Laura Myra

R Trebler G Berkowitz,

ABSTRACT

Winters, Jere The

requirements

and

Jin-Sook Haas, and

effects

and exercise Yoon, Daphne

of exercise

of riboflavin

intake

Heidi J Kalkwarf A Roe

Jayne

Chalfin

on riboflavin

clinical

and

training on

endurance

were

amined in 14 women, 50-67 y ofage, who participated wk, two-period crossover exercise study at two riboflavin 0. 1 5 ig/kJ

(0.6

exercised ergometer status

g/kcal)

20-25

and

min/d,

at 75-85% was

0.22

of their

assessed

by

g/kJ

6 d/wk,

(0.9

for 4-wk maximal

measuring

the

Subjects

KEY WORDS

Riboflavin,

women,

ygen

uptake,

erythrocyte

anaerobic

exercise,

glutathione

studies and

nutrient

requirements,

reductase,

maximal

ox-

threshold

have demonstrated physiologically

that the capacity to aerobic

exercise

(2,

3), exercise

is increasingly

promoted

Investigations required

of the dietary for the optimal

have been carried out since the onstrated that work performance 526

intake

to adapt

for older

of riboflavin

example,

studies

takes

of B vitamins

young vitamin

on work

male subjects, C to 30-35%

[including

0.038

performance.

and

continuous

changes

the onset

showed

that

and

for

ofblood

lactate

in

and of intakes an

in the maximal

8-wk

oxygen

accumulation

AT). The anaerobic threshold intensity (oxygen consumption blood lactate accumulation and

in gas exchange

( 1 4),

exercise

riboflavin)]

declines

(OBLA or anaerobic threshold, (AT) is defined as the exercise amount, or VO2) above which the associated

They

(0. 16 jg/kcal

in significant

(VO2max)

van der Beck ( 1 2) and the effect of marginal in-

restriction of specific B vitamins ofthe Dutch recommended daily

g/kJ

resulted

period, capacity

and

occur

has been

in the lungs

described

during

as an indicator

ofendurance (15, 16). These studies as well as previous studies performed by our group on the effects of exercise on riboflavin requirements in younger women motivated the current study, which was designed to determine whether riboflavin requirements are similarly inby exercise

training

in older

women

and

whether

ribo-

is retained

women.

Thus, the nutritional requirements of older women who exercise have become an important area of investigation. Both human and animal studies have demonstrated that riboflavin plays an important role in exercise-induced biochemical adaptations (4-7). Previous studies in our laboratory have shown that physical exercise increases riboflavin requirements of younger individuals (8-10). However, whether or not exercise training of older individuals increases their riboflavin requirements was not investigated before this study. vitamins

For

flavin intake affects the physiological adaptation for exercise in this sex and age group. Physical performance was evaluated both through determination of maximal oxygen uptake and AT.

by aging individuals (1). Because regular exercise by aging women confers multiple health benefits relative to risk reduction for coronary artery disease and retardation of age-related osteoporosis

deficiency.

Forty years later, van Dokkum van der Beck et al (1 3) reinvestigated

glutathione

Introduction Previous

vitamin

endurance.

creased

biochemically

clinical

reported by Keys et al (1 1) in 1945 showed that severe and prolonged restriction of B vitamins [from 0.069 zg/kJ (0.287 g/ kcal) to 3. 1 ng/kJ (0.0 1 3 zg/kcal) riboflavin] over a 27-wk period resulted in a steady decline in physical performance, notably in

Riboflavin

reductase activity coefficient (EGRAC) and urinary riboflavin excretion. Physical performance was evaluated by using a walking treadmill test to determine maximal oxygen capacity (VO2max) and anaerobic threshold by gas exchange (ATGE). Exercise significantly affected riboflavin status as EGRAC increased (P < 0.001) and riboflavin excretion decreased (P < 0.01) in both groups. VO2max increased significantly with exercise (P < 0.01). However, changes in VO2max (L/min) and ATGE with exercise training were not different in the two groups. Riboflavin requirements ofolder women increased with exercise training, but increased riboflavin intake did not enhance improvements in endurance. Am J C/in Nutr 1992;56:526-32.

older

Davies,

on a cycle

rate.

erythrocyte

ex-

in a 10intakes,

g/kcal).

periods

heart

adaptation

and

other

B

performance

ofphysical

1940s, when was impaired

it was first demby states of sub-

Am J C/in Nuir

work

l992;56:526-32.

