = ae-carotene 37 = / -carotene. Reprinted with permission from R. Rouseff et al. [41].

Fig. 2.23. Reversed-phase gradient HPLC profiles of carotenoids in human plasma. A human volunteer was given an oral dose of 5,6-epoxy-/l-carotene (9.1 /imol). Plasma was analysed for carotenoids before (a) and 6h after (b) the oral dose. Peak identification 1, bilirubin 2, lutein 3, zeaxanthin 4, /1-cryptoxanthin 5, 5,6-epoxy-/l-carotene 6, lycopene 7, /1-carotene. The detection wavelength was 445 nm. AU, absorbance unit. Reprinted with permission from A. B. Barua [50],...

Fig. 2.24. C30 chromatograms of carotenoids extracted from human serum (a) xanthophylls fraction, 7 93 (v/v) MTBE-methanol mobile phase (b) a- and / -carotenes fraction, 11 89 (v/v) MTBE-methanol mobile phase (c) lycopene fraction, 38 62 (v/v) MTBE-methanol mobile phase. Tentative peak identifications (a) 1, 13-c/s-lu- lutein 2, 13 r/.vlutein 3, a//-/ra s-lutein 4, zeaan-thin 5-7, unidentified P,e-carotenoids and 8, / -cyrptoanthin (b) 1-2, unidentified ae-carotene isomers 3, 15-eH -/f-carotenc 4, 13-cw-/ -carotene 5, all-trans-a-carotene 6, all-trans-P-carotene and 7, 9-ci.v-/3-carotene and (c) 1-11 and 13, c/s-lycopene isomers and 12, all-trans-lycopene. Reprinted with permission from C. Emenhiser el al. [51].

Fig. 2.26. Reversed-phase HPLC separation of (a) Sobrasada sausage extract and (b) saponified Sobrasade sausage extact in an ODS column at maximum absorbances at each point in time. Peak identification 1 - 2, 4 - 6, 8, 12, 14-17 = unidentified free 3 = capsorubin 7 = violaxanthin 9 = capsanthin 10 = anteraxanthin 11 = cw-capsanthin 13 = lutein and zeaxanthin 18 = cantaxanthin, internal standard 19 = cryptoxanthin 20, 24, 25, 28 = unidentified monoester 21 = /J-cryptoxanthin 22 = capsorubin monoester 23, 26, 27, 29 = capsanthin monoester 30, 31 = lutein-zeaxanthin monoester 32 = /1-carotene 33 = cis-f)-carotene 34, 37, 39, 41, 43 = capsanthin diester 35 = capsorubin diester 36, 38, 40, 42, 44 = unidentified diester. Reprinted with permission from J. Oliver et al. [56],...

Fig. 2.34. Sample chromatogram of light-adapted Z. marina leaf sample. Peak identification 1 = neoxanthin, 2 = violaxanthin, 3 = antheraxanthin, 4 = lutein, 5 = zeaxanthin 6 = chlorophyll b, 1 = chlorophyll a, 8 = / -carotene. Reprinted with permisson from P. J. Ralph et al. [76].

Fig. 2.124. HPLC absorbance chromatograms of phytoplankton samples collected from the study site on 24 November 2000 (a), 25 November 2000 (b) and 1 December 2000 (c). Peak identification 1 = chlorophyll-a 2 = chlorophyll-b 3 = chlorophyll-c2 4 = peridin 5 = fucoxanthin 6 = neoxanthin 7 = 19 -hexanoylfucoxanthin 8 = diadinoxanthin 9 = alloxanthin 10 = lutein 11 = zeaxanthin 12 = /1-carotene. Reprinted with permission from C. K. Wong et al. [282],...

R. L. Chan, M. D. Chaplin, Identification of Major Urinary Metabolites of Nafarelin Acetate, a Potent Agonist of Luteinizing Hormone-Releasing Hormone , Drug Metab. Dispos. 1985, 13, 566-571. 

