Plasma selenium levels in healthy blood bank donors in the central-eastern part of Belgium

ARTICLE IN PRESS

Journal of Trace Elements in Medicine and Biology 21 (2007) 225–233 www.elsevier.de/jtemb

APPLIED METHODOLOGY

Plasma selenium levels in healthy blood bank donors in the central-eastern part of Belgium Rudy Van Cauwenbergha, Harry Robberechta,b,, Veerle Van Vlaslaerb, Annie De Smetc, Marie-Paule Emondsc, Nina Hermansa a

Laboratory for Nutrition and Functional Food Science, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium b Plantijnhogeschool, Kronenburgstraat 47, B-2000 Antwerp, Belgium c Rode Kruis Vlaanderen, Bloedtransfusiecentrum Vlaams-Brabant – Limburg, Centraal Donorlaboratorium, Herestraat 49/BTC, B-3000 Leuven, Belgium Received 9 January 2007; accepted 9 June 2007

Abstract Graphite furnace atomic absorption spectrometry, with Zeeman background correction and after improved matrix modification, was used to measure the plasma selenium content of healthy blood bank donors in the central part of Belgium. The mean plasma selenium concentration of 80 men and 80 women was 79.774.4 ng/mL with a range of 55.0–117.4 ng/mL. There was no gender difference observed. Plasma selenium level was significantly highest for the adult group, aged 45–64 years, compared to the others, except the young adults (18–24 years). The mean plasma selenium concentration measured corresponded well with literature data for Belgium. The obtained values were found to be in the medium range, compared with recent literature values for the European countries. r 2007 Elsevier GmbH. All rights reserved. Keywords: Plasma; Selenium; Blood bank donors; Belgium

Introduction Selenium (Se) is an essential component of a number of selenoproteins including the glutathione peroxidases (GSH-PX), a family of enzymes that protects against oxidative injury by catalysing the breakdown of hydrogen peroxide and lipid peroxides [1]. A large number of Corresponding author. Laboratory for Nutrition and Functional Food Science, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium. E-mail address: [email protected] (H. Robberecht).

0946-672X/$ - see front matter r 2007 Elsevier GmbH. All rights reserved. doi:10.1016/j.jtemb.2007.06.003

studies have shown an inverse relation between serum Se concentrations and the incidence of cancer [2,3] or cardiovascular disease [3–5]. Furthermore, Se is promising as an anti-poliferative, anti-inflammatory, antiviral and immune altering agent [6]. Although many questions still remain to be solved, there is an increasing interest in the clinical significance of this element [7]. The more interested reader is referred to Refs. [4–67] in a previous paper [8] or a more recent review [9]. In most studies, the Se status has been assessed by measuring the element either in serum or plasma,

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erythrocytes, platelet or whole blood, and by determining the GSH-PX activity in whole blood or platelets. Serum Se levels reflect the recent dietary intake, whereas erythrocytes accumulate Se and presumably reflect the intake over their 120 days span of life [10]. GSH-PX is thought to be a good indicator of the Se status, as it is decreased in Se deficiency [11]. Ne`ve [12] discussed in full detail the different methods for the determination of Se status. Procedures, concentration levels and health implications of measuring Se in various blood components were reported by Robberecht and Deelstra [3]. Whole blood analyses of Belgian blood bank donors were carried out and published more than 15 years ago [13,14]. Serum Se level data for the Antwerp region were published nearly 25 years ago [15] and more quite recently [16]. In the present work the short-term Se status of healthy blood bank donors from the central part of Belgium was investigated by measuring the plasma Se level. A Zeeman-effect background corrected graphite furnace atomic absorption spectrometer (ETAAS) was used with an optimized preparation step for matrix modification [16]. The Se levels measured are compared with the scarce older data for healthy adults in Belgium and with data from recent publications on Se plasma levels from other European countries for the same population groups.

Materials and methods Subjects Belgium blood bank donors offer their blood freely, but decreted by law, they have to be older than 18 years and less than 65 years of age and weighing more than 50 kg. The blood was collected in 3 mL Vacutainers tubes, containing 5.4 mg EDTA. Blood was stored overnight at room temperature. Plasma was removed and stored at 20 1C until assayed. The donor resided in the central and eastern Flemish part of Belgium. A medical questionnaire concerning disease, possible infections, risk behaviour, pregnancy, stay outside Europe, and medication has been filled in, on the basis of which some people are excluded. No information about smoking, alcohol abuse or nutritional supplementation by the donors was obtained.