Subjects

and

methods

Subjects Fourteen women aged 50-67 y were selected for the from a volunteer pool. Volunteers were excluded if they hematological or physical signs of health impairment; 2) normal electrocardiogram or elevation ofblood pressure

1

From the Division ofNutritional

3

Address

Cornell

with the National reprint

University,

Received Accepted Printed

requests

Dairy

in USA.

Board

Savage

and administered

Dairy Council.

to DA Roe, Division

Savage Hall, Ithaca,

May 9, 1991. for publication

an ab-

during

Sciences, Cornell University,

Hall, Ithaca, NY. 2 Supported in part by the National cooperation

study had 1)

October

ofNutritional

Sciences,

NY 14853.

23, 1991.

© 1992 American

Society

for Clinical

Nutrition

in

Downloaded from https://academic.oup.com/ajcn/article-abstract/56/3/526/4715393 by University of Otago Library user on 17 December 2018

Riboflavin requirements in older women13

RIBOFLAVIN

TABLE

AND

foods

Exercise group 2

HRibo

2 1

Baseline (2 wk)

Subgroup 1 II III IV

1

Period 1 (4 wk)

Period 2 (4 wk)

N E N E

E N E N

N N N N

5 LRibo, low-riboflavin group 0. 15 zg/kJ (0.6 g/kcal), riboflavin group 0.22 .tg/kJ (0.9 g/kcal), N, nonexercise,

rest

or exercise;

could

hinder could

3) a history their

of knee,

ability

influence

joint,

to exercise;

exercise

or 4)

performance

injury

used

medications

or vitamin

Participation

in the study was approved

was by informed

by Cornell

University’s

The

Human

Po-

Subjects

The study

was designed

as a 10-wk,

two-period

at two riboflavin intakes: (LRibo) group] and

crossover

0.15 zg/kJ 0.22 g/kJ

(0.6 (0.9

cx-

group)]. Subjects were first randomly amount that was maintained throughout were randomly placed into one of two

exercise sequences and randomization process

thus,

II received

subgroups I). Subjects

(Table

emerged from the in subgroups I and

0. 1 5 pg/U (0.6 tg/kcal) riboflavin, whereas those in III and IV received 0.22 ig/kJ (0.9 g/kcal) riboflavin.

subgroups Members metabolic

ofsubgroups II and IV exercised during period (exercise group 1), and members III exercised

during

the

second

4-wk

metabolic

(exercise group 2). None ofthe subjects exercised wk baseline period. In addition, all subjects were discontinue any regular strenuous activities but maintain

normal

daily

the first 4-wk of subgroups period

during the 2instructed to to otherwise

routines.

rate

throughout

the study

all subjects

received

the same

basic

daily

menu designed to provide 0.l5 tg/kJ (0.6 g,/kcal) riboflavin. During the baseline period, each subject’s diet was adjusted for weight maintenance. Individuals in the HRibo group (subgroups III and IV) received a daily supplement of 0.46 mg riboflavin (as

flavin

mononucleotide,

provide

0.22

g/kJ

FMN)

(0.9

g/kcal)

content

Handbook

of the basal

that

g/d)

group

data

no. 456

(17).

diet was later

riboflavin

was receiving and

the

from The

together mended

with dietary

US

needed

foods

were

for preparation.

to prevent

riboflavin

Pre-

loss

by light

preparation.

periods,

subjects

ergometer

(Table

2).

each

workout

model

ExIIIA;

Hempstead,

NY).

Resistance

exercised

at

75-85%

HRs

were

their

with

adjusted

moni-

a HR

Instruments

was

min/d, maximum

continuously

session

Computer

20-25

of

monitor

Corporation,

when

necessary

to

subjects at the specified training HR. The amount performed during each exercise session was monitored

maintain

confirm

improvements

Body

in physical

fitness

with

exercise

of to

training.

composition

Body

composition

correction

was assessed for

residual

technique

(2 1 ). Measurements

metabolic

period.

