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Monitoring Recovery from Iron Deficiency Using Total Hemoglobin Mass

WACHSMUTH, NADINE B.; AIGNER, THOMAS; VÖLZKE, CHRISTIAN; ZAPF, JÜRGEN; SCHMIDT, WALTER F.

Medicine & Science in Sports & Exercise: February 2015 - Volume 47 - Issue 2 - p 419–427
doi: 10.1249/MSS.0000000000000420
APPLIED SCIENCES
Free

Introduction: Using hemoglobin concentration ([Hb]) to diagnose borderline iron deficiency and monitor the progress of its treatment is difficult because of the confounding effects of plasma volume. Because hemoglobin mass (Hbmass) is not affected by plasma volume, it may be a more sensitive parameter. The aim of this study was to monitor Hbmass, iron storage, and maximal oxygen consumption (V·O2max) during and after oral iron therapy in subjects with severe and moderate iron deficiency.

Methods: Three groups of female recreational athletes were monitored for at least 22 wk, as follows: 1) severe iron deficiency group (SID) (n = 8; ferritin, <=12 ng·mL-1), 2) moderate iron deficiency group (MID) (n = 14; ferritin, <=25 ng·mL-1), and 3) control group (n = 8; ferritin, >25 ng·mL-1). Hbmass and iron status were determined before, during, and up to 12 wk after at least 10 wk of oral iron supplementation. In total, five V·O2max tests were performed before, during, and after the supplementation period.

Results: Hbmass increased markedly in the SID group (15.6% ± 11.0%, P < 0.001) and slightly in the MID group (2.2% ± 3.7%, P < 0.05) by the end of the supplementation period and remained at this level for the following 12 wk. [Hb] and Hbmass were similarly affected, but Hbmass was more closely related to mean corpuscular volume and mean corpuscular hemoglobin than [Hb]. The SID group incorporated 534 ± 127 mg of iron into ferritin and hemoglobin, whereas the MID group incorporated 282 ± 68 mg of iron. V·O2max increased only in the SID group by 0.20 ± 0.18 L·min-1 (P < 0.05) and was closely related to Hbmass (P < 0.01).

Conclusions: Hbmass is a sensitive tool for monitoring recovery from iron deficiency anemia and assessing the effectiveness of iron supplementation in individuals with severe or moderate iron deficiency.

Department of Sports Medicine/Sports Physiology, University of Bayreuth, Bayreuth, GERMANY

Address for correspondence: Nadine Wachsmuth, MSc, Department of Sports Medicine, University of Bayreuth, Bayreuth, Germany; E-mail: mailto:Nadine.Wachsmuth@uni-bayreuth.de.

Submitted for publication January 2014.

Accepted for publication May 2014.

Iron deficiency anemia frequently occurs in female athletes who undergo regular endurance training, and reduced endurance performance is closely connected to the severity of the anemia (12). Ferritin and hemoglobin concentration ([Hb]) are common parameters used to diagnose iron deficiency. Plasma ferritin concentration in healthy subjects is directly proportional to the size of body iron stores (42) and is therefore a reliable indicator for detecting depleted iron stores. However, the recommended thresholds for iron deficiency range from 10 to 35 ng·mL-1 in the literature (6,25,30,32,33,41) and the criteria for iron supplementation are not consistent (9,14).

The recommended [Hb] thresholds for iron deficiency anemia are 12.0 g·dL-1 for women and 13.0 g·dL-1 for men (46). In general, the interindividual [Hb] is highly variable even among subjects with high iron availability. This variability is indicated by the normal ranges of 12.0–16.0 g·dL-1 for females and 13.5–17.5 g·dL-1 for males (25). Individuals with an [Hb] value in the upper normal range can therefore lose 20%–30% of their body iron before [Hb] decreases below the threshold for anemia (7). Thus, milder iron deficiency in subjects with a normally high [Hb] who do not fulfill the anemia criteria can often remain undetected (7).

A true indication of iron deficiency anemia is a positive response to iron therapy (15). Ferritin and [Hb] are used to measure the effect of iron supplementation. However, both parameters are influenced by external factors. Because ferritin is an acute-phase protein, the ferritin concentration can be influenced, for example, by inflammation (47) and acute exercise (23). [Hb] can be influenced by posture (18), hydration status, acute exercise, and training status (16).

Therefore, a parameter that is not influenced by acute inflammation or fluid changes might be a sensitive parameter for observing small changes caused by iron supplementation. We suggest total hemoglobin mass (Hbmass) as such a parameter. To date, few data relating iron deficiency to Hbmass are available. In a case study of severe iron deficiency anemia, Hbmass was demonstrated to be a reliable tool when tracking the success of iron supplementation (13). Oral and intramuscular (i.m.) supplementation for up to 8 wk in moderately iron-deficient nonanemic subjects, however, did not result in enhanced Hbmass, whereas intravenous iron supplementation had positive effects (1,12,14).

