THEJOURNALOF BIOLOGICAL CHEMISTRY
4177-4182 1984
0 1984 by The American Society of Biological Chemists, Inc.
Inhibition of Oxidation of Methyl Linoleate in Solution by Vitamin E
and VitaminC*
Etsuo Niki, Tadatoshi Saito, Akira Kawakami, and Yoshio Kamiya
From the Department of Reaction Chemkty, Faculty of Engineering, University of Tokyo, Hongo, Tokyo 113, Japan
The oxidation of methyl linoleate in solution initiated
with azo compounds has been studied in the absence
and presenceof vitamin E and vitaminC. Both vitamin
E and vitaminC acted as a chain-breakingantioxidant
and they suppressed the oxidation and produced an
induction period. The inhibition rate constant for the
scavenging of peroxy radical was calculated at 37 OC
as A+ = 6.1 x 10' M" s-l and 7.6 x lo4 M" s-l for
vitamln E and vitaminC, respectively. It was suggested
that each vitamin E could trap two peroxy radicals,
whereas vitamin C could trap only one peroxy radical
under the reaction conditions employedin this study.
When both vitaminE and vitaminC were present, the
oxidation was suppressed quite efficiently and the ap-
parent inhibition rate constant was obtained as & =
4.0 X lo6 M - ~s-'. Furthermore, vitamin E remained
almost unchanged and only vitamin C was consumed
at the initial stage and vitamin E was consumed after
vitamin C was exhausted. It was concluded that vita-
min E trapped the peroxy radical and theresulting a-
chromanoxy radical reacted with vitamin C to regen-
erate vitamin E.
Oxidation of unsaturated compounds by molecular oxygen
has been the subject of extensive study from the late 1940's
from the viewpoint of the deterioration of rubber, plastics,
and foods (1). The oxidation of polyunsaturated fatty acids
in the lipids and its inhibition has received much attention
recently (2-9) in connection with its pathological effects such
as inflammation (lo), platelet aggregation (ll), asthma (12),
and anaphylaxis (13). Peroxidation of membrane lipids has
been implicated as one of the primary events in oxidative
cellular damage (14).
We have been studying the oxidation of polyunsaturated
fatty acids in vitro and its inhibition, aiming primarily at
elucidating the rate andmechanism of lipid peroxidation and
its inhibition (15-18). In the present study, we have studied
the inhibition of oxidation of methyl linoleate in solution by
vitamin E and vitamin C.
EXPERIMENTAL PROCEDURES
Materials-Methyl linoleate was obtained from Sigma Chemical
CO. and used as received. Prior to the oxidation, neither conjugated
diene nor peroxides were detected (17). Commercial AIBN' and
* This paper is Part VI of the series on Oxidation of Lipids. The
costs of publication of this article were defrayed in part by the
payment of page charges. This articlemust therefore be hereby
marked "advertisement" in accordance with 18 U.S.C. Section 1734
solely to indicate this fact.
The abbreviations used are: AIBN, 2,2'-azobisisobutyronitriile;
AMVN, 2,2'-azobis(2,4-dimethylvaleronitrile); BMP, 2,6-di-t~rt-bu-
tyl-4-methylphenol; LH, methyl linoleate; IH, antioxidant; Ri, rate, of
chain initiation; R,, rate of oxygen uptake after induction period or
in the absence of antioxidant; R*, rate of oxygen uptake during
induction period.
4177
Vol. 259, No. 7, Issue of April 10,pp.
Printed in i r . ~ . ~ .
(Received for publication, July 13,1983)
AMVN were recrystallized from methanol. Natural d-a-tocopherol
(vitamin E) was kindly supplied from Eisai Co. Commercial L-=or-
bic acid (vitamin C) was used as received.
Procedures-The rate of oxidation was determined either by fol-
lowing the oxygen concentration in solution or by measuring the
pressure decrease due to oxygen uptake. Oxygen concentration in
solution was monitored by Biological OxygenMonitor, Model YSI53
(Yellow Springs Instrument Co., Yellow Springs, OH). Oxygen pres-
sure was measured by an automatic recording gas absorption appa-
ratus described previously (15) using a sensitive pressure transducer,
Model PMS-5M (Toyoda Machine Works, Ltd.,Kariya, Japan).
The reaction mixture was prepared by dissolving an appropriate
amount of radical initiator into solvent followed by the addition of
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methyl linoleate. The homogeneous solution was then taken into a
reaction vessel. The total liquid volume wasmaintained at 5 ml. The
reaction vesselwas connected to an oxygen electrode or pressure
transducer and then it was immersed into a water bath which was
carefully temperature-controlled at 37 or 50 'C. The reaction mixture
was stirred with a magnetic stirrer.
Vitamin E and vitamin C were monitored at 290 and 275 nm,
respectively, with a high performance liquid chromatography, model
Tri-Rotar, Japan Spectroscopic Co. Finepak SIL C18 column was
used and methanol was delivered as an eluent at 1 ml/min.
RESULTS AND DISCUSSION
Table I shows the rate of oxidation of methyl linoleate in
solution at 37 "C initiated byAIBN or AMVN. The azo
initiators were used in order to obtain a constant andknown
rate of chain initiation, which isessential to the kinetic study.
The oxidation of methyl linoleate under mild conditions
can be represented by the following scheme(15, 19,20).
Initiation
A-N=N-A -kd
(1 - e)A-A
A. + 02 + AOO.
+ 2eA' + Nz
AOO. + LH -%0 AOOH + LOO.
Propagation
LOO. + LH kP
L. +
-
0 24
LOOH
LOO
+ L.
Termination
2 LOO'
In this scheme, A-N=N-A
-2k nonradical products
is the azo initiator, LH repre-
(6)
sents methyl linoleate, and L. and LOO. are alkyl and alkyl-
peroxy radicals generated therefrom, respectively. When the
rate of initiation is constant, that is, when the new chains are
started at a constant rate, the steady state treatment can be
applied and the rateof oxidation is given by Equation 7 (15,
4178 Inhibition of Oxidation by Vitamins E and C
TABLEI 100
Rate of oxidation of LH in tert-butyl alcohol/methuml at 37 "C
101bi7
Run [LH] '2:;' [ A W N ] [AIBN] ky T
i (kcl); (23)'' -.80
dp

