Concentrations of the watersoIubIe vitamins thiamin, ascorbic acid, and folic acid in serum and cerebrospinal of healthy individuals13 Chantal

ME

Tallaksen,

Thiamin,

ABSTRACT

acid, fluid

of back-pain.

CSF

concentration

thiamin

16.9

± 8.3

nmol

acid;

the

All subjects

thiamin

in CSF

serum

otherwise 133

into

CSF

for thiamin

concentrations

higher

results support the of water-soluble monophosphate,

high

CSF

KEY

WORDS

should

therefore

concentrations.

Am

than

the thiamin and serum

known

healthy

subjects,

serum,

acid,

existence vitamins

thiamin,

thiamin

folic

whole

ditional samples

acid,

monophosphate

Introduction is a central

brane synthesis and folic acid decreased in brain

coenzyme

conservation

(1). Ascorbic acid is an participates in nucleotide

important synthesis

folic

in the

(CSF)

has been

related

to the

presence

of neurological

treatment Although been

acid

of the

to the onset

mem-

antioxidant (1). Thus,

acid

acid

in healthy

ofdelirium

tremens disorders

concentration

concentrations subjects,

cerebrospinal

or psychiatric

(4). A low concentration in chronic alcoholism

(7). the folic

studied

concentration

ascorbic

acute head trauma has been reported

is necessary

for lipid

a

has

been

fluid (2)

(3).

and A

related

to

of thiamin in CSF (5, 6) and antiepileptic in serum

alcoholics

(2),

and and

CSF

have

other

cat-

egories of patients (3, 7, 8), the concentrations of thiamin and ascorbic acid in CSF have only been reported in small groups of individuals (9- 1 1 ), in children ( 12, 1 3), or without relating them found

to serum concentrations as unphosphorylated

Am J C/in Nuir

1992;56:559-64.

(4, 5, 12). Moreover, thiamin thiamin, thiaminmonophosphate, Printed

in USA.

both

blood,

© 1992 American

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and

Department

CSF samples from similar their

serum

female, 20 male) y) during myelograms

were

collected

of Neuroradiology).

were collected without patients (9 female,

informed

ethics

samples

aged 48.0 ± 14.7 at the outpatients’

consent.

The

from

y (1 ± SD; clinic (Ul-

Fourteen

ad-

corresponding blood 6 male). All subjects

protocol

was

approved

by re-

committee.

All subjects were suffering from severe back pain but were otherwise healthy. The myelography was part of the routine investigation. The final diagnoses were lumbar disc prolapse (n = 24), spinal stenosis (n = 6), degenerative changes (n = 3),

of these three water-soluble vitamins impair the nerve cellular functions. A de-

crease

in the

and and

concentration tissue might

decrease

in glucolysis

in the mitochondria

therefore

ascorbic acid, and folic acid in CSF subjects and to assess their respective

(11

patients range 2 1-83 1

gional

Thiamin

should serum. to measure

and methods

CSF, 3

gave

for energy

compounds

in CSF and was therefore

comim-

Subjects

to ensure

fluid,

tissues

are the only their respective

ratios.

Subjects

acid,

1992;56:559-64.

cerebrospinal

(9, 14). Both

compounds, of healthy

in human

monophosphate in CSF (6), and

simultaneously of this study

lev#{225}l Hospital, Ascorbic

thiamin-triphosphate

thiamin reported

in

are correlated vitamins. High

Nutr

is not

determined The purpose

be

jmol

ascorbic

be advocated

J Clin

portance

acid/L. The CSF± 4.3 for thiamin and 3.3 ± 0.8 for

and folic acid. However, low CSF concentrations with low serum concentrations for the three serum

The

thiamin/L, ± 58.8

and

(1). Thiamin and pounds previously

healthy.