We postulate that Hbmass may be an important additional parameter for assessing the success of iron therapy. Individual changes due to iron supplementation in subjects with mild or severe iron deficiency may be monitored more sensitively using Hbmass than using [Hb]. In this study, special focus was placed on 1) the magnitude of the individual response in subjects with various levels of iron deficiency, 2) the individual time span necessary for complete compensation of the iron deficiency anemia, and 3) the stability of iron status after iron supplementation. Furthermore, we estimated the amount of iron incorporated into the body because of iron supplementation. Finally, we focused on how the changes in Hbmass caused by iron repletion affected aerobic capacity (V·O2max).

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METHODS

Subjects.

Thirty female recreational athletes age between 19 and 41 yr were recruited to participate in the study. An initial screening identified 22 iron-depleted women. Of these women, eight exhibited severe iron deficiency (SID), with ferritin levels <=12 ng·mL-1 (25), and 14 exhibited moderate iron deficiency (MID), with ferritin levels between 13 and 25 ng·mL-1 (6). The other eight females with ferritin levels >25 ng·mL-1 formed the control group (C). None of the subjects had iron supplementation for at least 3 months before the beginning of the study. The anthropometric data of the subjects are presented in Table 1. After receiving explanation of the requirements and risks of the study, all subjects signed a consent form. The study was approved by the ethics committee of the Friedrich-Alexander-University, Erlangen-Nuremberg, Germany.

TABLE 1

TABLE 1

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Study design.

Data were collected from all three groups (SID, MID, and C) for at least 22 wk. Members of the SID and MID groups were supplemented with iron (100 mg·d-1, Ferro Sanol Duodenal©; Sanol GmbH, Monheim, Germany) for 10 wk. Subjects were instructed to take the iron supplements in the morning on an empty stomach together with a glass of orange juice. After this period, the iron supplementation continued for 4–8 wk if any of the following criteria were met: 1) the ferritin values remained <25 ng·mL-1 and 2) Hbmass had not reached a plateau (a plateau was defined as a <20-g difference in Hbmass between two consecutive measurements). Hbmass was measured before, during, and up to 12 wk after the iron supplementation period (Fig. 1). Duplicate measurements were performed before and after the supplementation period and at the end of the entire observation period. In addition, single Hbmass measurements were performed every second or third week during the supplementation period and once during the postsupplementation period. Simultaneously, venous blood samples were drawn to determine [Hb], hematocrit (Hct), C-reactive protein (CRP), and iron (ferritin and transferrin) status. All subjects performed five V·O2max tests before, during, and after the supplementation period (Fig. 1). Four of the subjects developed common cold, and one suffered from an ankle injury, which interfered with their daily activity for less than 2 wk but did not influence the test protocol.

FIGURE 1.

FIGURE 1.

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Measurement of Hbmass.

Hbmass was measured using the optimized carbon monoxide (CO)-rebreathing method, as previously described (37), with the modifications discussed in previous studies (17,34. Briefly, a CO bolus 0.7 mL·kg-1 body mass was inhaled, followed by 2.5 L of pure oxygen, and these gases were rebreathed through a glass spirometer (SpiCo®; Bayreuth, Germany) for 2 min. Capillary blood was collected immediately before (two capillaries) and 6 and 8 min after the rebreathing. Each sample was analyzed in three replicates for carboxyhemoglobin using a CO hemoximeter (OSM3; Radiometer, Denmark). The remaining CO in the system and the end-tidal CO concentration were determined using a CO analyzer (Draeger, Germany) to calculate the amount of CO that was not taken up during the inhalation and the amount exhaled after the test. The reliability of the method was derived from the use of all duplicate measurements throughout the study and was characterized by a typical error of 1.7% (approximately 9 g). Blood volume (BV), red cell volume (RCV), and plasma volume (PV) were calculated according to the following formulas (37):

The value 0.91 refers to the cell factor (4).

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Blood analyses.

Venous blood samples (9 mL) were drawn in the morning without any preceding severe exercise from a cubital vein after a 10-min period spent resting in a sitting position. Aliquots of 2 mL were placed into EDTA tubes to determine the [Hb], Hct, and derived erythrocyte indices [mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and mean corpuscular volume (MCV)] using a Cell-Dyn 3700 (Abbott). In addition, 7 mL of the venous blood was centrifuged, and the serum was used to measure the concentrations of ferritin (chemiluminescent microparticle immunoassay), transferrin, and CRP using an Architect ci8200 (Abbott). In total, 70–80 mL of blood was removed during the whole observation period, which corresponds to a loss of Hbmass of approximately 10 g within 22 wk.

