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Improved high performance liquid chromatographic method for determination of carotenoids in the microalga Chlorella pyrenoidosa.

Microalgae have become an important commercial source of carotenoids and microalgae-derived functional foods are consumed by people worldwide. Therefore, an HPLC method was developed to discern the variety and content of carotenoids in the microalga Chlorella pyrenoidosa. The microalga sample was powdered, extracted, saponified and subjected to HPLC analysis. A mobile phase of methanol-acetonitrile-water (84:14:2, v/v/v) (A) and methylene chloride (100%) (B) with the following gradient elution was developed: 100% A and 0% B in the beginning, maintained for 14 min, decreased to 95% A in 25 min, 75% A in 30 min, 74% A in 35 min, 45% A in 50 min and returned to 100% A in 55 min. A total of 32 carotenoids were resolved within 49 min by using a C30 column with flow rate at 1 mL/min and detection at 450 nm. An internal standard beta-apo-8'-carotenal was used to quantify all the carotenoids. All-trans-lutein was present in exceptionally large amount (125034.4 microg/g), followed by cis isomers of lutein (27975.3 microg/g), all-trans-alpha-carotene (2465.8 microg/g), zeaxanthin (2170.3 microg/g), cis isomers of beta-carotene (2159.3 microg/g), all-trans-beta-carotene (2155.0 microg/g), cis isomers of alpha-carotene (1766.7 microg/g), beta-cryptoxanthin (334.9 microg/g), neoxanthin and its cis isomers (199.7 microg/g), neochrome (65.2 microg/g), auroxanthin (38.5 microg/g) and violaxanthin and its cis isomers (38.1 microg/g).

Carotenoids↗

Blood concentrations of methionine, selenium, beta-carotene, and other micronutrients in a case-control study of arsenic-induced skin lesions in West Bengal, India.

Previous studies have suggested that susceptibility to arsenic toxicity could be influenced by micronutrients, in particular selenium, methionine, and beta-carotene. A case-control study was conducted in West Bengal, India, in a region known to have groundwater arsenic contamination, to determine whether differences in micronutrient status contribute to susceptibility to arsenic-induced skin lesions. Micronutrient status was assessed by blood levels of specific micronutrients and metabolic indicators. Blood was obtained from 180 cases with skin lesions and 192 controls. Blood assays measured micronutrients and carotenoids (folate, selenium, vitamin B12, vitamin B6, retinol, alpha-tocopherol, lutein/zeaxanthin, beta-carotene, lycopene, beta-cryptoxanthin) and metabolic indicators such as glucose, cholesterol, transthyretin, amino acids, and proteins potentially associated with methylation (cysteine, homocysteine, methionine, glutathione). The distributions of nutrient concentrations were similar in cases and controls. The median selenium concentrations in cases and controls were both 1.15 micromol/L, and there was little evidence of differences in other micronutrients. Odds ratios (ORs) for arsenic-induced skin lesions were estimated for each quartile of nutrient concentrations, using the quartile with the highest nutrient level as the referent group. There were no clear trends associated with deficiencies of any micronutrient or metabolic indicator. For decreasing quartiles of selenium, the OR estimates were 1.00, 0.67, 0.99, 0.80; P=0.81; for methionine, the OR estimates were 1.00, 0.83, 0.78, 0.72; P=0.29. For beta-carotene, the ORs were 1.00, 0.53, 0.51, 0.96, demonstrating no increased risk at the lower quartiles. The measured micronutrients and metabolic indicators investigated do not appear to modify the risk of developing arsenic-induced skin lesions. The lack of any trend of increasing risk with lower selenium, vitamin E, and beta-carotene concentrations has important implications for proposed therapeutic interventions. The emphasis of interventions should be on reducing arsenic exposure.

Arsenic↗

Is serum gamma-glutamyltransferase inversely associated with serum antioxidants as a marker of oxidative stress?

A series of studies in black and white women and men have suggested that serum gamma-glutamyltransferase (GGT) within its normal range might be an early marker of oxidative stress. If serum GGT is a marker of oxidative stress, it might have important implications both clinically and epidemiologically because measurement of serum GGT is easy, reliable, and not expensive. We examined the cross-sectional association between deciles of serum GGT and concentrations of serum antioxidants among 9083 adult participants in the third U.S. National Health and Nutrition Examination Survey. After adjustment for race, sex, age, and total cholesterol, serum concentration of GGT across all deciles was inversely associated with serum concentrations of alpha-carotene, beta-carotene, beta-cryptoxanthin, zeaxanthin/lutein, lycopene, and vitamin C (p for trend <.01, respectively). Vitamin E was not associated with serum GGT. All these associations were not materially different after additional adjustment for total energy intake, body mass index, smoking status, smoking amount, alcohol intake, and exercise. These associations were similarly observed among most subgroups. In conclusion, the current and previous studies strongly suggest that serum GGT level within its normal range may be an early marker of oxidative stress.

