Retinyl Acetate Research Articles

Plasma Lipid Concentrations in Preruminant Calves Fed Whole Milk with Whey Protein Isolate

The objective was to investigate the acute effects of retinol acetate added to whey protein isolate (WPI) on postprandial changes in plasma retinol (experiment 1) and the acute effects of milk fat added to WPI on triglyceride (TG), chylomicrons and very low density lipoprotein (VLDL), and fatty acid concentrations (experiment 2) in suckling calves at 1 and 6 wk of age. In experiment 1, 16 Holstein male calves were alloted to 2 equal groups. On the days of measurement, the calves were fed at 0900h whole milk [4% of body weight (BW)] mixed with vitamin A acetate (500,000 IU) with or without WPI (0.04% of BW). At 1 wk of age, significantly higher postfeeding concentrations of plasma retinol were observed in the calves fed milk with WPI. At 6 wk of age, no differences in the plasma retinol concentrations were observed between 2 groups. On the days of measurement in experiment 2, 16 male calves were fed at 0900h whole milk (4% of BW) with added milk fat prepared by centrifugation from whole milk (2% of BW) with or without WPI (0.04% of BW). The milk supplemented with fat was prepared on the day before the measurement. At 1 wk of age, significant higher postfeeding concentrations of plasma TG concentrations were obtained in the calves fed WPI than in the control calves, immediately after the meal or from 7h later onward. Plasma chylomicrons and VLDL concentrations at 1 wk of age were significantly higher in the WPI-fed group than in the control group at 8h postfeeding. In the calves with the WPI diet, plasma concentrations of myristic, palmitic, stearic, oleic, and linoleic acids at 1 wk of age were significantly higher than those in the control calves at 8h after feeding. However, chylomicrons and VLDL, and fatty acid concentrations did not differ between the 2 groups after feeding at 6 wk of age. Results indicate that WPI increases plasma lipid concentration of preruminant calves only at 1 wk of age. These data are interpreted to indicate that WPI enhances mainly lipid uptake in the intestines of neonatal calves.

Photodecomposition of Retinyl Palmitate in Ethanol by UVA LightFormation of Photodecomposition Products, Reactive Oxygen Species, and Lipid Peroxides

Photodecomposition of retinyl palmitate (RP), an ester and the storage form of vitamin A (retinol), in ethanol under UVA light irradiation was studied. The resulting photodecomposition products were separated by reversed-phase HPLC and identified by spectral analysis and comparison with the chromatographic and spectral properties of synthetically prepared standards. The identified products include 5,6-epoxy-RP, 4-keto-RP, 11-ethoxy-12-hydroxy-RP, 13-ethoxy-14-hydroxy-RP, anhydroretinol (AR), palmitic acid, ethyl palmitate, and four tentatively assigned cis and trans isomeric 15-ethoxy-ARs. AR was formed as a mixture of all-trans-AR, 6Z-cis-AR, 8Z-cis-AR, and 12Z-cis-AR with all-trans-AR predominating. 5,6-Epoxy-RP, 4-keto-RP, 11-ethoxy-12-hydroxy-RP, and 13-ethoxy-14-hydroxy-RP were also formed from reaction of RP with alkylperoxy radicals generated by thermal decomposition of 2,2'-azobis(2,4-dimethylvaleronitrile). Formation of these photodecomposition products was inhibited in the presence of sodium azide (NaN3), a free radical inhibitor. These results suggest that formation of 5,6-epoxy-RP, 4-keto-RP, 11-ethoxy-12-hydroxy-RP, and 13-ethoxy-14-hydroxy-RP from photoirradiation of RP is mediated by a light-initiated free radical chain reaction. AR and the isomeric 11-ethoxy-ARs were not formed from reaction of RP with alkylperoxy radicals generated from 2,2'-azobis(2,4-dimethylvaleronitrile), and their formation was not inhibited when NaN3 was present during the photoirradiation of RP. We propose that these products were formed through an ionic photodissociation mechanism, which is similar to the reported formation of AR through ionic photodissociation of retinyl acetate. RP and all its identified photodecomposition products described above (i) were not mutagenic in Salmonella typhimurium tester strains TA98, TA100, TA102, and TA104 in the presence and absence of S9 activation enzymes, (ii) were not photomutagenic in Salmonella typhimurium TA102 upon UVA irradiation, and (iii) did not bind with calf thymus DNA in the presence of microsomal metabolizing enzymes. These results suggest that RP and its decomposition products are not genotoxic; however, photoirradiation of RP, 5,6-epoxy-RP, and AR with UVA light in the presence of methyl linoleate resulted in lipid peroxide (methyl linoleate hydroperoxides) formation. The lipid peroxide formation was inhibited by dithiothreitol (DTT) (free radical scavenger), NaN3 (singlet oxygen and free radical scavenger), and superoxide dismutase (SOD) (superoxide scavenger) but was enhanced by the presence of deuterium oxide (D2O) (enhancement of singlet oxygen lifetime). These results suggest that photoirradiation of RP, 5,6-epoxy-RP, and AR by UVA light generated reactive oxygen species resulting in lipid (methyl linoleate) peroxidation.

