PROJECT N°47
Omega-3 alpha-linolenic fatty acid (ALA) in Chia seeds (Salvia Hispanica L.)
and its influence on human performances
Inès Bahlawane, Lavinia Kadar
European School Luxembourg II
6, rue Gaston Thorn, 8268 , Bertrange, Luxembourg
S6 DE
Abstract
This project, led by Lavinia Kadar and Inès Bahlawane under the
supervision of Eric Castanet (EE Lux2) is divided in two parts.
In the first part, we want to show that the very beneficial ALA in chia
seeds can be used in the human body. Therefore, a simulation of the
human digestive system will be made. The acid should be detectable
after the experiments (conducted in school lab).
The objective of the second part is, to find out if chia seeds have an
influence on human performances. We carried out a study where 19
students from 4th and 2nd year participated in a test, containing a
memorization part, in which the students had to remember a quantity
of numbers with time limit, pull-ups( with two attempts) and a 800m run.
Then the students were given the recommended quantity of chia seeds
on a daily basis for six weeks. After this they were tested again.
The most likely result would be an improvement in the long distance run
and in memorization. The improvement of the female participants
should be higher than the one of the male ones.
In conclusion, our project aims to show how beneficial the “super food” is in reality, and if Chia seeds
are a good alternative for vegetarians (who don’t eat fish) to replace the essential omega-3 fatty.
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1. Introduction
Chia seeds, an aliment of the ancient Maya, were recently rediscovered as a so-called super food. It
has been shown several times that the seeds do not only contain plenty of essential omega-3 fatty
acids but also the recommended balance of omega-6 fatty acids. The food we consume in western
countries contains too many omega-6 in comparison to omega-3 fatty acids. Approximately 60% of the
chia seed oil contains omega-3 α-linolenic fatty acid (ALA), ligated in fat molecules.
ALA is very beneficial because of its anti-inflammatory effects.
Normally, enzymes in the human body turn linoleic acid (LA) into arachidonic acid (AA), which
promotes inflammation, when eicosanoids series are metabolized.
In presence of ALA, those enzymes are attracted to turn ALA into eicosapentaenoic acid (EPA) which is
turned into docosahexaenoic acid (DHA), which eventually leads to AA being produced in a lower
quantity. DHA and EPA are two other omega-3 fatty acids, which have beneficial impacts on humans.
The anti-inflammatory series-3-eicosanoids are also produced from the omega-3 fatty acids.
EPA and DHA are contained in a higher quantity in fish than in chia seeds. It is after all more profitable
to eat fish than to consume chia. So are the seeds a good alternative to fish for vegetarians?
1. Part of the project: A simulation of the human digestive system
Chia seeds are composed of many different substances. External is a
layer of polysaccharide and water-soluble dietary fibres (which swell
when the seeds are put into water). The shell is made of cellulose and
hemicellulose. The fat molecules on the inside of the shell are covered
by lignin, a substance also found in wood.
Is it possible for the human digestive system to open up the lignin
before the small intestine, so that the ALA can be split from the glycerine
and used?
2. Part of the project: A study with students of our school
It is said that chia seeds provoke an improvement in the physical and mental condition of humans
which is also due to ALA. The effects should be greater for women than for men.
Are these effects sufficient to notice a difference in physical and mental condition when chia seeds are
ingested?
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2. Materials
The simulation of a human digestive system – materials
Can the omega-3 α-linolenic acid be extracted from the seed by breaking-up the Lignin?
To find out, under what conditions the enzymatic reactions were the best, we did a row of pretests
based on information found in the internet.
The reagent number 2.1.1. had the highest quantity of unsaturated fatty acids, which shows us, that
this digestion worked the best and that it is possible for our body to split the lignin, which encloses
the fat molecules.
Materials used for the Tests
1. Cavum oris
Reagents
Equipment
2. Ventriculus
Chia seeds (Salvia Hispanica L.)
