Serum α- and γ-tocopherols, retinol, retinyl palmitate, and carotenoid concentrations in captive and free-ranging bottlenose dolphins (Tursiops truncatus)

Concentrations of retinol, retinyl palmitate, β-carotene, α-carotene, cryptoxanthin, lutein, lycopene, α-tocopherol, and γ-tocopherol were measured in blood samples collected from 15 captive and 55 free-ranging bottlenose dolphins (Tursiops truncatus). From June 1991 to June 1994, blood samples were collected from captive animals residing at two locations; at Seven Seas (Brookfield Zoo, Brookfield, IL) and Hawk’s Cay (Marathon Key, FL). Blood samples were collected from free-ranging animals from June 1991 to June 1996. Retinol levels were not significantly different between captive dolphin groups. However, Seven Seas animals had higher (P<0.01) serum retinol concentrations compared to free-ranging animals (0.061 vs 0.041 μg/ml). Retinyl palmitate was not detected in the serum of captive or free-ranging dolphins. Alpha-tocopherol levels were significantly (P<0.05) higher for Seven Seas dolphins (16.4 μg/ml) than for Hawk’s Cay (13.0 μg/ml) and free-ranging dolphins (12.5 μg/ml). Gamma-tocopherol concentrations were similar among captive and free-ranging dolphins. Free-ranging dolphins showed levels of circulating carotenoids (lutein and β-carotene) while the captive animals did not. Additional carotenoids (lycopene, α-carotene and cryptoxanthin) were analyzed but not detected in any samples. Serum vitamin differences between captive and free-ranging dolphins may reflect the natural diet or indicate some potential biological or nutritional status significance.     

Annual variations in plasma retinol and alpha-tocopherol levels in gentoo and rockhopper penguins

Blood samples were taken throughout the year from captive gentoo Pygoscelis papua (n = 5) and rockhopper Eudyptes crestatus (n = 10) penguins to measure seasonal variations in retinol and alpha-tocopherol levels. Retinol levels ranged from .58 μg/ml to 1.09 μg/ml and alpha-tocopherol levels from 30.8 μg/ml to 50.7 μg/ml for the gentoo penguins. The rockhopper penguins' retinol levels ranged from .63 μg/ml to 1.14 μg/ml and alpha-tocopherol from 22.3 μg/ml to 40.8 μg/ml. Changes in body mass were used as an indicator of the start and duration of feather moult. Food consumption was recorded daily for the year, and the vitamin A and E intake was calculated. Blood vitamin levels of penguins on a supplemented diet were similar to those in the wild. © 1993 Wiley-Liss, Inc.     

Determination of vitamins D,A, and E in sera and vitamin D in milk from captive and free-ranging polar bears (Ursus maritimus), and 7-dehydrocholesterol levels in skin from captive polar bears

The difference between serum levels from 36 captive and 56 free-ranging polar bears (Ursus maritimus) for 25-hydroxyvitamin D (25-OH-D) was found not to be significant (mean ± SD = 348 ± 215 nmol/L [captive], 360 ± 135 nmol/L [free-ranging], t = 0.30, df = 52.8, P = 0.76), whereas the difference for retinol and α-tocopherol was significant (retinol, 1.37 ± 0.67 μmol/L [captive] 1.89 ± 0.63 μmol/L [free-ranging], t = 3.88, df = 72.4, P <0.001, α-tocopherol, 18.56 ± 18.56 μmol/L [captive], 48.76 ± 13.92 μmol/L [free-ranging], t = 7.85, df = 61.9, P < 0.001). Due to the high fat content in the polar bear diet, seal blubber may be the source of these fat-soluble vitamins. Six skin biopsies were analyzed from captive polar bears at the Denver Zoological Gardens for 7-dehydrocholesterol levels and found to contain 0.11 ± 0.03 nmol/cm². This finding also helps to support the contention that the source of vitamin D for polar bears may be ingestion and not cutaneous production. Vitamin D content in the milk from one captive sow in the den (0.14 nmol/g) and 10 free-ranging sows with cubs of the year out on the ice pack (0.0042 ± 0.0073 nmol/g) were also evaluated. It would be helpful to evaluate additional milk samples from denning and non-denning sows with cubs to see whether vitamin D content varies according to the stage of lactation. Zoo Biol 17:285–293, 1998. © 1998 Wiley- Liss, Inc.     