was significantly

These mechanism

thiamin-diphosphate

cerebrospinal a myelography

8.6 ± 3.9 nmol folic 8.3 acid,

Bell

ascorbic

and

monophosphate/L,

serum for each compound. of a saturated transport from

were

and 44.9 ± 13.2 nmol 2. 1 ± 0.8 for thiamin, 3.0 ± 1 .4 for ascorbic

amount

He/ge

monophosphate, in serum underwent

(1 ± SD) was

ascorbic acid/L, serum ratio was monophosphate,

and

Bghmer,

and folic acid were determined (CSF) in 3 1 outpatients who

because

folic

Thomas

fluid

is

Society

spondylolisthesis No malignancy The reference consisted male; serum

(n = 3), and no pathological findings (n = 1 1). was found. group for the serum thiamin concentrations

of 40 healthy mean

(±SD)

concentration

the group

±2

hospital

staff

members

age 43 ± 9 y, range was

X SD (range

defined

as the

for thiamin

[25

23-58

male,

y]. The

mean

15 fe-

reference

concentration

5-23

nmol/L,

of

range

for

thiamin monophosphate 2-12 nmol/L). For serum the reference group consisted of32 healthy hospital

ascorbic acid, staff members

I From the Norway. 2 Supported

Hospital,

Medical

Department,

by Vinmonopolet

Aker AS, Oslo,

University Norway.

3 Address reprint requests to CME Tallaksen, Aker University Hospital, 05 14, Oslo 5, Norway. Received November 19, 1991. Accepted for publication March 17, 1991.

for Clinical

Nutrition

Oslo,

Medical

Department,

559

TALLAKSEN

560

aged

21-5

1 y (35.5

summer

and

44.4-88.8

tuted

the

± 8 y). Measurements

reference

imol/L.

Two

the reference

range, hundred

group

Hospital for folic 24 nmol/L).

acid

were

defined healthy

adult

of the Central

serum

done

concentrations

was

subjects

Laboratory

consti-

of Ullev#{227}l

(reference

range

6-

3

ratio

thiamin

of 1:3 and

monophosphate

concentration,

collection

Two milliliters mL CSF were

heparinized whole blood, collected from each subject.

5 mL serum, All samples

and were

times

Concentration

ples. Ascorbic the

method

acid

were

treated

was assayed

described

similarly

in serum

by Zannoni

termined oratory,

in serum, whole Ullev#{227}lHospital,

globulin

as a ligand

to the serum

1 1.

-

Folic

acid

There

no

correlation

concentration

was

monophosphate

con-

the

con-

between

de-

blood, and CSF in the Central Labby using a radioassay with a lacto(Charcoal Boil Dual Count, DPC, Los An-

thiamin

amount

CSF

correlated

(Fig

of serum

(concentration

thiamin

and

serum

thiamin)

thiamin

monophosphate

(Fig

1 B, NS).

(unphosphorylated

similarly

with

thiamin)2

of serum

between

in CSF

total

serum

(concentration

correlation

concentration

to

was

was

19

4.56

+

sam-

and CSF according

et al (16).

the thiamin

the thiamin

of

the thiamin

of CSF thiamin

=

samples

concentration

2.6 ± 1.8 times

centration in CSF and serum with a correlation coefficient of 0.60, but the data also fit a second-order polynomial regression plot with the following equation (Fig 1A):

Thiamin and thiamin phosphate esters were determined in whole blood, serum, and CSF by HPLC as previously described CSF

than

1). The

was

in serum

higher

(Table 1). was a positive

of vitamins

(1 5). The

4: 12 (Table in CSF

whereas

to three

centration There

frozen within 2 h of collection (-20 #{176}C for the thiamin and folic acid samples, -70 #{176}C for ascorbic acid) until analysis within 1 mo. Assays

and thiamin monophosphate were the only thiamin detected in CSF (Table 1). Both compounds had a higher concentration in CSF than in serum, with a

respective

one

Sample

AL

Thiamin compounds significantly

in late

as for thiamin,

ET

- 0.12

However,

the

+ phosphorylated)

thiamin

with

the total

in

amount

in

lC):

Concentration

of thiamin

thiamin

+

monophosphate

in CSF

geles). =

Statistical

analysis

Results nificant the

are given

as mean

differences

Student’s

alyzed

were

paired

regression 0.05.

were

All tests

used were

For

calculated

t test,

by the least-squares

SD.

