The Journal of Nutrition

Nutrition and Disease

Almonds Reduce Biomarkers of Lipid
PeroxidationinOlder Hyperlipidemic Subjects
1,2
David J. A. Jenkins,
35
* Cyril W. C. Kendall,
3,5
Augustine Marchie,
3,5
Andrea R. Josse,
3,5
Tri H. Nguyen,
3,5
Dorothea A. Faulkner,
3,5
Karen G. Lapsley,
6
and Jeffrey Blumberg
7
3
Clinical Nutrition and Risk Factor Modication Center, and
4
Department of Medicine, Division of Endocrinology and Metabolism;
St. Michaels Hospital, Toronto, Ontario M5C 2T2;
5
Department of Nutritional Sciences; Faculty of Medicine, University of Toronto,
Toronto, Ontario M5S 3E2;
6
Almond Board of California, Modesto, CA 95354; and
7
Jean Mayer USDA Human Nutrition Research
Center on Aging, Tufts University, Boston, MA 02111
Abstract
Nut consumption has been associated with reduced coronary heart disease (CHD) risk. In addition to cholesterol-lowering
properties, almonds have been shown to lower oxidized LDL concentrations. However, little is known regarding their
effects on other markers of oxidative stress. The dose-response effects of whole almonds, taken as snacks, were
compared with low-saturated fat (,5% energy) whole-wheat mufns (control) in the therapeutic diets of hyperlipidemic
subjects. In a randomized crossover study, 27 hyperlipidemic men and women consumed 3 isoenergetic (mean 423 kcal/d
or 1770 kJ/d) supplements each for 1 mo. Supplements consisted of full-dose almonds (73 6 3 g/d), half-dose almonds
plus half-dose mufns (half-dose almonds), and full-dose mufns (control). Subjects were assessed at wk 0, 2 and 4. Mean
body weights differed #300 g between treatments, although the weight loss on the half-dose almond treatment was
greater than on the control (P , 0.01). At 4 wk, the full-dose almonds reduced serum concentrations of malondialdehyde
(MDA) (P 0.040) and creatinine-adjusted urinary isoprostane output (P 0.026) compared with the control. Serum
concentrations of a- or g-tocopherol, adjusted or unadjusted for total cholesterol, were not affected by the treatments.
Almond antioxidant activity was demonstrated by their effect on 2 biomarkers of lipid peroxidation, serum MDA and
urinary isoprostanes, and supports the previous nding that almonds reduced oxidation of LDL-C. Antioxidant activity
provides an additional possible mechanism, in addition to lowering cholesterol, that may account for the reduction in CHD
risk with nut consumption. J. Nutr. 138: 908913, 2008.
Introduction
The effectiveness of nuts in lowering serum cholesterol has been
demonstrated (19) and has been used to explain their marked
effect in reducing coronary heart disease (CHD)
8
in cohort
studies (1013). On the basis of these data, the FDA has allowed
a CHD risk reduction qualied health claim for nuts and nut
products that contain 1.5 oz (42) nuts (14). This ruling has done
much to reverse the view that nuts should be excluded from the
diets of those at risk for CHD. Furthermore, there is less concern
over the high fat content of nuts, because the sources of fat are
monounsaturated fatty acids (MUFA) and PUFA. In this respect,
there has recently been a liberalization of MUFA intakes in
guidelines of the AHA (15), the American Diabetes Association
(16), and the National Cholesterol Education Program Adult
Treatment Panel III (17).
Nevertheless, it seems unlikely that the ;5% reduction in
serum cholesterol with nuts (1) could explain more than part of
the average 30% reduction in CHD risk in cohort studies (10
13). In statin trials, a 30% reduction in CHD risk is associated
with a 30% reduction in LDL cholesterol (LDL-C) (18,19).
Additional mechanisms must therefore be sought to explain the
benecial effect of almonds.
The skins of almonds are rich sources of phenolic antioxi-
dants (2022) and nuts are also good sources of vitamin E (1).