Retinol, α- and γ-tocopherol and carotenoids in natural and vitamin A- and E-fortified dairy products commercialized in Spain

Abstract Natural and added forms of vitamin A (all-trans-retinol, retinyl esters and β -carotene) and vitamin E ( α -tocopherol, γ -tocopherol, tocopheryl acetate) were determined in commercially available dairy products that are frequently consumed. Retinyl esters, β -carotene and α -tocopherol are customarily found in natural dairy products, whereas retinyl and α -tocopheryl acetate occur in most vitamin-fortified products. Lutein, zeaxanthin and γ -tocopherol were found in egg- and oil-containing products, respectively. α -Tocopheryl acetate was found in all vitamin-fortified dairy products analysed, whereas retinyl acetate was only present in fluid milks but not in margarines. Minor amounts of retinyl acetate may occasionally be found in some non-fortified milks, suggesting contamination during processing. In commercially available dairy products in Spain, retinyl palmitate and α -tocopherol contents vary substantially, especially in high fat and vitamin-fortified dairy products. Compared with the label claims in vitamin-fortified products, over-fortification predominates, although over- and under-fortification of both vitamins A and E may occur simultaneously in fluid milks. The wide variation observed in commercially available natural and vitamin-fortified dairy products may be an important source of bias and imprecision in nutritional studies.

Use of the deuterated-retinol-dilution technique to monitor the vitamin A status of Nicaraguan schoolchildren 1 y after initiation of the Nicaraguan national program of sugar fortification with vitamin A

Nicaragua initiated a national program of vitamin A fortification of its domestic sugar supply starting with the 1999-2000 sugarcane harvest. This study was conducted to document any change in the vitamin A status of a cohort of children during the first year of the program. The vitamin A status of 21 Nicaraguan schoolchildren (mean age: 6.7 y; range: 5.3-9.3 y) was assessed in March 2000 and in March 2001. Total-body vitamin A stores and liver vitamin A concentrations were estimated with the deuterated-retinol-dilution (DRD) technique at a dose of 5 mg [(2)H(4)]retinyl acetate at baseline and 5 mg [(2)H(8)]retinyl acetate during the repeat test 1 y later. Plasma retinol and carotenoids were measured by HPLC. Median total-body vitamin A stores increased from 0.33 to 0.72 mmol (P = 0.0001), liver vitamin A concentrations from 0.52 to 0.78 mumol/g (P = 0.0003), and plasma retinol concentrations increased from 0.97 to 1.17 mumol/L (P = 0.01). The vitamin A status of Nicaraguan schoolchildren improved during the year after the initial distribution of vitamin A-fortified sugar in Nicaragua.

