Human Nutrition and Metabolism

Controlled High Meat Diets Do Not Affect Calcium Retention or Indices

of Bone Status in Healthy Postmenopausal Women1,2
Zamzam K. (Fariba) Roughead,3 LuAnn K. Johnson, Glenn I. Lykken* and Janet R. Hunt
Grand Forks Human Nutrition Research Center, Agricultural Research Service, U.S. Department of
Agriculture, Grand Forks, ND 58202-9034 and *Physics Department, University of North Dakota,
Grand Forks, ND 58202

KEY WORDS:

calcium

meat protein

postmenopausal

The high protein content of the Western diet is often cited

as a risk factor for osteoporosis or bone fractures (17). The
calciuretic effect of ingesting purified proteins can result in a
negative calcium balance (8 11). However, common protein
sources such as meat also provide phosphorus, which reduces
urinary calcium (12,13), and adding meat to the diet may have
little or no effect on calcium balance (14 16). Although
human balance results suggest that dietary phosphorus reduces
protein-induced hypercalciuria, Kerstetter and Allen (17) in a
review of 15 studies conclude that the presence of phosphorus
may not fully restore calcium balance and project that a
negative calcium balance of only 2530 mg/d could reduce
body calcium (primarily from bone) by 10% per decade (17).
Unfortunately, the balance methodology used in such studies (17) is insensitive because of the variability associated with
fecal collections and the inability to distinguish the endogenously excreted calcium from the unabsorbed fecal calcium.

osteoporosis

renal acid

Only four reports have addressed the effect of meat protein on

calcium isotope retention, as measured in blood or stools
(15,18 20). Spencer et al. (15,18) have concluded that meat
protein does not affect calcium retention, although this conclusion is based on studies with few subjects. Breslau et al. (19)
have reported that animal compared with vegetable protein
increases urinary calcium without influencing calcium absorption, but the phosphorus content of the diet was kept constant
and the calcium isotope was not administered with the diet.
Heaney (20) has reported that, although coingested phosphorus counterbalances the calciuretic effect of protein, it increases intestinal calcium excretion; thus, there is no net effect
of phosphorus on calcium balance. However, this finding relies
on a statistical correction for the variability in phosphorus
intake, rather than on a direct control of phosphorus in the
diet.
Cross-sectional studies have only contributed to the controversy about dietary protein and bone health. Dietary protein, assessed from self-reported food intakes, has been correlated positively (2123), negatively (24,25) or not at all
(26,27) with bone mineral density and/or fracture risks.
To test whether increasing dietary protein from meat adversely affects calcium retention and bone metabolism,
healthy postmenopausal women were studied using controlled
high and low meat diets for 8 wk each in a randomized
crossover design. Radiotracer and whole body scintillation

1
Supported primarily by the U.S. Department of Agriculture, with additional
support from the North Dakota Beef Commission.
2
Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does
not imply its approval to the exclusion of other products that may also be suitable.
The U.S. Department of Agriculture, Agricultural Research Service, Northern
Plains Area, is an equal opportunity/affirmative action employer and all agency
services are available without discrimination.
3
To whom correspondence should be addressed.
E-mail: froughea@gfhnrc.ars.usda.gov.

0022-3166/03 $3.00 2003 American Society for Nutritional Sciences.

Manuscript received 18 October 2002. Initial review completed 7 November 2002. Revision accepted 21 November 2002.
1020

