Terry R. Neidlinger

Supercritical fluid extraction and reversed-phase liquid chromatography methods for vitamin A and β-carotene heterogeneous distribution of vitamin A in the liver

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.

The effect of a low carotenoid diet on malondialdehyde-thiobarbituric acid (MDA-TBA) concentrations in women: a placebo-controlled double-blind study.

OBJECTIVE 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. METHODS 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. RESULTS 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 (p < 0.05) 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 (p < 0.05) reduced to the point where they were similar to the MDA-TBA values for Group C subjects. CONCLUSIONS 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.

Tangerine tomatoes increase total and tetra-cis-lycopene isomer concentrations more than red tomatoes in healthy adult humans

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.

Dietary Intakes and Serum Concentrations of Vitamin E and Total Carotenoids of Healthy Adults with Severe Physical Disabilities are Lower than Matched Controls

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.

Relationship of holo-free and transthyretin-bound plasma retinol-binding protein levels with liver vitamin A in rats

Abstract 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.

Assay of human transthyretin-bound holo-retinol-binding protein with reversed-phase high-performance liquid chromatography

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.

Effect of low-calorie diets on plasma retinol-binding protein concentrations in overweight women.

The concentrations of total protein, albumin and retinol-binding protein, a major transport protein for vitamin A, are significantly decreased by protein-calorie malnutrition. Weight-loss diets, sometimes involving severe energy deficits over prolonged periods of time, are common in the United States. The effect, if any, of prolonged low calorie weight-loss diets with normal intakes of protein on albumin, total protein and retinol-binding protein concentrations (and potentially on vitamin A metabolism) had not been extensively studied. We measured total protein, albumin, apo + holo retinol-binding protein and holo-free- and holo-transthyretin-bound retinol-binding protein concentrations during the course of a nutritionally adequate weight-loss diet (50% calorie restriction). We found that this type of dieting did not affect total protein, albumin or apo + holo, holo-free or holo-transthretin-bound retinol-binding protein concentrations significantly. This suggests that protein intake is more critical than caloric intake for retinol-binding protein status.