C
M d/ml mM mM Mfs Mfs (MS)-* .IU
1 1.21 3/0 10.3 2.66 3.29 124 1.67 m
2
3
0.906 3.5/0
0.604 4/0
10.2
40.2
2.63
10.4
2.31
3.95
88
38
1.57
2.03
2 60
9)

4 0.604 4/0 20.5 5.29 2.77 52 1.99 C

5 0.604 4/0 10.9 2.81 2.36 84 2.33 u
6 0.604 4/0 10.2 2.63 1.88 71 1.92 5 40
7 0.604 4/0 3.99 0.096 0.211 220 1.13
*
br
8
9
0.604
0.604
6/2
3/1
10.0
9.94
2.58
2.56
2.00
1.53
78
60
2.06
1.58 ::
10 0.336 8.90 2.30 1.39 61 2.73 20
3/1
11 0.336 6/2 8.87 2.29 1.31 57 2.58
a Determined as described in the text.
* Kinetic chain length, (kcl), = Rp/Ri. 0
e Oxidizability constant,b/(24)'/2 = R,/[LH]Ri'".

.~ ,.,,._ . ? .~,,.- - ". ..." , ..

1.. .
.1. , . -" I..I. . 0 20 40 60 80 120 100
."
. , ,

Time/min
FIG. 2. Inhibition of oxidation of methyl linoleate in tee-
butyl alcohol byvitamin E at 37 OC. rAMvNl= 0.01 M.

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Curve 1 2 3 4 5
_____
[Vitamin E ] / ~ M 045 22 67 112
tds 0 2280 3600 5340 6420

6000 I I I

. 5000k
4000

Time
FIG. 1. Rate of oxygen uptake in the oxidation of methyl 1000
linoleate in the absence (curve 2 ) and presence (curve 2 ) of
antioxidant.
0
I 1
19,201, 0 1000 2000 3000
[vitamin EI/Rif 5