Instrumentation and methods A Perkin Elmer model 4100 ZL Zeeman atomic absorption spectrometer, equipped with a PE AS-70 autosampler, was used for all measurements. THGA

Table 1. Temperature programme for the determination of selenium in plasma Step

1 2 3 4 5 6

Temperature (1C) 110 130 550 1200 2000 2500

Time (s)

Gas flow (mL/min)

Ramp

Hold

1 5 5 10 0 1

10 28 5 20 5 5

250 250 250 250 0 250

Read

x

graphite tubes (L’Vov platforms, Perkin Elmer) were used for measurement with longevity of about 300 injections of plasma samples. The manufacturer’s recommendations for wavelength, spectral band width and lamp current parameters were followed. An adapted programme for Se determination was used, as summarized in Table 1. The detection limit, calculated following the IUPAC recommendations [17] and using peak area was 5 ng/mL in the original sample. The accuracy of the method was checked by analysing Seronorm reference materialTM (level 2). Our value of 130 ng/mL was found to be within the ‘‘recommended range’’ for Se (121–137 ng/mL). The relative standard deviation at 130 ng/mL (n ¼ 30) was 4.7%. Concentrations were calculated using standard addition at levels of 0, 10, 20 and 30 ng/mL. The diluting scheme was as follows: 200 mL of plasma, 200 mL of modifier, 100 mL Triton-X (5%) and finally 500 mL of the standard. From this volume 30 mL was injected so that this contained 1.8 mg Pd and 1.2 mg Mg(NO3)2 as modifiers [16]. Linearity proved to be valid up to 300 ng/mL.

Results and discussion Selenium concentrations in healthy Belgian adults Table 2 summarizes the measured plasma Se concentrations of healthy blood bank donors in the central part of Belgium. The overall mean value was 79.774.4 ng/mL. The range for all samples was 55.0–117.4 ng/mL. The Se content tended to be somewhat lower in females than in males, but at a statistically nonsignificant level (p40.05). Therefore, males and females are considered as one group to study influences of age. Here the group (45–64 years) revealed higher concentration levels, compared to the two younger groups. However, no statistically difference was observed compared with the values from the group (18–24 years).

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The scarce literature data on serum/plasma levels for an adult Belgian population are summarized in Table 3. As can be seen, values obtained in this study, are very similar to those obtained for other locations in Belgium. Blood donors in the Brussels area (1985–1986) revealed identical plasma Se levels [22]. Ne`ve [12] classifies plasma Se concentrations into three categories: ‘‘low’’, below 50–60 ng/mL; ‘‘high’’, above 100–120 ng/mL; and ‘‘normal’’, between these two ranges. The results of the present study belong to the ‘‘normal’’ category. Our results fit into published profiles of Se concentrations as a function of age. Data show a very low Se level during the first months of life with a gradual increase with age, as described for Belgium and The Netherlands [18] and discussed in our previous paper in this journal [16]. Highest values were obtained in healthy adults, while the elderly people revealed significantly lower concentrations. Here we observed that the eldest group (45–64 years) revealed the highest Se level, which was quite remarkable, but not significantly higher than the mean value, obtained for the youngest group of adults (18–24 years). Table 2. Plasma Se levels (ng/mL) of healthy blood bank donors in the central-eastern part of Belgium Group (years)

Sex

18–24 25–34 35–44 45–64 Overall mean

Males Females Males Females Males Females Males Females –

Number (n) 20 20 20 20 20 20 20 20 160

Concentration levels X7SD

Range

77.0711.4 82.4712.9 78.4710.5 75.3710.0 77.0711.4 75.579.0 86.679.2* 85.279.2* 79.774.4

55.0–95.9 61.3–106.3 63.1–93.7 58.9–100.1 61.0–99.4 59.7–90.4 72.8–117.4 67.5–105.0 55.0–117.4

*po0.05.

Table 3.