The analysis of the zero-order and the second derivative spectra of retinol acetate, tocopherol acetate and coenzyme Q 10 and estimation of their analytical usefulness for their simultaneous determination in synthetic mixtures and pharmaceuticals

The aim of the present work was to develop a simple and rapid method of retinol acetate, tocopherol acetate and coenzyme Q 10 determination in pharmaceuticals without involving any preparation operations like separation or masking. The values of second derivative amplitude at 212 nm for tocopherol, 351 nm for retinol and 222 nm for coenzyme were used for construction of calibration graphs. Beer’s law is obeyed in the concentration range 0.5–20, 0.5–7.5 and 0.5–30 μg ml −1 for retinol acetate, tocopherol acetate and coenzyme, respectively. The elaborated procedures were successfully applied to the simultaneous determination of studied compounds in their binary synthetic mixtures and in commercial preparations with high reliability and repeatability. Spectral properties of retinol acetate allows to determine its contents in ternary mixture which includes Vitamin E and coenzyme Q 10.

An optimum level of vitamin A supplements for Atlantic halibut ( Hippoglossus hippoglossus L.) juveniles

Six diets with different levels of retinyl acetate were fed in triplicate to Atlantic halibut larvae for 13 weeks. The dietary levels used were 0, 0.25, 0.75 (National Research Council (NRC) recommendation), 2.5, 25 and 250 mg retinol equivalents (RE) kg −1 diet (in dry weight). Any sign of deficiency or toxicity was recorded and analysed. No differences in mortality or weight gain were observed among the groups, although the groups fed 0 or 0.25 mg RE kg −1 diet had external haemorrhages and a reduced growth in length. Levels of retinol (ROH), didehydro retinol (dd-ROH), retinyl esters (ROHes) and didehydro retinyl esters (dd-ROHes) were analysed using HPLC equipped with a diode array detector. Between the groups fed 0–0.75 and 2.5–250 mg RE kg −1 diet, there was an overall change in retinoid levels from being indifferent to increasing exponentially. Groups fed 2.5 mg RE kg −1 diet had the highest specific activity in two brush border membrane (BBM) enzymes, indicating that the fish fed this diet had a highly developed or differentiated intestinal mucosa. Furthermore, DNA levels were lower in the groups fed 0.75–25 mg RE kg −1 diet (dry weight), while the other groups of fish showed higher proliferation of cells. This proliferation was confirmed by immunohistochemistry using proliferating cell nuclear antigen (PCNA). The results indicated that the optimum level of retinoids in the diets for Atlantic halibut juveniles was closer to 2.5 mg than the recommended 0.75 mg RE kg −1 diet. Our recommendation is to use 2.5 mg RE kg −1 diet but not more because retinoids are shown to be toxic in excess.

Liquid chromatographic method for the simultaneous determination of different lipid-soluble antioxidants in human plasma and low-density lipoproteins

We describe a reverse phase HPLC method, employing a simple methanol:water gradient as mobile phase, for the determination of several lipophilic antioxidants, such as retinol, γ-tocopherol, α-tocopherol, lycopene, α-carotene and β-carotene among others, using UV detection. Additionally, this method allows the simultaneous separation of probucol, an hypocholesterolemic drug with antioxidant properties. Retinol acetate and α-tocopherol acetate were added to samples as internal standards. A NovaPack ODS C18, 150×3.9 mm, 0.4 μm column was used and the flow rate was set constant at 1 m/min, which allowed the separation of all the desired antioxidants in a total run time of 35 min. A photodiode array detector was used because of its advantages to study the purity of the peaks, however, any programmable multiwavelength UV/VIS detector could be employed given the good resolution of the peaks. The analytical recoveries of the studied compounds were >96% and the detection limits were: retinol 0.050 μg/ml, γ-tocopherol 0.137 μg/ml, α-tocopherol 0.906 μg/ml, lycopene 0.022 μg/ml, α-carotene 0.008 μg/ml, β-carotene 0.015 μg/ml and probucol 1.503 μg/ml. The intra- and inter-assay coefficients of variation were calculated by using two human plasma samples with different levels of lipophilic antioxidants. The simplicity, rapidity and economy, make this method suitable for the routine measurement of plasma and low-density lipoproteins antioxidants, and may also be used in large scale epidemiological studies. The method has been used to measure antioxidants in samples from patients undergoing treatment with probucol, showing there is a good correlation between the probucol content in LDL and that in total plasma.

