1472 Am J C/in Nuir l987;45:l472-9. Printed in USA.

1987 Am erican Society for Clinical Nutrition

Dosage form and formulation effects on the
bioavailability of vitamin E, riboflavin, and vitamin B-6
from multivitamin preparations1-3
Kamlesh M Thakker, PhD; Harry S Sitren, PhD; Jesse F Gregory III, PhD;
Gerald L Schmidt, PharmD; and Thomas G Baumgartner, PharmD
ABSTRACT The effects of formulation factors and pharmaceutical dosage form on the bio-
availability of RRR-a-tocopherol (d-a-tocopherol), riboflavin, and pyridoxine hydrochloride were
studied after administration of two capsule formulations and a tablet to 12 normal humans. Absorption
of RRR-a-tocopherol was increased from the Aquabiosorb soft elastic gelatin (SEG) capsule for-
m ulation compared with the modified stai*lard-SEG capsule and the commercial tablet. There were
no significant differences in bioavailability of riboflavin and pyridoxine hydrochloride between the
SEG formulation and the tablet albeit a trend toward consistent absorption was seen from the SEG
formulation. The modified-SEG formulation exhibited significantly lower bioavailability for these
water-soluble vitamins. The enhanced bioavailability of vitam in E and the trend towards faster and
m ore consistent absorption of riboflavin and vitamin B-6 from the SEG form ulation m ay be related
to the surfactant vehicle employed and the attendant wetting properties. The results also suggest
ethnic differences in vitam in bioavailability. Am J Clin Nuir l987;45:1472-9.
KEY W ORDS RRR-a-tocopherol, riboflavin, pyridoxine HO, pyridoxal, pyridoxal phosphate,
bioavailability, dosage form effects, formulation effects, soft elastic gelatin capsule
Introduction
Multivitamin formulations are routinely
used by a large section of the American pop-
ulation either therapeutically or prophylacti-
cally. These may be physician prescribed or,
as is quite common with these nutritional
products, self-prescribed. In recent times it has
become increasingly apparent that excessive
absorption of ingested vitamins can cause tox-
icity. The reverse problem, namely incomplete
or suboptimal biological availability of the in-
gested vitamins from a multivitamin formu-
lation, has not received sufficient attention (1).
Since pharm aceutical product form ulations
containing multivitamins are usually expen-
sively priced, bioavailability differences be-
tween brands and/or generic preparations
would be especially im portant from the point
of view of consumer economics. Bioavailabil-
ity must be considered in evaluating the effi-
cacy of multivitamin products prescribed for
therapeutic or prophylactic use.
Since the mechanism of enteral absorption
of some of the vitamins is via specialized
transport processes such as carrier mediation
and/or facilitated diffusion across a window
site in the gastrointestinal tract, a theoretical
basis for a better absorption profile from a soft
elastic gelatin capsule (SEG) dosage form
com pared with a tablet dosage form does exist.
In practice, improved bioavailability from the
SEG dosage form has only recently been doc-
umented for vitamin preparations (1, 2) even
though such documentation for drugs such as
digoxin (3), tetracycline (4), and chemother-
apeutic and nonsteroidal antiinflammatory
drugs (5) already exists. Therefore, it is im-
perative to obtain data on the biological ab-
sorption of vitamins from multivitamin for-
mulations and to ascertain whether the par-
From the Departments of Pharmaceutics (KT), Food
Science and Human Nutrition (HSS, JFG), and Pharmacy
Practice (GLS, TGB), University of Florida, Gainesville,
FL
2Supported in part by grants from the Division of
Sponsored Research, University of Florida, from the RP
Scherer Corp. and from the Am erican Society of Hospital
Pharmacists Education and Research Foundation.
3Address reprint requests to Dr Kamlesh M Thakker,
20730 Dearborn Street, Chatsworth, CA 91311.
Received October 10, 1985.
Accepted for publication June 5, 1986.

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BIOAVAILABILITY OF M ULTIVITAM INS 1473
t ic u la r p h a rm a c e u t ic a l d o s a g e fo rm u s e d h a s
a n y in flu e n c e o n t h e b io a v a ila b ilit y . W it h
p re s e n t a c c e n t o n lifestyle vita m in s , m u ltivi-
t a m in p ro d u c t s s u c h a s S t re s s t a b s # { 1 7 4 } (Le d e rle
La b o ra to rie s , P e a rl R iv e r, NY ), w h ic h c o n t a in
vita m in s E , fo lic a c id , a n d b io t in in a d d it io n
to the B -c o m p le x g ro u p o fv it a m in s , a re q u it e
p o p u la r w it h t h e v it a m in -u s e r p o p u la t io n in
t h e Un it e d S t a t e s . A p re v io u s s t u d y e x p lo re d
t h e e ffe c t o f fo rm u la t io n fa c t o rs o n t h e b io -
a v a ila b ilit y o f t h e o il-s o lu b le v it a m in s A a n d
E a n d t h e w a t e r-s o lu b le v it a m in rib o fla v in (2 ).