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V·O2max tests.

An incremental test on a cycle ergometer (Excalibur; LODE®, Netherlands) was performed to determine V·O2max. After a 3-min warm-up at 50 W, the load was increased by 17, 17, or 16 W every minute (50 W per 3 min) until volitional exhaustion. The respiratory gases were analyzed breath by breath using the MetaMax II spirometric system (Cortex®, Germany).

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Iron absorption.

To calculate the total body iron incorporation during and after the supplementation period, the following formula was used:

The “pre” indices indicate the baseline values, and the “post” indices correspond to the actual values obtained during and after iron supplementation. The following conversion was used: 3.38 mg = the amount of iron stored in 1 g of hemoglobin (40). The studies of Walters et al. (45) and Cook (7) indicate that a close relation exists between ferritin concentration and body iron stores and that 1 μg·L-1 of ferritin corresponds to approximately 8 mg of tissue iron. In total, 19 of the 200 cases exhibited CRP >0.3 mg·dL-1, and the ferritin values of those cases were excluded from the calculation of the iron stores.

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Lean body mass.

To estimate lean body mass (LBM), body fat percentage was determined. Skinfolds were measured using a caliper at the musculi biceps and triceps brachii and at the subscapular and suprailiac sites twice before the supplementation period, once after the supplementation period, and once again after 12 wk, and fat mass was calculated according to Berres et al. (2):

The typical error of this method was 1.8% in our study. The LBM was calculated as the difference between the body mass and the absolute fat mass.

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Statistics.

The data are presented as the mean ± SD. For statistical analysis, the Statistical Package for the Social Sciences for Windows (version 18; SPSS Inc., Chicago, IL) was used. To compare the values before and after iron supplementation in identical subjects, a paired t-test was applied (21); to compare the means of different groups, an unpaired t-test that included Bonferroni correction was used. To evaluate any causal relation between two variables (e.g., V·O2max and Hbmass), linear regression analysis was performed.

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RESULTS

Because of the classification of the subjects into groups of severe and moderate iron deficiency, the SID group exhibited the lowest baseline ferritin (5.4 ± 1.9 ng·mL-1, range between 3 and 8 ng·mL-1) and [Hb] (11.6 ± 1.2 g·dL-1, between 9.1 and 12.5 g·dL-1) values (Fig. 2a and b). Hbmass, relative to both body mass and LBM, was significantly lower in the SID group than that in the MID and C groups (Table 2). The hematological parameters used to characterize iron deficiency were reduced (MCH and MCV) or elevated (transferrin) compared with those for the MID and C groups (Table 2). However, only one subject showed lower values than the normal range (25) for MCH (25 pg) and MCV (75 fL).

TABLE 2

TABLE 2

FIGURE 2.

FIGURE 2.

At the beginning of the study, except for lower ferritin concentration (between 13 and 25 ng·mL-1) and increased transferrin concentration in the MID group, similar hematological values were found in the MID and C groups (Table 2). In the MID group, [Hb] ranged from 12.2 to 14.0 g·dL-1 and one subject showed an MCV below the normal range (78 fL).

During the iron supplementation period, two SID subjects reached the criteria to stop the supplementation (ferritin >=25 ng·mL-1 and a plateau in Hbmass) after 10 wk, five subjects finished after 14 wk, and one subject met the criteria after 18 wk. At the end of the iron supplementation period, [Hb], ferritin concentration (Fig. 2a and b), and all other markers of the iron status were normalized in this group and did not differ from the values in the C group (Table 2). Twelve weeks after the supplementation, the ferritin level was markedly reduced again and the transferrin was increased whereas all of the other parameters remained constant (Table 2).

In the MID group, the ferritin (Fig. 2b) and transferrin concentrations were shifted into the normal range during the supplementation period and [Hb] (Fig. 2a), MCH, and MCHC exhibited slight but significant increase (Table 2). No change was observed during the postsupplementation period.

In the SID group, Hbmass significantly increased until the end of the supplementation period by 15.6% ± 11.0% (77.0 ± 37.7 g) (Fig. 2c) and reached the level of the C group. Hbmass remained at this level for the following 12 wk. In the MID group, Hbmass slightly increased (2.2% ± 3.7% or 16.1 ± 18.0 g, with P < 0.05) and remained at this elevated level for the following weeks (Table 2). Compared with the C group, Hbmass in the MID group demonstrated significant increase after the supplementation period (P < 0.05) and tendency toward higher values after 12 wk (P = 0.07). The considerable variability in the change of Hbmass within the groups (Fig. 2c) indicates the very different individual responses during and after the iron supplementation, as presented in Figure 2d and e. The individual change in Hbmass after the iron supplementation period ranged from 34.8 (6.0%) to 155.9 g (39.0%) in the SID group and from -5.2 (-1.0%) to 52.5 g (10.2%) in the MID group.