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Selenium concentrations in healthy adults from other European countries Most of the literature deals with plasma Se levels. No difference was made in this review between plasma and serum, as done by most authors [25–27]. Only one publication [28] claimed that serum concentrations cannot be equated with plasma values. In one paper [29] the word ‘‘plasma’’ was mentioned in the summary, while in the analytical methods the word ‘‘serum’’ is used. Moreover, since no Se values are mentioned in the article, it is not included in the following compilation. Recent literature data for Europe on Se in plasma (serum) are compiled in Table 4. Only data published from 2000 onwards are included. Age, gender, and other specifications on the sampled population, as far as could be traced and useful in interpreting the results, are presented. Also values for a healthy control group, as mentioned in the various studies on pathological conditions, are taken up in the compilation. Sometimes no control group is mentioned and concentration levels can therefore not be included [60]. Concentration levels, as presented in figures with barplots are also not included [61]. Values found in literature, expressed in mmol/L, are recalculated to ng/mL, using a conversion factor 78.96. However, concentration levels expressed as ng/g [62] are omitted. Plasma or Se levels from healthy people with confining factors, like lactation [63], pregnancy [64] or alcohol-consumption [65] are not included in the reference list. Similarly, situations which may influence the serum Se level, like stress as response to laparostomy and laproscopy [66] are not included. The interested reader, who would like to obtain more information about factors influencing blood Se concentrations, is referred to a literature review [3], or to the following more recent references for Se values in

Literature data on serum (S) or plasma (P) selenium levels (ng/mL) in healthy adults in Belgium

Specification

Sample

n

Sex

Specification Age (years): mean (range)

X7SD

Range

Ref.

Antwerp Antwerp Brussels Brussels Brussels Brussels Brussels Ghent Ghent Ghent

P S P P P P P P P S

10 26 72 6 74 145 – 110 54 27

M+F 13M+13F M+F 3M+3F M+F M+F – M F –

Healthy lab staff 38.7 (23–69) Blood donors 3275 (27–40) Blood donors; 1980–1981 Blood donors; 1985–1986 Healthy controls 33 (20–58) 34 (19–52) Pooled serum

95711 84.379.4 84.5712.6 85 97.177.9 83.7711.8 80 97712 97712 95

72–116 51.4–121.7 65–110 – – – 65–110 67–123 72–119 91–100

[18] [16] [19] [20,21] [22] [22] [23] [15] [15] [24]

M ¼ males, F ¼ females.

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Table 4. adults

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Recent literature data (from 2000 onwards) on serum (S) or plasma (P) selenium levels (ng/mL) of healthy European

Country (city or region) Belgium (central part) Croatia (Zagreb) (–) Czech Republic (Moravia) Denmark France (Grenoble) Germany (Rostock) (Jena) (Stuttgart) Poland (non-polluted area) (vicinity of a smelter) (Bydgoszcz) (Bydgoszcz) (Bydgoszcz) (Gdansk) (Gdansk) (central part) (Lodz) (Wroclaw-Southwest) (Wroclaw-Southwest) (Wroclaw-Southwest) (Wroclaw-Southwest) Portugal (Azores’ archipelago) (Azores’ archipelago) (Azores’ archipelago) (Azores’ archipelago) (Azores’ archipelago) (Azores’ archipelago) Spain (Canary Islands) (Barcelona) (Canary Islands) (Canary Islands) Slovenia Soviet Union Sweden Switzerland Turkey (Ankara) (Ankara) (Ankara) (Aegean area) UK (Hull) a

Specification (age in years: mean; range) (18–64) (3578) Healthy population Above 50 years (18.2710.0) Postmenopausal (55.470.6) Healthy subjects Reference population Blood donors (37.4711.7) (40.7)(22–66) (40.7)(22–66) (24–60) (28.4) (43.079.9) (19–70) (19–70) Low selenium area (5979)(34–74) Smokers Non-smokers Smokers Non-smokers (41710) (42710) Fishing population (20–60) Fishing population rural population (20–60) Rural population (38.4720.2)(6–75) (41.3712.4)(19–69) (35.979.0) (37.979.0) Young blood donors Different areas (20–53) (38*) Healthy blood bank donors (30.7715.4)(20–71) (46712)(22–60) (37.8718.8)(18–72) (40.6713.6) (30.373.5)