Th e s e a u t h o rs fo u n d a n e n h a n c e d b io a v a il-
a b ilit y o f a ll t h re e v it a m in s fro m t h e e x p e ri-
m e n t a l S EG c a p s u le s , w h ic h c o n t a in e d a
n e w s u rfa c t a n t liq u id v e h ic le (A q u a b io s o rb # { 1 7 4 } ,
R P S c h e re r No rt h A m e ric a , S t P e t e rs b u rg ,
FL), c o m p a re d w it h t h e c o m m e rc ia l d o s a g e
fo rm s a v a ila b le in A u s t ra lia . In t h is s t u d y , v i-
t a m in b io a v a ila b ilit y fro m t h e c o m m e rc ia l
t a b le t p re p a ra t io n (S t re s s t a b s 6 0 0 # {1 7 4 }) is c o m -
p a re d w it h t w o e x p e rim e n t a l S EG -c a p s u le
fo rm u la t io n s , t h e n e w s u rfa c t a n t liq u id v e h ic le
fo rm u la t io n a n d a m o d ifie d S EG -c a p s u le fo r-
m u la t io n (m o d ifie d b y t h e in c lu s io n o f a n o n -
s u rfa c t a n t a b s o rp t io n e n h a n c e r t o t h e s ta n d a rd
o le a g in o u s v e h ic le ).
Ma t e ria ls a n d m e t h o d s
The com m ercial tablet S tresstabs 600 (L ot 753 510,
Lederle Laboratories, Pearl River, NY) was used as the
reference standard. Two SEG-capsule form ulations were
used: A quabiosorb SEG capsules (L ot E-l29l8, RL-3420,
RP S cherer N orth A m erica, S t Petersburg, FL ) and m od-
ified-form ulation SEG capsules (L ot E-l299l, R L -349 1,
RP S cherer N orth Am erica). All three form ulations were
analy z ed using official pharm aceutical f orm ulary m ethods
(6). The labeled content of 2-am bo-a-tocopherol (d!-a-
tocopherol acetate) (v itam in E)w as 30 IU while the assayed
content was 33 IU for the tablet, 36.5 IU for the SEG-
capsule form ulation, and 39 IU for the m odified-SEG for-
m ulation. The labeled contents for riboflavin (vitam in B-
2) and pyridoxine hydrochloride (vitam in 8-6) were 15
m g and 5 m g, respectively, while the assayed contents w ere
17 m g and 7.8 m g for the tablet, 1.9 m g and 5.3 m g for
SEG capsule, and 18.7 m g and 5.3 m g for the m odif ied-
SEG capsule.
The bioavailability study was conducted as a random -
ized 3-way crossover. Twelve healthy m ale volunteers
(eight A m erican Caucasians, one British Caucasian, one
Ghanian African, and two Indian Asians) within 10% of
their ideal weights were selected to participate in the study
from a prestudy physical exam ination, which included a
m edical history rev iew , a SM AC-25 biochem ical screening,
CBC, and urine analysis. Volunteers on chronic m edication
or vitam in and/or m ineral therapy were excluded. All
subjects were requested to abstain from any vitam in prod-
ucts for 4 wk before and from alcohol for 24 h before
adm inistration ofthe m ultivitam in form ulation. They w ere
also instructed to not eat f ood 10 h before the dose ad-
m inistration (ie, af ter 2200 the night before an 0800 dose
adm inistration). W ater was allowed ad libitum until 2 h
before the dosing. The dose was adm inistered with 200
m L of water at 0800. W ater was then withheld until 2 h
af ter dosing and f ood until 4 h after dosing. A baseline
blood sam ple (10 m L ) was drawn at 0 h (im m ediately
before dosing) into green-top heparinized tubes. Blood
sam ples (10 m L ) were drawn at 0.5, 1, 2, 3, 4, 6, 8, 12,
and 24 h postdosing. A ll blood sam ples were centrifuged
im m ediately and the plasm a was frozen at -20#{176}Cuntil
analy z ed for RRR-a-tocopherol, riboflavin, pyndoxal, and
py ridox al phosphate. The sam elunch and dinnerwas eaten
by each subject on the dose adm inistration day s. A wash-
over period ofl d was allowed between each dose adm in-
istration. The volunteer subjects (ages 21-31 yr) were stu-
dents or em ployees of the College of Pharm acy or De-
partm ent of Food S cience and Hum an Nutrition and the
study protocol and appropriate consent form s were ap-
prov ed by the Institutional R ev iew B oard ofJ Hilhis M iller
Health Center.