Alanine Transaminase↗

Serum nutrient markers and skin prick testing using data from the Third National Health and Nutrition Examination Survey.

BACKGROUND: Diet nutrients and allergic sensitization both affect the development and severity of asthma, but the interrelationship between oral intake and allergic sensitization is not well characterized, although previous work suggests that a higher vitamin E intake may be protective against allergic sensitization. OBJECTIVE: To determine whether serum levels of dietary antioxidants, lipids, and other nutrients are associated with the presence of allergen skin sensitization. METHODS: Cross-sectional logistic regression analysis of 30 serum nutrient levels in relation to 10 skin test allergens in adults and children in the Third National Health and Nutrition Survey. RESULTS: In adults, allergic skin sensitization was less common in participants with higher serum levels of vitamin E: the adjusted odds ratio (OR) per SD difference in vitamin E level was 0.93 (95% CI, 0.87-0.99). Two of the carotenoids, beta-cryptoxanthin and alpha-carotene, were also inversely associated with allergic skin sensitization (OR, 0.89; 95% CI, 0.83-0.95; and OR, 0.95; 95% CI, 0.91-0.99, respectively). Among adults, serum lycopene had a positive association with allergen sensitization. In youths, however, vitamin A was associated with an increased risk, and high-density lipoprotein cholesterol was associated with a decreased risk of allergic sensitization. The use of vitamin or mineral supplements had little effect on the magnitude of all reported associations. CONCLUSION: Overall, there were no strong or consistent associations between serum levels of potentially antiallergic nutrients, and the occurrence of allergic skin sensitization in these data. However limited, findings suggest that increases in vitamin E, carotenoids, and high-density lipoprotein cholesterol may reduce the risk of allergic skin sensitization.

Adolescent↗

Estimated phytochemical content of the dietary approaches to stop hypertension (DASH) diet is higher than in the Control Study Diet.

The Dietary Approaches to Stop Hypertension (DASH) diet substantially lowers blood pressure and reduces blood lipid levels. The DASH diet menus were designed to reach beneficial levels of fiber, potassium, magnesium, and calcium, and therefore contain more fruits, vegetables, and whole grains relative to the control menus, and consequently more phytochemicals. Using the US Department of Agriculture food composition databases, the polyphenol, carotenoid, and phytosterol contents of the diets used in the DASH study were estimated. When compared with the control diet, the DASH diet is higher in flavonols, flavanones, flavan-3-ols, beta-carotene, beta-cryptoxanthin, lycopene, lutein+zeaxanthin, and phytosterols. Flavone levels are similar, whereas isoflavones are present in a small amount in the DASH diet. The roles of these compounds in disease risk reduction are becoming recognized. It therefore is possible that the health benefits of the DASH diet are partially attributable to the phytochemicals and might extend beyond cardiovascular disease risk reduction.

Blood Pressure↗

Nonsupplemented children of Latino immigrants have low vitamin E intakes and plasma concentrations and normal vitamin C, selenium, and carotenoid intakes and plasma concentrations.

OBJECTIVE: This study sought to determine and evaluate the intakes and plasma concentrations of vitamin E (alpha-tocopherol), gamma-tocopherol, vitamin C, selenium, and carotenoids (alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein/zeaxanthin, and lycopene) of nonsupplemented boys and girls, 4 to 8 years old, of Latino immigrants living in rural Nebraska. DESIGN: Dietary intakes of the Latino children were estimated. Their plasma samples (fasting) were analyzed for vitamin E, vitamin C, selenium, and carotenoid concentrations. Data were evaluated by sex. SUBJECTS: The subjects were a convenience sample of 4- to 8-year-old (n=29), apparently healthy, nonsupplemented children of Latino immigrants living in rural Nebraska. STATISTICAL ANALYSIS: Sex differences in parameter values were determined using general linear models; Pearson r was used for determining correlations. RESULTS: No significant differences in parameter values were observed by sex, with the exception of plasma lutein/zeaxanthin concentration. The majority (69%) had plasma vitamin E (alpha-tocopherol) concentrations<0.516 mg/dL (12 micromol/L), which is indicative of vitamin E inadequacy, and over half (59%) reported consuming less than the Estimated Average Requirement for vitamin E. All subjects had plasma vitamin C and selenium concentrations indicative of adequacy (>0.41 mg/dL or >23 micromol/L, and >6.32 microg/dL or >0.8 micromol/L, respectively), and consumed at least the Recommended Dietary Allowances for these nutrients. The plasma carotenoid concentrations of the children may be useful as norms. CONCLUSIONS: These children of Latino immigrants who did not take supplements had low plasma vitamin E (alpha-tocopherol) concentrations but normal plasma vitamin C and selenium concentrations. IMPLICATIONS: Dietetics professionals and others involved in health care need to work with Latino immigrant parents so that their children consume adequate amounts of vitamin E.