Exercise

by densitometry

volume

by

were

the

(20)

with

nitrogen

taken

at the

di-

washout end

of each

tests

Exercise

tests

conducted to determine VO2max and by at the end of each metabolic period. The test was performed before the baseline period because

gas exchange

baseline

were

(ATGE)

ofthe

need

tested

at approximately

to screen

individuals

for the

the same

time

study.

ofday

Each

subject

for all three

was

exercise

tests. The

exercise-test

protocol

by Wasserman

et al (14)

an incremental,

maximal

Quinton

Instrument

h. After

a 2-mm

proximately

every

10). Weight During

the

Quinton

subject’s ofeach

until

test

monitored Instrument

from

limit

each

Co). lead

system

(Quinton

exercise during

configuration

to best

tests.

electrocardiogram

a 12-lead

monitored

their cupouches

manner

exercise

the

1% apDuring

carried and waist

In subsequent

was

a CM5

the grade

subject’s

with

of 5.5 k/ ofO%,

of tolerance.

all three

of Q65;

speed

for in this

across

electrocardiogram mm

the

consisted

(Quinton grade

tests, subjects backpacks

compensated

first exercise

was continuously

test

by increasing

third exercise loss in weighted loss was

and

at a treadmill

increased

the workload

recommended

(22)

at a constant

period

minute

to that

treadmill

Co. Seattle)

was steadily

the second and mulative weight

similar

Wasserman

walking

warm-up

workload

2000;

was and

the

(Datamedix

tests

the

last

10 s

ST219

would

calculated

2

cycle ergometer

riboflavin

by direct

fluoro-

prepared indicated

(0.6 g/kcal,

TABLE Daily

Department

estimated

confirmed

0. 15 ig/kJ

diet, at that the

or 0.95

Week

Warm-up5

workouts

for the

4-wk

Stage 1 (75% HR max)t Time

exercise

period

S tage 2 (85% HR max)t

Warm down5

(mm)

zg/kJ

1 2

3 3

20 25

0 0

1 1

supplements of 200 g folic acid, and 3 ig vitamin D. The basic diet

3

3

20

5

1

4

3

18

7

1

HRibo

(0.9 g/kca1, or 1 .41 zg/d) All subjects received daily 250 mg Ca (calcium lactate),

diet

diet was initially

basal

metric analysis (1 8) on random samples ofthe least once during each study period. Analysis

LRibo

their

and dried

canned

Perishable

riboflavin.

The riboflavin content ofthe by using the nutrient-composition ofAgriculture

such

sources;

or batch. until

taken

food

(HR)

approximate

Throughout

single

lot

(Exersentry,

(9, Diets

were

a cycle

tored

from

same portions

exercise on

heart

zg/kcal) g/kcal)

[high-riboflavin (HRibo) allocated to a riboflavin the study. Next, subjects

four

protocol

rect

design

ercise study [low-riboflavin

Exercise

study

Committee. Study

during

527

were the

steps

exposure

work

any vitamin ofthe study.

consent.

cautionary

During

that

status.

tential subjects were instructed to discontinue using and mineral supplements 4 wk before the beginning

from

in preweighed

6 d/wk

HRibo, highE, exercise.

or back

provided

were

frozen

WOMEN

group

was

receiving

0.22

riboflavin.

these supplements allowances (RDA)

provided for women

the 1980 recomaged 50 y (19).

5

0 resistance.

t HR max,

maximal

heart

rate.

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LRibo

protocol

OLDER

All foods design5

Riboflavin group

I and

IN

1

Experimental

that

EXERCISE

TREBLER

528 oscilloscope;

Detamedix,

displayed (Beckman

MMC

struments

Inc,

Subjects valve during

the

dioxide

VE/VO2

‘ and

CO2

recorded

every

(the

exercise

Horizon

ventilatory

and

R (the

and

PETCO2

Subjects

alveolar

gas,

were

offive

plotted

encouraged

to walk

maximal

HR

until

(HRmax

did

not

attain

as a true

predicted

and

was

and

end

in y) and

measure

was measured

of

by using

Because all

scribed by Wasserman et al (14) and nique is based on predictable changes acid-base

buffering

intensity.

The

that

criteria

during

were

protein

by using

method

was

and

of

determined reaction

Crude

method

ri-

Urinary

the

carbamidodiacetyl

intake

the

discarded.

nitrogen

(26).

for

with unusual

in

nitrogen

balance

urea

nitrogen

urinary

of Weinsier

and

Butter-

Statistical

analysis riboflavin

position

status,

data

were

error

physical

analyzed

terms

performance,

by using

(28). The comparisons

concerning

riboflavin

interaction,

subject

within

group,

interaction,

period,

and

statistical analysis was performed for exercise by period interactions ysis

was

repeated

parisons

were

or paired

with

analyzed

t tests.

the

Main

by riboflavin

interaction.

If after

there was fairly strong or carry-over effects,

period

by using effects

1 data either

and

only.