Vitam in E in plasm a was determ ined using a newly
developed rapid, sensitive, and specific reversed-phase
HPL C (with a Cl8 colum n and 99:1 m ethanol:distilled-
deionized water at 2 m L/m in flow rate) procedure with
UV detection at 292 nm , capable of sim ultaneous deter-
m ination of plasm a R R R -a-tocopherol and retinol and
their esters. The m ethod involves plasm a protein precip-
itation with 4 vol ofacetonitrile(containing 80 g/dL reti-
nyl acetate as internal standard) to 1 vol of plasm a, m ixing
with a vortex m ixer for 1 m m , and then centrif uging for
3 m m after which 100 iL of the supernatant layer was
injected into the chrom atograph. The peak-height ratio
R R R -a-tocopherol:internal standard was used for quan-
titation. A ty pical linear equation for the calibration curv e
for R R R -a-tocopherol is Y = 0.2097 X + 0.0799 (r
= 0.998) and for retinol is Y = 0.01364 X + 0.0058 (r
= 0.999). Concentration and peak -height ratio were lin-
early related over the ranges inv estigated for both RRR-
a-tocopherol (0.335-2.68 m g/dL) and retinol (20-100 M g/
dL). W ithin-day coefficient of variation for the sam e
plasm a sam ple assay ed 10 tim es was 2.9% and 4.1% for
retinol (80 g/dL) and R R R -a-tocopherol (0.75 m g/dL ),
respectively. Between-day coefficients of variation for the
sam e plasm a sam ple assay ed 16 tim es over 1 m o was 2.7%
(retinol, 80 g/dL) and 4.0% (R R R -a-tocopherol, 0.75 m gI
dL). The efficiency of recovery from a sam ple of plasm a
containing 100 tg/dL retinol, 80 g/dL retinyl acetate,
and 2.68 M gJdL of R R R -a-tocopherol varied with the ratio
and ty pe of protein precipitant used. W ith 2 and 3 vol of
m ethanol, the recov eries f or v itam in A , retiny l acetate,
and vitam in E were 107, 31.5, and 12% and 146, 66.7,
and 20% , respectively. W ith 2,3, and 4 vol of acetonitrile,
the recoveries of vitam in A, retinyl acetate, and vitam in
E were 75.4, 72.6, and 62.8% ; 98.8, 70.0, and 76.7% ; and
95.2, 87.4, and 93.9% ; respectively. Consequently, 4 vol
of acetonitrile to 1 vol of plasm a was considered to be the
optim al ratio. The lower lim it of detection in the plasm a
w as 10 g/dL for retinol and 0.1 m g/dL for RRR-a-
tocopherol. T he standard solutions w ere prepared in ace-
tonitrile and were stable even af ter 2 m o except for retinyl
acetate which was stable for 2 wk. The assay can easily be
adapted for pediatric sam ples as well as for routine analysis
of vitam in A and E levels in disease states such as fi-

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nilIcant differences between products, subjects, and weeks
at a known K -r at io of a t o er r or (10). T r end analyses
were conducted using t he RSQUARE procedure in
SAS (10).
Re s u l t s
Vitamin E
T h e m e a n vitam in E le v e l o f the 12 subjects
is plotted in Figure 2 as a function o f t im e f o r
t h e t h re e products adm inistered. T a b le 1 lists
t h e p h a r ma c o k i n e t i c mo d e l i n d e p e n d e n t b i o -
availability param eters C M A X (m axim um
p la s m a c o n c e n t ra t io n achieved; m easured
graphically), TM A X (t im e re q u ire d t o attain
m axim um p la s m a c o n c e n t ra t io n ; m easured
graphically), a n d A UC O T O 2 4 (a re a u n d e r t h e
plasm a concentration vs tim e curve f ro m t im e
o t o 2 4 h ; m easured b y t h e trapezoidal rule)
a s w e ll a s t h e B A S E L IN E le v e ls a n d M A X -
B Y M IN (t h e m axim um increase norm alized
f o r t h e baseline levels) ratio. T h e descriptive
s t a t is t ic s p ro v id e d in T a b le 1 in c lu d e t h e m ean,
S D, and ra n g e f o r t h e v a ria b le s a s a function
o f t h e p ro d u c t .
T h e results o f t h e A N O V A s h o w t h a t t h e re
are significant d if f e re n c e s in p la s m a R R R -a -
t o c o p h e ro l B A S E L IN E b e t w e e n s u b je c t s (p
0
.0
0
0
A B
3
1
2
1
3
#{ 212} 8 08
FIG 1. Chromatograms of blank plasma from a subject
and blank plasma spiked with 100 gzg/dL retinol, 80 pgJ
dl retinyl acetate, and 2.68 mg/dL RRR-a-tocopherol.
Peak 1 is retinol, 2 is retinyl acetate, and 3 is RRR-a-
tocopheroL Detector sensitivity 0.05 absorbance units full
scale (aufs) at 292 nm; retention times: 3, 4, and 6.5 mm
for com p ound s 1, 2, and 3, respectively.
lip op r ot einem ia and cystic fibrosis. Figure 1 shows a rep-
resentative chromatogram of a plasma sample.
Riboflavin content was analyzed b y spectrofluorometry
as detailed b y Rivlin (7) except that t he method was au-
tomated with a continuous-flow analyzer system (8). Flu-
orescence was detected with a Perkin-Elmer Fluorescence
Spectrophotometer Model 204-A (Perkin-Elmer Corp,
Eden Prairie, M N), with excit at ion wavelength of 430 nm
and em ission of 510 nm . Blank values were determined
for each sample after sodium hydrosulfite reduction of
riboflavin. T he method was linear ov er t he usual range of
plasma riboflavin concentrations (0-100 ngjmL). Plasma
pyridoxal and pyridoxal phosphate concentrations were
determined using HPLC separation and postcolumn fluo-
rogenic derivatization with semi-carbazide (9). Pyridoxine,
w hich is negligible in plasma, was not measured.