FIG. 3. Plot of induction period as a function of [vitamin

E]/& in theoxidation of methyl linoleate in solution at 37 "C.
The line corresponds to slope = 2.0. 0, initiated with AMVN; 0,
where Ri is the rate of chain initiation, kp and 2kt are the rate initiated with AIBN.
constants for the rate-controlling chain propagation (Reac-
tion 4) and termination steps (Reaction 6 ) , respectively. The with decreasing rate of chain initiation, suggesting that the
rate constant ratio kp/(2kt)"is generally referred to as the peroxy radicals formed in Reaction 5 propagate the chains by
oxidizability of the substrate. attacking anothermethyl linoleate substrate intermolecularly
Methyl linoleate is oxidized by a straightforward mecha- (Reaction 4) before they decay by mutual interactions and
nism: it has been found that at the initial stage of the terminate chains (Reaction 6 ) .
oxidation of methyl linoleate, the rates of oxygen uptake, The rate of chain initiation was determined by the conven-
substrate disappearance, and peroxide and conjugated diene tional inhibitor method (20, 21,22).
formation agreed well with each other (15). Hence, the rate Ri = 2e&[A-N=N-A] = n[IHJ/tu (8)
of oxidation wasfollowed quantitatively in this study by
following the rate of oxygen uptake. The oxidation of methyl where IH is the chain breaking antioxidant, t u is the induc-
linoleate initiated by AMVN or AIBN proceeded smoothly tion period producedby the addition of an inhibitor, and n is
without any induction period and a constant rate of oxygen the stoichiometric number of radicals trapped by each inhib-
uptake was observed. As shown in Table I, the rate of chain itor. The value of n was taken as 2 (20).
initiation was varied from 1 X lo-' to 1 X lod7M/S and the The oxidizability kJ(2k)" of methyl linoleate calculated
kinetic chain length, that is, the number of propagation per from Equation 7 is also shown in Table I, the average being
one initiation,was always much larger than one andincreased 0.0181 and 0.0224 in tert-butyl alcohol and in the mixture of
 Inhibition of Oxidation by Vitamins E and C 4179
TABLE I1
Inhibition of oxidation of 0.604 M LH in solution at37 "cby vitamin E and vitamin c
Run Solvent" [AIBN] [AMVN] l@R, [Vitamin E] [Vitamin C] tm Rm ' (kcl)mb h ldR, (kcl),' :$%
"1 s-l (Ma)"
mM mM MfS PM IrM 8 Mfs
12
0.272 A
0.958 3.99 980 14.0 146 4.40 0.198 207 1.06
13
0.815 A
0.958 3.99 1680 5.05 0.183
53 7.12 191 0.98
14
0.815 A
0.967 4.03 0.191 3.89 752140 7.25 197 1.02
15
0.997 A
0.958 3.99 5.78 2260 60
0.256 4.62 1.37 267
1.50
16 A
0.958 3.99 3040 3.37 35
0.186 5.90 1.00 194
1.71
17 A
0.958 3.99 0.178 5.03 353560 3.37 186 0.95
40 1620 0.997
18 B
0.958 3.99 1.27 247
5.04 0.237
19 B 25.8 10.0 41 1860 30.1 2.07
12 1.08 80 2.13
20 B 25.8 10.0 55 2.07 243078 26.1 2.01
10 0.95
21 B 26.1 10.1 1.59 0.53110 7.9 5460 20.7 61 1.63
22 B 10.0 25.8 1.38 0.45
166 7.7 6750 19.9 54 1.42
0.213 23
0.997 B
0.958 3.99 1.35 264
1680 0.253 4.10
2.35 0.99724 B
0.958 3.99 1.16 2020227 0.217 3.79
5.16 0.99725 B
0.970 4.04 1.25 1840
243 0.236 4.64
10.3 0.99726 B
0.970 4.04 1.18 1960229 0.222 3.99
25.8 27
0.997 B
0.958 3.99
284 28
0.997 B
0.958 3.99
A, 4 ml of tert-butyl alcohol; B, 3 ml of tert-butyl alcohol + 1 ml of methanol.
'Kinetic chain length during induction period.
Kinetic chain length after induction period.
Oxidizabilityconstant &.J(2k)* = R,/[LH]Ri".

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TABLE111
Inhibition rate constant &for vitamin E
Temper- Reference
Peroxy radical Technique ature
"C "1 s-l