Antioxidants have been proposed to reduce CHD through their
ability to decrease oxidative damage to lipids, proteins, and
lipoproteins (23). Oxidized LDL is considered to be more
atherogenic than native LDL (24). Because almonds have been
shown to reduce serum concentrations of oxidized LDL (1), the
antioxidant activity of nuts may provide a further mechanism
for their cardioprotective effects. In addition to their antioxidant
capacity, dietary polyphenols appear also to possess additional
1
Supported by the Canada Research Chair Endowment of the Federal
Government of Canada, USDA Agricultural Research Service under Cooperative
Agreement no. 1950-51000-065-02A, and the Almond Board of California. The
contents of this publication do not necessarily reect the views or policies of the
U.S. or Canadian governments, nor does mention of trade names, commercial
products or organizations imply their endorsement.
2
Author disclosures: D. Jenkins and C. Kendall have sat on the Scientic
Advisory Board and have been on the speakers panel for the Almond Board of
California. D. Jenkins, C. Kendall, and J. Blumberg have received honoraria and
grants from the Almond Board of California. K. Lapsley is employed by the
Almond Board of California. A. Marchie, A. Josse, T. Nguyen, and D. Faulkner, no
conicts of interest.
8
Abbreviations used: CHD, coronary heart disease; LDL-C, LDL cholesterol;
MDA, malondialdehyde; MUFA, monounsaturated fatty acids.
* To whom correspondence should be addressed. E-mail: cyril.kendall@
utoronto.ca.
908 0022-3166/08 $8.00 2008 American Society for Nutrition.
Manuscript received 26 November 2007. Initial review completed 23 December 2007. Revision accepted 19 February 2008.

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cardioprotective functions, such as altering hepatic cholesterol
uptake, triglyceride assembly and secretion, plasma lipoprotein
processing, and inammation (25).
We have therefore assessed the effect of almonds on antiox-
idant vitamins and lipid markers of oxidative damage, including
serum malondialdehyde (MDA) and urinary isoprostane output,
to determine whether the antioxidant property of almonds may
be a further reason for their cardioprotective effects. The lipid
data from this almond dose-response study have been published
previously and demonstrated reductions in both LDL-C and
oxidized LDL (1).
Methods
Subjects. Healthy hyperlipidemic men and postmenopausal women
were recruited by newspaper advertisement and from patients attending
the Risk Factor Modication Center of St. Michaels Hospital, a
University of Toronto teaching hospital. Of the 43 subjects who started
the study, 16 withdrew during or after completing 1 or 2 study phases.
Three quit for reasons directly related to the study (food allergies, n 2;
abdominal discomfort, n 1). The majority of subjects (n 13)
withdrew for unrelated reasons. Twenty-seven subjects (15 men and 12
postmenopausal women) completed all 3 phases of the study. Four
subjects were $75 y but were healthy and interested in the study. The 27
subjects who completed the study were (mean 6 SD) 64 6 9 y (range,
4886 y) and had a BMI of 25.5 6 4.0 kg/m
2
(range, 20.531.5 kg/m
2
)
and a mean baseline LDL-C of 4.32 6 0.63 mmol/L (range, 2.775.32
mmol/L). The 16 subjects who withdrew from the study had similar
characteristics to those who completed: 9 men, 7 postmenopausal
women; age 62 6 8 y (range, 5174 y); BMI 25.6 6 4.0 kg/m
2
(range,
21.337.1 kg/m
2
); and baseline LDL-C 4.24 6 0.92 mmol/L (range,
3.196.40 mmol/L). All subjects had elevated LDL-C levels on initial
assessment at recruitment (.4.1 mmol/L) despite the lower values in
some subjects at baseline, and triglyceride concentrations ,4.0 mmol/L.
No subjects used tobacco and none had clinical or biochemical evidence
of diabetes or liver or renal disease. Of the 27 participants who com-
pleted the study, 3 men and 5 women were taking the following medi-
cations: a hypolipidemic agent (statin) (n 2), b-blocking agents (n 3),
angiotensin-converting enzyme inhibitors (n 3), angiotensin II receptor
blockers (n 1), thiazide diuretics (n 2), levothyroxine (n 2), and
hormone replacement therapy (n 2). Medications had been stable for
at least 2 wk prior to the study. Medication dosages were kept constant
throughout the study.