T he st at ist ical analy sis for comparing t he t hr ee products
w as d one using t he analysis of variance (A NO V A ) pro-
cedure fr om t he Statistical Analysis System package (SAS)
(10). M ult ip le comparison of means was achiev ed b y t he
Wailer-Duncan K -r at io t test as an integral part of t he
A NO V A procedure. T he latter is useful in comparing sig-
Q 9
I
C0j
0
A
0
o 00
0
00 0
coo 0 0
A A
A A
A
i 1Q5 14.0 I7 5 21.0
Tune (hrs)
F I G 2 . Me a n RRR-a-tocopherol plasma levels at each
t im e for 12 normal m ales. Each subject ingested 30 IU of
RRR-a-tocopherol acetate contained in three multivitamin
preparations in a crossover fashion. (0) tablet, (0) soft
elastic gelat in (SEG ) cap sule, and (Li) modified-SEG for-
mulation.
1474
THAKKER ET A L

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TABLE 1
Descriptiv e statistics for RRR-a-tocopherol plasma
concentration (mg/dL)
0
0
0
0
A
A
A 00
0
0
A0
40.0
37.5
35.0
E
30.0
a
27.5
22.5
20O
2
7.5
l5
12.5
I0
7.5
0.0 3.5 7.0 10.5 14.0 17.0 21.0
Time (hrs)
FIG 3. M ean riboflav in plasma lev els at each tim e for
12 normal males. Each subject ingested 15 mg of riboflavin
contained in three multiv itainin preparations in a crossov er
fashion. (0) tablet, (0) soft elastic gelatin (SEG) capsule,
and (Lx) m odified-SEG formulation.
0
0
A
I
BIOAVAILABILITY OF M ULTIVITAM INS
1475
Tablet
M inimum
V aiiable M ean SD v alue
M aximum
v alue
BASELINEt 0.73 0.27 0.41 1.44
CM AXf 0.96 0.37 0.66 1.89
TM AX 15.33 7.92 6.00 24.00
M AXBYM INfl 1.33 0.22 1.09 1.86
AUCOTO24II 19.62 7.36 13.42 37.40
Soft elastic gelatin (SEG) capsule
BASELINE 0.69 0.21 0.40 1.13
CM AX 1.23 0.45 0.75 1.97
TMAX 15.33 6.57 8.00 24.00
M AXBYM IN 1.90 1.01 1.27 4.88
A UCOTO24 21.39 7.07 14.26 34.46
M odified-SEG formulation
BASELINE 0.63 0.17 0.37 1.01
CM A X 0.89 0.24 0.53 1.38
TM A X 13.50 6.67 6.00 24.00
M AXBYM IN 1.47 0.40 1.11 2.50
AUCOTO24 15.90 3.45 10.86 21.60
#{ 149} pj l2subjects.
t Concentration at tim e 0.
f M aximum plasma concentration.
Tim e (h) required to attain CM A X .
IIRatio of m axim um to m inim um concentration (nor-
maliz ed for baseline level).
#{ 182} A rea under the plasma concentration v s time curv e
from tim e 0 to 24 h.
= 0.0001) but not products (p = 0.161), in-
dicative of an intrinsically wide interindividual
variation in vitamin E status across the pop-
ulation studied.
The multiple comparison of means test
shows that the TMAX was not significantly
different among the three products. However,
CMAX and MAXBYMIN were both signifi-
cantly higher for the SEG-capsule formulation
compared with the tablet and the modified-
SEG formulation (p < 0 . 0 5 ). The overall
ranking for CMAX, MAXBYMIN, and
AUCOTO24 is SEG capsule> tablet> mod-
ified-SEG formulation.
R iboflavin
The mean riboflavin absorption kinetic data
for the 12 subjects are shown in Figure 3. Table
2 lists the mean, SD, and range for BASE-
LINE, CMAX, TMAX, MAXBYMIN, and
AUCOTO2 4 . The ANOVA results indicate
that differences between products are statisti-
cally nonsignificant for the pharmacokinetic
mo d e l - i n d e p e n d e n t bioavailability parameters
of BASELINE, TMAX, CMAX (although a
trend toward differences between subjects is
indicated, p = 0.0802), and MAXBYMIN.