The BV exhibited slight increase in the SID group, and this increase was significant compared with the level in the C group (P < 0.05). The erythrocyte volume increased in a manner similar to that of Hbmass, whereas PV remained unchanged over the entire observation period (Table 2).

To demonstrate the dependence of the individual Hbmass response on the baseline ferritin value, a regression analysis was performed (Fig. 3a). At ferritin levels between 10 and 25 ng·mL-1, a moderate increase in Hbmass was found. If the baseline ferritin values were less than 10 ng·mL-1, an exponential increase in Hbmass was found and the lowest ferritin value (3 ng·mL-1) was related to the greatest response (156 g). Significant but weaker relations were demonstrated for the Hbmass response versus the initial [Hb] (r = 0.60; P < 0.01) and initial MCH (r = 0.52; P < 0.05), and a nonsignificant relation was observed with the initial MCV (r = 0.38; not significant).

FIGURE 3.

FIGURE 3.

Changes in Hbmass and changes in [Hb] were moderately correlated (r = 0.67, P < 0.01) (Fig. 3b); closer relations were found for changes in Hbmass with changes in MCV (r = 0.90, P < 0.001) (Fig. 3c) and with changes in MCH (r = 0.87, P < 0.001). Regression analyses of changes in [Hb] versus changes in MCV (r = 0.69) and changes in MCH (r = 0.71) also showed significant (in both cases, P < 0.01) but weaker relations when compared with those for Hbmass. The normal variability of Hbmass and [Hb] during the observation period expressed as the coefficient of variation of the control group was 2.25% for Hbmass and 3.10% for [Hb].

In the SID group, 1715 mg of iron was stored in hemoglobin at the beginning of the intervention, and this value significantly increased to 1975 mg after the iron supplementation period. By calculating the amount of additional iron that was stored in both Hbmass and ferritin during the supplementation period, it is possible to estimate the quantity of iron absorbed from the supplements. Although both treatment groups replenished their ferritin stores during the intervention (SID, +235 ± 105 mg; MID, +202 ± 149 mg iron), the SID group also stored approximately the same amount of iron in hemoglobin. In total, the SID group incorporated 495 ± 205 mg of iron in both the ferritin and hemoglobin stores; in contrast, the amount of iron incorporated was 234 ± 141 mg in the MID group and -43 ± 148 mg in the C group (Fig. 4).

FIGURE 4.

FIGURE 4.

The V·O2max value was slightly greater than its initial value in the SID group after the supplementation period, whereas no changes occurred in the MID and C groups (Table 2). There was a significant relation between the changes in Hbmass and the changes in V·O2max (y = 2.3 x – 4.3; r = 0.65; P <= 0.01), thereby indicating that a change of 1 g in Hbmass caused a change in V·O2max of 2.3 mL·min-1 (Fig. 3d).

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DISCUSSION

This study describes the time course of the recovery of Hbmass in severely and moderately iron-deficient recreational athletes during oral iron supplementation period. The main findings are the very individual responses in the magnitude and time course of Hb production in the SID group (in the range of 35–156 g over 10–18 wk) and in the MID group (a range of -5 to 52 g over 10 wk). Close relations between the Hbmass response and MCV and MCH were found. The changes in the main iron stores (i.e., ferritin and hemoglobin) were quantified, as follows: 495 mg in the SID group and 230 mg in the MID group.

In a case study of a severely iron-deficient athlete, Hbmass was monitored to evaluate the effect of iron supplementation via i.m. iron injections combined with oral iron supplementation and Hbmass increased by 77% within 7 wk (13). In the case of an athlete with normal iron status, however, i.m. injections failed to have any effect (1). In another study, i.v. injections of iron in iron-deficient, nonanemic elite athletes yielded 5% increase in Hbmass within 8 wk whereas no effect was observed in a similar group during oral iron administration (14). Similar findings of no changes in Hbmass were reported by Friedmann et al. (12), who monitored iron-deficient, nonanemic elite junior athletes (ferritin, 16 μg·L-1; [Hb], 13.8 g·dL-1) during a 12-wk iron supplementation period (100 mg of Fe twice per day). To date, no study has monitored the effects of oral iron supplementation on Hbmass in severely iron-deficient subjects.

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Individual responses.

In this study, iron supplementation was a successful treatment for the initially iron-deficient subjects in both groups and led to normalization of all iron status markers. Even after 2 wk of supplementation, all of the SID subjects exhibited increase in Hbmass, which ranged between +2% (11 g) and +22% (76 g) and was followed by very individual further elevation of between 6% and 39%. This quick response at the beginning of oral iron supplementation is described in the literature (47) but has not been precisely quantified.