Transport across liquid membranes containing vitamin A (retinol acetate)

The role of the surface activity of vitamin A has been studied in the light of the liquid membrane hypothesis of drug action. Transport of relevant amino acids such as serine, threonine, arginine, and histidine and various ions such as calcium, sodium, and potassium in the presence of liquid membranes generated by vitamin A has been studied. The data on the modifications in the permeability of relevant amino acids and ions indicate that the liquid membranes generated by vitamin A may also play a significant role in its physiological action.

Correlation of serum retinol and its relation with lipid prolile in Indian cancer patients

The present study was undertaken to investigate the relation of retinol with lipid profile of patients with cancers of breast, stomach, oesophagus, colon, gallbladder, pancreas, lung and cervix. Serum retinol was assayed in 120 patients and 40 healthy normal control by reverse phase HPLC using CLC-ODS C-18 columns and retinyl acetate as an internal standard.Significant decease in serum cholesterol and LDL was observed in patients with cancers of esophagus, colon, stomach, pancreas and gallbladder respectively.Retinol was reduced in all forms of cancers but pronounced decrease was observed in oesophagus, gallbladder, pancreas, stomach and colon. Serum Retinol in all patients was directly correlated with total cholesterol and LDL. These findings suggest that therapeutic modalities of this vitamin could be planned for these patients, as vitamin A is known to act as an antioxidant for prevention of certain cancers.

Beta-Carotene-vitamin A equivalence in Chinese adults assessed by an isotope dilution technique.

The present study was carried out to determine the conversion factor of synthetic (2)H-labelled beta-carotene to vitamin A in Chinese adults by using a stable-isotope dilution technique. Fifteen healthy volunteers aged 50-60 years were recruited for a 55 d experiment. The volunteers (nine males and six females) were each given a physiological dose of [(2)H8]beta-carotene (6 mg) in oil on the first day of the experiment, and a reference dose of [(2)H8]retinyl acetate (3 mg) in oil was given on the fourth day. Serum samples were collected at 0, 3, 5, 7, 9, 11, and 13 h on the first and the fourth days of the study, daily for 10 d, and then weekly from days 14 to 56. beta-Carotene and retinol were extracted from serum and isolated by HPLC, and their enrichments were respectively determined by using GC-electron capture negative chemical ionisation-MS and LC-atmospheric pressure chemical ionisation interface-MS. Four of the subjects exhibited beta-carotene to vitamin A conversion factors of >29.0:1 on a molar basis and were termed 'poor converters'. In the eleven normal converters (seven males and four females), the calculated conversion factors of beta-carotene to retinol ranged from 2.0:1 to 12.2:1 with an average of 4.8 (sd 2.8):1 on a molar basis, and from 3.8:1 to 22.8:1 with an average of 9.1 (sd 5.3):1 on a weight basis. The 52 d post-intestinal absorption conversion was estimated to be about 30 % of the total converted retinol.

Expression of the Peroxisome Proliferator-Activated Receptors (PPARs) in the Hepatic Stellate Cells

Hepatic stellate cells (HSCs) have an important role in maintaining vitamin A (retinoid) homeostasis [1,2]. Peroxisome proliferator-activated receptors (PPARs) are members of the steroid/retinoid nuclear hormone receptor superfamily of ligand-activated transcription factors, form a heterodimer with retinoid X receptor (RXR), and play an important role in lipid metabolisms. Several reports [3-5] have suggested that expression of PPAR-gamma was reduced with the acquisition of activated phenotype (termed as activation) such as lack of cytoplasmic lipid droplets containing vitamin A in subcultured HSC. Previously, we showed that the subcultured HSCs restored the cytoplasmic lipid droplets emanating vitamin A autofluorescence by the addition of retinyl acetate to culture medium [6]. However, it remains unclear whether or not PPAR-gamma expression is involved in formation of vitamin A-containing lipid droplets and maintaining of vitamin A homeostasis in HSCs. In this study, we examined roles of PPARs, particularly PPAR-gamma, in accumulation of vitamin A-containing lipid droplets in cultivated HSCs.