Reagents
HCl (0,1M)
Distilled water
Dry Pepsin
Dry α-Amylase
Sample of the cavum oris
14 test tubes
Equipment
14 Ziplock bag
Mortar
Micropipette
Water bath
Water bath
Spattle
Thermometer
2 beaker glasses
2 Pipettes
Pipette
Scales
3. Duodenum
Reagents
4. Intestinum
Dry pankreatin
Reagents
none
Dehydrated Bile
Equipment
none
Destilled Water
NaOH (0,1M)
Equipment
12 test tubes
Pipette
Water bath
14 glass batons
PH-meter
Balance
2 Spatulas
3
3. Methods
The simulation of a human digestive system – Methods
(Quantities of the test tube 2.1.1, this method was repeated for each test tube with different
quantities of the reagents; see below)
1. Digestion in cavum oris
a. 4g of chia seeds has to swell in 100ml destilled water for 30 min
b. Pestle 5,5ml of swelled chia seeds
c. Add 0,1ml of Amylase solution (2g dry Amylase + 50ml H2O)
d. Heat up to 37 °C for 20 seconds
2. Digestion in ventriculus
a. Put 0,5ml pepsin solution (1g dry Pepsin + 30ml HCl) in a zipbag
b. Heat up to 37 °C in water bath
c. Add sample from digestion in cavum oris
d. Knead for 30 min by 37 °C
3. Digestion in duodenum and intestinum
a. Test the pH value of pancreatin
b. Add 1 drop NaOH (0,1mol/L) to adjust the pH value to 7.1-8.2
c. Hydrate 1g dry Bile with 15ml H2O
d. Hydrate 1g dry pancreatin with 25ml H2O
e. Put 0,6ml Bile solution and 0,3ml Pancreatin solution in a test tube
f.
Add sample from digestion in Ventriculus
g. Stir with a glass baton
h. Heat up to 37 °C for 4 hours
i.
Freeze to stop the enzymatic reaction (for a week)
Quantities for the tests
Chia s.: 4g Chia + 100ml H2O (5min)
Test tubes with different quantities of enzymes
AmyA: 1g dry Amylase + 50ml H2O
1.1.1: 5,5ml Chia s.+0,1ml AmyA +0,5ml Peps. s. + 1 drop
AmyB: 2g dry Amylase + 50 ml H2O
NaOH +0,6ml Bile s.+0,3ml Pankr. s.
Peps. s.: 1g dry Pepsin + 30ml HCl (0,1M)
1.1.2: 5,5ml Chia s.+0,1ml AmyA +0,5ml Peps. s. + 1 drop
Bile s.: 1g dehydrated Bile + 15ml H2O
NaOH +0,3ml Bile s.+0,3ml Pankr. s.
Pankr. s.: 1g dry Pankreatin + 25ml H2O
1.1.3: 5,5ml Chia s.+0,1ml AmyA +0,5ml Peps. s. + 1 drop NaOH +0,3ml Bile s. +0,6ml Pankr. s.
1.2.2: 5,5ml Chia s.+0,1ml AmyA+0,2ml Peps. s. + 1 drop NaOH +0,3ml Bile s. +0,3ml Pankr. s.
1.2.3: 5,5ml Chia s.+0,1ml AmyA +0,2ml Peps. s. + 1 drop NaOH+0,3ml Bile s.+0,6ml Pankr. s.
2.1.1: 5,5ml Chia s.+0,1ml AmyB +0,5ml Peps. s. + 1 drop NaOH +0,6ml Bile s.+0,3ml Pankr. s.
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2.1.2: 5,5ml Chia s.+0,1ml AmyB +0,5ml Peps. s. + 1 drop NaOH +0,3ml Bile s.+0,3ml Pankr. s.
2.1.3: 5,5ml Chia s.+0,1ml AmyB +0,5ml Peps. s. + 1 drop NaOH +0,3ml Bile s.+0,6ml Pankr. s.
2.2.2: 5,5ml Chia s.+0,1ml AmyB +0,2ml Peps. s. + 1 drop NaOH +0,3ml Bile s.+0,3ml Pankr. s.