EFFECT OF VITAMIN E SUPPLEMENTATION AND PARTIAL SUBSTITUTION OF POLY- WITH MONO-UNSATURATED FATTY ACIDS IN PIG DIETS ON MUSCLE,AND MICROSOME EXTRACT a-TOCOPHEROL CONCENTRATION AND LIPID OXIDATION

The experiment was organized in a 3×2 factorial arrangement with three dietary fat blends and a basal (20 mg kg^?1 diet) or supplemented (220 mg kg^?1) level of α-tocopheryl acetate. Dietary vitamin E and monounsaturated to polyunsaturated fatty acid ratio (dietary MUFA/PUFA) affected muscle α-tocopherol concentration (α-tocopherol [log μg g^?1]=0.18 (±0.105)+0.0034 (±0.0003)·dietary α-tocopherol [mg kg^?1 diet] (P<0.0001)+0.39 (±0.122)·dietary MUFA/PUFA (P<0.0036)). An interaction between dietary α-tocopherol and dietary MUFA/PUFA exists for microsome α-tocopherol concentration (α-tocopherol [log μg g^?1]=1.14 (±0.169) (P<0.0001)+0.0056 (±0.00099)·dietary α-tocopherol [mg kg^?1 diet] (P<0.0001)+0.54 (±0.206)·dietary MUFA/PUFA (P<0.0131)?0.0033 (±0.0011)·dietary α-tocopherol [mg kg^?1)]×dietary MUFA/PUFA (P<0.0067)), and hexanal concentration in meat (hexanal [ng·g^?1]=14807.9 (±1489.8)?28.8 (±10.6) dietary α-tocopherol [mg·kg^?1] (P<0.01)?8436.6 (±1701.6)·dietary MUFA/PUFA (P<0.001)+24.0 (±11.22)·dietary α-tocopherol·dietary MUFA/PUFA (P<0.0416)). It is concluded that partial substitution of dietary PUFA with MUFA lead to an increase in the concentration of α-tocopherol in muscle and microsome extracts. An interaction between dietary α-tocopherol and fatty acids exists, in which at low level of dietary vitamin E inclusion, a low MUFA/PUFA ratio leads to a reduction in the concentration of α-tocopherol in microsome extracts and a concentration of hexanal in meat above the expected values.     

Vitamin E status and metabolism in adult and aged aryl hydrocarbon receptor null mice

The aryl hydrocarbon receptor (AhR) is involved in regulation of mechanisms for detoxification of xenobiotics, as well as vitamin A metabolism. Vitamin E is a fat-soluble nutrient whose metabolism is initialized via the cytochrome P450 system. Thus, AhR absence could alter hepatic regulation of α-tocopherol metabolism. To test this hypothesis, we assessed vitamin E status in adult (2-5 m) and old (21-22 m), wild-type and AhR-null mice. Plasma α-tocopherol concentrations in AhR-null mice (2.3±1.2 μmol/L, n=19) were lower than those of wild-type mice (3.2±1.2, n=17, P=.0131); those in old mice (3.2±1.2, n=20) were higher than those of adults (2.2±1.0, n=16, P=.0075). Hepatic α-tocopherol concentrations were not different between genotypes, but were nearly double in old (32±8 nmol/g, n=20) as compared with adult mice (17±2, n=16, P<.0001). Hepatic Cyp3a concentrations in AhR-null mice were greater than those in wild-type mice (P=.0011). Genotype (P=.0047), sex (P<.0001) and age (P<.0001) were significant modifiers of liver α-tocopherol metabolite (α-CEHC) concentrations. In general, Cyp3a concentrations correlated with hepatic α-tocopherol (r=0.3957, P<.05) and α-CEHC (r=0.4260, P<.05) concentrations. Since there were no significant genotype differences in the hepatic α- or γ-tocopherol concentrations, AhR-null mice did not have dramatically altered vitamin E metabolism. Since they did have higher hepatic α-CEHC concentrations, these data suggest metabolism was up-regulated in the AhR-null mice in order to maintain the hepatic tocopherol concentrations similar to those of wild-type mice.     