±

and

statistical

by the

first-order

analysis,

linear

done

two

(an-

5.32

+

concentration

(concentration

tailed.

Ascorbic Serum trations

and

had a thiamin whole

blood

reference

range

jects

a thiamin

had

erence

monophosphate

(results

established

not

concentration

shown)

by this method

concentration

above

(Table

slightly

below

nmol/L

Concentration

the

lower

8.6

±

CSF

16.9

±

Ratio

2.07

± 0.76

±

24 nmol/L

3.9 8.3t

SD. Serum reference for folate.

Significantly Significantly

greater greater

than than

ranges:

5-23

4.0 28.1

±

8.28

±

nmol/L

±

ascorbic acid, concentrations5

of CSF 46.8

=

and folate

thiamin

(paired

than

the serum

con-

polynomial

acid

(concentration

of serum

ascorbic

acid)2

2. 1 (concentration

of serum

ascorbic

acid)

- 0.006

in serum

subjects,

and the

and cerebrospinal

Thiamin + monophosphate

2-12 I test).

Downloaded from https://academic.oup.com/ajcn/article-abstract/56/3/559/4715412 by St Bartholomew's & the Royal London School of Medicine and Denistry user on 25 March 2018

nmol/L

for thiamin

fluid (CSF)

in healthy

Ascorbic

acid

Folic acid

.tmol/L

12.7 ± 4.2 44.9 ± 14.Ot 3.71 ± 0.93

for thiamin,

higher

fit a second-order following equation:

ascorbic

nmol/L

1.5 7.9ff 4.34

serum values, P < 0.001 thiamin, P < 0.001.

monophosphate)

ref-

nmo//L

Serum

thiamin

in

the lower 1). Four sub-

Thiamin monophosphate

Thiamin

of serum

thiamine

was two to four times

+

of thiamin, thiamin monophosphate, the CSF and the corresponding serum

ofserum

(Table 1). The data plot (Fig 2) with the

range.

TABLE 1 Concentrations ratios between

monophosphate)2

ascorbic acid was slightly below our reference concenin 14 of 3 1 subjects (Table 1). The CSF ascorbic acid

concentration

All subjects

thiamin

acid

centration regression

serum

t t

+

of serum

thiamine

- 0.09

Thiamin

1

concentration

of serum

second-order was set at

Results

5

+

and

regression

method) or polynomial ( I 7). The level of significance

(concentration

sig-

t test

Student’s

-5.28

monophosphate,

nmo//L

53.8

±

14.6

± 7.4

133 3.04

± 58.8t

44.9

±

l3.2t

1.37

3.27

±

0.83

45-91

mol/L

±

36.6

for ascorbic

acid, and 6-

WATER-SOLUBLE A

VITAMINS

IN SERUM C

40-

. 35.

AND

CSF

561

60-

70

.

-

. 30-

60-

.

25-

-j

r=0.68 S

r=0.73

S

-C 500

0

E

.

E

20

S

40-

S

ILC

15-

30-

5$

0

10 -

S. S.

S

20

-

10

-

S

5-

0-

0-

5

0

10

25

0

10

T (nmol/L)

Serum B

20

15

20

Serum

T+TP

FIG.

1.(Continued)

folic

acid

30

(nmol/L)

50-

40

Concentration

-

S

of CSF

2.8 (concentration

=

S S

of serum

folic

acid)2

S

S

S

+

S

4.

1

(concentration

of serum

folic

acid)

0.06

-

Discussion Li.

S

20-

r=0.25

C,)

The

results

show

folic acid, and ascorbic serum concentrations.