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ABSTRACT Calcium balance is decreased by an increased intake of purified proteins, although the effects of
common dietary sources of protein (like meat) on calcium economy remain controversial. We compared the effects
of several weeks of controlled high and low meat diets on body calcium retention, using sensitive radiotracer and
whole body scintillation counting methodology. Healthy postmenopausal women (n 15) consumed diets with
similar calcium content (600 mg), but either low or high in meat (12 vs. 20% of energy as protein) for 8 wk each,
in a randomized crossover design. After 4 wk of equilibration of each diet, calcium retention was measured by
extrinsically labeling the 2-d menu with 47Ca, followed by whole body scintillation counting for 28 d. Urinary and
blood indicators of bone metabolism were also determined for each diet. Calcium retention was not different during
the high and low meat dietary periods (d 28, mean pooled SD: 17.1 and 15.6%, 0.6%, respectively; P 0.09).
An initially higher renal acid excretion in subjects consuming the high meat compared with the low meat diet
decreased significantly with time. The diets did not affect urinary calcium loss or indicators of bone metabolism.
In conclusion, under controlled conditions, a high meat compared with a low meat diet for 8 wk did not affect
calcium retention or biomarkers of bone metabolism in healthy postmenopausal women. Calcium retention is not
reduced when subjects consume a high protein diet from common dietary sources such as meat. J. Nutr. 133:
1020 1026, 2003.

MEAT PROTEIN AND CALCIUM RETENTION

counting methodology enabled sensitive measurement of total

calcium retention, a net function of differences in absorption,
urinary excretion, intestinal excretion or any other routes of
calcium loss.
SUBJECTS AND METHODS

4
Abbreviations used: BMI, body mass index; cAMP, cyclic adenosine monophosphate; GFR, glomerular filtration rate; IGF-1, insulin-like growth factor 1;
PTH, parathyroid hormone.

TABLE 1
Ingredient differences and nutrient composition of controlled
high and low meat diets consumed by healthy
postmenopausal women for 8 wk each in a
crossover design1,2
High meat
Diet ingredient differences, g/d
Pork
Turkey breast
Beef round
Ham
Chicken breast
Pasta, cooked
Rice, cooked
Pretzels
Croissant
Bagel
Chow mein noodles
Oven-fried potatoes
Margarine, soft
Nutrient composition
Protein, % of energy
g/kg body weight
g/d
Fat, % of energy
Carbohydrate, % of energy
Calcium, mg
Phosphorus, mg
Sodium, mg
Potassium, mg
Sulfur, mg

31
30
117
34
40

20
1.62 0.15
117
31
50
596 19
1679 108
3601 278
2887 339
1789 125

Low meat

70
39
8
5
18
8
26
4
12
0.94 0.09
68
31
60
617 20
1266 90
3243 158
2249 290
1222 55

1 Based on the participants average energy intake of 9.6 MJ (2286

272 kcal). Protein, carbohydrate and fat were calculated from U.S.
Department of Agriculture food composition data. Minerals (mean SD)
were analyzed as described in the text.
2 Supplemented with a daily chewable multivitamin tablet containing vitamin A (800 g), ascorbic acid (60 mg), cholecalciferol (10 g),
-tocopherol (30 IU), pyridoxine (2 mg), cyanocobalamin (6 g), thiamin
(1.5 mg), riboflavin (1.7 mg), niacin (20 mg), folic acid (400 g) and
pantothenic acid (10 mg).

scintillation counting (29), with adjustments to disregard 47Sc activity. The 47Ca isotope was obtained by neutron activation (University
of Missouri, Columbia, MO) of stable 46Ca (as calcium carbonate,
30.89% enriched; Oak Ridge National Research Laboratory, TN).
The custom-made scintillation counter (30) detects gamma emissions
with 32 crystal NaI(T1) detectors (10 10 41 cm each), arranged
in two planes above and below a bed.
After 4 wk of dietary equilibration, the 2-d menu was labeled with
148 kBq (4 Ci) 47Ca (4 g elemental calcium). Because of the
concern that ingested calcium from some dietary sources may not
form a common absorptive pool (31), both diets were designed with
milk as the primary source of calcium. For each meal, the tracer was
mixed with milk and allowed to equilibrate overnight. The specific
activity (ratio of 47Ca to elemental calcium) was constant for all
meals for each individual. The energy provided by the radiolabeled
meals was constant between the two diets for each participant. All
labeled meals were consumed at the research center.
The initial total body activity was determined from the whole
body count 13 h after the first labeled meal (before any isotope was
excreted), divided by the fraction of the total activity in the first
meal. Whole body calcium retention was monitored for 28 d. Activity
was corrected to the midpoint of the 2nd d of labeled meals and
adjusted for background and minor fluctuations in the measurement
of a 47Ca standard distributed in water (32). The precision of the
whole body counting measurements was 1.4%.
Analyses. The subjects provided total 48-h urine collections during wk 3, 5 and 8 of each dietary period. Blood samples were drawn
at wk 4 and 8 of each dietary period. Calcium in the urine and