Oral a-tocopherol Supplementation Inhibits Lipid Oxidation in Established Human Atherosclerotic Lesions

Background: Much experimental evidence suggests that lipid oxidation is important in atherogenesis and in epidemiological studies dietary antioxidants appear protective against cardiovascular events. However, most large clinical trials failed to demonstrate benefit of oral antioxidant vitamin supplementation in high-risk subjects. This paradox questions whether ingestion of antioxidant vitamins significantly affects lipid oxidation within established atherosclerotic lesions. Methods and results: This placebo-controlled, double blind study of 104 carotid endarterectomy patients determined the effects of short-term α-tocopherol supplementation (500 IU/day) on lipid oxidation in plasma and advanced atherosclerotic lesions. In the 53 patients who received α-tocopherol there was a significant increase in plasma α-tocopherol concentrations (from 32.66±13.11 at baseline to 38.31±13.87 (mean±SD) μmol/l, p&lt;0.01), a 40% increase (compared with placebo patients) in circulating LDL-associated α-tocopherol (p&lt;0.0001), and their LDL was less susceptible to ex vivo oxidation than that of the placebo group (lag phase 115.3±28.2 and 104.4±15.7 min respectively, p&lt;0.02). Although the mean cholesterol-standardised α-tocopherol concentration within lesions did not increase, α-tocopherol concentrations in lesions correlated significantly with those in plasma, suggesting that plasma α-tocopherol levels can influence lesion levels. There was a significant inverse correlation in lesions between cholesterol-standardised levels of α-tocopherol and 7β-hydroxycholesterol, a free radical oxidation product of cholesterol. Conclusions: These results suggest that within plasma and lesions α-tocopherol can act as an antioxidant. They may also explain why studies using &lt;500 IU α-tocopherol/day failed to demonstrate benefit of antioxidant therapy. Better understanding of the pharmacodynamics of oral antioxidants is required to guide future clinical trials.     

Stability of individual carotenoids, retinol and tocopherols in human plasma during exposure to light and after extraction

We have modified gradient HPLC procedures for simultaneous quantification of retinol, γ-tocopherol, α-tocopherol, lutein/zeaxanthin, β-cryptoxanthin, trans-lycopene, cis-lycopene, α-carotene and β-carotene in 200-μl aliquots of human plasma. The photosensitivity of these analytes in plasma exposed to fluorescent lighting for up to 72 h was investigated and most were stable under these conditions. The stability of these analytes held in darkness at −20°C, 4°C or room temperature for up to 48 h after extraction from plasma was also investigated. Variability in measurement of most analytes was greater at room temperature than at 4°C or −20°C. There were statistically significant variations in the measured concentrations of some analytes in samples kept cold. However, the magnitude of these variations was small and of little biological significance, particularly over the first 24 h.     

Carotenoids in fish. XXVI. Pungitius pungitius (L.) and Gasterosteus aculeatus L. (Gasterosteidae)

The author investigated the presence of various carotenoids in the different parts of the body of Pungitius pungitius (L.) and Gasterosteus aculeatus L. by means of columnar and thin-layer chromatography. The investigations revealed the presence of the following carotenoids:

in Pungitius pungitius. α-carotene, β-carotene, β-cryptoxanthin, mutatochrome, zeaxanthin and astaxanthin;

in Gasterosteus aculeatus: β-carotene, β-cryptoxanthin, β-carotene epoxide, neothxanthin, canthaxanthin, mutatochrome, lutein, phoenicoxanthin, zeaxanthin, taraxanthin, tunaxanthin, astaxanthin, astaxanthin ester and α-doradexanthin. The total carotenoid content ranged from 2.229 to 138.504 µg/g wet weight.

