3. Results
Table 1 shows baseline characteristics of 180 subjects in relation with four types of diet. The proportion of
men was higher in diet III and IV, which included wine. In the same way, the percentage of current cigarette
smokers was higher among these categories. Cigarette smoking data showed striking differences between sub-
jects who drank wine 7.4 and 5.5 gday of tobacco in diets III and IV, respectively versus those who did not,
in diets I and II tobacco consumption was approxi- mately 2.2 gday. Lipid profile was characterized by the
highest levels of plasma total cholesterol and HDL cholesterol in diets III and IV. Blood levels of triacyl-
glycerol were higher in diets with alcohol consumption than in the other diets, but the differences were not
significant. No significant difference between the four groups was observed for age, blood pressure, BMI and
WHR.
Table 2 presents nutrient protein, fat, carbohydrate, food dairy products, meat, egg, fish, vegetable and
fruit, alcohol total alcohol and wine and catechin intakes. Subjects in diet I and II did not drink wine and
those in diet I and III did not consume vegetable and fruit or very marginally. Total energy intake was higher
among subjects who drank alcohol and was approxi- mately twice the energy intake of diet I and II. The
highest value 4207 kJ was observed in diet III. The highest mean values of total energy were due to alcohol
intake mainly. Energy supplied by alcohol accounted for 17.6 and 17.3 of total energy intake for diet III
and IV, respectively. The highest amount of alcohol was provided by wine consumption ranging from 86 to
88 in diet III and IV, respectively. Diet I was charac- terized by the lowest meat, egg and fish intake, and by
the highest consumption of dairy products. The amount of dairy products per MJ 59.7 gMJ was in this diet
twofold higher than in diet II 29.6 gMJ and fourfold- higher than in diet IV 14.0 gMJ. Comparisons of
absolute mean values of catechin intake showed large differences between diet III and IV on the one hand
and diet I and II on the other hand. When catechin intake was based on MJ consumed, the differences were
smaller. Diet I supplied the lowest amount of dietary catechin 1.4 mgMJ when compared with diet II 8.1
mgMJ and the difference was more significant when compared with diet IV which included wine consump-
tion 20.6 mgMJ.
BMI was not significantly different in the four diets. In contrast, total energy intake was significantly higher
in diets III and IV than in diets I and II. After subtraction of energy supplied by alcohol in the corre-
sponding groups, and after adjustment for sex, smoking habits, age and physical activity, no significant differ-
Table 1 Descriptive characteristics of 180 subjects in relation to four types of diet
Diet groups P
I
a
n = 24 II
b
n = 75 III
c
n = 22 IV
d
n = 59 41.7
43.9 Percent of male
e
81.8 67.8
B 0.001
NS 27.1
31.8 Physical activity
f
22.4 25.0
B 0.01
Smoking 27.3
24.7 41.7
Ex-smoker 40.7
15.6 Current smoker
36.4 16.7
33.9 B
0.05 2.3 6.3
Cigarette consumption gday
g
7.4 12.4 2.2 5.5
h
5.5 10.9 NS
49.9 7.4 46.1 7.7
Age year 48.2 7.9
47.7 8.7 NS
BMI kgm
2 i
24.7 4.8 24.8 4.8
24.4 3.3 24.5 3.1
WHR
i
NS 0.87 0.06
0.88 0.06 0.87 0.07
0.87 0.09 130.9 16.3
129.5 15.1 NS
131.3 18.1 130.2 19.4
Systolic blood pressure mmHg
i
NS 81.5 11.6
Diastolic blood pressure mmHg
i
80.4 10.1 80.8 8.2
80.7 9.1 5.44 0.88
k
5.84 0.92 Total cholesterol mmoll
i
5.87 1.03 6.16 1.02
B 0.05
HDL cholesterol mmoll
i
1.33 0.34
k
1.48 0.36 1.50 0.27
1.58 0.35 B
0.05 1.35 0.95
1.08 0.37 1.21 0.43
Triacylglycerol mmoll
i,j
NS 1.27 0.38
a
Without vegetable, fruit and wine.
b
With vegetable and fruit.
c
With wine and without vegetable and fruit.
d
With wine, vegetable and fruit.
e
x
2
-Test.
f
Intense physical activity for at least 20 min, three times a week or more.
g
One cigarette = 1 g of tobacco, Kruskal–Wallis test.
h
X S.D..
i
Adjusted for sex.
j
Processed after log transformation.
k
Significantly different from diet IV, PB0.05 ANOVA+Scheffe test.