All

standard

evidence the analother

corn-

Student’s

interactive

on

by carry-over

exercise

by period

based

were those

riboflavin

exercise,

exercise

body-corn-

t tests

of interest

carry-over, group,

and

Student’s

effects

t tests

were

con-

besides

the

technique

de-

analysis

of the

ficulties

with

it was

greater

of subjects.

endurance

occurs

from

were

con-

specific

samples

samples

by using

urea

of the

and

P < 0.05 and P < 0. 15, respectively. One subject was excluded from the final analysis of the cxercise-test data because she was unable to participate in the last exercise test. Two subjects were excluded from the statistical

Wasserman (22). in gas exchange

used

three

every cxof ‘O2 vs

the highest This value

majority

a noninvasive

an

to determine during

because

volume

determined Urinary

for three

(27).

The of

Estimates

- age

study, from equation

for the

for

was (25).

calculated

worth

above

at the

exhaustion.

value

value

gravity

made volume

aliquot

Urine

of thiosemicarbazide

measurements

of 02

analysis.

a modification

the presence

ratio

her maximum.

their

HRmax

HRmax

ATGE ‘ another

the

of specific

Urine

end-tidal;

HR. VO2max was predicted for each test by using HR attained among the three tests for that subject.

from

by using

All ofthe

HRmax

subjects

creasing

preparing

VCO2,

,

basis

time

220

=

the VO2rnax for each subject in the test, was calculated from a regression

The

before

Morell

tension

breaths. against

several

TO2max,

measured

and urea nitrogen

Slater

for oxygen

usually

as a guide

the

were

pooled

were

confirmed

gravity

mixing

exchange

[the highest

.

than

week.

excretion

L/min),

equivalents respiratory

R 1 15 were used during the test whether the subject was approaching

was

ofeach

riboflavin

VO2

were

at the end

values

ventilation

collections

days

breathing

test.

of a predicted

tests, ercise

In-

urine

tinuous

measured

by using

on a breath-by-breath

as an average

variables

were

as 15-s averages:

in expired 5

Beckman

data

VE (minute

determined

15

gas-exchange

MMC

L/min),

PETO2 were

#6;

system

AL

Twenty-four-hour

and

test

two-way

gas response

and recorded

VE/VCO2

(14)]

System

a Hans-Rudolph

by the

respectively,

,

mmHg

the

test

dioxide),

“CO2/’O2).

recorded

stress

boflavin

following

output,

carbon

and

Universal

through

technology

(carbon

was

CA).

The

exercise

HR

ET

This techthat result

exercise

of in-

1) a non-

as follows:

linear increase in VE and VCO2, 2) a systematic increase in VE/ VO2 without an increase in VE/VCO2 and 3) a systematic increase in PETO2 without a decrease in PETCO2 (14, 22). Before

sidered

significant

with

body-composition

data

the densitometric

because

of technical

dif-

measurements.

Results Nutrient

intake

and

body

composition

,

the ATGE estimations the identity of the above

were

generally

indicators marily

of the used

currence time

ATGE

times

against

time.

ject and were as distinguished

AT that tration

then

equations

were

criteria

time

the

mean

subject

equation

was

measuring an exercise

changes

in blood

re-

estimated

was of V02

were generated

priof oc-

set of criteria

and

for each

Prediction

sensitive

two

estimated

to each

ATGE

converted regressed

for each

subAT,, of the

lactate

concen-

test.

status status

activity

and

was

assessed

urinary

blood samples were collected at the end of each week. The

by erythrocyte

riboflavin

excretion.

glutathione Fasting

before breakfast on the erythrocyte glutathione

revenous

same day reductase

coefficient (EGRAC) was determined according to the of Sauberlich et al (24) and was calculated as the ratio

of the total dinucleotide

The

are the most

based on data from the treadmill tests. The from the ATGE , is a more direct measure

Riboflavin

activity method

.

averaged

a prediction

during

ductase

these

ATGE

with

involves

Riboflavin

and

23),

according

of the

(L/min)

22,

the

were

of occurrence

to discern

(14,

to estimate

The

to ‘O2

made, the plots were coded to conceal Because the last two criteria listed

easier

ATGE

of the

corded.

were subjects.

stimulated (FAD)

activity in vitro

in the presence to basal

unstimulated

of flavin

adenine

activity.