Ascorbic Acid↗

Cocoa flavanol-enriched snack bars containing phytosterols effectively lower total and low-density lipoprotein cholesterol levels.

BACKGROUND: Dietary intervention studies incorporating phytosterol-enriched margarine spreads have reported significant decreases in total and low-density lipoprotein (LDL) cholesterol in populations with both normal lipid levels and those with hypercholesterolemia. There is emerging support for more diverse and lower-fat phytosterol-enriched matrixes. Controversy exists, however, over whether phytosterol-enriched foods affect serum fat-soluble vitamins. OBJECTIVE: We investigated whether a flavanol-rich cocoa snack food containing phytosterols would decrease total and LDL cholesterol levels in subjects with hypercholesterolemia and significantly affect serum fat-soluble vitamins and carotenoids. DESIGN: A randomized, double-blind parallel arm study design was used. Subjects were randomized to one of two dietary treatments: a cocoa flavanol-enriched snack bar containing 1.5 g phytosterol (n=32), or a control product containing no phytosterols (n=35). Subjects consumed two servings per day. RESULTS: Consumption of the phytosterol-enriched snack bars but not control bars for 6 weeks was associated with significant reductions in plasma total (4.7%; P<0.01) and LDL cholesterol (6%; P<0.01), and the ratio of total to high-density lipoprotein cholesterol (7.4%; P<0.001). There were no changes in high-density lipoprotein cholesterol, triglycerides, or lipid-adjusted lycopene, beta-cryptoxanthin, lutein/zeaxanthin, alpha-carotene levels, or levels of serum vitamins A or E. A significant reduction in lipid-adjusted serum beta-carotene was observed in the phytosterol but not the no-phytosterol-added group (P<0.05). CONCLUSIONS: This study supports the use of a novel phytosterol-enriched snack bar to effectively reduce plasma total and LDL cholesterol levels in a population with hypercholesterolemia. The data suggest that the incorporation of this snack food into a balanced diet represents a practical dietary strategy in the management of serum cholesterol levels.

Cacao↗

A cross-sectional analysis demonstrated the healthy volunteer effect in smokers.

OBJECTIVE: This cross-sectional descriptive analysis sought to determine if a healthy volunteer effect can be demonstrated among smokers selected to participate in a dietary intervention trial. STUDY DESIGN AND SETTING: Body mass index (BMI), body fat, physical activity, dietary intake, and plasma concentration of antioxidant nutrients and carotenoids were assessed cross-sectionally, at the time of enrollment into a dietary intervention trial, among 136 adult smokers. RESULTS: Mean BMI was below national age- and gender-specific averages as was prevalence of overweight and obesity. Physical activity was reported to average 15.4 h/wk. Compared with other sample populations of smokers, our smokers reported lower total fat and cholesterol intakes, higher vitamin C and beta-carotene intakes, and generally equal vitamin E intakes. Plasma ascorbic acid, alpha-tocopherol, alpha- and beta-carotene, and beta-cryptoxanthin concentrations were higher than those of smokers surveyed by NHANES III. CONCLUSION: These findings suggest that a "healthy volunteer effect" can be described among adult smokers. Future dietary intervention trials among smokers should cautiously estimate sample size because smokers electing to participate may report healthier dietary patterns than other smokers. Screening criteria regarding baseline micronutrient status of smokers should be evaluated given that intervention effects may be dependent on overall health status.

Adolescent↗

Xanthophyll biosynthesis: molecular and functional characterization of carotenoid hydroxylases from pepper fruits (Capsicum annuum L.).

