The measurement of vitamin A (VA) and iron status is very important in the assessment of nutritional deficiencies. The objective of this research was to develop a sandwich ELISA technique for the simultaneous measurement of ferritin, soluble transferrin receptor, retinol binding protein, and C-reactive protein (CRP) as indicators for VA and iron status. The inclusion of CRP as marker of infection allows for more accurate interpretation of VA and iron status. This is accomplished in a 30-microL serum or plasma sample using an ELISA with different capture and detection antibodies and different dilutions of the sample. Commercially available clinical serum controls were used for calibration purposes. The developed assays were compared to commercially available traditional tests. Regression coefficients comparing both assays were better than 0.84 (P < 0.001). Using a limited sample set, the sandwich ELISA assay produced very similar specificity and sensitivity compared to traditional methods when common cutoff values were applied. Intra- and interassay variability was between 5 and 14% for all tests. The cost of the materials for all 5 measurements decreases to less than $1/sample if a large number of samples is analyzed. Due to the low cost, high throughput, and comparability to traditional tests, this procedure has several advantages for assessing VA and iron status in population surveys.

The Biomarkers of Nutrition for Development (BOND) project is designed to provide evidence-informed advice to anyone with an interest in the role of nutrition in health. The BOND program provides information with regard to selection, use, and interpretation of biomarkers of nutrient exposure, status, function, and effect, which will be especially useful for readers who want to assess nutrient status. To accomplish this objective, expert panels are recruited to evaluate the literature and to draft comprehensive reports on the current state of the art with regard to specific nutrient biology and available biomarkers for assessing nutritional status at the individual and population levels. Phase I of the BOND project includes the evaluation of biomarkers for 6 nutrients: iodine, folate, zinc, iron, vitamin A, and vitamin B-12. This review of vitamin A is the current article in this series. Although the vitamin was discovered >100 y ago, vitamin A status assessment is not trivial. Serum retinol concentrations are under homeostatic control due in part to vitamin A’s use in the body for growth and cellular differentiation and because of its toxic properties at high concentrations. Furthermore, serum retinol concentrations are depressed during infection and inflammation because retinol-binding protein (RBP) is a negative acute-phase reactant, which makes status assessment challenging. Thus, this review describes the clinical and functional indicators related to eye health and biochemical biomarkers of vitamin A status (i.e., serum retinol, RBP, breast-milk retinol, dose-response tests, isotope dilution methodology, and serum retinyl esters). These biomarkers are then related to liver vitamin A concentrations, which are usually considered the gold standard for vitamin A status. With regard to biomarkers, future research questions and gaps in our current understanding as well as limitations of the methods are described.

A variety of bonded phase parameters (endcapping, phase chemistry, ligand length, and substrate parameters) were studied for their effect on column retention and selectivity toward carotenoids. Decisions were made on how each of these variables should be optimized based on the separation of carotenoid and polycyclic aromatic hydrocarbon test probes. A column was designed with the following properties: high absolute retention, enhanced shape recognition of structured solutes, and moderate silanol activity. These qualities were achieved by triacontyl (C30) polymeric surface modification of a moderate pore size (approximately 20 nm), moderate surface area (approximately 200 m2/g) silica, without subsequent endcapping. The effectiveness of this "carotenoid phase" was demonstrated for the separation of a mixture of structurally similar carotenoid standards, an extract of a food matrix Standard Reference Material, and a beta-carotene dietary supplement under consideration as an agent for cancer intervention/prevention.

The relative solubility, stability, and absorptivity of lutein and @-carotene were determined in 18 organic solvents. The solubility of both carotenoids was greatest in tetrahydrofuran, while hexane exhibited the least solubility for lutein; methanol and acetonitrile exhibited the least solubility for @-carotene. Stability was monitored for 10 days at room temperature by measuring absorbance changes at the wavelength maximum. In the majority of the solvents, initial absorbance decreased by less than 10% during the 10-day period. Degradation was greatest for both carotenoids in cyclohexanone. The relative absorptivities were determined by calculating the carotenoid concentration in a reference solvent using a reference absorptivity, and then Beer's Law was applied to the measured absorbance of the same carotenoid concentration in other organic solvents. Absorbance maxima and relative absorptivities were in good agreement with available literature values.

Changes in seven plasma carotenoids were measured in 30 men for 11 d after ingesting a single dose of pure beta-carotene or a high carotenoid vegetable. A controlled, low-carotenoid diet was fed in a crossover design. Maximum plasma concentrations of beta-carotene occurred 24-48 h after dosing with beta-carotene (12 or 30 mg) or carrots (270 g). A large intake of broccoli (600 g) or tomato juice (180 g) did not change any plasma carotenoids. We concluded that 1) normal subjects vary widely, three to fourfold, in efficiency of carotenoid absorption; 2) peak plasma response to beta-carotene in a capsule occurs at 24-48 h; 3) a large single intake of carrots produces a small increase in plasma beta-carotene but single intakes of broccoli or tomato juice do not change plasma carotenoids; and 4) plasma response to pure beta-carotene is greater than the response to a similar amount of beta-carotene in carrots.