The param eter AUCOTO24, although not
significantly different among products, is nev-
ertheless significantly different among subjects
( p = 0 . 0 0 3 1 ) , i n d i c a t i n g wi d e i n t e n n d i v i d u a l
differences in the extent of riboflavin absorp-
tion (s e e Discussion). Al t h o u g h t h e me a n
AUCOTO24 was slightly higher for the tablet
compared with the SEG capsule and the
mo d i f i e d - SEG f o r mu l a t i o n , t h e SD wa s a l s o
much higher (Table 2) thereby suggesting that
the two SEG capsule formulations had a more
reproducible release pattern than did the tablet
dosage form. This is consistent with the site-
specific mechanism of absorption of riboflavin
favoring the faster dissolved and available
product (the two capsule formulations) over
the slower dissolved and available tablet for-
mulation. Indeed, the slower release of ribo-
flavin from the tablet favors nonsaturation of
the absorption flux and consequently the
greater extent of absorption reflected in the

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TABLE 2
Descriptiv e statistics for riboflav in plasma concentration
(ng/mL)*
40 .o
0
130
120
A0
0
-90
A
o80
C ) 0
0
0
60 A
0
0
3(J A
20L
l0
0 I I
I i
0.0 3.5 7.0 10.5 140 17.5 210
Tim e (hrs)
FIG 4. M ean py ridoxal plasm a levels at each tim e for
12 norm al males. Each subject ingested 5 m g pyridoxine
hydrochloride contained in three m ultivitam in prepara-
tions in a crossov er fashion. (0) tablet, (0) soft elastic
gelatin (SEG) capsule, and (Es) m odified-SEG formulation.
8
1476 THA K K ER ET AL
Tablet
M inim um
V ariable M ean SD v alue
M axim um
v alue
BA SELINEt 7.00 1.28 5.00 9.00
CM AXf 42.92 19.40 13.00 80.00
TM AX 1.38 0.80 0.50 3.00
M AXBYM INII 6.31 2.67 1.86 11.43
AUCOTO24II 378.22 124.40 186.50 587.75
Soft elastic gelatin (SEG) capsule
BASELINE 6.75 0.87 5.00 8.00
CM AX 41.08 22.13 11.00 78.00
T MAX 1.17 0.54 0.50 2.00
M AXBYM IN 6.08 2.83 1.82 11.14
AUCOTO24 325.56 98.97 202.75 492.75
M odified-SEG formulation
BASELINE 7.58 3.32 5.00 17.00
CM AX 31.75 13.59 13.00 55.00
TM AX 1.17 0.65 0.50 2.00
M AXBYM IN 4.63 2.58 1.86 11.00
A UCOTO24 328.02 90.28 190.25 480.75
* n = 12 subjects.
t Concentration at time 0.
M aximum plasma concentration.
Tim e (h) required to attain TM A X .
H Ratio of m axim um to m inim um concentration (nor-
maliz ed for baseline level).
#{182} Area under the plasma concentration -time curv e from
tim e 0 to 24 h.
h ig h e r m e a n AUC O TO 2 4 v a lu e s . Th e g r e a t e r
S D o f t h e m e a n AUC O TO 2 4 v a lu e s fo r t h e
t a b le t is t h e n e t e ffe c t o f t h e in t e r a c t io n b e -
t w e e n t h e t r a n s it t im e a n d d is s o lu t io n a n d
a v a ila b ilit y a t t h e o p t im a l in t e s t in a l s e g m e n t
(1 1 , 1 2 ). A s im ila r o b s e r v a t io n c a n b e m a d e
fo r t h e e x t e n t o f absorption o f v it a m in E fr o m
t h e m o d ifie d -S E G fo r m u la t io n c o m p a r e d t o
t h e S E G c a p s u le a n d t a b le t (Ta b le 1 ).
Pyridoxal
B oth pyridoxal a n d py ridoxal p h o s p h a t e r e -
s p o n d r a p id ly t o a n oral dose o f v it a m in B -6
a n d t h u s m a y b e used a s in d ic a t o r s o f a b s o r p -
t io n in c o n t r o lle d e x p e r im e n t s s u c h a s t h is .
P yridoxal in p la s m a is d e r iv e d fr o m h y d r o ly s is
o f p y r id o x a l p h o s p h a t e w h ic h is fo r m e d fr o m
t h e oral d o s e o f p y r id o x in e . Th e p lo t o f m e a n
p y r id o x a l le v e ls a t e a c h t im e fo r t h e t h r e e
products is show n in Figure 4 . Ta b le 3 lis t s t h e
m ean, S D, a n d r a n g e fo r B A S E LIN E , C M A X ,
TMAX, MAXB YMIN, a n d AUC O TO 2 4 .
Th e ANO VA r e s u lt s s h o w s ig n ific a n t d iffe r -
e n c e s a m o n g p r o d u c t s fo r t h e p a r a m e t e r s o f
B AS E LINE (in t e r in d iv id u a l d iffe r e n c e s p r o b -
a b le , p = 0 . 0 5 1 ) b u t n o t TMAX a n d MAX-
B YMIN. C MAX is s ig n ific a n t ly d iffe r e n t b e -
t w e e n p r o d u c t s (p = 0 . 0 3 6 5 ) a n d s u b je c t s (p
=0 . 0 0 3 2 ) a s is AUC O TO 2 4 fo r b o t h p r o d u c t s
(p = 0 . 0 2 4 4 ) a n d s u b je c t s (p = 0.0001).
W a le r -Du n c a n K-r a t io t t e s t fo r t h e c o m -
p a r is o n o f m e a n s in d ic a t e s t h a t t h e m e a n
C MAX fo r S E G c a p s u le s is significantly h ig h e r
t h a n t h a t fo r t h e m o d ifie d -S E G fo r m u la t io n
(p <0 . 0 5 ). Th e m e a n AUC O TO 2 4 also is
s ig n ific a n t ly h ig h e r fo r t h e S E G c a p s u le
a n d t h e t a b le t fo r m u la t io n s c o m p a r e d w it h
t h e m o d ifie d -S E G fo r m u la t io n (p < 0.05).