To dissect the mechanism by which carotenoid hydroxylases catalyze xanthophyll formation, we have cloned two pepper cDNAs encoding beta-cryptoxanthin and zeaxanthin biosynthetic enzymes. Using an in vitro system, we find that both enzymes are ferredoxin dependent and that their activity is strongly inhibited by iron chelators such as o-phenanthroline or 8-hydroxyquinoline. This suggests the transfer of a reducing equivalent from NADPH to the hydroxylase via ferredoxin and the involvement of an iron activated oxygen insertion process. Based on sequence analysis, the putative histidine clusters involved in the iron coordination were identified and their roles evaluated. Following site-directed mutagenesis of the identified histidine residues hydroxylase activity was totally inactivated. Collectively, our data indicate that carotenoid hydroxylases belong to a new class of diiron proteins structurally related to membrane fatty acid desaturases. Mechanistically, both types of enzymes exploit iron activated oxygen to break the C-H bond with concomitant formation of double bond or oxygen insertion. We propose that the same mechanism operates for beta-carotene ketolase and probably for other carotenoid oxygenases as well.

Amino Acid Sequence↗

Preparation and value assignment of Standard Reference Material 968c Fat-Soluble Vitamins, Carotenoids, and Cholesterol in Human Serum.

Standard Reference Material 968c Fat-Soluble Vitamins, Carotenoids, and Cholesterol in Human Serum provides certified values for retinal, delta-, gamma-, and alpha-tocopherol, trans- and total beta-carotene, and cholesterol in human serum. Values are also reported for 16 additional compounds including lutein, zeaxanthin, alpha- and beta-cryptoxanthin, lycopene, alpha-carotene, retinyl palmitate, and 25-hydroxyvitamin D. The certified values for the fat-soluble vitamins and carotenoids in SRM 968c were based on the agreement of results from the means of at least two liquid chromatographic methods used at the National Institute of Standards and Technology (NIST) and from the medians from an interlaboratory comparison study among institutions that participate in the NIST Micronutrients Measurement Quality Assurance Program. The assigned values for cholesterol in the SRM are the means of results obtained using the NIST definitive method, gas chromatography-isotope dilution mass spectrometry.

Calibration↗

Serum oxidative and antioxidant parameters in a group of Italian patients with age-related maculopathy.

OBJECTIVES: The purpose of this study was to measure the oxidative and antioxidant biochemical parameters in the serum of Italian patients with age-related maculopathy (ARM) and in a similar age control group from the same area, in order to determine the weight of oxidative status as risk factor in the early stage of macular degeneration onwards. DESIGN AND METHODS: Forty-eight ARM patients (19 early and 29 late form) and 46 normal subjects, similar for age, sex and life-style, were studied. A series of serum and/or plasma antioxidants (vitamins C, E, A, total and individual carotenoids, zinc, total plasma antioxidant capacity--TRAP) and oxidative parameters (reactive oxygen metabolites--ROM, oxidized-low-density lipoprotein antibodies-anti-Ox-LDL) were evaluated in both groups, also with regard to age and disease stage. RESULTS: Levels of vitamins C, E, total carotenoids and beta-cryptoxanthine were lower in late ARM than in early ARM (p<0.05). Of the serum carotenoids investigated, only lycopene was lower in the two ARM forms than in controls (p<0.05). The main biochemical parameters, TRAP, zinc, anti-Ox-LDL and ROM were similar in the two groups. CONCLUSIONS: A deficit of antioxidants (vitamins C, E and carotenoids) seems to be associated with ARM in Italian patients, particularly the advanced form, it is also suggested that in ARM patients macular susceptibility to oxidative damage is not related with age.

Age Factors↗

Carotenoid mixtures protect multilamellar liposomes against oxidative damage: synergistic effects of lycopene and lutein.

Antioxidant activity of carotenoids in multilamellar liposomes assayed by inhibition of formation of thiobarbituric acid-reactive substances was in the ranking: lycopene> alpha-tocopherol > alpha-carotene > beta-cryptoxanthin > zeaxanthin = beta-carotene > lutein. Mixtures of carotenoids were more effective than the single compounds. This synergistic effect was most pronounced when lycopene or lutein was present. The superior protection of mixtures may be related to specific positioning of different carotenoids in membranes.

Antioxidants↗

Carotenoids inhibit DNA synthesis in human aortic smooth muscle cells.

