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Objective Studies show low correlations between dietary intake and serum concentrations of lycopene, which make it difficult to assess the effectiveness of dietary interventions with this phytonutrient. We hypothesized that 1) combining food-frequency questionnaires (FFQs) and 3-d diet records (3D-DRs) by the triads method would improve the validity of this relation and 2) correcting dietary information for differences in lycopene absorption from food matrices would further improve validity. Methods We measured dietary intakes of lycopene from 49 adults by 3D-DR and FFQ. Serum lycopene was measured by high-performance liquid chromatography with diode array detection. Cholesterol and triacylglycerol concentrations were measured spectrophotometrically. Lycopene-containing foods were given absorption factors based on literature and laboratory values. Associations between dietary and serum lycopene were modeled using multiple regression. The triads method was used for validation of relations among FFQ, 3D-DR, and serum lycopene. Results Raw data showed low correlations between dietary and serum lycopene ( r = +0.15 for 3D-DR, +0.35 for FFQ). Mathematical modeling showed that the 3D-DR and FFQ methods must be used to collect accurate dietary information for lycopene. Validity coefficients calculated by the triads method were +0.34 for 3D-DR and +0.78 for FFQ. Correcting for absorption increased the validity coefficient to +0.72 for 3D-DR and from +0.45 to +0.66 for serum lycopene. Conclusion The relation between dietary intake and serum concentrations of lycopene and other carotenoids can be improved by collecting 3D-DR and FFQ data and by adjusting dietary information for nutrient absorption.

The vitamin A activity of beta-carotene is variable and surprisingly low in women. The reasons for this are not well understood. The vitamin A activity of beta-carotene in men is still uncertain. Contributions of dietary factors compared with individual traits are largely unknown.Our objective was to measure the intrinsic variability in the vitamin A activity of beta-carotene among healthy, well-fed men living in a controlled environment.We used a double-tracer test-retest design. We dosed 11 healthy men orally with 30 micromol hexadeuterated (D6) retinyl acetate (all-trans-19,19,19,20,20,20-[2H6]retinyl acetate) and then with 37 micromol D6 beta-carotene (19,19,19,19',19',19'-[2H6]beta-carotene) 1 wk later. Doses were taken with breakfasts containing 16 g fat. We measured D6 retinol, D6 beta-carotene, and trideuterated (D3) retinol (derived from D6 beta-carotene) concentrations in plasma. Areas under the plasma concentration x time since dosing curves (AUCs) were determined for D6 retinol, D6 beta-carotene, and D3 retinol.All men had detectable D6 retinol concentrations in plasma. The mean (+/-SE) absorption of D6 beta-carotene in all subjects was 2.235 +/- 0.925%, and the mean conversion ratio was 0.0296 +/- 0.0108 mol retinol to 1 mol beta-carotene. Only 6 of 11 men had sufficient plasma concentrations of D6 beta-carotene and D3 retinol that we could measure. The mean absorption of D6 beta-carotene in these 6 subjects was 4.097 +/- 1.208%, and the mean conversion ratio was 0.0540 +/- 0.0128 mol retinol to 1 mol beta-carotene.The vitamin A activity of beta-carotene, even when measured under controlled conditions, can be surprisingly low and variable.

The purpose of the study was to evaluate the effect of a low carotenoid diet (83 micrograms Beta-carotene) on malondialdehyde-thiobarbituric acid (MDA-TBA) concentrations of nine pre-menopausal women.Subjects lived on the metabolic research unit of the Western Human Nutrition Research Center (WHNRC), where diet, exercise and other activities were controlled. Five subjects (Group C, control group) consumed a low carotenoid diet and received an additional 0.5 mg/day of Beta-carotene while four subjects (Group P, placebo group) received only the low carotenoid diet during days 1 to 60 (period 1). All subjects received 0.5 mg/day of Beta-carotene during days 60 to 100 (period 2), plus three capsules/day mixed carotenoid supplement (Neo-Life Company) during study days 100 to 120. Changes in MDA-TBA concentrations were analyzed during the study periods and between the groups.At the start of the study (day 1), no significant difference in the MDA-TBA concentration was observed between the control (Group C) and the placebo (Group P) subjects. During period 1 (days 2 to 60), when Group P subjects consumed the low carotenoid diet without supplementation, the MDA-TBA values for Group P rose markedly and were significantly (p0.05) higher than the MDA-TBA values for Group C subjects who were receiving carotenoid supplementation. During period 2 (days 60 to 100) when both groups received carotenoid supplementation, the MDA-TBA values of Group P subjects were significantly (p0.05) reduced to the point where they were similar to the MDA-TBA values for Group C subjects.These findings provide evidence to support the beneficial effects of carotenoids in preventing lipid peroxidation in the cells. Further studies are needed to identify the exact mechanism by which carotenoids prevent lipid peroxidation and the amount needed for normal activity.