Pulse radiolysis 5.0 X 10s 26

Pulse radiolysis 2.3 X lo7 27
Chemiluminescence 60 3.3 x 108 28
Oxygen consumption 30 2.35 X 10' 23
Chemiluminescence 37 1.8 x 106 29
and tinh 37 5.1 X lo6 This work
Oxygen consumption 60 2.0 xo
bl 30
Chemiluminescence 1.5 25 X IO6 16
Pulse radiolysis 8 X 104 31

tert-butyl alcohol and methanol (3:l by volume), respectively.
Vitamin E has been known to act as an antioxidant both
in vivo and in vitro by scavenging chain propagating peroxy
radicals (16,18,23,24). Fig. 1illustrates the patternof oxygen
uptake curve in the oxidation of methyl linoleate and shows
the effect of addition of antioxidant on the oxygen uptake. In
the presence of antioxidant (curve 2 in Fig. l), the rate of
oxygen uptake issuppressed. When all the antioxidant is
consumed, the induction period is over and the oxidation
proceeds at the same rate as in the absence of inhibitor. In
this paper, the rate of oxygen uptake during and after the
induction period are designated as Rinh and R,, respectively.
In the presence of antioxidant, the chains are terminated
by Reactions 9 and 10 instead of Reaction 6, where IH
represents the antioxidant and I' the radical generated from
the antioxidant by donating a hydrogen atom to chain-prop-
agating free radicals.
by reacting with each other to give dimers, but in the presence
of peroxy radical the chromanoxy radical should react pre-
dominantly with the peroxy radical.Since, as mentioned later
in the text, n is close to 2, Reaction 10 should give LOOI.'
When all the 1. radicals are destroyed by Reaction 10, the
rate of oxidation during the induction period is givenby
Equation 11 (23).
R i a h =$[LH]Ri
- (11)
nkhUH1
Table I1 and Fig. 2 show some representative data for the
oxidation of methyl linoleate inhibited by vitamin E. Metha-
nol was added as a solvent in order to increase the solubility
of vitamin C. It is noteworthy that therate of oxidation after
the induction period is the same as that in the absence of
vitamin E, that is, the lines in Fig. 2 after theinduction period
are parallel with each other.

LOO' + IH
(n - 1)LOO' + 1.
- kh LOOH + I'

nonradical
products
(9)
(10)
Equation 8 suggests that the induction period is propor-
tional to the ratio of the inhibitor concentration to the rate
of initiation, [IH]/Ri. Fig. 3 shows this plot for AIBN- and
-D AMVN-initiated oxidations inhibited bv vitamin E. A satis-
factory straight line was obtained andthe slope, n, is very
Theconstant n is, as mentioned above, the stoichiometric close to 2.0, or strictly speaking, for vitamin E and
factor for the entioxidant. Vitamin E reacts withperoxy
radical by donating phenolic hydrogen atom to yield chro- z ~ ~ h ~ ~ has~ been h suggested, it has
been
manow radical, which can be observed by electron spin
resonance analysis (18).The chromanoxy radicals can decay
identifiedquantitatively. We are currentlyworkinn to isolateand
identify the-product of &action 10.
-
4180 Inhibition of Oxidation by Vitamins E and C

0 20 40 60 80 100 120
0 20 40 60 80 100
Time/min
Time/min
FIG.4. Inhibition of oxidation of methyl linoleate in tert-
butyl alcohol/methanol(3:1 by volume) by vitamin C at 37 "C, FIG.6. Inhibition of oxidation of methyl linoleate in tert-
[AMVN] = 0.01 M. butyl alcohol/methanol (3:l by volume) by vitamin E and
vitamin C, [AMVN]= 0.01 M.

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Curve 1 2 3 4 5
Curve I 2 3 4
IVitamin C]/pM 0 41 55 110 166
tinh/s 0 1860 2430 5460 6750 [Vitamin E]/pM 0 0 30 30
[Vitamin C]/pM 0 55 0 55
tinh/S 0 1710 2420 3940

I linoleate at 30 "C by Howard and Ingold (25)as k, = 62 M"

s-'. It was also estimated as 230 I "'s-' at 50 "C (17). The
interpolation of these values gives k, = 100 M-' s" at 37 "C.
The rate constant k,,,h calculated from Equation 12 is included
in Table 11. The average value isobtained as & = 5.1 x lo6
I"'s" for vitamin E.3 As compared in Table 111, this is in
fair agreement with those values reported previously.
Fig. 4 shows the effect of vitamin C in the oxidation of
methyl linoleate in tert-butyl alcohol and methyl alcohol(3:l
by volume). Some pertinent results are also shown in Table
11. It can be seen that vitamin C also acts as a chain breaking
antioxidant and produces a distinct induction period. The
rate of oxygen uptake after the induction period is again equal
to that in the absence of antioxidant. Fig. 5 shows the plot of
i induction period t b h as a function of [vitamin c]/&It gives
a fairly straight line and the slope is closeto 1, implying that