Quiescent, serum-starved human aortic smooth muscle cells were restimulated with 20% foetal calf serum in Dulbecco's modified Eagle medium, in the presence and absence of beta-carotene, canthaxanthin, zeaxanthin, lycopene, lutein or beta-cryptoxanthin, at final concentrations up to 23 microM. Concentration-dependent inhibition of DNA synthesis, measured by [methyl-3H]thymidine incorporation, was observed for the carotenoids, except for canthaxanthin and lutein which had no effect. Lycopene was the most potent of the carotenoids tested. The results suggest that antiproliferative effects of dietary carotenoids might be of significance in vivo.

Aorta↗

Relation of elevated serum alanine aminotransferase activity with iron and antioxidant levels in the United States.

BACKGROUND & AIMS: Oxidative stress is thought to play a role in liver injury. Hepatic iron may promote liver injury, whereas antioxidant vitamins and minerals may inhibit it, but few clinical studies have examined such relationships. We analyzed the associations of serum iron measures and antioxidant concentrations with abnormal serum alanine transaminase (ALT) activity in a large, national, population-based study. METHODS: A total of 13,605 adult participants in the third U.S. National Health and Nutrition Examination Survey (NHANES III), 1988-1994, underwent phlebotomy. Exclusions included excessive alcohol consumption, hepatitis B and C, and iron overload. RESULTS: Elevated ALT levels were found in 3.1% of the population. In univariate analysis, factors associated with abnormal ALT levels (P < 0.05) included higher transferrin saturation and iron and selenium concentrations, and lower vitamin C, alpha and beta carotene, and lutein/zeaxanthin concentrations. In multivariate logistic regression analyses, elevated ALT level was associated positively with increasing deciles of transferrin saturation (odds ratio [OR] per decile, 1.10; 95% confidence interval [CI], 1.03-1.18) and iron concentration (OR, 1.13; 95% CI, 1.06-1.21). Abnormal ALT level was associated negatively with increasing deciles of alpha carotene (OR, 0.82; 95% CI, 0.72-0.94), beta carotene (OR, 0.91; 95% CI, 0.86-0.96), beta cryptoxanthin (OR, 0.91; 95% CI, 0.84-0.99), lutein/zeaxanthin (OR, 0.90; 95% CI, 0.84-0.96), and a variable combining the 5 carotenoid measures (OR, 0.89; 95% CI, 0.83-0.95). Vitamin C was associated inversely, but only at the highest concentrations. CONCLUSIONS: In this large, national, population-based study, the risk for apparent liver injury was associated with increased iron and decreased antioxidants, particularly carotenoids.

Alanine Transaminase↗

High plasma levels of alpha- and beta-carotene are associated with a lower risk of atherosclerosis: results from the Bruneck study.

BACKGROUND AND PURPOSE: A large number of studies have contributed to the hypothesis that carotenoids, vitamins A and E are protective against atherosclerosis by acting as antioxidants. The aim of this study was to assess the relationship between plasma levels of carotenoids (alpha- and beta- carotene, lutein, lycopene, zeaxanthin, beta-cryptoxanthin), vitamins A and E, and atherosclerosis in the carotid and femoral arteries. METHODS: This prospective and cross sectional study involved a randomly selected population sample of 392 men and women aged 45-65 years. Carotid and femoral artery atherosclerosis was assessed by high-resolution duplex ultrasound. RESULTS: alpha- and beta- carotene plasma levels were inversely associated with the prevalence of atherosclerosis in the carotid and femoral arteries (P=0.004) and with the 5-year incidence of atherosclerotic lesions in the carotid arteries (P=0.04). These findings were obtained after adjustment for other cardiovascular risk factors (sex, age, LDL (low density lipoproteins), ferritin, systolic blood pressure, smoking, categories of alcohol consumption, social status, C-reactive protein). Atherosclerosis risk gradually decreased with increasing plasma alpha- and beta-carotene concentrations (P=0.004). No associations were found between vitamin A and E plasma levels and atherosclerosis. CONCLUSIONS: This study provides further epidemiological evidence of a protective role of high alpha- and beta- carotene in early atherogenesis.

Adult↗

Effects of plant stanol esters supplied in low-fat yoghurt on serum lipids and lipoproteins, non-cholesterol sterols and fat soluble antioxidant concentrations.