We developed supercritical fluid procedures for extracting vitamin A and beta-carotene from vitamin supplements and calf liver tissue. The SF extracts could be injected onto an HPLC column without further pretreatment. Samples were analysed by RP-HPLC using diode array detection or by spectrophotometry. Recoveries were very good. SF extracts from a vitamin preparation of uniform composition had an R.S.D. of 4%. Extracts from calf liver supplements were predictably more heterogeneous. The SF extraction method is less labor intensive then traditional liquid-liquid procedures for extracting vitamin A and carotenoids from tissues.

Millions of people worldwide have severe chronic physical disabilities (e.g., polio, spinal cord injury). We hypothesized that these people are at risk for poor antioxidant nutrient intakes and status, because of their typically low energy expenditures and low socioeconomic status. We surveyed antioxidant nutrient intakes and concentrations in subjects with physical disabilities and age matched non-disabled controls. Only one subject with physical disabilities ate five or more servings of vegetables per day. The group had low intakes of vegetables (minus potatoes) and citrus fruits, and trends for low intakes of all antioxidant nutrients. Serum a-tocopherol and total carotenoid concentrations were lower in subjects with physical disabilities, while retinol and individual carotenoids showed non-significant lower trends. Since adequate antioxidant nutrients are associated with decreased chronic disease risk, nutrition professionals might improve the health of patients with physical disabilities by periodically monitoring their intakes and status of antioxidant nutrients.

Vitamin A containing (holo) free and transthyretin-bound (TTR) retinol binding protein (RBP) concentrations in plasma from rats with variable vitamin A status were determined by high performance liquid chromatography. Two different methods were used: (1) molecular exclusion with a TSK 2000 column and (2) reverse phase using a Protesil Octyl 300 column. Holo TTR-RBP peak areas were positively correlated to liver vitamin A concentrations (r = 0.79 for molecular exclusion 0.81 for reverse phase) in rats with marginal and normal vitamin A status. This correlation was higher than the correlation of serum retinol to liver vitamin A in these rats (r = 0.58). The correlation of holo-TTR-RBP to liver vitamin A was also higher than its correlations to plasma vitamin A. Therefore, plasma concentrations of holo—TTR-RBP may be influenced by marginal vitamin A liver stores to a greater extent than plasma retinol is. This suggests that holo—TTR-RBP protein concentrations may be the better indicator of marginal vitamin A nutritional status in rats. The correlations of both holo—TTR-RBP and serum retinol decreased sharply at high liver vitamin A concentrations. Neither method is suitable for measuring sub-toxicity in rats.

We describe a reversed-phase high-performance liquid chromatographic method for the determination of vitamin A-transporting (holo) transthyretin-bound (TTR) retinol-binding protein (RBP) concentrations in serum or plasma. Holo-TTR-RBP and free retinol derived primarily from free RBP are consistently observed with this chromatographic method. Holo-TTR-RBP concentrations determined by this method are highly correlated to holo-TTR-RBP concentrations measured by chromatography. This method has the advantage of using less expensive columns and having peak areas which are more proportional to their true concentrations in plasma, as determined by comparison to purified protein spectrophotometry and radial immunodiffusion. The percentage of RBP circulating as holo-TTR-RBP decreased significantly as the total concentration of RBP or retinol increased. Because purified holo-TTR-RBP did not dissociate under these chromatographic conditions, this suggests that more vitamin A circulates as holo-free RBP or free retinol in the blood of people with high serum RBP.

Lycopene, or the foods that contain it, may prevent prostate cancer. Studies suggest that some cis-lycopene isomers are more bioavailable than the trans-lycopene isomer. We hypothesized that tangerine tomatoes, which predominantly contain the tetra-cis isomer, should be a good source of bioavailable lycopene. We fed lunches containing 300 g tangerine or red tomato sauce per day to 21 healthy adults in a double-blind crossover design. We collected blood at baseline and after each treatment and washout period. We measured tetra-cis, other cis, and trans lycopene, as well as other carotenoids, by reversed-phase high-performance liquid chromatography. Both tomato sauces increased lycopene concentrations in blood, but the tangerine tomato sauce caused a greater increase of total and tetra-cis-lycopene. The cis isomer(s) may also have facilitated absorption of the trans-lycopene isomer. Indices of oxidative damage decreased as serum lycopene concentrations increased. Our results suggest that total lycopene concentrations can be increased by substituting tetra-cis-lycopene-rich tangerine tomatoes for common red tomatoes in the diet.

Vitamin A deficiency is a health problem in Southeast Asia that can be corrected by feeding orange fruits and vegetables such as mango. Pumpkin is a traditional Korean food that is easy to store and is already believed to have health benefits. We extracted carotenoids from pumpkin by liquid-liquid extraction and by supercritical fluid extraction. We measured carotenoids by reversed-phase chromatography with diode array detection. The major carotenoid in pumpkin (> 80%) is beta-carotene, with lesser amounts of lutein, lycopene, alpha-carotene and cis-beta-carotene. Pumpkin is a rich source of beta-carotene and might be useful for preventing Vitamin A deficiency.