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Postmenopausal women for this study were recruited through

public advertising (television, newspapers) and by direct mailings.
The women were selected after an interview and blood analysis and
qualified to enter the study with the following conditions: they were
age 50 75 y; it was at least 3 y since last menses (follicle stimulating
hormone 40 IU/L); they were nonsmokers; they had no apparent
underlying disease; they had bone mineral density indicating a femoral neck Z score 2, as determined by dual-energy X-ray absorptiometry (Hologic QDR 2000, Waltham, MA); they had normal
thyroid, liver and kidney functions; they were willing to discontinue
any nutritional supplements as soon as their applications were received; they did not regularly use any medications (except for hormone replacement therapy); and had body mass index (BMI)4 32
kg/m2.
The study was approved by the University of North Dakotas
Radioactive Drug Research Committee and Institutional Review
Board, and by the USDA Radiological Safety Office. The study was
explained verbally and in writing by the investigators and written
informed consent was given by each woman.
Of 18 women enrolled, one was dismissed for illness, and two were
excluded for noncompliance. The remaining 15 (14 white, one
Asian; 6 using hormone replacement therapy) were age 60.5 7.8 y
(mean SD), with BMI of 26.5 4.0 kg/m2 and femoral bone
mineral density of 0.707 0.121 g/cm2. As estimated from 3-d food
records, their calcium and protein intakes before the study were 774
279 mg/d and 71 14 g/d, respectively.
Diets. Weighed high and low meat diets using ordinary foods in
a 2-d menu cycle (Table 1) provided similar amounts of calcium
(600 mg/d) and energy (9.6 0.9 MJ, or 2286 222 kcal), with
20 and 12% of energy as protein (1.62 0.15 and 0.94 0.09 g/kg
body weight), and 297 and 45 g/d of meat (10.5 and 1.6 oz/d),
respectively (Table 1). The amount of calcium in the diet was chosen
to mimic typical intakes by postmenopausal women in the United
States (599 16 mg/d) (28). Furthermore, the effects of dietary
protein on calcium retention were expected to be more pronounced
when calcium intake was inadequate. To keep the diets isoenergetic
without substantially increasing the phytate content, added vegetable
fat and low fiber complex carbohydrates were used as substitutes for
meat. To maintain body weights, energy intakes were adjusted by
proportionally changing the amounts of all foods. Coffee, tea and
artificially sweetened, noncola carbonated beverages (containing citric acid rather than phosphoric acid) were individualized, limited to
two total servings daily and kept constant. Similarly, salt consumption was individualized and kept constant during the two dietary
periods; mean sodium intake was 3759 and 3782 mg during the low
and high meat dietary periods, respectively. City water and chewing
gum were consumed as desired, after analyses indicating minimal
mineral content. The participants were given a list of approved
over-the-counter medications, toothpaste and dental adhesives that
contained minimal amounts of calcium or other minerals. All diet
ingredients except water were weighed to 1% accuracy, prepared and
provided to the participants. The participants consumed the food
quantitatively, with the aid of spatulas and rinse bottles, consuming
one meal at the research center on weekdays, and the remaining foods
elsewhere. Because the study was scheduled during the winter months
in North Dakota (latitude 47.5 N), all participants received a daily
multivitamin supplement that included 20 g of cholecalciferol (see
footnote Table 1).
Calcium retention measurements with 47Ca. Dietary calcium
retention was measured with a 47Ca radiotracer and whole body

1021

ROUGHEAD ET AL.