Study protocol. Three 1-mo diet phases taken in a randomized cross-
over design with each phase separated by a minimum 2-wk washout
period were completed by 27 subjects. The 3 phases consisted of a mufn
phase (control) and 2 almond phases: 1 full-dose almond and the other
half-dose almond plus half-dose mufn (half-dose almond). During all
study phases, the background diet that the subjects followed was their
own self-selected low-fat therapeutic diet. Subjects were counseled on
strategies to facilitate weight maintenance, including holding exercise
constant throughout the study, and were questioned by the dietitian at
each clinic visit to ensure they had not deviated from their usual exercise
routine during the previous 2 wk.
After overnight fasts (1214 h), body weight, blood samples, and
blood pressure were obtained at the start and at wk 2 and 4 of each 4-wk
diet phase. A 24-h urine sample was also collected at the end of wk 4.
Seven-day weighed diet records were obtained prior to baseline (Table 1)
and at wk 4 (Table 2) of each phase. Subjects were instructed to weigh all
foods consumed with the self-taring electronic food scales provided
during the week when diets were recorded.
The Ethics Committee of the University of Toronto and St. Michaels
Hospital approved the study. All subjects gave informed consent. The
clinical trial registration number is: NCT00507520.
Diets. Before the study, all subjects had been instructed to follow a
therapeutic National Cholesterol Education Program Step 2 diet (,7%
energy fromsaturated fat and ,200 mg/d dietary cholesterol) for at least
2 mo prior to the study. Achievement of the dietary goals at baseline was
assessed by 7-d food records (Table 1). Subjects added 1 of 3 supplements
to their diet: whole rawunblanched almonds (73 63 g/d); mufns (147 6
6 g/d); and half portions of almonds (37 6 2 g/d) plus mufns (75 6 3
g/d) as described previously (1). All subjects took each supplement and
the intake level was based on subjects estimated daily energy require-
ment (26). The mufns were made from whole-wheat our with corn oil
sufcient to provide the same amount of SFA, PUFA, and ber as the
almonds, and with skim-milk powder and egg white to provide a similar
level of protein, although the mufn protein was 46% of animal origin.
MUFA from almonds balanced the starch from mufns. The macronu-
trient composition of the mufns as a percentage of energy was 14.7%
protein, 53.3% available carbohydrate, 32.1% fat, 4.3% SFA, 7.6%
MUFA, and 18.9% PUFA with 18 g/1000 kcal (4.3 g/1000 kJ) dietary
ber and 6 mg/1000 kcal (1.4 mg/1000 kJ) cholesterol. Nonhydro-
genated corn oil was the only fat used in mufn preparation to avoid the
addition of trans fatty acids to the diet. Mufn supplements were
provided at biweekly intervals and were stored in the freezer until the
day before use. Subjects were instructed to reduce total food intake,
especially starchy foods (breads, bagels, nonstudy mufns, and breakfast
cereals), to allow supplements to be taken as snacks without increasing
total energy intake and to keep the background diet constant across all 3
phases. Detailed dietary counseling was undertaken prior to and at wk
1 and 2 of each phase. During the study, we asked subjects to not
consume any additional nuts or nut products or alter consumption of
dietary ber or vegetable protein foods. Compliance was assessed from
7-d diet records (Table 2), a supplement checklist on which subjects
recorded supplements consumed and return of uneaten supplements,
which were weighed and recorded.