Dietary intakes of carotenoids are highly variable in human populations as are serum carotenoid concentrations. However, there are few controlled data relating carotenoid intake to concentration. Most of the data that are available are from measurements of the absorption and decay of large pharmacologic doses of carotenoids, and are therefore of unknown physiologic relevance. Our objective was to determine the half-life (t(1/2)) of the most abundant carotenoids in blood serum from healthy adult women living under controlled conditions. As part of two carotenoid isotopic studies, we measured serum concentrations of beta-carotene, alpha-carotene, lutein, zeaxanthin, beta-cryptoxanthin and lycopene in 19 healthy young adult women that were fed controlled low carotenoid diets for approximately 10 wk. All other nutrients (vitamins A, E and C) were provided at 100-150% of the 1989 U.S. recommended dietary allowance levels. Exercise and activities were controlled throughout the studies to simulate usual activity patterns. Carotenoid concentrations were measured by reversed-phase HPLC. Serum carotenoid concentration decreases during depletion followed first-order kinetics. The half-lives determined in decreasing order were as follows: lutein (76 d) > alpha-carotene (45 d) = beta-cryptoxanthin (39 d) = zeaxanthin (38 d) = beta-carotene (37 d) > lycopene (26 d). Half-lives were unrelated to physical or demographic characteristics such as body mass, body fat, racial background or age in these relatively homogeneous groups. Carotenoids decreased by similar first-order mechanisms, although the rates differed for individual carotenoids.

BACKGROUND Blood beta-carotene and vitamin A responses to oral beta-carotene are variable in humans. Some individuals are characterized as responders and others as low- or nonresponders. A better understanding of the conditions that produce the variability is important to help design public health programs that ensure vitamin A sufficiency. OBJECTIVE Our objective was to assess variability in absorption and conversion of beta-carotene to vitamin A in vivo in humans by using a novel double-tracer ?hexadeuterated (D(6)) beta-carotene and D(6) retinyl acetate approach. DESIGN Eleven healthy women were housed at the US Department of Agriculture Western Human Nutrition Research Center metabolic unit for 44 d, where they consumed diets adequate in vitamins and minerals except for carotenoids. After an adaptation period, the women were given 30 micromol D(6) retinyl acetate orally, followed 1 wk later with 37 micromol D(6) beta-carotene (approximately equimolar doses). Time-dependent plasma concentration curves were determined for D(6) retinol, D(6) beta-carotene, and trideuterated (D(3)) retinol (derived from D(6) beta-carotene). RESULTS Mean (+/-SE) absorption of D(6) beta-carotene was 3.3 +/- 1.3% for all subjects. The mean conversion ratio was 0.81 +/- 0.34 mol D(3) retinol to 1 mol D(6) beta-carotene for all subjects. However, only 6 of the 11 subjects had plasma D(6) beta-carotene and D(3) retinol concentrations that we could measure. The mean absorption of D(6) beta-carotene in these 6 subjects was 6.1 +/- 0.02% and their conversion ratio was 1.47 +/- 0.49 mol D(3) retinol to 1 mol D(6) beta-carotene. The remaining 5 subjects were low responders with

We measured immunologically active (apo + holo) retinol-binding protein (RBP), vitamin A-carrying (holo) free RBP, and transthyretin-bound (TTR) holo-RBP in serum from 34 retrospective cases of fluctuating acute renal failure. All subjects had high serum creatinine concentrations caused by renal failure. Apo + holo, holo-TTR-RBP, and (especially) holo-free RBP all correlated poorly but significantly with serum creatinine concentration. Therefore, the use of any form of RBP to measure vitamin A status may be of limited value in subjects with high creatinine concentrations in serum. However, molecular-exclusion HPLC may be able to distinguish increases in RBP concentration associated with renal failure from those caused by altered vitamin A status, because renal failure causes abnormalities in the number and retention times of chromatographic peaks as well as their areas.

Dietary intakes are usually estimated by either a food frequency test, or by food records. We hypothesized that dietary intake estimates for fat soluble vitamins might be more accurate if information from both food frequency tests and food records were used. We estimated dietary intakes in 10 healthy adults by 4 food frequency questionnaires and ten 3-day food records collected over a year. Serum antioxidant nutrient concentrations (vitamins A, E, and the carotenoids) were measured by HPLC throughout the year. Few changes in intake occurred over the year. Estimates of nutrient densities correlated more often than estimates of nutrient intakes to serum nutrient concentrations. Nutrient density estimates derived from the average of food frequency and food record estimates were significantly correlated with serum nutrient concentrations more often than dietary estimates derived from either food frequency or food records alone. We suggest that nutrient density estimates derived from a combination of food frequency and food records may be useful for studies of free-living individuals.