S 10

relation vitamins,

-

the CSF

concentrations

oftotal

serum

;

02

0

4

6

TP (nmol/L)

Serum

FIG 1 . A: relationship between the concentrations of thiamin (T) in cerebrospinal fluid (CSF) and in serum in 31 healthy subjects. B: relationship between the concentrations ofthiamin monophosphate (TP) in CSF and in serum in 31 healthy subjects. C: Relationship between the concentrations of total thiamin (T + TP) in CSF and in serum in 31 healthy subjects.

Are of the

concentration in CSF was two (Table 1 ). There was a positive trations polynomial

in CSF

and

serum,

regression

plot

the

in serum and whole normal range (Table

to four times correlation which

(Fig

is shown

3) with

the

blood 1). The

higher than in serum between the concenin a second-order following

equation:

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the

CSF. were

outpatients

influenced and

thiamin,

and

CSF

status?

reliably

(18).

can concentrate

barrier in CSF

(thiamin methods

any

disease

at the time

reliable

that

ofthe

vitamin intakes, the results.

that

by the

of vitamins

biological

measurements

study. were

not

markers in blood

of the tissue concentration for the three it has been postulated that the concennot represent the brain tissue concenHowever,

it is from

its vitamins,

to nerve provide measurements

as reflected

from

state

concentrations It is known

for the three low at low serum

the amount free

the nutritional

and

vitamin

blood brain Thiamin tration lytical

times higher than that there is a cor-

state

their diets, including their may therefore influence

serum

compartment concentrations multaneous

nutritional

will also influence

cells give a truer picture vitamins, and, similarly, tration in the CSF may

CSF All the folic acid concentrations (results not shown) were within

have

However, known,

trations acid

Thus,

concentrations,

present in the All subjects may

acid is two to four They show, moreover,

between serum and CSF concentrations and that the CSF concentrations were

concentrations.

Fo/ic

that

S

0

cells thus

(8, 19). is usually

+ thiamin used do not

and

the

serum

the CSF

accessible to investigations. a quantitative assessment, in serum may reveal damage given

as the

monophosphate) discriminate

total

that

the

is the closest

thiamin

CSF and sito the concen-

because the anabetween unphosphor-

562

TALLAKSEN

ET

actual source for CSF ified by this study.

70

The data

S

60

c?

AL

55

indicates

r=0.84

with

serum,

E

(14, was

40

have

21-23). reported

:

.

2

S

10

,

#{149}

0

the presence

HC1, but the increase

#{149}

I

10

20

Serum

folic

acid

)

(nmol/L

The

ofa

in serum

ylated and phosphorylated analysis of both compounds

acid

in

thiamin (5, 10, 20, 21). A concomitant gives supplementary information -

centrations using

than

reported

by assay.

assay,

reported

L for CSF

total

advantage

ofdetermining

(9). reports-Rindi

72 nmol/L

a microbiological

crobiological

importance previous

(22)

of ascorbic

conclusion,

but at a lower

However,

thiamin

(6) 60 ±

(23)-but

show

Botez

(10)

Wyatt

et al (12),

(102

concentrations

in children both

lower

± 21 also

2

con-

nmol/L),

using

component

previous crease

physiological serum continue to increase,

tion

thiamin

Our

method

compounds

of the

reason the

is not

in the CSF.

agrees acid

for the high

in CSF

that

reach

of pre-

an upper

serum

concentration

meostatic

(23-25). results

.

with

transport

nervous

system

trations

of ascorbic

(45 imol/

for ascorbic acid into a saturable and a non-

also

show

low serum system

against

that

low

concentrations

is thus

short-time

unable

CSF

concentrations

and that

to protect

reduction

the

in serum

the hocentral

concen-

acid.

has the and

clear.

in CSF, high

from

serum

to CSF transport

suggested

mechanism,

in rabbits

as suggested

infancy

and

S

to

S

in our

23).

into

supports

presence

in Table

#{149}S S

.