Analyses. Samples stored at 270C were used for determination of
serum vitamin A (retinol), a- and g-tocopherol, MDA, and urinary
isoprostanes. Following extraction with hexane, a- and g-tocopherol
and retinol were determined via reverse phase HPLC at UV 292 nm for
tocopherols and 540 nm for retinol according to Bieri et al. (27). MDA
was determined by reverse phase HPLC according to Volpi and Tarugi
(28), in which a thiobarbituric acid-MDA conjugate derivative was
injected onto a C18 column and uorometrically quantied at excitation
515 nm and emission 553 nm. MDA concentration was calculated from
calibration curves of authentic standard, with a linear relationship of
R
2
. 0.995. F
2a
-isoprostanes were measured in duplicate by GC/MS af-
ter isolation using HPLC according to the method described by Sacheck
et al. (29) and modied from that of Walter et al. (30). Briey, samples
were thawed and deuterated, and prostaglandin F
2
was added as an
internal standard. Pentaurobenzyl esters of isoprostanes were prepared
and puried by HPLC, silylated, and analyzed by GC/MS with a mass
selective detector operated in negative chemical ionization mode. Results
were obtained as mg/L and converted to pmol/L using a mean molecular
weight for isoprostanes of 354 atomic mass units. We determined urinary
TABLE 1 Baseline calculated macronutrient intakes
of all subjects
1
Variable Baseline
Energy, kcal (kJ) 1789 6 92 (7489 6 385)
Total protein, % 18 6 1
Vegetable protein, % 7 6 0.4
Available carbohydrate, % 54 6 1
Total dietary fiber, g/1000 kcal (g/1000 kJ) 16 6 1 (3.8 6 0.2)
Total fat, % 26 6 1
SFA, % 8 6 0.4
MUFA, % 10 6 1
PUFA, % 5 6 0.3
Dietary cholesterol, mg/1000 kcal (mg/1000 kJ) 100 6 8 (23.9 6 1.9)
Alcohol, % 2 6 0.6
1
Values are means 6 SEM, n 27; all % values reported as a percentage of energy.
Almonds and lipid peroxidation 909

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creatinine in the routine clinical laboratory at St. Michaels Hospital in
Toronto (Synchron LX 20, Beckman Coulter).
Study supplements were analyzed using AOAC methods for fat,
protein, and ber. Available carbohydrate was calculated as total carbo-
hydrate minus ber (31). The fatty acid composition was determined by
GC. Dietary macronutrient intakes (Tables 1 and 2) were assessed on the
7-d diet records, with an in-house computer program using the USDA
database (32). The percentage gures for soluble and insoluble ber were
derived from published data (31).
Power calculation. The power calculation was based on the ability to
detect a 4.5% difference in LDL-C in the primary analysis of the full-
dose almonds vs. the control assuming a 10% SD of effect (a 0.05 and
a b-1 0.8) and an anticipated drop-out rate of 25%. To satisfy these
specications, 40 subjects were required and 42 subjects were recruited.
However, the drop-out rate was 37%, permitting only a 5.6% difference
in LDL-C to be detected as signicant.
Statistical analyses. The results are expressed as means 6 SEM. The
data were analyzed using SAS software throughout (SAS Institute, 2004,
SAS 9.1 for Windows). Mixed models were constructed (SAS: Proc
Mixed) to assess the effect of diet on serum and urinary measures at wk
4. All models contained a subject effect to control for variance in the
outcome by accounting for the crossover nature of the study. All models
initially included diet, total cholesterol, subject sex, diet sequence, and
the interactions between sex and diet and sex and cholesterol. Nonsig-
nicant (.0.10) effects were removed sequentially until the model con-
tained signicant interactions or signicant main effects. The exception
was that the main effects of interest (sex and diet) were retained in all
reduced models. The nal model included subject as a random factor,
sex, diet sequence, interactions between sex and diet and sex and se-
quence, and baseline serum measures as a covariate. Diet sequence and
interactions were removed from the nal models if they were not
signicant. When diet was signicant, post hoc contrasts were used to
compare outcomes between the 3 diets. ATukey adjustment was used for
multiple means comparisons.
Results
Compliance with supplement consumption was good at 97.8 6
0.7% to 99.5 6 0.6% for all 3 treatments. Baseline dietary data
are shown in Table 1 and treatment dietary data are shown in
Table 2. Body weight did not differ at baseline and weight loss
did not differ between the full-dose almonds and the control,
although the half-dose almond treatment resulted in a weight
loss that was signicantly greater than the control (Table 3;
previously published data) (1). Blood lipid data have also been
published previously (Table 3) (1). The serum total cholesterol
concentration was signicantly lower in subjects while taking
both the half- and full-dose almond treatments compared with
the control treatments. Subjects taking the full-dose almond
treatment also had signicantly lower serum LDL-C and higher
serum HDL-C concentrations compared with the control
treatment (Table 3). The serum triglyceride concentration was
not affected by the treatments (Table 3).