     

Antioxidant activity of palm oil carotenes in peroxyl radical-mediatedperoxidation of phosphatidyl choline liposomes

Abstract

The antioxidant efficacy of α-carotene and comparison with β-carotene in multilamellar liposomes prepared from egg yolk phosphatidyl choline (EYPC) exposed to the lipid soluble 2,2′-azobis (2,4-dimethyl valeronitrile) (AMVN) was investigated. Lipid peroxidation was measured as thiobarbituric acid reacting substances (TBARS)at 532 nm or as hydroperoxide formation at 234 nm after separation of phosphatidyl choline hydroperoxide (PCOOH) by high-pressure liquid chromatography (HPLC). Lutein and zeaxanthin, the hydroxyl derivatives of α- and β-carotenes, and the chain breaking antioxidant α-tocopherol were also included in the study.AMVN being a lipid soluble, non polar azo initiator penetrates into the hydrophobic interior of the phospholipid bilayer, forming peroxyl radicals which peroxidate the phospholipid leading to PCOOH accumulation. All the carotenoids tested at 1 mol% relative to EYPC significantly suppressed the formation of PCOOH compared to control samples.In this system, α-carotene retarded PCOOH formation better than β-carotene. Similarly, lutein was a better antioxidant than is zeaxanthin. But lutein and zeaxanthin were more effective antioxidants than α- and β-carotenes, respectively. After 1 h of incubation of the carotenoid with AMVN, α-, β-carotene, lutein and zeaxanthin limited PCOOH formation by 77%, 68%, 85%and 82%, respectively, while α-tocopherol elicited 90%reduction.AMVN incubated with EYPC for 2 h induced the formation of TBARS compared to control (P <0.001). α-Carotene significantly suppressed the TBARS formation by 78% whilst β-carotene, lutein, zeaxanthin and α-tocopherol elicited 60%, 91%and 80% reductions, respectively. Increasing the concentration of the carotenoid >1 mol% to EYPC did not significantly increase protection of the membrane against free radical attack.Our findings suggest that α-carotene is a better antioxidant than is β-carotene in phosphatidyl choline vesicles. It may, therefore, be useful in limiting free radical mediated peroxidative damage against membrane phospholipids _{in vivo}.     

Response to different sources of vitamin E orally injected and to various doses of vitamin E in calf starter on the plasma vitamin E level in calves around weaning

Calves often face a lower plasma vitamin E level than the recommended level (3 µg/ml for adult cows) after weaning, a level which has been related to a good immune response. Two experiments were performed to determine the most effective source and level of vitamin E to be included in a calf starter to maintain the plasma vitamin E level above the recommended level after weaning. Experiment 1 (Exp 1) and experiment 2 (Exp 2) included a total of 32 and 40 calves, respectively, from 2 weeks before weaning until 2 weeks after weaning. In Exp 1, calves were orally injected a daily dose of different vitamin E sources including, no α-tocopherol (0 dose; Control), 200 mg/d of RRR-α-tocopherol (ALC), 200 mg/d of RRR-α-tocopheryl acetate (ACT), or 200 mg/d of all-rac-α-tocopheryl acetate (SYN). In Exp 2, a dose response study was carried out with 0, 60, 120, and 200 mg/kg of ALC in a pelleted calf starter. Final BW (100 ± 16 and 86 ± 11 kg) and average daily gain (956 ± 303 and 839 ± 176 g/d in Exp 1 and 2, respectively; mean ± SD) were unaffected by either source or level of α-tocopherol. In Exp 1, the plasma RRR-α-tocopherol level was affected by α-tocopherol source (P < 0.001), week (P < 0.001), and interaction between them (P < 0.001). At weaning time, the plasma RRR-α-tocopherol was 2.7, 2.1, 1.1, and 0.8 μg/ml in ALC, ACT, SYN, and Control, respectively. In Exp 2, the plasma α-tocopherol level was affected by ALC dose (P = 0.04), week (P < 0.001), and a tendency for an interaction between them was observed (P = 0.06). At weaning, a 36, 31, and 28% reduction in plasma α-tocopherol level was observed compared to the beginning of the experiment with 0, 60, and 120 mg/kg of ALC, respectively; however, with 200 mg/kg of ALC, a 9% increase in the plasma α-tocopherol level was observed. In addition, 200 mg/kg of ALC was able to maintain plasma α-tocopherol after weaning higher than the recommended level. The results showed that the ALC was the most efficient source of α-tocopherol supplementation to be used in a calf starter. In addition, the 200 mg/kg of ALC in the calf starter was the only effective dose to maintain the postweaning plasma vitamin E concentration at the recommended level after weaning and α-tocopherol similar to that observed before weaning.     