Group during

means

for age,

baseline,

nutrient

exercise,

and

intake,

in Table 3. Differences between with regard to age, height, body and

percent Attempts

for weight weight

body were

during

ofall

subjects

the remainder

ofthe

study.

to one

subject

who

wished

weight

loss during

baseline balance between

exercise nitrogen

to period

declined

the

the LRibo weight,

composition are presented

and

HRibo

groups

fat-free

mass

(FFM),

energy

intake

subject’s

the baseline

period,

yet the body

2.3 ± 1.9 kg (1 ± SD) Additional

energy

to maintain

study,

subjects

and nonexercise balance. The 2 was 0.7

body

periods

fat were not significant. made to adjust each

maintenance

FFM during ofFFM and

and

nonexercise

was

her

weight.

did

not

throughout

only

appear

effect

or an exercise

Riboflavin Mean

provided EGRACs exercise

the

to lose

periods based on estimates overall change in FFM from

± 1.6 kg. On average,

crude

was nearly neutral or positive. The difference nonexercise and exercise periods was nearly 0.07) and there was no exercise and riboflavin

(P < effect. The overall loss in percent 2 was 3.0 ± 2.9%. Again, there

provided

Despite

body

fat from

baseline

was no evidence

by riboflavin-interactive

nitrogen in FFM significant interactive to period

for an exercise effect.

status values

for EGRAC

in Table 4. The than the LRibo periods; however,

and

urinary

riboflavin

excretion

are

HRibo group had significantly lower group during both nonexercise and both the LRibo and HRibo groups

Downloaded from https://academic.oup.com/ajcn/article-abstract/56/3/526/4715393 by University of Otago Library user on 17 December 2018

chamber

=

Horizon,

size).

MA).

by a computerized

Anaheim

breathed

(large

and

Sharon,

as 1 5-s averages

WINTERS

RIBOFLAVIN TABLE

AND

EXERCISE

IN OLDER

WOMEN

3

Group

means

for age, body

composition,

and nutrient

intake5 Baseline

7)

56±5 160.8 ± 67.0 ± 37.2 ± 41.9 ± +1.54 ±

(g/d)

=

showed

64.9

±

2.5

34.4

± 3.0

42.4

± 6.9

7.6

1.33

+2.18

13.0

65.3

±

33.8

± 3.1

42.9

± 6.6

+1.70

± 2.52

7849 ± 1051 1875 ± 251 68.8 ± 6.5 1.16 ± 0.15

57±6 159.7 ± 7.8 62.9 ± 9.5 34.5 ± 6.2 38. 1 ± 2.6 +0.39 ± 1.83

(g/d4

± 3.22

7849 ± 1051 1875 ± 251 68.8 ± 6.5 1.16 ± 0.15

-

-

-

-

61.7 ± 8.7 33.3 ± 6.1 38.4

61.3 39.5

0.79

±

± 7.7

31.7 ± 7.7

± 3.5

+3.28

8092 ± 691 1933 ± 165 70.5 ± 4.9 1.77 ± 0.14

for the high-riboflavin

significant

during

increases

values

pletion

of riboflavin

HRibo

group group

Mean

urinary

approximately during

showed

within

by exercise times

a significant

± 4.2

+2.34

8272 ± 837 1976 ± 200 7 1.3 ± 5.0 1.81 ± 0.18

throughout

1.20),

exercise

group

tissue

de-

whereas

those

for

the

range.

There

was

no

acceptable

nary

±

1.02

8272 ± 1976 ± 7 1.3 ± 1.81 ±

833 199 5.0 0.18

decline

ofthat

exercise

LRibo

group

and

both

groups

riboflavin

excretion

for the urinary excretion were within an acceptable

the study

(29).

However,

was

for the

periods

in urinary

group

with

of riboflavin for range (> 0.080

four

ofseven

LRibo

Maximal

excretion

Nonexercise

Urinary riboflavin (mg/d) High-riboflavin group

EGRAC riboflavin

(mg/d)

1.224

±

0.17

±

1.070

±

0.66

±

SD. EGRAC, erythrocyte glutathione t Significantly greater than high-riboflavin S

Significantly

different

from

nonexercise:

1.283 0.14

±

0.031 0.49

1.109 0.46

±

fP




the

excretion three

both

in EGRAC

for the LRibo (EGRAC

were

riboflavin

t

-

12.5

7)

body fat and fat-free mass data 6 during nonexercise period.

EGRAC

mg/d)

-

-

SD.

±

Mean

-

6.5 13.9

7539 ± 544 1801 ± 130 66.9 ± 4.0 1.1 1 ± 0.08

t Percent n

Exercise

Nonexercise

0.0005. 0.0001, §P