Measures of oxidative damage. Serum MDA, a biomarker of
lipid peroxidation, decreased during the full-dose almond
intervention and this differed from the change during the
control period (P 0.040). The changes during the treatments
were: control, from 1.6 6 0.1 to 1.7 6 0.2 mmol/L; half-dose
almonds, from 1.4 6 0.1 to 1.6 6 0.3 mmol/L; and full-dose
almonds, from 1.6 6 0.1 to 1.3 6 0.1 mmol/L. The baseline
MDA concentration was a signicant predictor of the treatment
effect (P 0.002). The overall diet effect approached signif-
icance (P 0.051). Subjects 24-h urine volumes did not differ at
the ends of the 3 treatments. In 24-h urine collections, the
isoprostane concentrations during the control, half-dose, and
full-dose almond treatments were 63.0 615.0, 48.6 610.2, and
53.7 6 11.3 nmol/L, respectively. After creatinine correction,
urinary isoprostane outputs during the half- (0.7 60.2 nmol/mmol)
and the full-dose almond (0.8 6 0.2 nmol/mmol) treatments were
TABLE 3 Subjects body weight and serum lipids
before and after control, half-almond, and
full-almond treatments
1
Variable Week Control
2
Half-almond Full-almond
Body weight, kg 0 71.0 6 2.4 71.1 6 2.4 71.2 6 2.5
4 71.2 6 2.5
a
70.9 6 2.4
b
71.0 6 2.5
ab
Total cholesterol, mmol/L 0 6.54 6 0.16 6.47 6 0.15 6.60 6 0.16
Treatment 6.44 6 0.15
a
6.25 6 0.15
b
6.21 6 0.15
b
LDL-C, mmol/L 0 4.34 6 0.14 4.30 6 0.12 4.45 6 0.13
Treatment 4.22 6 0.13
a
4.10 6 0.12
ab
4.01 6 0.12
b
HDL cholesterol, mmol/L 0 1.43 6 0.09 1.38 6 0.08 1.40 6 0.08
Treatment 1.41 6 0.08
b
1.43 6 0.08
ab
1.45 6 0.09
a
Triglycerides, mmol/L 0 1.69 6 0.13 1.75 6 0.16 1.66 6 0.13
Treatment 1.80 6 0.12 1.58 6 0.11 1.64 6 0.11
1
Values are means 6 SEM, n 27. Means in a row with superscripts without a
common letter differ, P , 0.05.
2
Mean of wk 2 and wk 4 measurements.
TABLE 2 Calculated macronutrient intakes for the nal week of the control, half-almond and
full-almond treatments
1
Variable Control Half-almond Full-almond
Energy, kcal (kJ) 1900 6 89 (7950 6 372) 1951 6 94 (8163 6 393) 2034 6 98 (8510 6 410)
Total protein, % 17.5 6 0.5 17.6 6 0.5 17.4 6 0.4
Vegetable protein, % 7.2 6 0.3
b
7.8 6 0.3
ab
8.7 6 0.3
a
Available carbohydrate, % 54.5 6 1.1
a
48.4 6 1.1
b
44.8 6 1.2
b
Total dietary fiber, g/1000 kcal (g/1000 kJ) 16.3 6 1.1 (3.9 6 0.3) 16.5 6 0.9 (3.9 6 0.2) 16.4 6 0.9 (3.9 6 0.2)
Total fat, % 26.3 6 1.0
c
32.1 6 1.0
b
36.0 6 1.0
a
SFA, % 7.0 6 0.4 7.5 6 0.4 7.2 6 0.4
MUFA, % 9.0 6 0.5
c
14.5 6 0.5
b
18.9 6 0.6
a
PUFA, % 8.0 6 0.3 8.0 6 0.3 8.2 6 0.2
Dietary cholesterol, mg/1000 kcal (mg/1000 kJ) 89.7 6 7.3 (21.4 6 1.7) 96.8 6 6.2 (23.1 6 1.5) 79.3 6 5.3 (19.0 6 1.3)
Alcohol, % 1.8 6 0.5 1.9 6 0.5 1.8 6 0.6
1
Values are means 6 SEM, n 27. Means in a row with superscripts without a common letter differ, P , 0.05.
910 Jenkins et al.

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lower than during the control treatment (1.1 60.3 nmol/mmol)
(P , 0.03). The 24-h urinary isoprostane concentrations were
higher in women (83.9 615.0 nmol/L) than in men (31.4 613.6
nmol/L; P 0.016) as were the creatinine-corrected outputs
(1.5 60.2 vs. 0.4 60.2 nmol/mmol; P 0.001). The diet 3sex
interaction was not signicant for urinary isoprostane output
(P 0.415).