Sample n

P S S S S P S P P S S P P P P P P P S S S S S S S S S S S P S S S S S S P S S P S

160 62 – – 41 26 75 – 99 93 76 22 28 20 65 81 98 210 11 5 5 12 45 57 24 40 23 32 395 152 72 114 10 556 124 110 17 10 18 30 18

Sex

80M+80F 26M+36F – – M F – – 45M+54F M MF 18M+8F F 9M+11F M F F – M M F F M F M F M F 187M+208F 81M+71F M F F F F – 15M+2F – 14M+4F 5M+25F F

Concentration level X7SD

Range

79.774.4 64.2711.5 – 96.1718.3 77.4711.7 75.872.4 106.5716.5 80.5723.7 66.377.9 54.8710.9 58.0710.9 72.6716.1 66.1713.1 82.1 76.7713.2 70.4714.7 54712 53.7714.3 47.4725.5 73.7732.7 57.2728.7 80.5733.6 98716 88715 110726 90721 85722 80717 74.7725.2 82.2717.5 81.5726.6 72.7724.2 66715 – 70.0a 112.7732.3 69.375.8 57.9711.5 46.077.9 109.1734.2 110710.0

55.0–117.4 52.3–78.6 58–79 – – – – – 52.9–81.3 29.0–78.3 35.5–89.8 – – – – – – 14.6–110.1 – – – – 65–137 58–131 – – – – 7.9–182.3 50–145 26.5–182 21.7–140 32–88 68.7–137.4 46–154 87.3–143.5b – 44.6–76.6 – – –

Ref.

This study [30] [31] [32] [33] [34] [35] [36] [37] [38] [38] [39] [40] [41] [42] [42] [43] [44] [45] [45] [45] [45] [46] [46] [47] [47] [47] [47] [48] [49] [50] [50] [51] [52] [53] [54] [55 [56] [57] [58] [59]

Median value. 5th–95th percentile.

b

children [67–72], adolescents [68,73,74] or elderly people [75–77]. Values of plasma Se concentration for healthy blood bank donors in the central-eastern part of Belgium fall within data recently published for other European regions, like Denmark [33], some parts of Poland [39,42,45], different regions of Spain [49,50] and Sweden [53].

The lowest Se values in Europe were found in Eastern Europe, Croatia [30,31], several parts of Poland [38,43–45], Slovenia [51] and Turkey [56]. This corresponds to older literature data for these parts of Europe, as discussed somewhere else [8]. The highest levels in Europe were found in Germany [35], Portugal [46,47], Switzerland [54], the UK [59] and quite remarkably also for the Aegean area of Turkey [58].

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All data are however much lower than those observed in the USA or seleniferous regions. Daily intake of Se is most of the time responsible for the variation in blood elemental level [3]. For Lanzarote, one of the Canary Islands of Spain, healthy individuals established a significantly higher Se level, compared to the other small islands. This could be attributed to differences in soil and/or dietary habits of the population. No relationship with socio-economic status, educational level, smoking habits, physical exercise or beer consumption was found. However, individuals who consumed wine more than three times a week showed higher Se concentrations than individuals with lower consumption. Also, individuals with consumption above seven units of spirit drinks a week had the highest mean Se concentration [50]. The fact that the geographical location is of paramount importance can be the best illustrated by the situation in Poland, where Se status is quite well studied by various groups. Results by Hac et al. [42], are quite similar to those obtained by Zachara et al. [78], but differ quite well from results for a population of northwestern Poland, as observed by Trzcinka-Ochocka et al. [38] and also for a Se-low area in the central part of that country [43]. Another research group in the same country proved that frequent consumption of ham, tea and honey may be effective to improve the Se concentration in the sera of Se-inadequate subjects [79]. Still for the same country, males living near a polluting smelter did not reveal variation in serum Se levels [38].

Selenium blood level as a function of sex The absence of a gender influence on plasma Se concentration in this study was also demonstrated by our research group in a previous study on serum level in an Antwerp population [16] and on serum level of two healthy groups of adults in Greece [8,80]. Results from literature are quite confusing. If there is an effect of gender most of the time (probably due to larger intake amount) a higher concentration levels is found in men. For the USA [81,82] this was very significant (po0.0001). For that country it was even proven that the plasma Se level fluctuated during the female life cycle [83]. This suggests that stage of life cycle and the possible effect of hormone fluctuations, especially estrogen status, should be considered when assessing female Se status and establishing dietary Se recommendation.