Serum Vitamin A Esters Are High in Captive Rhesus (Macaca mulatta) and Marmoset (Callithrix jacchus) Monkeys

We showed previously that hepatic vitamin A concentrations of captive rhesus monkeys (Macaca mulatta) are subtoxic to toxic, with livers exhibiting stellate cell hypertrophy and hyperplasia. Although marmoset (Callithrix jacchus) livers are also high in vitamin A, no stellate cell irregularities were observed. To further characterize the effects of high dietary vitamin A from preformed sources, stored serum samples were analyzed from monkeys used for biomedical research and housed at the Wisconsin Primate Research Center. The monkeys had been fed commercially available monkey diets, providing vitamin A (as retinyl acetate) at levels exceeding NRC recommendations by a factor of four. The serum from both rhesus and marmoset monkeys had total serum vitamin A (retinol, retinyl esters and metabolites) within the expected range for both species, i.e., 1.44 +/- 0.34 and 1.41 +/- 0.72 micromol/L serum for rhesus and marmoset monkeys, respectively. However, high serum retinyl ester concentrations as a percentage of total serum vitamin A were present in both species, 12 +/- 5.1% (range, 5.5-23%) for rhesus and 27 +/- 14% (range, 10-57%) for the marmosets. Serum retinol concentrations were normal, i.e., 1.21 +/- 0.28 (rhesus) and 0.92 +/- 0.43 micromol/L (marmoset), compared with published values.

Photoreaction, Phototoxicity, and Photocarcinogenicity of Retinoids#

Sunlight is a human carcinogen. Many retinoid-containing cosmetics are used to protect damages caused by sunlight irradiation. Since retinol is thermally unstable and retinyl palmitate (RP) is relatively more stable, RP is also widely used as an ingredient in cosmetic formulations. In general, little is known about the photodecomposition of retinoids and the toxicity of retinoids and their photodecomposition products on the skin's responses to sunlight. This review focuses on the update information on photoreactions, phototoxicity, and photocarcinogenicity of the natural retinoids including retinol, retinal, retinoid acid (RA), retinyl acetate, and RP. #This article is not an official guidance or policy statement of US Food and Drug Administration (FDA) or National Toxicology Program (NTP). No official support or endorsement by the US FDA and NTP is intended or should be inferred.

Supplementation with Micronutrients in Addition to Iron and Folic Acid Does Not Further Improve the Hematologic Status of Pregnant Women in Rural Nepal

Iron deficiency is one of the main causes of anemia during pregnancy, although other micronutrient deficiencies may play a role. We examined the effects of daily antenatal and postnatal supplementation with four combinations of micronutrients on maternal hematologic indicators in a double-masked randomized controlled community trial. Communities, called sectors, were randomly assigned to supplementation with folic acid (400 microg), folic acid plus iron (60 mg), folic acid plus iron and zinc (30 mg) and folic acid plus iron, zinc and 11 other micronutrients, each at the approximate recommended daily allowance for pregnancy all given with vitamin A as retinol acetate (1000 microg retinol equivalent), or vitamin A alone as the control group. Hemoglobin (Hb) and indicators of iron status were assessed at baseline and at 32 wk of gestation. At 6-wk postpartum, Hb assessment was repeated using a finger stick. Severely anemic women (Hb < 70 g/L) were treated according to WHO recommendations. Folic acid alone had no effect on maternal anemia or iron status. Hb concentrations were 14 g/L, [95% confidence limits (CL), 8.3-19.2], 10.0 g/L (CL, 5.2-14.8) and 9.4 g/L (CL, 4.7-14.1) higher in the groups receiving folic acid plus iron, folic acid plus iron and zinc and folic acid plus iron, zinc and multiple micronutrients, respectively, relative to the control. Anemia in the third trimester was reduced by 54% with folic acid plus iron, by 48% with folic acid plus iron and zinc and by 36% with folic acid plus iron, zinc and multiple micronutrients supplementation, relative to the control (P < 0.05). Thus, the combinations of folic acid plus iron and zinc and folic acid plus iron, zinc and multiple micronutrients provided no additional benefit in improving maternal hematologic status during pregnancy compared with folic acid plus iron. The level of compliance and baseline Hb concentrations modified the effect of iron.