Table 2 Food, nutrient, alcohol and catechin intakes evening meal in 180 subjects according to four types of diet
P Diet groups
I
a
n = 24 II
b
n = 75 III
c
n = 22 IV
d
n = 59 2531 948
h,i
4207 2092 Total energy kJ
e
3587 1307 2281 2079
f,h,i
B 0.001
Total alcohol g 0.4 1.7
0.3 1.9 22.0 15.2
20.6 13.6 –
0.3 1.7 236.3 140.8
Wine ml 228.7 149.8
– –
– 86
88 Percent of alcohol from wine with regard to total
– alcohol
Percent energy without alcohol 20.7 7.9
23.3 7.0 18.2 6.3
20.8 8.2 Protein
NS 38.4 13.7
36.3 13.8 Fat
40.3 15.4 41.9 16.1
NS 40.9 15.3
40.4 12.2 39.9 17.8
38.9 15.0 Carbohydrate
NS Percent alcohol from total energy
0.5 3.4 0.1 0.5
17.6 13.3 17.3 10.3
Density gMJ
of 29.6 35.1
25.8 24.9 Milk, cheese
e
14.0 17.1 59.7 72.4
j
B 0.001
34.2 26.1 30.8 13.2
23.0 23.0 30.9 19.5
Meat, egg, fish
e
NS 88.2 57.1
– Vegetable, fruit
54.1 40.1 1.3 4.0
– 19.3 24.1
h,k
65.6 35.5 3.0 2.4
h,i,k
71.2 41.7 Catechin mg
e
B 0.001
8.1 9.3
h,k
Catechin mgMJ
e,g
17.9 11.1 1.4 1.0
h,i,k
20.6 10.9 B
0.001
a
Without vegetable, fruit and wine.
b
With vegetable and fruit.
c
With wine and without vegetable and fruit.
d
With wine, vegetable and fruit.
e
Kruskal–Wallis test.
f
X S.D..
g
Based on MJ consumed.
h
Significantly different from diet III, PB0.001 Kruskal–Wallis+Mann–Whitney test. Mann–Whitney significance level adjusted for the number of pairs.
i
Significantly different from diet IV, PB0.001, Kruskal–Wallis+Mann–Whitney test. Mann–Whitney significance level adjusted for the number of pairs.
j
Significantly different from diet IV, PB0.01 Kruskal–Wallis+Mann–Whitney test. Mann–Whitney significance level adjusted for the number of pairs.
k
Significantly different from diet II, PB0.001 Kruskal–Wallis+Mann–Whitney test. Mann–Whitney significance level adjusted for the number of pairs.
ences in energy intake were observed between the four diets the average energy intake was, 2266 kJ in diet I,
2554 kJ in diet II, 3164 kJ in diet III and 2771 kJ in diet IV.