Th e o v e r a ll r a n k in g b a s e d o n C MAX a n d
AUC O TO 2 4 is S E G c a p s u le > t a b le t > m o d -
ifie d -S E G fo r m u la t io n .
Pyridoxal phosphate
P yridoxal phosphate ordinarily is t h e p r i-
m a r y c ir c u la t in g fo r m o f v it a m in B -6 . It is t h e

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TABLE 3
Descriptive statistics for py ridoxal plasm a concentration
(pm ol/m L)*
0
0
A
8
A
0
0
A
0 0
95
9 0
85
8 0 0
0 o
7 5 0
. 000
0
-6 5 A
9 A
o OAA
60
. 0
A
50
. 0
A
4o
35
3 0 -
0.0
BIOAVAILABILITY OF M ULTIVITAM INS 1477
Tablet
M inim um
V ariable M ean SD v alue
M axim um
v alue
BASELINEt 25.00 14.48 9.00 65.00
CM AXf 157.25 79.24 53.00 343.00
TM AX 1.25 0.45 1.00 2.00
M AXBYM INII 6.90 2.78 3.11 11.62
AUCOTO24II 1175.35 521.89 571.50 2478.50
Soft elastic gelatin (SEG) capsule
BASELINE 24.92 10.87 9.00 47.00
CM AX 171.75 73.79 59.00 342.00
TM AX 0.92 0.42 0.50 2.00
M AXBYM IN 7.38 2.89 4.22 14.87
AUCOTO24 1161.81 471.99 478.50 2289.80
M odified-SEG form ulation
BASELINE 21.33 9.73 12.00 46.00
CM AX 117.50 50.04 46.00 210.00
TM AX 1.00 0.67 0.50 3.00
M AXBYM IN 6.10 2.96 2.26 12.27
AUCOTO24 987.42 461.44 358.00 2246.30
* n = 1 2 subjects.
t Concentration at time 0.
f M aximum plasm a concentration.
Tim e (h) required to attain CM AX.
I I Ratio of m axim um to m inim um concentration nor-
maliz ed for baseline level.
#{ 182} A rea under the plasma concentration -time curv e from
time 0 to 24 h.
m a jo r c o e n z y m ic fo r m o f v it a m in B -6 , is
fo r m e d p r im a r ily in t h e liv e r fr o m d ie t a r y v i-
t a m in B -6 , a n d c o m p r is e s o v e r h a lf o f t h e t o t a l
b o d y p o o l o f v it a m in B -6 (1 3 ). Th e c o n c e n -
t r a t io n o f p y r id o x a l p h o s p h a t e in p la s m a is
c o n s id e r e d t o b e r e lia b ly in d ic a t iv e o f v it a m in
B -6 n u t r it io n a l s t a t u s (1 3 -1 5 ).
F ig u r e 5 d e p ic t s t h e fo r m a t io n o f p y rid o x a l
p h o s p h a t e (P LP ) a s a fu n c t io n o f t im e fo r t h e
t h r e e p r o d u c t s a n d Ta b le 4 lis t s t h e m e a n , S D,
a n d r a n g e fo r t h e ir p h a r m a c o k in e t ic -m o d e l
in d e p e n d e n t b io a v a ila b ilit y p a r a m e t e r s o f
C MAX, TMAX, a n d AUC O TO 2 4 a n d fo r
B AS E LINE a n d MAXB YMIN. Th e ANO VA
r e s u lt s in d ic a t e a h ig h ly s t a t is t ic a lly s ig n ific a n t
d iffe r e n c e a m o n g s u b je c t s (p = 0 . 0 0 0 1 ) b u t
n o t p r o d u c t s fo r t h e p a r a m e t e r B AS E LINE .
Th e p a ra m e te rs TMAX a n d MAXB YMIN a r e
n o t s ig n ific a n t ly d iffe r e n t a m o n g t h e t h r e e
p r o jd u c t s . C MAX is s ig n ific a n t ly d iffe r e n t b e -
t w e e n p r o d u c t s (p = 0 . 0 1 4 7 ) a n d s u b je c t s (p
= 0 . 0 0 0 1 ), a s is t h e AUC O TO 2 4 b e t w e e n
p r o d u c t s (p = 0 . 0 4 5 9 ) a n d s u b je c t s (p
= 0 . 0 0 0 1 ).
Th e W a lle r -Du n c a n K-r a t io t te s t in d ic a t e s
t h a t t h e m e a n C MAX a n d AUC O TO 2 4 a r e
s ig n ific a n t ly h ig h e r fo r t h e S E G -c a p s u le fo r -
m u la t io n t h a n t h e m o d ifie d -S E G fo r m u la t io n
(p <0 . 0 5 ). Th e ov erall r a n k in g o n t h e b a s is
o f C MAX a n d AUC O TO 2 4 is S E G c a p s u le
> t a b le t > m o d ifie d -S E G fo r m u la t io n , w h ic h
is s im ila r t o t h e r e s u lt s o b t a in e d fo r p la s m a
p y r id o x a l le v e ls .