1022

y 1ek1t 2ek2t
where y represents 47Ca retention as a percentage of the administered
dose, t represents the time (in h) and coefficients 1 and 2 represent
the turnover of the radiotracer (%) at rates k1 and k2, respectively.
Because of the early delay in isotope elimination related to the
gastrointestinal transit of unabsorbed isotope, calcium retention data
for d 25 were not included in the model. The percentage of 47Ca
absorbed was separately estimated from the y-intercept of the linear
portion (d 9 28) of a semilogarithmic retention plot [ln (% 47Ca
retained) vs. time].
Diet and sequence effects were evaluated using repeated-measures
ANOVA followed by Tukeys contrasts (41). Variances in the data
were expressed as pooled SD from the ANOVA. When data were not
normally distributed, they were logarithmically transformed and both
transformed and geometric means are reported. Using two-tailed
probabilities, P 0.05 was considered significant. The diet sequence
did not significantly affect any of the reported variables.

RESULTS
Whole body retention and intestinal absorption of calcium. Calcium retention was not adversely affected by the
high meat diet; rather, calcium retention was similar, if not
slightly improved, compared with that of the low meat diet
(Fig. 1, Table 2). As indicated by the two-component exponential model, about 74% of the calcium tracer was cleared
rapidly from the body. This represented early elimination of
the unabsorbed isotope as well as short-term endogenous urinary and fecal excretion, with biological half-lives of 1.7 and
1.5 d, associated with the high and low meat diets, respectively
(Table 2). The remaining calcium tracer was eliminated less
rapidly with biological half-lives of 56 and 40 d in subjects
consuming the high compared with the low meat diets, respectively (P 0.04; Table 2). Thus, loss of calcium tracer
from the body was slightly but significantly greater in subjects
consuming the low meat compared with the high meat diet.
Other measures from the exponential model of calcium retention were not affected by diet (Table 2).
Whole body calcium retention was not affected by diet at

FIGURE 1 Whole body retention of a calcium radiotracer (47Ca)

administered to healthy postmenopausal women at the 4-wk midpoints
of controlled high and low meat diets consumed for 8 wk each in a
crossover design. (A) Whole body calcium retention, as percent of dose
(means SEM, n 15) over time. Calcium retention was not different in
subjects consuming the high and low meat diets at all weekly time points
tested (see Table 2). (B) Two-component exponential models that fit the
data for 14 of the 15 subjects. These group models (y 74.5e0.0188t
25.7e0.0006t for high meat and y 74.0e0.0211t 26.0e0.0008t for low
meat, where t is expressed in h) use the mean coefficients from the
modeled retention curves for each individual. Because of the variability in
the early elimination of the isotope, calcium retention data for the first 2 to
5 d were omitted before the modeling of the data. The retention curves,
modeled separately for individuals consuming each diet, had coefficients
of determination (R2) of 0.97 to 0.99. The high protein diet was associated
with a slightly longer biological half-life of the longer-term component of
calcium retention (P 0.05; see Table 3).