Oil-based products enriched with plant stanol esters can lower low-density lipoprotein (LDL) cholesterol concentrations by 10-14%. Effectiveness of low-fat products, however, has never been evaluated, although such products fit into a healthy diet. We therefore examined the effects of plant stanol esters emulsified into low-fat yoghurt (0.7% fat) on fasting concentrations of plasma lipids and lipid-soluble antioxidants, which may also change by plant stanol consumption. Sixty non-hypercholesterolemic subjects first consumed daily three cups (3 x 150 ml) of placebo yoghurt for 3 weeks. For the next 4 weeks, 30 subjects continued with the placebo yoghurt, while the other 30 subjects received three cups of experimental yoghurt. Each cup provided 1 g of plant stanols (0.71 g sitostanol plus 0.29 g campestanol) as its fatty acid ester. LDL cholesterol (mean+/-S.D.) increased by 0.06+/-0.21 mmol/l in the placebo group, but decreased by -0.34+/-0.30 mmol/l in the experimental group. The difference in changes between the two groups of 0.40 mmol or 13.7% was highly significant (P<0.001; 95% confidence interval for the difference, (-)0.26 -(-)0.53 mmol/l). Effects were already maximal after 1 week. HDL cholesterol and triacylglycerol concentrations did not change. Total tocopherol levels increased by 1.43 micromol/mmol LDL cholesterol (14.0%, P=0.015). beta-carotene levels, however, decreased by -0.02 micromol/mmol LDL cholesterol (-14.4%, P=0.038). Decreases in absolute beta-carotene concentrations were found in all apoB-containing lipoproteins. LDL-cholesterol standardised phytofluene levels decreased by 21.4+/-25.7% (P<0.001), while other plasma carotenoid (lutein/zeaxanthin, beta-cryptoxanthin, lycopene and alpha-carotene) levels did not change significantly. We conclude that low-fat yoghurt enriched with plant stanol esters lowers within 1 week LDL cholesterol to the same extent as oil-based products. LDL-cholesterol standardised concentrations of tocopherol increased. The observed decrease in beta-carotene levels, as found in many other studies, appears not to be limited to the LDL fraction.

Adolescent↗

Serum carotenoids and atherosclerosis. The Rotterdam Study.

High circulating levels of carotenoids have been thought to exhibit a protective function in the development of atherosclerosis. We investigated whether aortic atherosclerosis was associated with lower levels of the major serum carotenoids in alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein, lycopene, and zeaxanthin-in a subsample of the elderly population of the Rotterdam Study. Aortic atherosclerosis was assessed by presence of calcified plaques of the abdominal aorta. The case-control analysis comprised 108 subjects with aortic atherosclerosis and controls. In an age- and sex-adjusted logistic regression model, serum lycopene was inversely associated with the risk of atherosclerosis. The odds ratio for the highest compared to the lowest quartile of serum lycopene was 0.55 (95% CI 0.25-1.22; p(trend)=0.13). Multivariate adjustment did not appreciably alter these results. Stratification by smoking status indicated that the inverse association between lycopene and aortic calcification was most evident in current and former smokers (OR=0.35; 95% CI 0.13-0.94; p(trend)=0.04). No association with atherosclerosis was observed for quartiles of serum concentrations of alpha-carotene, beta-carotene, lutein, and zeaxanthin. In conclusion, this study provides evidence for a modest inverse association between levels of serum lycopene and presence of atherosclerosis, the association being most pronounced in current and former smokers. Our findings suggest that lycopene may play a protective role in the development of atherosclerosis.

Aged↗

High-performance liquid chromatography for the characterization of carotenoids in the new sweet orange (Earlygold) grown in Florida, USA.

High-performance liquid chromatography with photodiode array detection was developed for the separation and identification of carotenoids from a new sweet orange, Earlygold. Carotenoid pigments were extracted using hexane-acetone-ethanol and saponified using 10% methanolic potassium hydroxide. More than 25 carotenoid pigments were separated within 40 min using a ternary gradient (acetonitrile-methanol, methyl tert-butyl ether and water) elution on a C30 reversed-phase column. The carotenoid pattern of Earlygold was generally similar to the early season Hamlin but with some quantitative differences, especially with violaxanthin. Major carotenoids including violaxanthin, lutein, beta-cryptoxanthin, antheraxanthin, luteoxanthin, zeaxanthin, beta-carotene, and alpha-carotene were identified based on on-line spectral data obtained by a photodiode array detector, and comparison to the spectra of the standards and reported values. A numerical notation, the ratio of the peak heights between absorption bands, was also calculated to compare to the literature values.

Carotenoids↗