0 1000 2000 3000 4000 5000 6000

1
700G
each vitamin C scavenges only one peroxy radicalunder the
present reaction conditions. The rateconstant &, for vitamin
C was also calculated from Equation 12 as shown in Table I1
and the average value was obtained as k,,,b = 7.5 X lo' M-'
[vitamin CI/Ri/s
S-1.
FIG.5. Plot of induction period against [vitamin C]/Rdin The above results show that both vitamin E and vitamin C
the oxidation of methyl linoleate in tert-butyl alcohol/metha- act as a chain-breaking antioxidant. It has been observedthat
no1 (3:l by volume) at 37 "C inhibited by vitamin C. The dotted the combination of vitamin E and vitamin C has a synergistic
line corresponds to slope = 1.0. effect on the inhibition of oxidation (32-35). Nakamura and
Hishinuma (36) have observed that the vitamin E level is
BMP is the same, since the rate of initiation was calculated maintained even in vitamin E-deficient rats and have sug-
from Equation 8 assuming n is 2.0 for BMP. The result of gested the contribution of some repair component. Similar
BMP will be shown in Table IV.
Equations 8 and 11 give Equation 12. Itmust be pointedout that during theinductionperiodthe
antioxidant decreases gradually and the inhibited rate of oxidation,
k , = b[LH]/Rats (12) R a , increases accordingly. The value of R s that must be used in the
calculationof k,from Equation 12 is the initial rate at thevery s t a r t
Since [LH] is known and R b h and t a are determined experi- of the induction period. In this study, some average initial rate of
mentally, the inhibition rate constant & can be calculated oxygen uptake was used, andthe values of k ,thus obtainedmay be
smaller than its true value. However, it may be noteworthy that the
if k, is known. As shown in Figs. 1 and 2, both Rinhand t b h value of & calculated from the quantity of oxygen absorbed after
can be determined accurately from the oxygen uptake record. some specific time during the induction period (23)was similar to
The absolute rate constant kp was determined formethyl those shown in Table11.
 Inhibition of Oxidation by Vitamins E and C 4181
TABLEIV
Inhibition of oxidation of 0.60 hf LH by vitamin E and vitamin C at 50 "C in tert-butyl alcohol/mthanol(3:1 by
volume)
RUn [AMVN] [BMP] [Vitamin E] [Vitamin C] tad 1P RM (kc1)L Id Rp (kcl),'
mM mM mM mM 8 M/s M/S
29 34.6 0 10.2
0.482 30 10.3
31
1.0932 10.2 31.1 9.10 3.8 5,820 1.10
10.2 33 32.4
0.595 9.460.62 0.58,340 0.16
10.3 34
Induction period.
* Kinetic chain length during induction, period.
Kinetic chain length after induction period.

FIG. 7. Disappearance of vitamin

E (0)and vitamin C ( 0 ) in the oxi- 2
A

i
dation of methyl linoleate at 37 OC
intert-butylalcohol/methanol (3:l 2 0.5

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by volume). [LH] = 0.60 M, [AMVN]
= 0.010 M, [vitamin E] = 0.595 mM,
[vitamin C1 = 0.620 mM ( A ) and 1.03 LI

0 50 100 150 0 50 100 150 200 250

Time/nin Timelmln

C to regenerate vitamin E. They havemeasured therate

constant for this reaction as 1.55 X 10' M" s-'. We (18, 39)
have also shown by ESR study that a-chromanoxy radical