Dis c u s s io n
Th e in t e s t in a l a b s o r p t io n o f d r u g s c a n b e
m a r k e d ly in flu e n c e d b y fo r m u la t io n fa c t o r s a s
w e ll a s b y t y p e o f o ra l d o s a g e fo r m u tiliz e d fo r
a d m in is t r a t io n (1 6 ). W e ll-fo r m u la t e d h a rd
g e la t in a n d S E G c a p s u le s , b e c a u s e t h e y d o n o t
u n d e r g o h ig h -c o m p a c t io n fo r c e s d u r in g t h e ir
m a n u fa c t u r e (u n lik e t a b le t s ), w o u ld b e e x -
p e c t e d t o h a v e m o r e r a p id in v iv o r e le a s e p r o -
file s (1 6 ). F u r t h e r m o r e , c o m p o u n d s fo r m u -
t I I I I I -
3.5 7.0 05 14.0 17.5 21.0
Time (hrs)
FIGS. M ean py ridoxal phosphate plasma lev els at each
time for 12 norm al m ales. Each subject ingested 5 m g
py ridoxine hy drochloride contained in three m ultivitam in
preparations in a crossov er fashion. (0) tablet, (0) soft
elastic gelatin (SEG) capsule, and (A) m odified-SEG for-
m ulation.

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TABLE 4
Descriptiv e statistics for py ridoxal phosphate plasma
concentration (pm ol/m L)*
Tablet
M ean
45.92
101.08
9.83
2.45
1985.70
SD
29.37
71.40
7.87
0.84
1337.92
M inimum
v alue
9.00
30.00
1.00
1.40
622.70
V ariable
BA SELINEt
CM A X f
TM A X
M A X BY M INII
AUCOTO24
BASELINE
CM AX
TMAX
M AXBYM IN
A UCOTO24
M aximum
v alue
92.00
299.00
24.00
4.27
5621.00
132.00
305.00
24.00
4.00
6185.00
76.00
178.00
24.00
4.38
2943.80
1478 THA K K ER ET AL
Soft elastic gelatin (SEG) capsule
47.83 33.00 11.00
102.75 69.82 34.00
10.17 7.37 1.00
2.37 0.78 1.42
2075.45 1422.87 699.20
M odified-SEG formulation
BA SELINE 38.58 17.61 18.00
CMAX 80.17 37.47 35.00
TM AX 12.25 6.15 3.00
M AXBYM IN 2.27 0.91 0.88
A UCOTO24 1598.24 628.58 684.20
*p l2subjects.
t Concentration at tim e 0.
t M aximum plasma concentration.
Tim e (h) required to attain CM A X .
II Ratio of m axim um to m inim um concentration (nor-
maliz ed for baseline level).
#{ 182} A rea under the plasma concentration -time curv e from
tim e 0 to 24 h.
lated in SEG capsules because they represent
liquid fills, as opposed to hard gelatin capsules
which encapsulate a dry-pow der blend, often
tend to be better absorbed (1-5).
In this study, the absorption of the oil-so!-
uble vitamin, vitamin E, was significantly en-
hanced when formulated in the SEG-capsule
formulation but not in the m odified-S E G for-
mulation. Such an increased oil-soluble vita-
min absorption phenomenon from SEG cap-
sules at higher dosages than that studied here
has been reported before (2). It is also known
that surfactants enhance t h e d i s s o l u t i o n r a t e
of drugs from dosage forms (by means of their
w etting properties) when used in concentra-
tions below their critical m icelle concentra-
tion. Above their critical micelle concentra-
tion, however, surfactants form micelles, so!-
u b i l i z i n g t h e drug and thereby causing an
overall decrease in the amount of released and
absorbed drug (16). Alternatively, surfactants
can alter membrane perm eability, thereby af-
fecting absorption.
This study show s that the modified-SEG
formulation was not equivalent to the SEG-
capsule formulation with respect to the ab-
sorption ofR R R -a-tocopherol, riboflavin, and
pyridoxine. Interestingly, the bioavailability of
the water-soluble vitamins showed no statis-
tically significant differences betw een the tablet
and the SEG-capsule formulation. However,
there was a consistent trend towards increased
absorption of pyridoxine, as reflected by
plasma pyridoxal and consequent pyridoxal
phosphate from the SEG-capsule product
com pared to the tablet.
A brief discussion of probable ethnic differ-
ences (as evidenced by the significant between-
subject variability in means) in the baseline
status and in the absorption of the oil-soluble
and water-soluble v i t a mi n s assay ed in this
study, is appropriate. For the vitam in E study
there was a wide distribution of BASELINE
means with subjects H , C, and B at one end
with highest means, subject L with the lowest
mean, and everyone else in the middle with
overlapping means. Subject C is African
(G hanian), B is British, L is Indian (Asian),
and H is American. The parameters M A X -
BYMIN and AUCOTO24 were also signifi-
cantly higher for the American subject H (cor-
relating to his higher cholesterol level). For the
water-soluble vitamins, the mean extent of
absorption AUCOTO24 and the mean CMAX
parameter show a wide distribution with sub-
jects C (African, Ghanian) and K (Indian,
Asian) positioned at the lower and upper end
of the overall distribution curve, respectively.