any of the weekly time points tested. On d 28, 17.1 and 15.6%
(0.6) of the calcium tracer was retained from the high and
low meat diets, respectively (P 0.09; Table 2, Fig. 1).
Because the tendency for a slight difference in retention while
consuming the two diets was apparent as early as 5 d after
isotope administration (Fig. 1A), this possible divergence in
the calcium retention observed on d 28 cannot be clearly
attributed to differences in excretory and absorption pathways.
Percentage calcium absorption was not different in subjects consuming the high compared with low meat diets (29.9
vs. 28.4%, Table 2), with about 178 and 175 mg calcium
absorbed daily, respectively. Calcium retention also was not
different in the two diets in women using hormone replacement therapy (n 6). Although the mean calcium retention
in subjects consuming the two diets tended to be slightly lower
in these women, compared with the women not using hormone replacement (n 9), this difference was not significant
(d 28: 14.4 and 18.0%, 2.0, respectively; P 0.08). At least
12 subjects per group would have been needed (with 90%
power) to detect a 5 percentage point difference in calcium
retention between the hormone replacement therapy users and
nonusers.
Biomarkers of bone metabolism. Diet did not affect biomarkers of bone formation (serum bone-specific alkaline
phosphatase, osteocalcin and IGF-1) or of bone resorption
(serum tartrate-resistant acid phosphatase, urinary N-telopeptides) (Tables 3 and 4). Several other indices of bone and
mineral metabolism, such as intact PTH (and urinary cAMP,
an indicator of PTH secretion), estradiol, 25-hydroxy-vitamin

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acid-digested diet aliquots (33) was determined by inductively coupled argon plasma emission spectrophotometry. Mean (SD) measurements were 98 4% of certified values for calcium in a standard
reference material (Typical Diet, 1548b, U.S. National Institute of
Standards and Technology).
Urinary ammonium was determined colorimetrically (34)
(Raichem; Hemagen Diagnostics, San Diego, CA). Titratable acidity
was determined in undiluted urine by titrating to pH 7.40 with 0.1
mol/L NaOH. Free organic acids were measured by the method of
Van Slyke and Palmer (35) as modified by Lemann et al. (36).
Urinary sulfates were determined turbidometrically (37). Enzymelinked immunoassays were used to determine serum bone-specific
alkaline phosphatase (Metra Biosystems, Mountain View, CA), estradiol (Abbott Laboratories, Abbott Park, IL) and urinary cyclic
adenosine monophosphate (cAMP; Biomedical Technology,
Shoughton, MA). Serum tartrate-resistant acid phosphatase activity
was determined using -naphthylphosphate and diazotized-2-amino5-chlorotoluene as substrates (38). Creatinine clearance was calculated from serum and urinary creatinine, which were measured using
alkaline picric acid (39). Serum parathyroid hormone (PTH), calcitonin, osteocalcin (Diasorin, Stillwater, MN) and 25-hydroxy vitamin D (INSTAR, Stillwater, MN) were determined by radioimmunoassay. Serum insulin-like growth factor 1 (IGF-1; Diagnostic
Systems Laboratory, Webster, TX) and urinary N-telopeptides (Ostex, Seattle, WA) were determined by enzyme-linked immunosorbant assays. Plasma-ionized calcium was measured with an electrode
(Nova 8 Electrolyte Analyzer, Waltham, MA) (40).
Statistics. Individual 47Ca retention data were modeled with a
two-component exponential equation:

MEAT PROTEIN AND CALCIUM RETENTION

TABLE 2
Whole body retention of a calcium radiotracer (47Ca)
administered to healthy postmenopausal women at the 4-wk
midpoints of controlled high and low meat diets consumed
for 8 wk each in a crossover design1,2
Variable

1, % of dose
2, % of dose
ln (T1), d
ln (T2), d

Low meat

74.5
25.7
0.51
(1.7, 1.02.8)
4.0
(56.2, 26352)

74.0
26.0
0.38
(1.5, 1.02.8)
3.7*
(40, 24112)

28.6
21.5
18.9
17.1
29.9

26.8
20.6
17.8
15.6
28.4

Pooled

SD

3.1
2.9
0.3
0.4
3.4
2.4
2.1
2.1
2.7

1 Values are arithmetic means or (geometric means, range), n 14.

All times are relative to the time of administration of the isotope.
* Different from the high meat period, P 0.04.
2 A two-component exponential model fit the data (R2 of 0.97 to
0.99 for models of individuals consuming each diet). One subjects data
could not be modeled using the two-component exponential equation.
1 and 2 represent the percentage of the isotope having a biological
half-life of T1 and T2, respectively.
3 Intestinal calcium absorption, as % of dose, was estimated by
using the linear portion (d 9 28 after 47Ca administration) of a semilogarithmic retention plot [ln (% retention vs. time)] and extrapolating
back to the time of tracer administration.