R
reacts quite rapidly with vitamin C, glutathione, and thio-
phenol.
In order to verify the synergistic interaction between vita-
min E and vitamin C, the oxidation of methyl linoleate in the
O e4
0.3 presence of both vitamin E and vitamin C was studied. Fig. 6
shows the oxygen uptake in the oxidation of methyl linoleate.
As described above, in the absence of an inhibitor, the oxi-
0.2 dation proceeds smoothly without any induction period (curve
1). The addition of vitamin E (curve 3) and vitamin C (curue
2) suppressed the oxidation and gave a clear induction period.
0.1
Vitamin E gave a slower rate of oxidation during the induction
period than vitamin C, apparently because of a higher inhi-
bition rate constant and n for vitamin E than for vitamin
C. When both vitamin E and vitamin C were added, the
0
induction period was lengthened to the sum of the induction
0 50 100 150 period observed wheneither vitamin E or vitamin C was used
Time/min
alone. What is interesting is that the rate of oxidation was
the same throughout the induction period as the inhibited
FIG. 8. Effect of addition of vitaminC on the rateof disap-
pearance of vitamin E during the oxidation of methyl linoleate rate observed whenvitamin E was used alone.
at 37 OC in tert-butyl alcohol/methanol(3:1by v/v). About 0.5 Table I1 also shows some data for the oxidation of methyl
mM of vitamin C was added at the time indicatedby an arrow. [LH] linoleate in the presence of both vitamin E and vitamin C. It
= 0.60 M, [AMVN] = 0.010 M, [vitamin E] = 0.390 mM. is noteworthy that therate constant obtained in this system,
= 4.0 X lo6 M" s-', is much larger than that for vitamin
observation has been reported elsewhere (37): C and close to that observed forvitamin E.
Packer et al. (26) have shown more directly that a-chro- The rateof consumption of vitamin E and vitamin C during
manoxy radical generated from vitamin E reacts with vitamin the oxidation of methyl linoleate was measured. The results
are shown in Table IV and Fig. 7. When either vitamin E or
'After we submitted this paper, we found an interesting paper by vitamin
Barclay andhis co-workers (38),where they observed a synergism of C wasused alone, they disappeared linearly with
inhibitors vitamins C and E in the autoxidation of linoleic acid and time. However,when both vitamin E and vitamin C were
methyl linoleate in micelles. used, vitamin C was consumed first and vitamin E remained
4182 Inhibition of Oxidation by Vitamins E and C
almost constant. Vitamin E was consumed after vitamin C Vane, J. R. (1978) Biochim. Biophys. Acta 5 2 3 , 250-262
disappeared. Similar result was observed when vitamin C was 12. Packer, C. W. (1979) AUergy Clin. Immuml. 6 3 , 1-17
added during the induction period. Fig. 8 shows that vitamin 13. Corey, E. J., Barton, A. E., and Clark, D. A. (1980) J. Am. Chem.
E decayed linearly with time at first but it was not consumed Soc. 102,42784279
14. Plaa, G. L., and Witachi, H. (1976) Annu. Reu. Phurmacol.
but remained constant when vitamin C was added. To~icol.16, 125-141
These results strongly suggest that vitamin E, which has 15. Yamamoto, Y., Niki, E., and Kamiya, Y. (1982) BuU. Chem. Soc.
higher k+,h than vitamin C , scavenges the peroxy radical more Jpn. 55,1548-1550
quickly than vitamin C , but thevitamin E radical reacts with 16. Niki, E., Tanimura, R., and Kamiya, Y. (1982) Bull. Chem. Soc.
vitamin C to regenerate vitamin E. This sequence may con- Jpn. 55,1551-1555
17. Yamamoto, Y., Niki, E., and Kamiya, Y. (1982) Lipids 1 7 , 870-
tribute to the regeneration and maintenance of vitamin E 877
level in tissue. 18. Tsuchiya, J., Niki, E., and Kamiya, Y. (1983) BuU. Chem. Soc.
Jpn. 56,229-232
vitamin E
L o o x vitamin E'
LOOH 3c vitamin C'
vitamin C
19. Howard, J. A. (1972) Adv. Free Radical Chem. 4.49-173
20. Barclay, L.R.C., and Ingold, K. U. (1981) J. Am. Chem. Soc.
103,6478-6485
Finally, it must be mentioned that the results and discus- 21. Boozer, C. E., Hammond, G. S., Hamilton, C. E., and Sen, J. N.
sion given above relate to the interactions of vitamin E and (1955) J. Am.Chem. Soc. 77,3233-3237
vitamin C in homogeneous solution. Vitamin E is oil-soluble, 22. Niki, E., Kamiya, Y., and Ohta, N. (1969) BuU. Chem. Soc. Jpn.
while vitamin C is water-soluble and, in biological systems, 11,3220-3223
these two vitamins must be located in separate phases. The 23. Burton, G. W., and Ingold, K. U. (1981) J. Am. Chem. Soc. 1 0 3 ,
6472-6477
discussion given in this paper for the homogeneous solution 24. Machlin, L. J. (ed) (1980) Vitamin E Marcel Dekker, New York
can not be directly applied to the biological systems. However, 25. Howard, J. A., and Ingold, K. U. (1967) Can. J. Chem. 45,793-
h u n g et al. (34) have suggestedthe interaction of vitamin E 802
and vitamin C in liposome systems which are more biologi- 26. Packer, J. E., Slater, T. F., and Willson, R.L. (1979) Nature

