Low antioxidant intake and status have been shown to be associated with an elevated risk for various diseases. Data on the status of antioxidant vitamins, selenium and coenzyme Q10 of younger female seniors are scarce. The aim of this study was to assess the status of these antioxidants, as well as influencing factors such as dietary intake, anthropometric data and educational level in female seniors (60–70 years) in Germany.
Dietary intake of alpha-tocopherol, beta-carotene and ascorbic acid was determined by a 3-day diet record. Serum concentrations of alpha-tocopherol, beta-carotene, ascorbic acid, selenium and coenzyme Q10 were measured. Anthropometric measures, socioeconomic and educational status were assessed.
In total, 178 elderly women without severe diseases in the region of Hannover, Germany, were included in the study. The mean (±s.d.) age and BMI of the women was 63.2 (2.73) years and 25.6 (3.77) kg/m2, respectively. The study participants were generally better educated than the overall German female population.
Dietary intake of the ascorbic acid and alpha-tocopherol was below RDA in six and 75% of the women, respectively. In comparison to estimated desirable serum concentrations of alpha-tocopherol, ascorbic acid, beta-carotene and selenium, lower concentrations were found in 23, 1, 6, and 39% of the women, respectively. Ascorbic acid (r=0.205, P=0.009) and beta-carotene (r=0.173, P=0.025) intake were significantly associated with serum concentrations. Beta-carotene concentrations were influenced by the type of diet, BMI, and school education (R2=0.128, P<0.001). Serum selenium was positively associated with alcohol intake (r=0.229, P=0.003). Neither employment nor vocational training was predictive for the serum concentrations of antioxidant vitamins, selenium or coenzyme Q10.
Poor status of selenium and alpha-tocopherol is highly prevalent even among younger, well-educated female seniors, whereas ascorbic acid and beta-carotene status seems sufficient in most women.
The study was supported by grants from the Stoll Vita Foundation, Waldshut-Tiengen, Germany.
Epidemiological data suggest that a higher antioxidant intake and/or higher plasma concentrations of antioxidants may protect against chronic diseases such as coronary artery disease (CAD) (Rimm et al., 1993; Osganian et al., 2003) and cancer (Hansson et al., 1994; Mark et al., 2000). Therefore, a sufficient status of antioxidants (ascorbic acid, alpha-tocopherol, beta-carotene, selenium, and coenzyme Q10) may help to optimize antioxidant defence. This has been confirmed by several prospective observational studies which have shown that the use of vitamin supplements is associated with a lower risk for CVD (Morris and Carson, 2003). However, results of randomized clinical trials do not support a protective role of antioxidants in CAD (Hennekens et al., 1996; Rapola et al., 1997; Duffield-Lillico et al., 2003) or cancer (ATBC, 1994; Omenn et al., 1996; Albanes et al., 2000; Malila et al., 2002). Thus, it is concluded that the antioxidant status is only a marker of a diet rich in fruit and vegetables whereas the underlying mechanisms are likely to be multiple (Bazzano et al., 2003).
Finally, an insufficient dietary intake of antioxidants may be associated with an increased risk of degenerative diseases. The reported dietary intakes of antioxidant vitamins and minerals of women have been shown to be below recommended values which could contribute to subsequent health risks (Lewis et al., 2003). Furthermore, elderly subjects are at higher risk for poor antioxidant status due to several reasons. They exhibit a reduction in energy requirement and thus may have a decreased food intake compared with younger adults. Although energy requirement decreases with age, the need for antioxidants generally stays the same (Food and Nutrition Board, 2000). Moreover, elderly individuals have a higher incidence of chronic diseases and often use therapeutic drugs both of which can impair the appetite and/or affect nutrient utilization (Schumann, 1999; Ekmekcioglu, 2001). Therefore, we hypothesized that even well-educated younger female seniors may exhibit an insufficient antioxidant status. Although a great number of studies on the antioxidant status have been published (Palli et al., 1999; Murphy and Cashman, 2002; Neuhouser et al., 2003), there are only few data available for elderly people. Especially information on the selenium and coenzyme Q10 status of elderly people is scarce. The aim of our study was to evaluate the status of antioxidant vitamins, selenium and coenzyme Q10 and to assess associated factors such as dietary intake, BMI and sociodemographic factors in younger seniors without severe diseases. Correlations between the measured variables have been utilized to show potential links between the associated factors.
Subjects and methods
As described previously (Wolters et al., 2003), 178 women aged 60–70 years were recruited by advertisement in several newspapers in the region of Hannover, Germany. Subjects with severe chronic diseases, cancer or a history of gastrointestinal resection as well as smokers and vitamin and mineral supplement users were excluded. All subjects gave written informed consent. The study was conducted in accordance with the Helsinki Declaration of 1964 as amended in 1983 and 1996.