D and plasma ionized calcium, were also unaffected (Tables 3

and 4). Although urinary hydroxyproline concentration was
greater when subjects consumed a high meat diet than when
they consumed a low meat diet (P 0.001), this was likely
attributable to the greater dietary collagen content (39),
rather than to bone resorption (Table 4).
Urine composition. Urinary acidity adapted with time in
subjects consuming the two diets. Although the urinary pH

was lower after 3 wk when women consumed the high compared with the low meat diet (5.68 and 6.02), this difference
decreased with time, from 0.34 to 0.02 and 0.03 pH units at 3,
5 and 8 wk, respectively (P 0.05; Table 4). Similarly, a
greater urinary titratable acidity when women consumed the
high compared with the low meat diet was moderated by time
with differences of 15.6, 11.1 and 4.9 mEq/d, at 3, 5 and 8 wk
(P 0.05; Table 4), respectively. This continually diminishing difference suggests that further adaptation may have occurred beyond 8 wk. Urinary free organic acid excretion,
representing compounds such as citric, lactic and acetic acids,
was marginally, but not significantly, higher (P 0.07) when
women consumed the high meat diet (Table 4), and this
difference was also smallest at 8 wk.
As expected, the higher exogenous substrates provided by
the high compared with the low meat diet (Table 1) significantly increased urinary ammonium, sulfate, creatinine and
phosphorus at all time points (Table 4). The high compared
with the low meat diet increased creatinine clearance (1.38
and 1.21 mL/s, respectively, P 0.05; Table 4), without
affecting blood creatinine concentrations (80 9 mol/L).
This likely reflected a difference in the substrate load, rather
than in the glomerular filtration rate (GFR) of these healthy
women (see Discussion).
Despite differences in urinary acidity and excreta, urinary
calcium loss between 3 and 8 wk was unaffected by diet.
Urinary calcium was 2.45 and 2.38 0.88 mmol/d in subjects
consuming the high and low meat diets, respectively, without
detectable change over time (Table 4).
DISCUSSION
These results indicate that high vs. low meat diets consumed for several weeks did not reduce whole body calcium
retention (Table 2, Fig. 1) or adversely affect biochemical
indicators of bone turnover (Tables 3 and 4). Although the
slightly greater retention when women consumed the high
meat diet (significantly longer biological half-life; Table 2, Fig.
1) was of questionable practical importance, it emphasizes that
calcium was at least as well retained in subjects consuming the
high compared with the low meat diet, and that sufficient

TABLE 3
Blood biomarkers of bone metabolism in healthy postmenopausal women consuming
controlled high and low meat diets for 8 wk each in a crossover design1
Biomarker
Tartrate-resistant acid phosphatase, nkat/L
Bone-specific alkaline phosphatase, U/L
Serum ionized Ca,2 mmol/L
ln (intact PTH),3 pmol/L
Calcitonin, mol/L
ln (25-OH vitamin D),4 nmol/L
Osteocalcin, g/L
Insulin-like growth factor 1, g/L
ln (estradiol),5 pmol/L

High meat

Low meat

72
18.1
1.21
0.32
(1.4, 0.73.5)
13.4
4.63
(103, 51210)
4.01
12.7
4.84
(127, 53300)

73
18.3
1.21
0.35
(1.4, 0.53.4)
11.9
4.64
(104, 68268)
3.94
12.8
4.81
(123, 65254)

Pooled

SD

4
1.0
0.02
0.23
3.6
0.16
0.9
1.0
0.40

1 Values are arithmetic means or (geometric means, ranges) and pooled SD from measurements at wk 4 and 8 of each dietary period, except for
serum IGF-1, which was measured only at midpoint of each dietary period (wk 4), n 15. None of the variables was affected by diet.
2 To convert the values for calcium to mg/L, multiply by 40.08.
3 To convert PTH to pg/mL, multiply by 9.5.
4 To convert 25-OH vitamin D to ng/mL, multiply by 0.401.
5 To convert the values for estradiol to pg/mL, multiply by 0.272.