Moreover, both these individuals were very
consistent with respect to intraindividual
variations in absorption of these vitamins, ie,
subject K was consistently high regardless of
product and subject C was consistently low
regardless of product.
In sum m ary, it can be concluded that the
absorption of oil-soluble vitamins such as vi-
tam in E can be significantly enhanced from
m ultivitam in f o r mu l a t i o n s b y f o r mu l a t i n g t h e
vitamin in a surfactant-based SEG capsule.
Similar behavior would also be expected for
other oil-soluble vitamins such as vitamin A
when formulated in the surfactant base as has
been reported previously (2). Further, a con-
sistent, albeit statistically nonsigni#{241}cant, trend
towards increased absorption of pyridoxine
(and consequent pyridoxal and pyridoxal

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BIOAVAILABILITY O F MULTIVITAMINS 1479
p h o s p h a t e f o r ma t i o n ) a n d t o a lesser extent
riboflavin a b s o rp tio n w a s n o tic e d with the
S E G -c a p s u le f o r mu l a t i o n a s c o m p a re d w ith
the tablet. The modified-SEG formulation,
however, w a s significantly le s s e ffe c tiv e c o m -
p a re d w ith th e ta b le t a n d th e S E G -c a p s u le for-
mulation.
Ethnic proclivities in the absorption and/or
m e ta b o lis m a n d d is p o s itio n kinetics of th e s e
vitamins are implicated, and th e s e n e e d to be
e v a lu a te d in a s e p a ra te s tu d y with a larger
s a m p le of n o rm a l s u b je c ts of d iffe re n t ra c ia l
and ethnic b a c k g ro u n d s . Such p re d is p o s itio n
could have w id e -ra n g in g im p lic a tio n s o n
m in im u m d a ily requirements and c o n s e q u e n t
d o s in g re g im e n s .
References
1. Horwitt MK, Elliot WH, Kanjanaggulpan P, Fit ch
C D. Serum concentrations of a-tocopherol after
ingestion of various vitamin E preparations. A m J
Clin Nut r l984;40:240-.5.
2. Bateman NE, Uccellim DA . Effect of formulation on
the bioavailability of retinol, d-a-tocopherol and r i-
boflavin. J Pharm Pharmacol l984;36:46l-4.
3. Lindenbaum J. Greater bioavailability of digoxin so-
lut ion in cap sules. C lin Pharmacol T her l977 ,278-
82.
4. G und er t -R em y U, Bi l z e r W , Lehmann M P, Weber
E. Biologische Verfugabarkeit verschiedener Darrei-
chungsformen. Die pharmazeutische industrie 1974;
36:574-6.
5. Albert KS, Sedman AJ, Wilkinson P, St oll R G , M ur -
r ay W J, Wagner J. Bioavailability studies of acet -
am inop hen and nitrofurantoin. J Clin Pharmacol
1974; 14:264-70.
6. US Pharmacopeia XXI-National formulary XVI, 21st
rev. Rockville, M D: US Phar m acop eial C onv ent ion,
Inc. 1985.
7. R.ivlin RS. Riboflavin in biological fluids and tissues.
In: R iv lin R S, ed . Riboflavin. New Y or k Plenum
Press, 1975:51-79.
8. K ir k JR . Automated method for t he analy sis of r i-
boflavin in m ilk with application to ot her selected
foods. J Assoc Off Anal Chem l974;57: 1085-8.
9. Gregory if, Litherland SA . Efficacy of t he r at bioassay
for t he determination of biologically available vitamin
B-6. J Nut r l986;l 16:87-97.
10. SA S Institute. SAS users guide: statistics, Cary, NC :
SAS Institute, 1982.
11. Thakker KM. Predicting t he dose-dependent bio-
availability of hydrocortisone and chlorothiazide in
humans. J Pharm Sci 1983;72:577-8.
12. Wood JH, Thakker KM. Michaelis-Menten kinetics
in t he absorption of drugs-examples and im p lica-
t ions. Eur J Clin Pharmacol l982;23:l83-8.
13. Lumeng L, Li T-K, L w A . T he interorgan transport
and metabolism of vitamin B-6. In: Reynolds RD,
Leklem JE, eds. Vitamin B-6: it s role in health and
disease. New York: Alan R Liss, Inc, 1985:35-54.
14. L um eng L , R y an M P, L i T -K . Validation of t he d i-
agnostic v alue of plasma PLP measurements in v i-
tamin B-6 in t he human. Am J Clin Nutr l97225:
629-42.
15. Shane B. Vitamin B-6 and blood. In: Sauberlich H,
Brown M, eds. Hum an vitamin B-6 requirements
Washington, DC: National Academy of Sciences,
1978:18.
16. G ib ald i M . Biopharmaceutics and clinical pharma-
cokinetics 3rd ed. Philadelphia, PA : Lea & Febiger,
1984.

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