A 3-day diet record assessed energy and nutrient intake of the women. Since the German Food Code and Nutrient Data Base (BLS II.2) does not contain data on selenium and coenzyme Q10 content of food (BgVV, 1994), assessment of dietary intake from food was not possible for these nutrients. The calculated nutrient intake of the sample was compared with the recommended dietary allowance (RDA) for the particular age and sex group (Food and Nutrition Board, 2000).
Anthropometric data were determined on the same day blood samples were drawn. Questionnaires asking for sociodemographic data were sent to the study participants and returned on the day of the blood sampling (Wolters et al., 2003).
Blood samples were drawn after an overnight fast and centrifuged at 2665 g for 10 min at 19°C. The serum samples for ascorbic acid evaluation were deproteinized at once by the addition of trichloro acetic acid and recentrifuged. Serum aliquots for analytical measurements were stored at −4°C and transported to the laboratory (Department of Clinical Chemistry of the University of Giessen) within 5 h where the vitamins, serum cholesterol and triglycerides were determined as described previously (Wolters et al., 2004). Determination of coenzyme Q10 were performed via gradient reversed phase high-performance liquid chromatography and has been specified in detail elsewhere (Wolters and Hahn, 2003). Since alpha-tocopherol and coenzyme Q10 are transported in association with lipoproteins, the blood concentrations of these nutrients are correlated with the cholesterol concentrations. Thus, serum concentrations of alpha-tocopherol and coenzyme Q10 are also indicated as adjusted values for total cholesterol (mmol/mol total cholesterol) (Wolters and Hahn, 2003).
Serum aliquots for selenium analysis were stored at −20°C and transported to the laboratory of Dr Rurainski, Ettlingen, Germany, for selenium determination at the end of the blood sampling. Serum selenium was determined using electrothermal atomic absorption spectroscopy with Zeeman background correction (Rukgauer et al., 1997). The sample was diluted with Triton X-100 containing 0.1% (w/v) and introduced into the electrothermal atomizer. The reliability was checked by analysing certified reference material. Intra-assay CV was 6% and the detection limit was 0.04 μmol/l. The calibration curve was linear from 0.05 to 3.54 μmol/l.
Data were analysed using SPSS 12.0. (SPSS Inc., Chicago, IL, USA). Results are shown as mean±standard deviation. Normal distribution of data was checked using the Kolmogorov–Smirnov test. Bivariate correlations were analysed using the Pearson method to identify associations among normally distributed variables, while Spearman correlation coefficients were calculated in the case of a skewed distribution. We used partial correlations to identify associations between dietary intake and plasma concentrations adjusted for energy intake and BMI. In order to assess for differences in antioxidant status between employed and unemployed women as well as between those living alone and those living in a family, the independent-sample t test was used. For analysis of variance, data with skewed distribution were normalized by log10-transformation. ANOVA (one-way procedure) was used to test if different educational levels had an influence on the determined blood variables. Linear stepwise regression analysis was used to detect factors influencing antioxidant concentrations. P values <0.05 were considered statistically significant.
Table 1 shows the characteristics of the 178 women as well as the dietary intake of macronutrients and antioxidant vitamins. Eight (4.5%) women indicated consuming a vegetarian diet. In all, 153 (86%) women had already retired from their professional occupation. The study participants were generally better educated than the overall German female population: 21.6% had the qualification for university entrance compared with only 8% of women aged 60 years and older in the general German population (Federal Statistical Office, 1994).
Hormone replacement therapy (HRT) was used by 71 (40%) women. A total of 25 (14%) women were hypertensive and 10 (5.6%) women reported having diagnosed hyperlipoproteinemia.
Dietary intake of ascorbic acid and alpha-tocopherol was below RDA (Food and Nutrition Board, 2000) in 10 (5.7%) and 130 (74.7%) women, respectively. A total of 40 (23%) women consumed less than 3 mg beta-carotene daily. Absolute beta-carotene intake as well as body weight-adjusted and energy-adjusted beta-carotene intake of women with a BMI of 25 kg/m2 or above was significantly lower than in women with normal weight.