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Retention, % dose
d7
d 14
d 21
d 28
Absorption,3 % dose

High meat

1023

ROUGHEAD ET AL.

1024

TABLE 4
Renal acid excretion and urinary biomarkers of bone metabolism in healthy postmenopausal women consuming
controlled high and low meat diets for 8 wk each in a crossover design1,2
High meat

Calcium, mmol/d
Phosphorus, mmol/d
Sodium, mmol/d
Potassium, mmol/d
Ammonium, mmol/d
ln (sulfate), mmol/d

ln (cAMP),4 nmol/mg creatinine

ln (N-telopeptide),4,5 nmol
BCE/mmol creatinine
Hydroxyproline, mol/d

wk 3

wk 5

wk 8

wk 3

wk 5

wk 8

2.61
22.6
117.3
42.2
42.1
3.1
(22, 1230)
8.7

5.68a
40.2a
2.40
(11, 522)
1.11
(3, 19)
3.82
(46, 22143)
76.6

2.25
19.3
116.6
43.9
40.6
3.1
(22, 1532)
9.2

5.88ab
37.4ab
2.33
(10, 626)
0.92
(3, 15)
3.71
(41, 18116)
72.8

2.47
20.3
120.9
45.6
42.5
3.1
(22, 1230)
8.8
1.38
5.89ab
35.4ab
2.26
(10, 323)
1.01
(3, 17)
3.79
(44, 27103)
71.5

2.43
15.2
109.0
32.4
32.3
2.6
(14, 917)
7.9

6.02b
24.6c
1.96
(7, 215)
1.16
(3, 125)
3.92
(50, 13128)
63.4

2.22
15.2
100.6
31.7
32.7
2.7
(15, 822)
8.1

5.90ab
26.3c
2.15
(9, 417)
0.76
(2, 0.210)
3.90
(49, 29143)
59.7

2.5
19.2
100.0
30.9
35.0
2.6
(14, 523)
7.8
1.21
5.92ab
30.5bc
2.23
(9, 342)
1.03
(3, 114)
3.83
(46, 2785)
64.5

Pooled

SD

Diet effect

0.88
5.7
15.9
5.5
6.3
0.2

NS
0.001
0.0001
0.0001
0.0001
0.0001

1.0
0.18
0.23
7.4
0.56

0.0001
0.05
0.01
0.0001
NS

0.47

NS

0.33

NS

13.4

0.001

1 Values are arithmetic means or (geometric means, ranges) and pooled SD, n 15. To convert the values to mg/d, multiply by 40.08 for calcium,
by 31.0 for phosphorus, by 23.0 for sodium, by 39.0 for potassium, and by 0.131 for hydroxyproline. To convert the values for ammonium to g/d,
multiply by 18.0; to convert the values for creatinine to g/d, multiply by 0.113.
2 Urinary measurements are from total 48-h urine composites collected during wk 3, 5 and 8 of each dietary period [except creatinine clearance,
which was measured at the end of each dietary period (wk 8)].
3 Significant diet time interaction (P 0.05). Values followed by a common letter are not significantly different as determined by Tukeys
contrasts (P 0.05). NS, not significant.
4 Because urinary creatinine excretion was higher when subjects consumed the high meat diet, the cAMP and N-telopeptide data were tested with
and without correction for urinary creatinine, with no change in the statistical results.
5 BCE, bone collagen equivalents.