Serum Retinol,alpha‐tocopherol,and lipids in four species of adult captive pinnipeds

The sera of adult aquarium‐held pinnipeds from four species (family Phocidae: harbor seals (Phoca vitulina) and gray seals (Halichoerus grypus); family Otariidae: northern fur seals (Callorhinus ursinus) and California sea lions (Zalophus californianus)) were analyzed for vitamin A (retinol), vitamin E (α‐tocopherol), total cholesterol, triglycerides, phospholipids, and fatty acids. Each subject animal was healthy at the time of blood collection, was fasted for at least 12 hr prior to sampling, and was maintained on a constant diet and supplement regime throughout the study. Retinol values for the four species ranged from 0.16 to 0.92 μg/mL, with the lowest concentrations seen in the harbor seals and the highest in the northern fur seals. Vitamin E values ranged from 10.55 to 43.58 μg/mL, with northern fur seals showing the highest and gray seals the lowest levels. Vitamin E/lipid ratios (cholesterol, triglyceride, phospholipid, and total lipids) were also examined. A significant correlation was seen between vitamin E and total lipids (P<0.05) and phospholipid (P<0.01). Statistical analysis of the retinol, tocopherol, triglyceride, and phospholipid levels showed significant differences between phocid and otariid seals. Otariids had significantly lower tocopherol and phospholipid values (19.36 μg/mL, 4.29 mg/mL) and the phocids had significantly lower retinol and triglyceride levels (0.29 μg/mL, 124 mg/dL). There was no significant difference in serum cholesterol. Zoo Biol 22:83–96, 2003. © 2003 Wiley‐Liss, Inc.     

Effect of α-Tocopherol, β-Carotene, Monogalactosyldiglyceride and Phosphatidylcholine on Light-Induced Degradation of Chlorophyll a in Acetone

The effect of α-tocopherol, β-carotene, monogalactosyldi-glyceride and phosphatidylcholine on red light induced degradation of chlorophyll a was studied in acetone at 4°C. Monogalaclosyldi-glyceride was ineffective up to a molar ratio of monogalactosyldi glyceride to chlorophyll of 1:10. α-Tocopherol, β-carotene and phosphatidylcholine inhibited chlorophyll degradation. Maximal protection by α tocopherol and β-carotene was similar (76%) but on a molar basis a tocopherol was less effective. Protection by phosphatidylcholine was less than by a tocopherol and α-carotene but the lipid was effective at a lower ratio of chlorophyll to protectant. Inhibition by phosphatidylcholine was independent of the degree of unsaturation of the fatty acids. Effects of β-carotene and α-tocopherol were additive at suboptimal concentrations, but addition did not increase the maximal protection of 76% by these substances alone. Phosphatidylcholine increased the effectiveness of α-tocopherol and β-carotene independent of their concentrations. It is suggested that interactions between lipids participate in the mechanism protecting chlorophyll a against photooxidation in the chloroplast membrane.     