Serum concentrations of antioxidants, cholesterol and triacylglycerol are shown in Table 2. Ascorbic acid and beta-carotene intake were significantly correlated with corresponding serum concentrations (r=0.205, P=0.009, n=166; Figure 1 and r=0.173, P=0.025, n=173; Figure 2, respectively), whereas alpha-tocopherol intake was not associated with serum levels. One-way ANOVA showed a significant linear trend (P=0.014) between ascorbic acid serum concentrations and quintiles of intake (Figure 3). No significant associations were found between serum ascorbic acid, alpha-tocopherol and beta-carotene and typical food sources such as fruits and vegetables or nuts. Beta-carotene concentrations were significantly inversely correlated with the BMI (r=−0.342, P<0.001, n=177; Figure 4). In comparison to estimated desirable serum concentrations of >30 μmol/l of alpha-tocopherol, >50 μmol/l of ascorbic acid, and >0.4 μmol/l of beta-carotene as indicated by Gey (1998) and Weber et al. (1997) lower concentrations were found in 41 (23.2%), two (1.2%) and 11 (6.2%) women, respectively. Linear regression analysis revealed that type of diet, BMI and school education influenced beta-carotene serum concentrations (R2=0.128, P<0.001). Cholesterol-adjusted alpha-tocopherol concentrations were weakly influenced by age (R2=0.036, P=0.008), and coenzyme Q10 concentrations by the type of diet (R2=0.026, P=0.022).
Since the content of selenium and coenzyme Q10 in food items is not included in nutrient data bases, it was not possible to calculate the intake of these nutrients. However, no significant associations were found between the serum selenium concentrations and the intake of various food groups, such as meat or vegetables, nor with the intake of micronutrients such as zinc and iron. No significant associations were found between serum selenium concentrations and age, BMI, protein, or fibre intake, whereas the daily alcohol intake correlated significantly with selenium concentrations (r=0.229, P=0.003, n=163). Serum selenium concentrations were not associated with serum cholesterol, triacylglycerol or total plasma homocysteine. In this study, 65 (38.9%) women exhibited serum selenium concentrations below 1.1 μmol/l and 153 (91.6%) below 1.5 μmol/l.
Likewise to the results of selenium, no association was found between coenzyme Q10 and food groups such as meat. Serum coenzyme Q10 concentrations were not related to age, BMI, macronutrient, fibre or alcohol intake. The cholesterol-adjusted Q10 serum concentrations were significantly correlated with equally adjusted tocopherol levels (r=0.188, P=0.013, n=175), whereas a significant inverse association between the adjusted Q10 and beta-carotene concentrations was determined (r=−0.191, P=0.011, n=175).
Some factors that are known to have influence on the here-described outcome variables were looked at in more detail: Table 3 shows the serum concentrations of vegetarians and omnivores for selenium and coenzyme Q10. Women who used HRT had significantly lower serum total cholesterol (P=0.037) and LDL cholesterol (P=0.001) and significantly higher cholesterol-adjusted coenzyme Q10 concentrations (P=0.017) as shown in Figure 5. Women with hyperlipoproteinemia had significantly higher coenzyme Q10 serum concentrations than those without (P=0.001), but this difference disappeared if cholesterol-adjusted coenzyme Q10 concentrations were considered.
Neither employment nor vocational training was predictive for the serum concentrations of antioxidant vitamins, selenium or coenzyme Q10.
The results show that low serum alpha-tocopherol and selenium concentrations are highly prevalent even among well-educated and well-nourished German women aged 60–70 years. It has to be considered that the study participants were recruited by advertisement in newspapers and were probably particularly motivated. Therefore, it can be assumed that they take more care of their health and diet than the overall population. According to the Food and Nutrition Board, a dietary intake of 15 mg of alpha-tocopherol for women aged 51 years and older is recommended (Food and Nutrition Board, 2000). Nearly 75% of the women did not reach the recommended intake for alpha-tocopherol. Thus, one-quarter of the sample had serum alpha-tocopherol concentrations below the desirable threshold of 30 μmol/l (Weber et al., 1997; Gey, 1998). The mean alpha-tocopherol concentration of 34.4 μmol/l in our study was similar to those found in younger European women in different countries (Olmedilla et al., 2001).
Studies indicate that serum selenium concentrations of around 1.1 μmol/l are associated with maximum activity and concentration of plasma selenoproteins (Hill et al., 1996; Duffield et al., 1999), while a serum concentration of around 1.5 μmol/l has been suggested as desirable because it may be optimal for cancer protection (Combs, 2001; Thomson 2004). Nearly 40% of the elderly women in our study exhibited a low selenium status (below 1.1 μmol/l) and more than 90% did not reach the desirable selenium level of 1.5 μmol/l. Serum selenium below 1.2 μmol/l has been reported to be associated with reduced immune function in adult UK subjects (Broome et al., 2004). Other scientists have reported even lower mean selenium concentrations than in our sample in elderly adults, especially in institutionalized elderly people (Navarro-Alarcon et al., 1999; Bates et al., 2002). A suboptimal selenium status has also been observed in other European countries (Murphy and Cashman, 2002; Borawska et al., 2004). While we found no association between selenium status and different food groups and a positive association between selenium status and alcohol intake, a Polish study reported an inverse association with the intake of the sum of beer, wine and liquor (Borawska et al., 2004). However, a positive relation of selenium status and alcohol intake was also found in women in the 3rd National Health Nutrition Examination Survey with 7517 female subjects (Kafai and Ganji, 2003). Meat and meat products are an important selenium source. We did not find a direct association between the intake of this food group and selenium. However, alcohol consumers have been shown to have a higher intake of meat (Villegas et al., 2004; Ruf et al., 2005). Thus, in our study, the diet of women with a higher selenium status may reflect a typical pattern of higher alcohol consumption and higher meat and selenium intake.