Only two of the eight subjects in the SID group met the criteria (ferritin >=25 ng·mL-1 and increase in Hbmass <20 g within the previous 3–4 wk) to stop supplementation after 10 wk, thus indicating that 10 wk was insufficient in most cases for complete recovery from severe iron deficiency. This time evolution of recovery is consistent with current guidelines, which recommend at least 3 months of oral iron supplementation (31). In contrast with the subjects studied by Friedmann et al. (12) and Garvican et al. (14), the MID subjects also exhibited small but significant mean increase in Hbmass. In two of the 14 subjects, the increase in Hbmass reached approximately 10%, which is far greater than the mean response to physiological stimuli, e.g., training at sea level (38) or even at altitude (19,43). The fact that MCV and MCH increased by 3 fL and 2 pg, respectively, in both of these subjects emphasizes recovery from iron deficiency.

The changes in Hbmass in response to iron supplementation differed among individuals and were dependent on the severity of iron deficiency, which was indicated by the initial ferritin value (Fig. 3a). In this study, relatively small responses in Hbmass were observed for initial ferritin values between 10 and 25 ng·mL-1. At lower values, however, exponential increase occurred. These data quantify the effects that occur during iron supplementation when the threshold for severe iron deficiency of 10–12 ng·mL-1 (5,20,32) is passed. In contrast, four of the eight subjects with ferritin concentration less than 10 ng·mL-1 had initial [Hb] that was slightly greater than 12.0 g·dL-1. In clinical practice, [Hb] thresholds are—in combination with ferritin—used to classify the degree of iron deficiency; for example, the World Health Organization (46) uses thresholds of 12.0 g·dL-1 for females and 13.0 g·dL-1 for males. The individual normal [Hb], however, exhibits considerable interindividual oscillations, ranging between 12.0 and 16.0 g·dL-1 in women and 13.5 and 17.5 g·dL-1 in men (25). Thus, a subject with a high normal [Hb] that decreases [Hb] because of iron deficiency (e.g., [Hb] decreases from 14.5 to 12.5 g·dL-1) is not classified as an anemic subject but may benefit from iron supplementation, which is particularly important for elite athletes. In this study, one female subject exhibited low ferritin concentration of 14 ng·mL-1 and normal [Hb] of 14.0 g·dL-1 and was therefore classified as nonanemic. To fall below the anemic threshold, she had to lose approximately 90 g of hemoglobin. During the supplementation period, however, she responded with increase of 8.3% (52.7 g) in Hbmass, proving to be anemic “for her” at the beginning of the study.

As described previously, in addition to limitations in the exact diagnosis of iron deficiency anemia using [Hb], the time course and magnitude of recovery after iron supplementation may also not be precisely quantified by [Hb]. As demonstrated in Figure 3b, significant relation existed between Hbmass and [Hb], although this relation was due only to the data of the SID group, as no relation existed within the MID group. Because the relations between the changes in Hbmass versus MCV and MCH were stronger than those for [Hb] versus MCV and MCH, we suggest Hbmass as a more sensitive parameter to monitor recovery from iron deficiency. This opinion is supported by the lower coefficient of variation for Hbmass than that for [Hb] in the control group, indicating lower biological and/or methodological variability for Hbmass.

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RCV and red cell hemoglobin content.

In the SID group, the percentage increase in RCV (10.0%) was less than the increase in Hbmass (15.2%). This difference was due to the increased incorporation of hemoglobin into premature erythrocytes (+11.1%, MCH from 27.8 to 30.9 pg), which exceeded the increase in MCV (+5.2%, from 84.5 to 88.9 fL); both effects are normal responses that occur when iron stores refill. Because PV did not change, BV was only slightly affected. Because of the increased erythrocyte count, the augmented red cell hemoglobin content and the almost unchanged BV the hemoglobin concentration increased in the SID group and also in the MID group, but to a much lower degree.

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Aftereffects.

Twelve weeks after the supplementation period, mean Hbmass remained constant in the two groups whereas the ferritin concentration significantly decreased in the SID group (Table 2). This indicates that in early iron deficiency, iron stores are depleted before erythropoietic activity is affected. As demonstrated previously, severe interference with erythropoiesis occurs with very low ferritin levels. This is demonstrated by the subject who lost the greatest amount of Hbmass (approximately 50 g), which was accompanied by reduction in ferritin to 4 ng·mL-1.

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Iron incorporation.