Table 3 shows the correlations between the evening meal recall and the 3-day food record for some nutri-
ents and foods. A positive correlation was found between the nutri-
tional pattern of the evening meal food recall and the 3-day food record. Pearson correlation coefficients for
nutrients ranged from 0.40 for carbohydrates to 0.60 for protein intake. All these relationships were statisti-
cally highly significant. For total alcohol, Pearson cor- relation coefficients were 0.70 P B 0.001 when the
evening meal recall was compared to the 3-day food record and 0.73 P B 0.001 when the evening meal
food recall was compared to the alcohol consumption assessed by the quantitative recall frequency question-
naire. The coefficients were similar for wine, 0.69 and 0.80, respectively. The relationship between the evening
meal and the 3-day food record was consistent. The evening meal supplied a good estimation of usual nutri-
tional habits. On average, evening meal contributed a third 34 of total energy intake during a day. This
proportion varied according to the various types of diet. A total of 35, 26, 42 and 38 of daily energy
intake was supplied by the evening meal in diet I, II, III and IV, respectively. Fig. 1 shows mean plasma concen-
tration of + -catechin according to different types of diet. The differences found in + -catechin concentra-
tions in plasma between the four diets were statistically significant overall test P B 0.001. The highest concen-
trations were found in diets III 577.4 mgl and IV 673.4 mgl and the lowest in diet I 133.9 mgl. Diet II
had an intermediate plasma level 456.1 mgl. After adjustment for age, sex, smoking, BMI, WHR and total
energy intake, the lowest concentration of + -catechin in plasma was found in diet I 131.6 mgl. + -Cate-
chin level was threefold higher in plasma when veg- etable and fruit 449.5 mgl were consumed diet II
and fourfold higher when only wine 598.5 mgl was consumed diet III. When consumption of vegetable,
Table 3 Pearson correlations between evening meal food recall and the 3-day
food record for some nutrients and food Nutrientsfood
P r
0.59 Energy
B 0.001
Protein B
0.001 0.60
B 0.001
Animal protein 0.52
B 0.001
0.49 Vegetable protein
Fat 0.45
B 0.001
B 0.001
0.55 Saturated fatty acids
B 0.05
Polyunsaturated fatty acids 0.31
B 0.01
0.35 Monounsaturated fatty acids
B 0.01
Carbohydrates 0.40
B 0.01
0.28 Oligosaccharide
B 0.001
Polysaccharide 0.54
Meat B
0.01 0.34
Fruitvegetable 0.42
B 0.01
B 0.01
0.35 Dairy products
0.69
a
–0.73
b
Total alcohol B
0.001 0.70
a
–0.80
b
Wine B
0.001
a
Between evening meal recall and 3-day record.
b
Between evening meal recall and weekly recall.
were, 125.8, 209.1, 190.1 and 225.2 mgl in diets I, II, III and IV, respectively.
The relationship between dietary catechin and + - catechin concentration in plasma was evaluated in the
entire population sample using the Pearson correlation coefficient r = 0.58, P B 0.001. + -Catechin concen-
tration in plasma depended on the amount of dietary catechin and on the length of the fasting period be-
tween the last food intake and the blood collection. In this study, the ratio plasma + -catechindietary cate-
chin was negatively related to the length of the fast Pearson correlation coefficient = − 0.20, P B 0.01.
The average fasting period was 11 h and 55 min rang- ing from 10 to 15 h. The length of the fasting period
was not significantly different between the four types of diet. The average difference was 44 min between diet I
the shortest fasting period, 11 h and 23 min and diet III the longest fasting period, 12 h and 7 min. After
adjustment for the length of the fasting period, differ- ences in the + -catechin concentration in blood re-
mained significant.
Table 4 shows the contribution of food to plasma concentration of + -catechin in a general linear
model. Vegetable and fruit and even more wine intake were the major determinants of plasma concentration
of + -catechin P B 0.001, whereas the contribution of dairy products, meat, egg and fish was marginal.
Forty six percent of the variance of + -catechin con- centration in plasma was explained by the intake of
dietary catechin. fruit and wine was combined, the + -catechin concen-
tration had the most elevated level 637.1 mgl. When plasma concentration of + -catechin was based on
MJ energy intake, differences between the four diets were smaller. Plasma concentrations of + -catechin
Fig. 1. Plasma concentration of + -catechin according to four types of diet among 180 subjects means with S.E.M..