In contrast to a study with British elderly subjects (Bates et al., 2002), plasma selenium was not correlated with plasma cholesterol, vitamin levels or anthropometric variables in our sample. An association of selenium status with blood lipids was reported previously (Navarro-Alarcon et al., 1999; Lee et al., 2003).
Gonzalez et al. (2004) observed a correlation between selenium status and plasma homocysteine. This was not seen in our sample.
Selenium and alpha-tocopherol work synergistically in inhibiting lipid peroxidation. Insufficient status of one of these nutrients or both may elevate the risk for oxidative stress and possibly for associated diseases (Thomson, 2004). Since selenium and alpha-tocopherol status was low in many women of our sample, there may be an elevated risk especially in those subjects with both antioxidants far below the desirable concentrations.
Compared with other European studies with younger participants, the mean ascorbic acid serum concentration was quite high in our sample (Olmedilla et al., 2001). The mean was more than twice as high as the estimated preventive threshold of >50 μmol/l and only two women had concentrations below this value. The beta-carotene blood values of our study population were in the recommended range as well. Serum levels of women aged 76±6 years in the Framingham collective (Vogel et al., 1997) and levels of younger European women were about half as high as in our study population with the exception of French women who reach levels nearly as high as in our sample (Preziosi et al., 1998; Olmedilla et al., 2001). As previously observed in other investigations, we found an inverse correlation between serum beta-carotene and the BMI (Palli et al., 1999; Wallström et al., 2001). Since beta-carotene is distributed between serum and adipose tissue with adipose tissue being the dominant storage tissue, a high fat mass leads to a higher absorption of beta-carotene by fat tissue compared with a low fat mass (Wallström et al., 2001). Another explanation may be that the women with a BMI below 25 kg/m2 ingested significantly more beta-carotene than those with a BMI above 25 kg/m2. This was true for the absolute beta-carotene intake as well as for energy-adjusted or body weight-adjusted beta-carotene intake in our study.
Since coenzyme Q10 acts as an antioxidant and is able to regenerate alpha-tocopherol (Crane et al., 1993; Kaikkonen et al., 2000), a positive correlation between both antioxidants can be assumed and was confirmed. There was a negative association between beta-carotene and cholesterol-adjusted Q10 concentrations. Since the coenzyme Q10 content of the diet was not assessed, it cannot be determined whether the inverse relationship results from different food intake patterns: the main beta-carotene sources are fruits and vegetables, while the main sources of coenzyme Q10 are meat, meat products and fish (Weber et al., 1997).
Our data reveal that the selenium and alpha-tocopherol status of well-nourished younger female seniors often seems to be insufficient. Nearly 40% of the sample failed to reach desirable serum selenium concentrations and nearly a quarter of the women had alpha-tocopherol levels below the desirable threshold. Healthy eating patterns may lead to high beta-carotene and ascorbic acid concentrations as seen in most of the participating women. Results of the SU.VI.MAX study with 13 017 men and women aged 35–60 years (Hercberg et al., 2004) indicate that supplementation with antioxidants (median 7.5 years) may result in a lower cancer incidence. A protective effect of antioxidants was observed only in men, but not in women, possibly due to the lower baseline status in men (Hercberg et al., 2004). This result indicates that only women with low antioxidant levels would possibly benefit from an additional intake of antioxidants.
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Guarantors: M Wolters and A Hahn.
Contributors: AH and MW originated and designed the study. SH coordinated the study and organised blood sampling and data collection. SG and NK were responsible for the analytical measurements. MW wrote the paper.
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Wolters, M., Hermann, S., Golf, S. et al. Selenium and antioxidant vitamin status of elderly German women. Eur J Clin Nutr 60, 85–91 (2006). https://doi.org/10.1038/sj.ejcn.1602271
- ascorbic acid
- coenzyme Q10
- nutritional status
- vitamin C
- vitamin E
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