Knowing the absolute Hbmass of a subject and the change in Hbmass during the iron supplementation period allows for exact quantification of the iron that is incorporated into hemoglobin molecules and, therefore, the success of iron therapy. A previous study (45) reported that the amount of nonhemoglobin body iron can be estimated using the equivalence of 1 ng·mL-1 of ferritin and 8 mg of iron. Consistent with Cook et al. (8), who found 309 ± 346 mg of iron in body stores in females, we found a very similar mean value for all three groups (315 ± 125 mg) at the end of the supplementation period. The total iron content can be calculated by accounting for the storage of 10% of the iron content in myoglobin and the involvement of 2% of the iron content in electron transport, antioxidant enzymes, and DNA replication (31). At the end of the supplementation period, the total iron content did not differ between the groups. The mean of all three groups was 2.6 g; 76% (1930 mg) was stored in hemoglobin, and 12% (315 mg) was stored in ferritin. These results closely coincide with previously reported values (29).

In the SID group, the increase in ferritin concentration paralleled the change in Hbmass, thus indicating similar time evolution for filling the two main iron stores of the body. Neglecting the baseline iron absorption rate of 1–3 mg·d-1 (36), which is counterbalanced by a similar daily excretion rate, approximately 210 mg of additional iron was resorbed from the supplemented iron dose during the first 2 wk and was equally distributed between the ferritin (95 mg) and hemoglobin (115 mg) stores. The amount of additional iron that was resorbed during the entire supplementation period was approximately 500 mg, and the amounts of that iron distributed to ferritin and hemoglobin were 235 mg and 260 mg, respectively (Fig. 4).

The amount of incorporated iron in the MID group (230 mg) was approximately 50% less than that in the SID group and was primarily used to fill the ferritin store (+200 mg) rather than for incorporation into hemoglobin. Although no previous data regarding iron incorporation into the hemoglobin of MID subjects are available, the increase in the ferritin iron store in this study is consistent with data from the literature, in which an 88- to 240-mg increase in ferritin iron is reported (31).

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Aerobic capacity.

Although it is well known that iron deficiency anemia impairs endurance exercise performance (e.g., ref. 3), the effect of iron supplementation on V·O2max and performance, particularly for borderline iron deficiency, is debated upon. Some studies have demonstrated significant increase in V·O2max (12,26,27) or endurance exercise performance (35) after oral iron supplementation, and some studies have failed to demonstrate such an effect on V·O2max (10,24) or performance (22,30,32,33). Furthermore, in this study, we did not detect any increase in V·O2max in the MID group. Similarly, in a very recent study, Garvican et al. (14) did not find any change in V·O2max or performance in nonanemic athletes after oral iron supplementation but did find significant improvement in a similar group after i.v. iron injection (an increase in V·O2max of 1.9%); thus, they concluded that i.v. supplementation was the superior therapy.

In the SID group, however, V·O2max clearly increased by 7.4%, which is consistent with previous data (39) and was closely related to the elevated Hbmass. The slope of the regression line indicates that a change in Hbmass by 1 g as a result of the iron supplementation is associated with an increase of 2.3 mL·min-1 in V·O2max. This number is less than a previously reported value (39) for elite athletes (4 mL·min-1). This difference might result from the subjects in our study being recreational athletes, for whom factors other than Hbmass, e.g., mitochondrial oxidative capacity, may be the primary limiting factors for V·O2max (44). The two subjects with the highest training level also showed the greatest response in V·O2max related to 1 g of Hbmass (5.9 and 5.2 mL·min-1). We therefore suggest that the recovery in Hbmass resulting from iron supplementation has a high effect on aerobic capacity, particularly in well-trained endurance athletes.

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Possible limitations of the study.

One limitation of the study is that menstrual blood loss was not considered in our calculations. However, the occurrence and intensity of menstruation were documented by questionnaires and did not obviously affect the data. The iron loss during normal menstruation is 0.5–1 mg·d-1 (11,28), and the mean iron incorporation in the SID group during the supplementation period was 8 mg; therefore, the effect of menstruation did not severely interfere with iron balance.

The dietary intake of iron was not controlled. The test subjects were advised to maintain their usual diet; therefore, we did not expect a significant dietary effect on our results.

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CONCLUSIONS

Hbmass is a very sensitive tool for monitoring the magnitude of iron deficiency anemia and the time evolution of recovery during and after iron supplementation periods. Because [Hb] is acutely and chronically influenced by changes in PV, small changes in Hbmass may not be detected. We therefore recommend that regular measurements of Hbmass be used in addition to [Hb] and ferritin measurements for the early detection of iron deficiency, when the optimal Hbmass is known, and for monitoring the effectiveness of iron supplementation.

This project was funded by the World Anti-Doping Agency (08E09WS).

The authors declare no conflicts of interest. The results of the present study do not constitute endorsement by the American College of Sports Medicine.

