Evaluation of reversed-phase liquid chromatographic columns for recovery and selectivity of selected carotenoids

Epler, K. S.; Sander, Lane C.; Ziegler, Regina G.; Wise, Stephen A.; Craft, Neal E.

doi:10.1016/0021-9673(92)85149-n

Cited by 111 publications

(44 citation statements)

References 42 publications

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“…In addition to spectrophotometers (UV-VIS) and fluorescence detectors, DAD's are increasingly used in pigment analysis (Wright et al 1991, Almela et al 1992, Epler et al 1992, Kraay et al 1992, van Heukelem 1992, Tsavalos et al 1993, Jeffrey et al 1997, Descy et al 2000. Initially DAD's were developed for the UV range (Meyer 1999).…”

Section: Discussionmentioning

confidence: 99%

Patterns of major photosynthetic pigments in freshwater algae. 2. Dinophyta, Euglenophyta, Chlorophyceae and Charales

Schagerl

Pichler

Donabaum³

2003

Ann. Limnol. - Int. J. Lim.

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Section: Discussionmentioning

confidence: 99%

Patterns of major photosynthetic pigments in freshwater algae. 2. Dinophyta, Euglenophyta, Chlorophyceae and Charales

Schagerl

Pichler

Donabaum³

2003

Ann. Limnol. - Int. J. Lim.

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“…Extracts were obtained by using methanol containing 6% (wt/vol) KOH at 60°C for 20 min. The extracts were partitioned against 10% (vol/vol) diethyl ether in petrol, and the carotenoid content was determined by highperformance liquid chromatography (HPLC) on a 25-cm C 18 Vydac 218TP54 column with methanol as the mobile phase at 20°C (10). Reference carotenoids were isolated from cyanobacteria (38) or pepper leaves (35).…”

Section: Methodsmentioning

confidence: 99%

Transformation of the Green Alga Haematococcus pluvialis with a Phytoene Desaturase for Accelerated Astaxanthin Biosynthesis

Steinbrenner

Sandmann

2006

Appl Environ Microbiol

211

118

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Astaxanthin is a high-value carotenoid which is used as a pigmentation source in fish aquaculture. Additionally, a beneficial role of astaxanthin as a food supplement for humans has been suggested. The unicellular alga Haematococcus pluvialis is a suitable biological source for astaxanthin production. In the context of the strong biotechnological relevance of H. pluvialis, we developed a genetic transformation protocol for metabolic engineering of this green alga. First, the gene coding for the carotenoid biosynthesis enzyme phytoene desaturase was isolated from H. pluvialis and modified by site-directed mutagenesis, changing the leucine codon at position 504 to an arginine codon. In an in vitro assay, the modified phytoene desaturase was still active in conversion of phytoene to -carotene and exhibited 43-fold-higher resistance to the bleaching herbicide norflurazon. Upon biolistic transformation using the modified phytoene desaturase gene as a reporter and selection with norflurazon, integration into the nuclear genome of H. pluvialis and phytoene desaturase gene and protein expression were demonstrated by Southern, Northern, and Western blotting, respectively, in 11 transformants. Some of the transformants had a higher carotenoid content in the green state, which correlated with increased nonphotochemical quenching. This measurement of chlorophyll fluorescence can be used as a screening procedure for stable transformants. Stress induction of astaxanthin biosynthesis by high light showed that there was accelerated accumulation of astaxanthin in one of the transformants compared to the accumulation in the wild type. Our results strongly indicate that the modified phytoene desaturase gene is a useful tool for genetic engineering of carotenoid biosynthesis in H. pluvialis.

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“…After addition of appropriate internal standards (a-tocopherol-acetate and ethyl-apo-b Hcarotenoate), extraction was performed with n-heptane and n-heptane/diethyl ether (1:1, v/v). Following evaporation of the solvents, the residue was dissolved in eluent and atocopherol, a-carotene and b-carotene were separated by reversed phase HPLC using 201 TP54 Vydac column (Separations Group, Hesperia CA, USA) and methanol/ tetrahydrofuran/ammonium acetate solution (1 g/L) (95:5:2, v/v) as mobile phase at a¯ow rate of 0.8 ml/min and a column temperature of 13 C Epler et al, 1992). Vitamin C concentration in trichloroacetic acid-treated plasma was determined¯uorimetrically as ascorbic acid plus dehydroascorbate as described by Vuilleumier & Keck (1989).…”

Section: Plasma and Ldl-analysesmentioning

confidence: 99%

Antioxidant fortified margarine increases the antioxidant status

et al. 1998

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Objective: To assess the effect of supplementation with an antioxidant forti®ed margarine on the body's antioxidant status and on parameters of oxidative damage to lipids. Design: Single blind, placebo controlled trial, two treatment groups balanced for sex, age and Quetelet Index. Setting: Unilever Research Laboratorium, The Netherlands. Subjects: Thirty-one healthy adult volunteers accomplished the study. Volunteers were recruited among inhabitants of the surrounding area of the research laboratory. Interventions: Volunteers consumed during the four weeks either 15 g/d of an antioxidant forti®ed margarine (providing 121 mg vitamin C, 31 mg vitamin E, 2.7 mg a-carotene and 5.3 mg b-carotene) or an ordinary margarine. Fasting blood samples were taken before and at the end of the study. Results: Consumption of the antioxidant forti®ed margarine signi®cantly increased the levels of the supplied antioxidants in plasma and LDL as compared to the changes found after consumption of the control margarine, with the largest increases found in LDL levels of a-carotene (15.5-fold increase, 95% CI: 8.4±27.8-fold) and bcarotene (4.3-fold increase, 95% CI: 2.2±7.9-fold). This increased antioxidant status in the antioxidant forti®ed margarine group resulted in a signi®cantly increased total antioxidant activity of LDL and resistance of LDL to oxidation (lag time and rate of oxidation) as compared to baseline but not in comparison to the changes found in the control group. Conclusion: Consumption of moderate doses of vitamin E, vitamin C, a-carotene and b-carotene, supplied in a full-fat margarine and consumed as part of a normal diet, effectively increases the blood levels of these antioxidants.

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Evaluation of reversed-phase liquid chromatographic columns for recovery and selectivity of selected carotenoids

Cited by 111 publications

References 42 publications

Patterns of major photosynthetic pigments in freshwater algae. 2. Dinophyta, Euglenophyta, Chlorophyceae and Charales

Patterns of major photosynthetic pigments in freshwater algae. 2. Dinophyta, Euglenophyta, Chlorophyceae and Charales

Transformation of the Green Alga Haematococcus pluvialis with a Phytoene Desaturase for Accelerated Astaxanthin Biosynthesis

Antioxidant fortified margarine increases the antioxidant status

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