Also, urinary creatinine outputs during the half-dose almond
(8.9 6 0.7 mmol/d) and full-dose almond (8.6 6 0.7 mmol/d)
periods were higher than during the control period (7.1 6 0.5
mmol/d; P # 0.03).
Vitamin E and A. The serum concentrations of a- or
g-tocopherol as well as those of other vitamins, adjusted or
unadjusted for total cholesterol, were not affected by the
treatments.
Discussion
This study provides support for the antioxidant property of nuts
as an additional mechanism of action which, together with
lowering cholesterol, may account for the cardioprotective prop-
erty of nut consumption. Reactive aldehydes formed as products
of free radical-mediated lipid peroxidation can modify apolipo-
protein B. These more negatively charged particles are avidly
bound to scavenger receptors and thus are more likely to be
taken up by the arterial wall (33). Consequently, arteriosclerotic
plaques contain increased levels of oxidized lipids (34). In this
study, the full dose of almonds decreased serum MDA and both
the full and half dose of almonds decreased urinary isoprostanes
as biomarkers of lipid peroxidation. Previously, almond consump-
tion was shown to reduce concentrations of oxidized LDL,
the more atherogenic form of the particle (24,35). The present
data suggest that this may be a part of an overall lipid anti-
oxidant effect of almonds.
Nuts, unsalted, raw, or dry roasted, possess cardioprotective
properties (19,11,14,36,37). The focus has been on serum cho-
lesterol reduction as the primary mechanism, but this is modest
by comparison with the often dramatic benets for CHD risk
reduction demonstrated in cohort studies (1013). Nuts are also
rich sources of antioxidants. This property has been demon-
strated clearly for almonds (2022). For example, almonds are
among the richest food sources of vitamin E as RRR-a-
tocopherol. Almonds also contain a variety of phenolic compounds,
localized principally in their skin, including avonols (isorham-
netin, kaempferol, quercetin), avanols (catechin, epicatechin),
avanones (naringenin), anthocyanins (cyanidin, delphinidin),
procyanidins (B2 and B3), and phenolic acids (caffeic acid,
ferulic acid, p-coumaric acid, protocatechuic acid, vanillic acid)
(20,38,39). Almond avonols and avanols have been shown to
be bioavailable and contribute to the antioxidant protection
against LDL-C oxidation in vitro and in vivo (1,21,40).
We predicted that the higher intake of vitamin E, MUFA, and
phenolic constituents with almond consumption, and the inter-
actions between these nutrients, would increase the status of
vitamin E and decrease the level of lipid peroxidation, specif-
ically reducing the biomarkers of oxidative damage, serum
MDA, and urinary isoprostanes. Although our intervention did
not increase serum a- or g-tocopherol, which was seen in a
previous study (2), serum MDA and urinary isoprostanes were
signicantly reduced. The contributing antioxidant compounds
of almonds are therefore likely to be avonoids and phenolics
found in high concentrations in their skins (2022). These com-
pounds may contribute to the antioxidant property of almonds
not only independently but also via synergy with other antiox-
idants, including vitamin E and other components of the anti-
oxidant defense network (e.g. with vitamin C, where synergy has
also been observed) (21). In addition, MUFA, which is found in
high concentration in almonds, is also considered to have
antioxidant properties (36,40,41).
Reductions in oxidized LDL and lower urinary isoprostane
outputs have been reported after feeding soy and other vegetable
protein diets (42,43). In the present study, baseline urinary
isoprostanes were not measured. We assumed that after 4 wk of
the diet, a steady state would be reached that would allow the
almond treatments to be compared with the 100% mufn as the
control.
The reduced creatinine-corrected isoprostane output on
almonds may have resulted from altered creatinine metabolism
or renal function on almonds. It is possible that increased
urinary creatinine output on almonds may have been due to
increased meat intake or increased muscle turnover. However,
animal protein intake was lower during the almond phases and
there is no evidence that nut consumption increases protein
turnover. Increased creatinine clearance on almonds is a further
possibility. Unfortunately, we do not have the corresponding
serum measurements for this assessment and there is no
preexisting literature on this specic topic.