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REFERENCES

1. Ashenden MJ, Fricker PA, Ryan RK, Morrison NK, Dobson GP, Hahn AG. The haematological response to an iron injection amongst female athletes. Int J Sports Med. 1998; 19 (7): 474–8.
2. Berres F, Ulmer H-V, Lamberty M. Calculating of total body fat from skinfold thickness by using an age corrected formula. In: Eur J Physiol. 1980; 384 (R 35 Suppl).
3. Celsing F, Ekblom B. Anemia causes a relative decrease in blood lactate concentration during exercise. Eur J Appl Physiol Occup Physiol. 1986; 55 (1): 74–8.
4. Chaplin H Jr, Mollison PL, Vetter H. The body/venous hematocrit ratio: its constancy over a wide hematocrit range. J Clin Invest. 1953; 32 (12): 1309–16.
5. Clark SF. Iron deficiency anemia: diagnosis and management. Curr Opin Gastroenterol. 2009; 25 (2): 122–8.
6. Clement DB, Asmundson RC. Nutritional intake and hematological parameters in endurance runners. Physician Sports Med. 1982; 10: 37–43.
7. Cook JD. Diagnosis and management of iron-deficiency anaemia. Best Pract Res Clin Haematol. 2005; 18 (2): 319–32.
8. Cook JD, Flowers CH, Skikne BS. The quantitative assessment of body iron. Blood. 2003; 101 (9): 3359–64.
9. Fallon KE. Screening for haematological and iron-related abnormalities in elite athletes-analysis of 576 cases. J Sci Med Sport. 2008; 11 (3): 329–36.
10. Fogelholm M, Jaakkola L, Lampisjärvi T. Effects of iron supplementation in female athletes with low serum ferritin concentration. Int J Sports Med. 1992; 13 (2): 158–62.
11. Fraser IS, Warner P, Marantos PA. Estimating menstrual blood loss in women with normal and excessive menstrual fluid volume. Obstet Gynecol. 2001; 98 (5 Pt 1): 806–14.
12. Friedmann B, Weller E, Mairbaurl H, Bartsch P. Effects of iron repletion on blood volume and performance capacity in young athletes. Med Sci Sports Exerc. 2001; 33 (5): 741–6.
13. Garvican LA, Lobigs L, Telford R, Fallon K, Gore CJ. Haemoglobin mass in an anaemic female endurance runner before and after iron supplementation. Int J Sports Physiol Perform. 2011; 6 (1): 137–40.
14. Garvican LA, Saunders PU, Cardoso T, et al. Intravenous iron supplementation in distance runners with low or suboptimal ferritin. Med Sci Sports Exerc. 2014; 46 (2): 376–85.
15. Garza D, Shrier I, Kohl HW 3rd, Ford P, Brown M, Matheson GO. The clinical value of serum ferritin tests in endurance athletes. Clin J Sport Med. 1997; 7 (1): 46–53.
16. Gillen CM, Lee R, Mack GW, Tomaselli CM, Nishiyasu T, Nadel ER. Plasma volume expansion in humans after a single intense exercise protocol. J Appl Physiol (1985). 1991; 71: 1914–20.
17. Gore CJ, Bourdon PC, Woolford SM, Ostler LM, Eastwood A, Scroop GC. Time and sample site dependency of the optimized CO-rebreathing method. Med Sci Sports Exerc. 2006; 38 (6): 1187–93.
18. Gore CJ, Scroop GC, Marker JD, Catcheside PG. Plasma volume, osmolarity, total protein and electrolytes during treadmill running and cycle ergometer exercise. Eur J Appl Physiol Occup Physiol. 1992; 65: 302–10.
19. Gore CJ, Sharpe K, Garvican-Lewis LA, et al. Altitude training and haemoglobin mass from the optimised carbon monoxide re-breathing method determined by a meta-analysis. Br J Sports Med. 2013; (47 Suppl 1): i31–9.
20. Heinrich H. Falsely low normal values for serum ferritin? Clin Chem. 1981; 27 (5): 768–9.
21. Hopkins W. A spreadsheet for analysis of straightforward controlled trials. A new view of statistics. 2012 [cited 2013 November 21]. Available from: http://www.newstats.org/otherrems.html.
22. Karamizrak SO, Islegen C, Varol SR, et al. Evaluation of iron metabolism indices and their relation with physical work capacity in athletes. Br J Sports Med. 1996; 30 (1): 15–9.
23. Kasprowicz K, Ziemann E, Ratkowski W, et al. Running a 100-km ultra-marathon induces an inflammatory response but does not raise the level of the plasma iron-regulatory protein hepcidin. J Sports Med Phys Fitness. 2013; 53 (5): 533–7.