Selenium blood level as a function of age Our results fit well into previous published profiles for Se blood concentration as a function of age in Belgium

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[15,18,84]. This profile was also observed in various other countries and resembles quite well with serum or plasma profiles for various other trace elements. This starts with very low Se level during the first months of life, with a gradual increase with age [18,84]. Highest values were observed in healthy adults, while the elderly revealed significantly lower concentrations [75–77]. A significant decrease of Se serum values was demonstrated in a group of 90 non-institutionalized nonagenarians/centianarians (91–110 years), when compared with 62 elderly subjects (60–90 years), in both males and females [85]. For the USA the same observation was published, where a low plasma Se level before diagnosis was associated with a 4–5-fold increased risk of prostate cancer [86].

Other factors, possibly influencing blood selenium level Various factors, that probably influence the blood Se concentration level of people residing in a given geographical area, as published in older literature data are previously reviewed in this journal [3]. The literature was screened for the effects of age, gender, smoking and drinking habits, race, state of lactation, environmental exposure and medical treatment. The differences are mostly found to be small on the absolute scale, with only nutrition seeming to be a major factor. For the recent literature some significant observations are mentioned here. A study in Nuremberg (Germany), where Se level in plasma was mentioned in the abstract and ‘‘serum’’ level in the article, did not publish the Se concentrations found. However, they claimed that smokers and non-smokers (27–66 years) in various combinations did not differ in Se level [29]. A similar observation was published for the USA, where the effect of smoking was small [87]. The authors claimed a significant reduction in serum levels of Se in male smokers but not in female ones. However, even though statistically significant a 4% reduction in male smokers does not provide a clear negative association between serum level of Se and smoking. They argued that the lower Se concentrations could be explained by low dietary intake rather than by smoking effect. This was also mentioned in an older publication for a seleniferous area in the USA [88]. To illustrate the various observations on effect of smoking: for the same country (Poland) no effect [78], a small effect [45], as well as an effect [64] was published in recent literature. In the Czech Republic a significantly lower whole blood Se level in smokers as opposed to non-smokers was observed [89]. The trend however, seemed to be lower values in smokers, even in the children either (or both) of whose parents were smoking [90].

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Unexpectedly, there was no evidence that children who themselves smoked, had lower levels. Maybe the amount of cigarettes is the most important factor. In Poland Lutz-Frackiewicz and coworkers [45] claimed that for people smoking over 20 cigarettes/day mean serum concentration is significantly lower in the group of smokers, compared to non-smokers. For men that smoke, the concentration was lower than that found for smoking women, however the difference was without statistical significance. In our study we are unfortunately not aware of the smoking attitude of the donors. Also for alcohol consumption literature data are quite controversial and also here the amount of alcohol may be of paramount importance influencing lower daily intake of Se. For a group of people in lower Silesean in Poland who drink an equivalent of 50 g or more of pure ethanol per week the serum Se level has been found to be lower compared to the group that does not consume that much alcohol [45]. For the Azores’archipelago, Portugal Se levels were higher in fishers of both genders than in subjects living in a rural population [47]. In the fishers, but not in the rural population Se was higher in the males, but it did not show significant variation with age. A quite remarkable observation was published by Gonzalez et al. [77], where an inverse relation was registrated between Se level and homocysteine concentration in blood of elderly people.

Conclusion Plasma Se levels of the healthy blood bank donors in the central-eastern part of Belgium are similar to concentrations found in previous studies on adults in this country. Compared to the reviewed values in the literature for healthy adults in Europe, the plasma Se content of the Belgian adults proved to be in the medium range. Gender does not seem to influence the observed plasma level, as was already published for most other countries. The observed variation in Se level in the plasma of our donors as a function of age was quite remarkable compared to literature data. This could be due to a limited number of cases in each age group, an insufficient severe screening for the background of the donors or most probably to differences in intake levels.

Acknowledgements H. Robberecht wishes to thank the University of Antwerp (UA) for research facilities on a freelance basis. He was allowed to do this work because of the

association intention of the Antwerp Highschools with the University of Antwerp.

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