An improved method for the determination of alpha-tocopherol in sheep liver

A method is described for the analysis of α-tocopherol in sheep liver, employing a direct attack on the tissue with alcoholic alkali in the presence of pyrogallol and hydroquinone. The alkali treatment is rapid and avoids drying, grinding, and solvent extraction. Combined with two-dimensional paper chromatography, it eliminates time-consuming freezing and column chromatographic techniques. Mean ± SE recoveries of α-tocopherol alone, or mixed with liver tissue, were 97.1 ± 0.05% and 87.1 ± 4.1%, respectively. The mean ± SE levels of α-tocopherol in the liver of sheep fed dry rations were 1.0 ± 0.1 μg/g, and in that of sheep fed green pasture, 18.5 ± 1.4 μg/g.     

a-Tocopherol Content and a-Tocopherol Transfer Protein Expression in Leukocytes of Children with Acute Leukemia

α-Tocopherol (a form of vitamin E) is a fat-soluble vitamin that can prevent lipid peroxidation of cell membranes. This antioxidant activity of α-tocopherol can help to prevent cardiovascular disease, atherosclerosis and cancer. We investigated the α-tocopherol level and the expression of α-tocopherol transfer protein (α-TTP) in the leukocytes of children with leukemia. The plasma and erythrocyte α-tocopherol levels did not differ between children with leukemia and the control group. However, lymphocytes from children with leukemia had significantly lower α-tocopherol levels than lymphocytes from the controls (58.4±39.0 ng/mg protein versus 188.9±133.6, respectively; p&lt;0.05), despite the higher plasma α-tocopherol/cholesterol ratio in the leukemia group (5.83±1.64 μmol/mmol versus 4.34±0.96, respectively; p&lt;0.05). No significant differences in the plasma and leukocyte levels of isoprostanes (the oxidative metabolites of arachidonic acid) were seen between the leukemia patients and controls. The plasma level of acrolein, a marker of oxidative stress, was also similar in the two groups. Investigation of α-TTP expression by leukocytes using real-time PCR showed no difference between the two groups. These findings suggest that there may be comparable levels of lipid peroxidation in children with untreated leukemia and controls, despite the reduced α-tocopherol level in leukemic leukocytes.     

Elicitors,SA and MJ enhance carotenoids and tocopherol biosynthesis and expression of antioxidant related genes in Moringa oleifera Lam. leaves

Moringa oleifera Lam. leaves are rich source of carotenoids (provitamin A) and α-tocopherol (vitamin E), and there is a scope for their further enhancement, through elicitor mediation, thereby a great potential for addressing these vitamins deficiency. In the present study, we report the efficacy of foliar administration of biotic elicitors, carboxy-methyl chitosan and chitosan, and signaling molecules, methyl jasmonate (MJ) and salicylic acid (SA) for enhancement of major carotenoids and α-tocopherol. Highest α-tocopherol content of 49.7 mg/100 g FW was recorded upon foliar application of 0.1 mM SA after 24 h of treatment, which represented a 187.5 % increase in comparison to the untreated control. Similarly, a maximum of 52.6 mg/100 g FW lutein, and 21.8 mg/100 g FW β-carotene content were observed in leaves after 24 h of treatment with MJ, which represented a 54.0 and 20.3 % increase in comparison to the untreated control, respectively. Among the major genes of carotenoid biosynthetic pathway, the expression of lycopene β-cyclase (LCY-β) was maximum influenced after treatment with elicitors and signaling molecules, compared to phytoene synthase and phytoene desaturase, suggesting the LCY-β-mediated enhancement in the production of β-carotene in elicitor treated M. oleifera leaves. Enhanced production of α-tocopherol under respective elicitor treatment was further supported by 2.0–2.7 fold up-regulation of γ-tocopherol methyl transferase, compared to untreated control. This is the first report on elicitor-mediated enhanced production of tocopherol and carotenoids in foliage of economically important food plant.     

