13 -cis-lutein

Canning at 121°C for 30 min was also responsible for the highest losses of carotenoids in carrot juice, reaching 60% for P- and a-carotene, whereas the lutein level decreased 50%, all accompanied by the formation of 13-c -p-carotene in the largest amount, followed by 13-cA-lutein and 15-cA-a-carotene. Canning (T x = 121°C, F = 5) of sweet com resulted in a decrease of lutein by 26% and zeaxanthin by 29%, accompanied by increased amounts of 13-cis- lutein, 13 -CM-lutein, and 13-c/i-zeaxanthin. ° The relative amounts of cis isomers of lutein, mainly the 13-cis, increased by 15% and of 13-di-zeaxanthin by 20% after com canning." ... 

Maize is exceptionally high in lutein at a concentration of about 20 mg/kg. In addition to the high content of lutein, maize also has high concentration of zeaxanthin (6-10 mg/kg), P-cryptoxanthin (2 mg/kg), p-carotene (1 mg/kg) and small concentrations of 15-cis-lutein, 13-cis-lutein, 13 -ds-lutein, 9-ds-lutein, 9 -cis-lutein and 9-c 5-zeaxanthin. 

Losses from 21 to 23% in P-carotene, a-carotene, and lutein contents were observed during storage of carrot juice under light (1500 lux) at 25°C for 12 wk. The losses were accompanied by increased concentrations of the 13-cis isomer type of P-carotene, a-carotene, and lutein during dark storage, while the formation of 9-cw-P-carotene, 9-cii-a-carotene, and 13-cii-lutein was favored under light storage. ... 

Astaxanthin, capsanthin, lutein, zeaxanthin, canthaxanthin, P-cryptoxanthin, echinenone, 15-cis-P-carotene, 13-cis- P-carotene, a-carotene, all-ircms P-carotene, 9-cis-P-carotene, 5-carotene, lycopene... 

All-trans p-carotene 13-cis p-Carotene 9-cis p-Carotene a-Carotene Lutein Lycopene... 

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.)...

All-trans-isomers of carotenoids in fresh and thermally processed materials are accompanied by small amounts of ds-isomers, called neocarotenoids. fS-Carotene is accompanied mainly by geometric isomers 9-cis, 13-cis- and 15,15 -cis-P-carotene. Lutein is accompanied mainly by 9-ds- and 9 -cis, 13-cis- and 13 -cis isomers less common are 15-cis and 15 -cis isomers. Neoxanthin is accompanied by 9-cis-, 9 -cis-, 13-cis- and 13 -cis isomers. Thermal processing can induce carotenoid trans to cis isomerisation. 

Capsorubin, violaxanthin, capsanthin-5,6-epoxide, capsanthin, 9-cw-capsanthin, 13-c -capsanthin, antheraxanthin, mutatoxanthin, cucurbitaxanthin A (capsolutein), zeaxanthin, 9-cis-zeaxanthin, 13-cis-zeaxanthin, p-apo-8 -carotenal, cryptoeapsin, P-cryptoxanthin, p-earotene, and cw-p-carotene Neoxanthin, violaxanthin, lutein, antheraxanthin, mutatoxanthin, P-carotene, neochrome, auioxanthin, luteoxanthin, and chltuophylls... 

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. 2.24. 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],...

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. 

Lutein, astaxanthin, -cryptoxanthin, and a- and / -carotene were separated on a C30 column (electrospray MS) using a unique 60-min 85/15 -> 10/90 methanol/ MtBE (1 mM ammonium acetate) gradient. Postcolumn derivatization with 2,2,3,3,4,4,4-heptafluoro-l-butanol dramatically increased the sensitivity of the method [354], Detection limits 100-fold lower than that obtained with UV detectors (i.e., l-2pmol injected) were reported for cc- and j8-carotene. The isomerization of franj-astaxanthin to 9-cis- and 13-cw-astaxanthin was monitored through separation on a 25°C Cjg column (A = 480nm) using an 85/5/5.5/4.5 methanol/ dichloromethane/acetonitrile/water mobile phase [355]. Separation was good and elution was complete in < 10 min. 

In an excellent article by Bell et al. [356], the retention of 12 carotenoids (zeaxanthin, lutein, echinenone, / -cryptoxanthin, and a-, 9-cis-a-, 15-cis-a-, 9 -cis-a-, 13-cir-a-, ] -, 3-cis-P-, and 15-c/j- -caiotene) was studied with respect to temperature (277 K to 323 K) on C,g, C30, and C34 columns (A = 450 nm). Methanol was used as a mobile phase on the C g column and 95/5 methanol/methyl r-butyl ether on the C30 and C34 columns. The authors noted that acetonitrile was a potential mobile phase modifier but that high acetonitrile levels ofien led to decreases in recoveries. The use of dichloromethane was discouraged since residual HCI due to natural solvent degradation was implicated in poor recoveries as well. The latter is supported by the fact that low molecular weight alkenes are ofien used as preservatives in dichloromethane. A series of van t Hoff plots (essentially Infc vs. 1 /T) were presented where C]g phase showed near linearity and C30 and C34 phases exhibited nonlinear relationships for most carotenoids. 

A total of 22 carotenoids were extraced fiom human plasma, 13 of which were identified (e.g., astaxanthin, lutein, echinenone, cis-lyeopene, carotenes). A 45-min separation was achieved on a C,g column (photodiode array detector, A = 300-600nm) using a 70/15/5/10 acetonitrile/methanol (50mM ammonium acetate)/ water/dichloromethane mobile phase [1125]. Good (but not necessarily baseline) resolution was obtained for all peaks. A linear range of 10-200 ng injected and detection limits of 5 ng injected were reported. 

Land plants including moss, fern, gymnosperm, and angiosperm majorly contain the carotenoids, (3-carotene 13, violaxanthin 16,9 -cis neoxanthin 19, and lutein 11 in chloroplasts for photosynthesis. Some unique carotenoids are found in specific organs, such as petals, fruits, and roots these carotenoids are not involved in photosynthesis. 

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