statistical power existed to detect a more meaningful difference had it occurred. Prospective power analysis estimated
that 12 subjects were needed to detect a difference of 5
percentage points in the percentage absorption (pooled SD of
2.5, 90% power, 0.05, two-tailed test). Data from 14
subjects did not show the hypothesized difference.
The calciuretic effect of dietary protein has been partially
attributed to an increased GFR (17,42). Increased protein
intake from isolated protein sources has been repeatedly reported to increase GFR (10,11,13,42,43) by 10 20%, as measured by creatinine clearance, a magnitude similar to the
present results with meat as the protein source (Table 4).
However, creatinine clearance is not a direct measure of GFR,
given that small quantities of creatinine are reabsorbed or
secreted in the renal tubules, and because a stable intake and
endogenous production of creatinine is assumed (39). The
latter assumption cannot be applied in the present study because meat was an exogenous source of creatinine. The assumption also may not be applicable with an increased intake
of isolated proteins, which may increase endogenous creatinine production (44,45). Unfortunately, previous studies of
increased GFR with increased intake of isolated proteins
(10,11,13,42,43) did not report total creatinine excretion. In
the present study, the apparent greater creatinine clearance
associated with the high meat diet was proportional to a
comparable increase in urinary creatinine excretion, without
affecting urinary calcium (Table 4).
The calciuretic effect of protein has also been related to an
increase in renal acid load, which in this study adapted over
time. The potential renal acid load when women consumed
the high meat diet was estimated to be about twice that when

women consumed the low meat diet (60 vs. 30 mEq/d) using
the method of Remer and Manz (46), which assumes constant
absorption of dietary minerals and electrolytes (without allowing for adaptation over time). The higher dietary acid load
during the high meat dietary period was reflected in the higher
initial renal acid excretion; however, this difference abated
between 3 and 8 wk (Table 4). Furthermore, despite this early
difference in urinary acid excretion and the continuing greater
excretion of sulfate and ammonium, urinary calcium excretion
was not different at any time point tested. Similarly, in previous studies, although addition of meat to the diet has been
associated with hypercalciuria in studies of 3 (47) or 15 d (48),
Spencer and co-workers (15) have reported adaptation that
almost completely reversed an initial hypercalciuria in a single
subject studied for 72 d. Urinary calcium excretion by postmenopausal women, measured during the last 18 d of 7-wk
controlled diets (in a crossover design), does not differ between high and low meat diet periods (14). The present results
concurred with these latter studies (14,15) and suggest that if
early hypercalciuria occurred in response to the high meat
diet, it was reversed by adaptation within 3 wk, despite a more
gradual adaptation in acid excretion (Table 4). In contrast to
meat protein, with increased intakes of isolated protein sources
for 75 (42) or 95 d (10), no adaptation in renal acid excretion
or hypercalciuria was observed. This difference in adaptation
may reflect differences in the protein source.
This studys results were consistent with previous observations that coingested phosphorus offsets the hypercalciuric
effect of protein, possibly through a PTH-mediated mechanism (49), and that net calcium balance is not reduced (14
16,18,50). Our calcium retention results were in contrast to

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Creatinine, mmol/d
Creatinine clearance, mL/s
pH3
Titratable acidity,3 mEq/d
ln (free organic acid), mEq/d

Low meat

MEAT PROTEIN AND CALCIUM RETENTION

ACKNOWLEDGMENTS
We gratefully acknowledge the insightful reviews of the study
protocol by Bess Dawson-Hughes and Robert Heaney. We are also
thankful for the invaluable assistance of the staff at Grand Forks
Human Nutrition Research Center, Carol Zito, Emily Nielsen,
Brenda Hanson, Debbie Krause, Angela Scheett, Jackie Nelson and
Sandra Gallagher. We thank Steve Morris of Missouri Research
Reactor. We are deeply indebted to the study participants.

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In summary, using controlled diets and whole body counting tracer methodology with supportive clinical chemistry, this
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bodys calcium status and thus bone health.

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