thia-

os

100

S

monophosthe

monophosphate,

The

by the data

data

150

(1), which

of thiamin

of thiamin

r=0.75

S

thiamin

is formed

diphosphate

5

200

concentra-

dephosphorylated

gradient

(14,

dea par-

v monophosphate

shown

and

from

55

with

a relative

thiamin

Thiamin

concentration

agrees

thiamin

monophosphate

is further

of an active

This

amount for

of thiamin and

represented

thiamin

dephosphorylation

The

with

a mi-

directly

reported

ofCSF

unphosphorylated

is not

found

et al (12)

a ratio of 38-62% in children.

phate

the

ofthiamin amount

tence been

monophosphate

(6, 9). Wyatt

with

in CSF

from

conin-

of 84 ± 5 1 nmol/

(12).

thiamin

phosphorylated

increase

The

that

amount

reports

childhood, monophosphate

mm.

show

total

ofthe

allel

(22).

250

results

ofthe

62%

the

are correlated

simultaneously.

Our

control

mechanism

limit in our data at 22 Mmol/L. Thus, at a serum concentration between 5.5 and 1 1 zmol/L, the active uptake process of ascorbic acid appears to become saturated. Ridge et al (1 1) came to a

saturable

.

that may be of diagnostic Our results agree with Baker

homeostatic

was not assessed

L). This agrees with the uptake process CSF shown in rabbits and involves both

and

studies

increase of CSF thiamin administration of 50 mg

concentrations

concentrations

similar

nmol/L

compared Kinetic

and ofa saturable

show however that, within the CSF concentrations

the CSF and the serum vious studies (11, 13).

1

40

30

FIG 2. Relationship between the concentrations of ascorbic cerebrospinal fluid (CSF) and in serum in 30 healthy subjects.

-

plot, which

concentrations

mechanism.

by the brain,

be clar-

dicating that the homeostatic barrier must be set at a high level for thiamin transport into CSF. Our results agree with those of Ridge and Fairhurst (1 1) and show slightly lower CSF concentrations than Brau et al’s (4) (36 ± 8.6 mg/L). The difference may be due to differences in the nutritional status ofthe groups. However, the ratio of2:4 between

S

20

0

transport

cannot

correlation

CSF

In humans, a threefold 6 h after an intravenous

Our results centrations,

30

in the

established

absorption

thiamin

polynomial

increase

ie, a saturated

ofthiamin

-5:

monophosphate

fit a second-order

a limited

in rabbits

50

-

thiamin

exis-

S

of a saturation

2, further

S S

as has S#{149}

supports

hypothesis. Alternatively,

concentration tulated by regulatory monophosphate

CSF

thiamin

of thiamin

monophosphate

diphosphate

in brain

Poloni et al (9). But, because we do mechanisms of the dephosphorylation in the

transport

locus

or in the

may

reflect

tissue, not

the

know the of thiamin

nerve

#{176} 0

as pos-

cell,

the

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50

Serum

FIG

3. Relationship

rebrospinal

fluid (CSF)

100

ascorbic

between the concentrations and in serum in 31 healthy

150

acid

(umol/L)

of folic acid subjects.

in ce-

WATER-SOLUBLE

VITAMINS

TABLE 2 Ratios between cerebrospinal fluid and serum concentrations of thiamin, subjects with low (n = 16) and high (n = 15) serum concentrations

5

0.75

±

AND

monophosphate,

CSF

total

Thiamin + thiamin monophosphate

8.16

±

4.30

10.7 5.5

±

4.5 l.4t

3.18

±

0.80

thiamin,

563

ascorbic

acid,

Ascorbic acid 3.04

±

and folic acid in

Folic acid 1.37

3.27

±

0.83

1.9 ± 0.7 2.2 ± 0.8

±

3.3 ± 0.6 3.1 ±0.9 4.0± 1.2 2.l±0.6t 3.6 ± 0.9 2.9 ± 0.7t

SD.

±

1

2.05

thiamin

SERUM

Thiamin monophosphate

Thiamin All subjects Thiamin 16 nmol/L Thiamin monophosphate 4.9 nmol/L Thiamin + thiamin monophosphate 21.3nmol/L Ascorbic acid 8.3mg/L Folic acid 12 nmol/L

IN

tP