Our data did not show a dose response for antioxidant
activity, because both the half-dose and full-dose almonds
reduced urinary isoprostanes equally, suggesting the possibility
of a threshold effect for this biomarker.
The components of nuts such as avanols, avonols, ava-
nones, anthocyanins, procyanins, and phenolic acids all found in
almonds may be analogous to the isoavones in legumes,
especially soy, and the lignans in ax and whole-grain cereals
that have also attracted attention for their antioxidant proper-
ties. These substances with antioxidant activity could contribute
to the cardioprotective effect currently ascribed to nuts (1013).
Increased intake of MUFA in the diet has also been associated
with reduced susceptibility of LDL to oxidation (41). Thus,
there are a number of components of almonds that may con-
tribute to the reduction of LDL oxidation and would be
predicted to reduce oxidative damage, as observed in this study.
Vitamin E did not appear to inuence blood levels despite an
increased intake on the full-dose almond diet by 18 mg/d. Our
results contrast with those of Jambazian et al. (2) who found
that, relative to baseline, feeding almonds for 4 wk at 28 and 56
g/d signicantly increased plasma a-tocopherol from 26.3 to
29.9 mmol/L. However, our baseline a-tocopherol levels were
.40% higher than those reported by Jambazian et al. (2), i.e.
;44 compared with 31.2 mmol/L. In NHANES III, serum
vitamin E concentrations for the 50th and 90th percentiles in
men were 25.9 and 43.6 mmol/L, respectively, and were 29.7 and
48.3 mmol/L, respectively, for women (44). Thus, our subjects
started the study in the top decile of a-tocopherol status of the
population. Signicantly higher concentrations of a-tocopherol
are difcult to achieve by diet alone. The reason for our subjects
higher prestudy vitamin E levels is not clear.
It is also possible that the antioxidant benet of almonds
demonstrated as a reduced concentration of oxidized LDL may
have been associated with other antioxidant systems not mea-
sured in the present study. Acute test meal studies have shown
that almond supplementation of a bread meal reduced post-
prandial destruction of serum protein thiol groups (45). Serum
protein thiols were not measured in the present study. Never-
theless, both serum MDA and urinary isoprostanes are consid-
Almonds and lipid peroxidation 911

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ered to be reliable biomarkers of oxidative stress and would be
expected to be favorably affected, because oxidative stress is
reduced by the provision of exogenous antioxidants (46,47).
The issue of antioxidants and CHDhas not been a simple one
(48). Early cohort studies demonstrated the protective effect of
vitamin E in preventing CHD (49,50). The theoretical basis for
oxidative damage to lipids, lipoproteins, and DNA was devel-
oped (35,46,51) with relevance to CHD and diabetes (5254).
However, antioxidants, including vitamin E, or carotenoids
given singly or in combination as supplements have not demon-
strated effectiveness in reducing the risk of cardiovascular dis-
ease in randomized controlled trials (5557). At the same time,
these trials did not undertake any assessment of biomarkers of
oxidative stress to determine the effectiveness of the dose or
duration of the supplements as antioxidants (58). It therefore
still remains possible that antioxidants may have a cardiopro-
tective role if a meaningful reduction in oxidative damage can be
achieved by dietary or pharmacologic interventions.
We conclude that the antioxidant potential of almonds was
demonstrated in this study as reductions in serum MDA and
urinary isoprostanes as markers of lipid peroxidation. However,
serum vitamin E concentrations did not increase despite an
additional 18 mg/d for the full-dose almonds. It is likely that the
antioxidant property of almonds we obtained was related to the
effect of avonoids and other phenolic antioxidants and the high
MUFA content of almonds (50% of calories). Flavonoids and
other phenolic antioxidants in almonds may reduce oxidative
damage directly as well as indirectly via synergy with vitamin E
and other components of the antioxidant defense network. The
antioxidant ingredients in nuts, combined with their favorable
effects on the blood lipid prole, may help to explain their
success in cohort studies in protecting from CHD (1013).
Acknowledgments
We acknowledge the statistical expertise of Mr. Edward Vidgen,
Toronto, and Dr. Laurel Duquette, the Department of Statistics,
University of Toronto.
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