Downloaded from http://www.jbc.org/ by guest on May 17, 2020

cally related and we have also found recently that this inter-
action can occur in the oxidation of phosphatidylcholine 27. Simic, M. G. (1980) in Autoxidation in Food and Biological Sys-
liposome.6
REFERENCES
1. Lundberg, W. 0.(ed) (1961) Autoxidation and Antioxidants, Vols.
I and 11, Interscience, New York
2. Pryor, W.A. (ed) Free Radicals in Biology (1976) Vol. I, (1976)
Vol. 11, (1977) Vol. 111, (1980) Vol. IV, i n d (1982) Vol.V,
Academic Press, New York
3. Del Maestro, R.F., Thaw, H. H., Bjork, J., Planker, M., and
Arfos, K. E. (1980) Acta Physiol. Scand. Suppl. 492,43-57
4. Oxygen Free Radicals and Tissue Damage (Chiba Foundation
Symp.) (1979) Excerpta Medica, Amsterdam
5. Hayaishi, O., and Asada, K. (eds)(1977) Biochemical and Medical
Aspects of Active Oxygen, Japan Scientific Societies Press,
Tokyo
6. Rodgers, M. A., and Powers, E. L. (eds) (1981) Oxygen and Oxy-
Radicals in Chemistry and Biology Academic Press, New York
7. Caughey, W. S. (ed) (1979) Biochemical and Clinical Aspects of
Oxygen Academic Press, New York
8. Simic, M. G., and Karel, M. (eds) (1980) Autoxidation in Food
and Biological Systems Plenum, New York
9. Yagi,K. (ed) (1982) Lipid Peroxides in Biologyand Medicine
Academic Press, New York
10. Kuehl, F. A., Jr., and Egan, R. W. (1980) Science (Wash. D.C . )
210,978-984
11. Salomon, J. A., Smith, D. R., Flower, R. J., Moncada, S., and
- 39. Niki, E., Tsuchiya, J., Tanimura, R., and Kamiya, Y. (1982)
'E. Niki, A. Kawakami, and Y. Kamiya, unpublished work.
( h n d . ) 278,737-738
tems (Simic, M. G., and Karel, M., ed) pp. 17-26, Plenum, New
York
28. Burlakova, Ye. B., Kukhtina, Ye. N., Ol'Khovskaya, I. P., Sary-
cheva, 1. K., Sinkina, Ye.B., and Khrapova, N. G. (1979)
Biophysics 24,989-993
29. Aristarkhova, S. A., Burlakova, E. B., and Kharapova, N.G.
(1972) Izv. A M . Nauk SSSR Ser. Khim. 2714-2718
30. Kharitonova, A. A., Kozlova, Z. G., Tsepalov, V. F., and Glad-
yshev, G. P. (1979) Kinet. KataL 20,593-599
31. Patterson, L. K. (1981) in Oxygen and Oxy-Radicals in Chemistry
and Biology (Rodgers, M.A. J., and Powers, E.L., eds) pp. 89-
95, Academic Press, New York
32. Tappel, A. L. (1962) in Autoxidation and Antioxidants (Lundberg,
W. O., ed) pp. 506-526, Wiley, New York
33. Cort, W. M. (1974) J. Am. Oil Chem. Soc. 51,321-325
34. h u n g , H.-W.,Vang,M. J., and Mavis, R.D. (1981) Biochim.
Biophys. Acta 664,266-272
35. Cort, W. M. (1982) in Ascorbic Acid: Chemistry, Metabolism, and
Uses (Seib, P. A., and Tolbert, B. M., eds) pp. 533-550, Amer-
ican Chemical Society, Washington, D. C.
36. Nakamura, T., and Hishinuma, I. (1978) in Tocopherol, Oxygen,
and Biomembranes (de Duve, C., and Hayaishi, O., e&) pp. 95-
108, Elsevier/North-Holland, Amsterdam
37. Nakano, M., Sugioka, K., Nakamura, T., and Oki, T. (1980)
Biochim. Biophys. Acta 619,274-286
38. Barclay, L. R. C., Locke, S. J., and MacNeil, J. M. (1983) Can. J.
Chem. 61,1288-1290
Chem. Lett. 789-792
Inhibition of oxidation of methyl linoleate in solution by vitamin E and vitamin C.
E Niki, T Saito, A Kawakami and Y Kamiya
J. Biol. Chem. 1984, 259:4177-4182.

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