Retinol and α-tocopherol concentrations in whole fish commonly fed in zoos and aquariums

Retinol (n = 17 spp.) and α-tocopherol (n = 9 spp.) concentrations in whole fish utilized for captive animal feeding programs were determined by high-performance liquid chromatography (HPLC) following routine storage and preparation after commercial purchase by two zoological institutions. Vitamin A activity was calculated from retinol values and ranged from 55 IU/100 g (immature herring) to >2,000 IU/100 g (salmon) on an as-fed basis. α-Tocopherol values, a measure of vitamin E activity, ranged from 0.9 IU/100 g (butterfish) to 12.3 IU/100 g (tilapia) on a wet basis. Vitamin levels in whole fish were intermediate to values previously quantified for muscle or liver tissues alone. Vitamin concentrations in fish livers were quantified separately in seven of these species; liver contributed 35–63% of total retinol measured and 8–34% of total α-tocopherol. Based on these analyses, whole fish commonly fed in zoos, aquariums, and marine zoological parks would appear to meet vitamin A requirements established for most species without additional supplementation, whereas levels of vitamin E quantified indicate a need for supplementation of diets for piscivores.     

Nutritional status of free‐ranging Mexican howler monkeys (Alouatta palliata mexicana) in Veracruz,Mexico: Serum chemistry; lipoprotein profile; vitamins D,A, and E; carotenoids; and minerals

The purpose of this work was to measure important nutritional status parameters for a group of free‐ranging Mexican mantled howler monkeys (A. palliata mexicana) and compare those data to published data for primates. The nutritional status of six free‐ranging Mexican mantled howler monkeys was examined using biochemical analysis. Blood samples were analyzed for serum chemistry; lipids; vitamins D, A, and E; carotenoids; and minerals. Serum chemistries were somewhat different from published values, but did not indicate clear abnormalities. Circulating lipids were not different from those in captive primates. Circulating vitamin D metabolites (83±16.3 for 25(OH)D ng/mL; 563±53.8 for 1,25(OH)₂D pg/mL) were similar to those in wild‐caught tamarins (Saguinus oedipus), lower than some published data for captive Cebidae and Callitrichidae, and higher than for Old World primates. Serum concentrations of retinol (16.5±1.64 μg/dl) were similar to those in captive spider monkeys (Ateles geoffroyi). Retinyl palmitate and retinyl stearate was present in howler samples and may have reflected recent dietary intake. Circulating α‐tocopherol (997±97.6 μg/dl) was similar to published values for other primates. Carotenoid levels in howlers were within the ranges reported for many primates. A significant finding was the presence of cadmium in samples that should be further studied. The number of individuals sampled was limited, and further investigation into the effects of seasonality is needed. However, this information provides new data for howler monkeys and for free‐ranging primates in general. Zoo Biol 22:239–251, 2003. © 2003 Wiley‐Liss, Inc.     

Photochemiluminescent detection of antiradical activity; IV: testing of lipid-soluble antioxidants

A new method for quantification of antiradical properties of pure lipid-soluble antioxidants and for measurement of integral antioxidant capacity in the lipid phase (ACL) of polycomponent systems, such as blood plasma or tissue homogenates, is developed. It is based on an antioxidant-sensitive inhibition of a photo-induced, chemiluminescence accompanied autoxidation of luminol. The sensitivity of the photochemiluminescent (PCL) assay lies within nmol quantities of substances, the measuring range for α-tocopherol is between 0.1 and 3 nmol. The interassay variability of the method is lower than 5%, the intraassay variability <2%. The antioxidant efficiency of γ-tocopherol was found to be 43% of α-tocopherol. The results of the PCL measurements on pure antioxidants and on lipid extracts from blood plasma were compared with the level of, ‘vitamin E’ (VE) determined as a sum of α- and γ-tocopherol by HPLC. Very good coincidence of both methods was observed for pure substances (r = 0.998, P<0.001). The ACL of human blood plasma was found to be 27.98 ± 0.68 μmol equivalents of α-tocopherol/l (mean ± mean error, n = 142), it is ∼ 25% more than the concentration of VE found in the same samples (22.09 ± 0.59 μmol/l). In this case, the correlation of both parameters was lower: r = 0.811, P<0.001. The animal experiments showed that synthetic antioxidants may not only increase the value of ACL of blood plasma but in the same time reduce the concentration of biological antioxidants, e.g. VE drastically. The prooxidant activity of synthetic antioxidants in vivo or the replacing of structured α-tocopherol from its position can be the cause. This important circumstance has to be considered during the testing of new antioxidants for clinical application.