Fig. 11 HPLC of carotenoids solvent-extracted from (A) raw and (B) thermally processed carrots. Column, 5-/um polymeric C1(J (250 X 4.6-mm ID) mobile phase, methyl tert-butyl ether/methanol (11 89), 1 ml/min absorbance detection, 453 nm. Tentative peak identifications (1) all-trans-lutein (2) 13-cis-a-carotene (3) a cis-a-carotene isomer (4) 13 -cA-a-carotene (5) 15-cis-/3-carotene (6) 13-cis-/3-carotene (7 and 8) cis-fi-carotene isomers (9) all-frans-a-carotene (10) 9-cis-a-carotene (11) all-frans-/3-carotene (12) 9-ci. -/3-carotene. (Reprinted with permission from Ref. 192. Copyright 1996, American Chemical Society.)...

HPLC is commonly used to separate and quantify carotenoids using C18 and, more efficiently, on C30 stationary phases, which led to superior separations and improved peak shape.32 4046 An isocratic reversed-phase HPLC method for routine analysis of carotenoids was developed using the mobile phase composed of either methanol acetonitrile methylene chloride water (50 30 15 5 v/v/v/v)82 or methanol acetonitrile tetrahydrofuran (75 20 5 v/v/v).45 This method was achieved within 30 minutes, whereas gradient methods for the separation of carotenoids can be more than 60 minutes. Normal-phase HPLC has also been used for carotenoid analyses using P-cyclobond46 and silica stationary phases.94 The reversed-phase methods employing C18 and C30 stationary phases achieved better separation of individual isomers. The di-isomers of lycopene, lutein, and P-carotene are often identified by comparing their spectral characteristic Q ratios and/or the relative retention times of the individual isomers obtained from iodine/heat-isomerized lycopene solutions.16 34 46 70 74 101 However, these methods alone cannot be used for the identification of numerous carotenoids isomers that co-elute (e.g., 13-ds lycopene and 15-cis lycopene). In the case of compounds whose standards are not available, additional techniques such as MS and NMR are required for complete structural elucidation and validation. 

Young, J.C. Abdel-Aal, E.S.M. Rabalski, I. Blackwell, B.A. 2007. Identification of synthetic regioisomeric lutein esters and their quantification in a commercial lutein supplement. J. Agric. Food Chem. 55 4965 972. 

Han, K. Park, J. Chung, Y. Lee, M. Moon, D. Robinson, J. Identification of enzymatic degradation products of luteinizing hormone releasing hormone (LHRH)/[D-Ala6]LHRH in rabbit mucosal homogenates. Pharm. Res. 1995, 12, 1539-1544. 

Zheng LM, Caldani M, Jourdan F. 1988. Immunocytochemical identification of luteinizing hormone-releasing hormone-positive fibres and terminals in the olfactory system ofthe rat. Neurosci 24 567-578. 

Zheng, L.M., Caldani, M. and Jourdan, L. (1988) Iramunocytochemical identification of luteinizing hormonereleasing hormone-positive fibres and terminals in the olfactory system of the rat. Neuroscience, 24, 567-578. 

All lutl alleles are recessive mutations (Fig. 3 A) that accumulate an additional carotenoid with an HPLC retention time and UV/visible absorption spectrum characteristic of a monohydroxy / , e-carotenoid (an a-carotene derived xanthophyll). The identification of the accumulating compound as zeinoxanthin ()3,e-caroten-3-ol) was confirmed by mass spectrometry (Pogson et al., 1996). The only difference between lutein and zeinoxanthin is the presence of an hydroxyl group on carbon 3 of the -ring in lutein. The decrease in lutein and its partial replacement by its immediate precursor, zeinoxanthin, defines lutl as a mutation... 

Kanamaru, K. S. Wang J. Abe T. Yamada K. Kitamura. Identification and characterization of wild soybean Glycine soja Sieb. et Zecc.) strains with high lutein content. Breeding Sci., 2006, 56, 231-234. 

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