Relief effect of vitamin A on the decreased motility of sperm and the increased incidence of malformed sperm in mice exposed neonatally to bisphenol A.

Administration of 50 microg of bisphenol A (BPA) for the first 5 days after birth resulted in a decrease in the percentage of moving sperm, and an increase in the incidence of malformed sperm, in the epididymides of mice at 10 weeks of age, although no marked changes were found in the testicular histology between BPA-treated and vehicle-treated control mice. The deteriorating effects of 50 microg of BPA were ameliorated by the concurrent administration of 100 IU of retinol acetate (RA). Neonatal treatment with 0.5 microg of BPA for 5 days resulted in an increase in the incidence of malformed sperm, whereas the BPA effect became more severe in mice nursed by mothers fed a vitamin A-deficient (VAD) diet only a few days before and after parturition. On the other hand, neonatal treatment with 20 microg of estrogen for the first 5 days after birth resulted in an increase in the number of estrogen receptor alpha (ERalpha)-positive cells in the epithelium of the vas deferens, whereas only a few epithelial cells showed weak ERalpha-positive signals in the vehicle-treated control mice at 18 days after birth. This increase, however, was suppressed by the concurrent administration of RA. Although five daily treatments with 50 microg BPA led to no significant increase in the number of ERalpha-positive cells, it may have been due to the weak estrogenic activity of BPA, as discussed. These findings clearly showed that in mice, neonatal exposure to a relatively large dose of BPA causes damage to the motility and morphology of sperm, but the BPA effect is, to some extent, inhibited by a supplement of VA, and enhanced under a VAD condition.

Antioxidants are necessary for myelination of dorsal root ganglion neurons, in vitro.

We have demonstrated that myelination of dorsal root ganglion (DRG) axons occurs in a fully defined, serum-free medium (B27). This implies that there may be components in B27 medium that support myelination. To determine which of the components in B27 were essential for myelination, we systematically removed components from B27 until myelination was lost. We added these components to a fully defined minimal medium (N2) that supports neuron survival but not myelination. When antioxidants were removed from B27, myelination was lost. However, the individual antioxidants did not induce myelination when added to N2 medium. Addition of ascorbic acid along with the B27 antioxidants was sufficient to induce myelination in N2 medium, which was enhanced by retinyl acetate. Removal of vitamin E from B27 caused a partial loss of myelination, and addition of vitamin E to N2 medium containing ascorbic acid induced partial myelination. Addition of serum to the B27 myelinating medium inhibited myelination completely. These results indicate that antioxidants are important for myelination, in vitro. Vitamin E may play an important role. Use of a serum-free medium may be beneficial for in vitro myelination studies because serum has unknown inhibitory effects.

Rapid determination by reversed-phase high-performance liquid chromatography of Vitamins A and E in infant formulas

A rapid, sensitive method has been developed for the simultaneous determination of retinol acetate, δ-, γ-, α-tocopherol and α-tocopherol acetate. We compare two experimental procedures for simultaneous direct solvent extraction of these vitamins without previous saponification. Method I: the fat milk sample was extracted with ethanol-hexane and injected directly into the chromatographic column. Method II: the power milk sample was extracted with ethanol-hexane and also injected directly into the column. Under optimum conditions the limits of detection for retinol acetate, δ-, γ-, α-tocopherol and α-tocopherol acetate were 0.33, 21.2, 32.9, 32.5 and 3.2 ng and the limits of quantification were 0.42, 25.3, 37.9, 36.8 and 6.3 ng, respectively. The precision results showed that the relative standard deviations of repeatability and reproducibility were between 0.74 and 5.7%.