, Crude values ANOVA analysis, overall test, P B 0.001;
, adjusted for age, sex, smoking habits, BMI, WHR and total energy intake ANCOVA analysis, overall test,
P B 0.001; b, plasma concentration of +-catechin based on MJ consumed, adjusted for age, sex, smoking habits, BMI and WHR ANCOVA analysis, overall test; P B 0.001, after log transformation. I, without vegetable, fruit and wine; II, with vegetable and fruit; III, with wine and
without vegetable and fruit; IV, with wine, vegetable and fruit. , Significantly different from corresponding bar of diet II, III and IV, P B 0.001 Scheffe test; †, significantly different from corresponding bar of diet II, IV, P B 0.001 and III, P B 0.05 Scheffe test; ‡, §, significantly different
from corresponding bar of diet IV, ‡ P B 0.001, § P B 0.01 Scheffe test.
Table 4 Contribution of food to plasma concentration of +-catechin mg
l
a,b
SEE n = 180
Partial R
2
b P
0.020 Dairy products
c
0.01 −
0.026 0.19
Meat–egg–fish
c
0.029 0.023
0.01 0.20
0.016 0.25
0.0001 Vegetable–fruit
c
0.126 0.015
0.32 0.0001
0.133 Wine
c a
Analysis performed with logcatechin.
b
General linear model.
c
Expressed as logdensity in gMJ for dairy products, meat–egg– fish, vegetable–fruit and in mlMJ for wine, coefficient of determina-
tion, R
2
= 0.46; PB0.001.
When compared to recent studies, the higher catechin levels found in our study can be explained by the fact
that each subject had a complete evening meal and not only a single beverage or food such as, wine [27], tea
[28,29], or chocolate [30]. High catechin levels, found in plasma 9 – 12 h after the meal might also be the conse-
quence of a higher initial level of catechin before the evening meal no flavonoid depletion diet, or of cate-
chin protection induced by other antioxidants present in blood, either natural, vitamin E, vitamin C, b-
caroten, selenium or non-natural such as terbutylhy- droquinone added during the plasma manipulation
phases before the analysis. As indicated by Chen [31] other components of the diet could have affected
plasma concentration and the elimination of flavonoids. Interspecies differences in metabolism of flavonoids
were significant and depended on several parameters such
as intestinal
microflora, able
to hydrolyze
flavonoid conjugates [32]. For human beings, 3 h after green tea intake, total
level of flavonoids such as, epicatechin, epicatechingal- late and epigallocatechingallate could reach levels up to
1100 ngml [33]. These results did not take into account catechin, anthocyanine flavonols such as quercitin and
metabolites, and flavonoids might reach higher levels if catechin, anthocyanine, flavonol such as quercitin and
metabolite compounds were quantified. Generally, a clear dose – response relationship was not achievable
when the subject sample was too small.
A recent comparison between detection limits for plasmatic catechin and derivatives by HPLC – UV,
HPLC Coulochem Electrode Array System, HPLC- fluorimetry and GCMS showed that only the last two
had the highest sensitivity close to 3 ng, whereas the others reached 20 ng [34]. Flavonoid levels from plasma
found after using different extraction and detection methods reached up to 10 of CV intra-day and inter-
day for HPLC – UV and HPLC fluorescence [25], for HPLC Coulochem Electrode Array System [35]. The
several stages of plasmatic catechin extraction may have impaired the results due to the potential loss
occurring during the various recuperation phases before the injection in HPLC or GC.
Significant individual variability could be observed and so far, no explanation has ever been given. Identi-
cal results had already been reported in several studies for flavonoids epicatechin, epigallocatechin and epigal-
locatechingallate. Flavonoid plasmatic levels measured in a small number of subjects around five or six within
the same period of time after food or beverage intake could be multiplied by six [35]. Finally, in our study,
the same methodology used to perform catechin mea- surement among 180 subjects, enabled comparisons of
plasma catechin levels between various diets.
Several reports [36 – 39] have shown a positive rela- tionship between alcohol consumption and WHR, but
4. Discussion
Kategori