24. Klingshirn LA, Pate RR, Bourque SP, Davis JM, Sargent RG. Effect of iron supplementation on endurance capacity in iron-depleted female runners. Med Sci Sports Exerc. 1992; 24 (7): 819–24.
25. Kratz A, Ferraro M, Sluss PM, Lewandrowski KB. Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Laboratory reference values. N Engl J Med. 2004; 351 (15): 1548–63.
26. LaManca JJ, Haymes EM. Effects of iron repletion on VO2max, endurance, and blood lactate in women. Med Sci Sports Exerc. 1993; 25 (12): 1386–92.
27. Magazanik A, Weinstein Y, Abarbanel J, et al. Effect of an iron supplement on body iron status and aerobic capacity of young training women. Eur J Appl Physiol Occup Physiol. 1991; 62 (5): 317–23.
28. Mettler S. Ferrum—a mineral in sports. Schweizerische Zeitschrift für Sportmedizin und Sporttraumatologie. 2004; 52 (3): 105–14.
29. Munoz M, Villar I, Garcia-Erce JA. An update on iron physiology. World J Gastroenterol. 2009; 15 (37): 4617–26.
30. Newhouse IJ, Clement DB, Taunton JE, McKenzie DC. The effects of prelatent/latent iron deficiency on physical work capacity. Med Sci Sports Exerc. 1989; 21 (3): 263–8.
31. Nielsen P, Nachtigall D. Iron supplementation in athletes. Current recommendations. Sports Med. 1998; 26 (4): 207–16.
32. Peeling P, Blee T, Goodman C, et al. Effect of iron injections on aerobic-exercise performance of iron-depleted female athletes. Int J Sport Nutr Exerc Metab. 2007; 17 (3): 221–31.
33. Powell PD, Tucker A. Iron supplementation and running performance in female cross-country runners. Int J Sports Med. 1991; 12 (5): 462–7.
34. Prommer N, Schmidt W. Loss of CO from the intravascular bed and its impact on the optimised CO-rebreathing method. Eur J Appl Physiol. 2007; 100 (4): 383–91.
35. Rowland TW, Deisroth MB, Green GM, Kelleher JF. The effect of iron therapy on the exercise capacity of nonanemic iron-deficient adolescent runners. Am J Dis Child. 1988; 142 (2): 165–9.
36. Santiago P. Ferrous versus ferric oral iron formulations for the treatment of iron deficiency: a clinical overview. ScientificWorldJournal. 2012; 2012: 1–5. doi: 10.1100/2012/846824.
37. Schmidt W, Prommer N. The optimised CO-rebreathing method: a new tool to determine total haemoglobin mass routinely. Eur J Appl Physiol. 2005; 95: 486–95.
38. Schmidt W, Prommer N. Effects of various training modalities on blood volume. Scand J Med Sci Sports. 2008; 18 (1 Suppl): 57–69.
39. Schmidt W, Prommer N. Impact of alterations in total hemoglobin mass on VO2max. Exerc Sport Sci Rev. 2010; 38 (2): 68–75.
40. Skikne BS, Flowers CH, Cook JD. Serum transferrin receptor: a quantitative measure of tissue iron deficiency. Blood. 1990; 75 (9): 1870–6.
41. Suominen P, Punnonen K, Rajamaki A, Irjala K. Serum transferrin receptor and transferrin receptor-ferritin index identify healthy subjects with subclinical iron deficits. Blood. 1998; 92 (8): 2934–9.
42. Valberg LS. Plasma ferritin concentrations: their clinical significance and relevance to patient care. Can Med Assoc J. 1980; 122 (11): 1240–8.
43. Wachsmuth NB, Völzke C, Prommer N, et al. The effects of classic altitude training on hemoglobin mass in swimmers. Eur J Appl Physiol. 2012; 113 (5): 1199–211.
44. Wagner PD. New ideas on limitations to VO2max. Exerc Sport Sci Rev. 2000; 28 (1): 10–4.
45. Walters GO, Miller FM, Worwood M. Serum ferritin concentration and iron stores in normal subjects. J Clin Pathol. 1973; 26 (10): 770–2.
46. World Health Organization. Iron deficiency anaemia assesment, prevention, and control. A guide for programme managers. 2001 [cited 2014 January 14]. Available from: http://www.who.int/nutrition/publications/micronutrients/anaemia_iron_deficiency/en/.
47. Worwood M. Serum ferritin. CRC Crit Rev Clin Lab Sci. 1979; 10 (2): 171–204.
Keywords:

IRON DEFICIENCY ANEMIA; IRON STORES; ERYTHROPOIESIS; V·O2MAX; IRON (II)-GLYCINE-SULFATE

© 2015 American College of Sports Medicine