Short-term (intestinal) and long-term (postintestinal) conversion of β-carotene to retinol in adults as assessed by a stable-isotope reference method

Quantitative information on the conversion of beta-carotene to vitamin A in humans is limited. We determined the short- and long-term conversion of labeled beta-carotene to vitamin A by using a stable-isotope reference method. [(2)H(8)]beta-Carotene (11,011 nmol, or 6 mg) in oil was given with a liquid diet (25% of energy from fat) to 22 adult volunteers (10 men, 12 women). Three days after the [(2)H(8)]beta-carotene dose, the volunteers each took a dose of [(2)H(8)]retinyl acetate (8915 nmol, or 3 mg) in oil with the same liquid diet. Blood samples were collected over 56 d. The 53-d area under the serum [(2)H(4)]retinol response curve (from the [(2)H(8)]beta-carotene dose) was 569 +/- 385 nmol. d, and the 53-d area under the serum [(2)H(8)]retinol response curve (from the [(2)H(8)]retinyl acetate dose) was 1798 +/- 1139 nmol. d. With the use of [(2)H(8)]retinyl acetate as the vitamin A reference, the [(2)H(4)]retinol formed from [(2)H(8)]beta-carotene (11,011 nmol) was calculated to be equivalent to 3413.9 +/- 2298.4 nmol retinol. The conversion factor of beta-carotene to retinol varied from 2.4 to 20.2, and the average conversion factor was 9.1 to 1 by wt or 4.8 to 1 by mol. This conversion factor was positively correlated with body mass index (r = 0.57, P = 0.006). The postabsorption conversion of beta-carotene was estimated as 7.8%, 13.6%, 16.4%, and 19.0% of the total converted retinol at 6, 14, 21, or 53 d after the [(2)H(8)]beta-carotene dose, respectively. The quantitative determination of the conversion of beta-carotene to vitamin A in humans can be accomplished by using a stable-isotope reference method. This approach provides in vivo metabolic information after a physiologic dose of beta-carotene.

Supplementing Hen Diets with Vitamins A and E Affects Egg Yolk Retinol and α-Tocopherol Levels

Laying hens were fed a basal diet supplemented with increasing levels of retinyl acetate and α-tocopheryl acetate to investigate the effects of vitamin A and E supplementation upon egg yolk retinol and tocopherol concentrations. The high concentration of added vitamin E caused a decline in egg production and poor feed conversion. Egg quality was not affected by vitamin A and E levels. Yolk retinol concentration was enhanced by added vitamin A, from 24.6 IU/g for eggs from the control group, to 33.6 and 37.7 IU/g of yolk when hens were fed 15,000 and 30,000 IU/kg of diet. Yolk α-tocopherol was significantly increased by dietary tocopherol supplementation, ranging from 10.9 μg/g (control group) to 160.6, 264.1, and 383.2 μg/g of yolk, respectively, when 200, 400 and 600 mg/kg of ration were added, respectively. Yolk α-tocopherol was increased by 24.9 and 44.0% with increasing vitamin A supplementation at 15,000 and 30,000 IU/kg of diet, respectively. When correlation coefficients and regression equations were calculated, it was found that yolk α-tocopherol decreased (P < 0.05) as supplemental vitamin A increased, indicating the adverse effect of dietary vitamin A on yolk tocopherol deposition. The nutritional value of eggs, related to retinol and tocopherol, can be improved by dietary manipulation of hens diet, but attention must be focused on their inter-relationship.

Effect of carbonyls on the reactivity of polyenes in autoxidation

AbstractAutoxidation of the biologically active polyenes β‐carotene, canthaxanthin, retinyl acetate, methyl retinoate, retinal and 3,7,11,11‐tetramethyl‐10,15‐dioxo‐2,4,6,8‐hexadecatetraenal (diketoretinal) in solid amorphous films was investigated. The course of the process was followed by electronic and IR spectroscopy. The overall activation energies were obtained. It was shown that insertion in the polyene molecule of a carbonyl group conjugated with polyene chain results in a drastic decrease in the reactivity towards molecular oxygen. The results are discussed and explained on the basis of radical stabilization energy, ES(R·), as a driving force of the autoxidation process. Semi‐empirical AM1 calculations of ΔHf of some retinyl polyenes and polyenyl radicals were carried out and the C—H bond strengths and ES(R·) values of corresponding polyenyl radicals were evaluated. It was shown that the incorporation of a carbonyl group in the polyene results in a decrease in the overall autoxidation rate due to the decrease in both the initiation and propagation rates. Copyright © 2003 John Wiley &amp; Sons, Ltd.