2.2.3: 5,5ml Chia s.+0,1ml AmyB +0,2ml Peps. s. + 1 drop NaOH +0,3ml Bile s.+0,6ml Pankr. s.
After unfreezing the seeds, the detection reaction for unsaturated fatty acids was performed to find
out which reagent reacted the best.
Detection of unsaturated fatty acids
a. Add the same quantity of bromine water (300 microliter) to each reagent
b. Observe if the bromine water was decolorized
c. To the decolorized reagents were added 300 microliter of bromine
water more
d. Observe which one of the reagents decolorized the most.
(e. Negative tests were made with undigested chia seeds and distilled water)
4. Results
The simulation of a human digestive system – Result
By using the optimal amount of enzymes, the optimal buffer and a stable temperature (37°) the
digestion was able to separate the unsaturated fatty acids from the lignin and the rest of the seeds.
Considering the fact that ≈56% of the unsaturated fatty acids contained in chia seeds is the very
beneficial alpha-linolenic acid and that the fatty acids could easily be detected we assume that the ALA
can be separated from the other components. Due to the separation in the intestine the beneficial
fatty acid can be taken in and used by the human body.
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The path of the digestive system from different components of chia seeds
Polysaccharides
Dietary
Fiber
Proteins
Fats
Cellulose
Mouth: Amylase
Split into double
sugar
/
/
/
Broken up
Stomach: HCl und
Peptids
/
/
Split into
Polypeptides
/
/
/
Fatty acids
Amino acids
and
Glycerin
Duodenal: Pancreatic
juice, Bile
Split into
monosaccharide’s
/
5. Methods
The study – methods
Are the effects of chia seeds sufficient to notice a difference in physical and mental condition of
humans?
Student participants (18):
-
8 girls (4 of S4 + 4 of S2)
-
10 boys (6 of S4 + 4 of S2)
Time interval:
6 weeks (including 2 weeks of Christmas vacations)
Amount of Chia seeds intake:
Recommended quantity (5g) of dry chia seeds swollen in 100ml of water for 30 min each
morning
Tests before and after the intake of chia seeds containing:
-
Memorization part: an exponentially increasing quantity of numbers (4 to 25) to be memorized
in a predetermined time (30 seconds)
-
Pull-ups (numbers counted) with two attempts, the better result counted
-
Long distance run: time measured (800m)
The tests persons were separated in three groups for the three different tasks. Each test was led by
one assistant person. The results were separately listed. The second and final test was in the same
period of the same weekday than the first test. The participants undertook the tests in the same order
both times. At the three stations, the same assistant person noted the results of each student, so that
the outcome is the most exact possible.
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6. Results
The study – Results
A clear improvement appeared in the memorization part, the students had an average of 6,5% better
result after the intake of chia seeds. The girls result showed an average of 0,8% higher improvement
than the boys, a result similar to our expectations. Amelioration also appeared in the pull-up part. In
average the probands achieved 0,47 pull-ups more in the second test than in the first one. In this part
the girls also showed better results (the girls improvement is higher by 0,4 pull-ups). In the last task,
the long distance run, no noticeable difference was visible. There was in average an aggravation of
0,11 seconds what could be due to the fact, that 2 of the 6 weeks of chia intake were Christmasvacations for the students. Against the expectations, the improvement of the boys was 12,85 seconds
better than the improvement of the girls. There is no clear reason to explain this observance.
The Memorization part of the study:
(y-axis: percentage of right answers)
70,0%
60,0%
50,0%
40,0%
Test 1
30,0%
Test 2
20,0%
Improvement
10,0%
0,0%
Girls
Boys
both
The Pull-ups part of the study:
(y-axis: number of pull-ups)
6
5
4
Test 1
3
Test 2
2
Improvement
1
0
Girls
Boys
both
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The long distance run part of the study:
(y-axis: time in seconds)
300
250
Test 1
200
Test 2
150
100
Improvement/
Aggravation
50
0
Girls
Boys
both
-50
7. Discussion and Conclusion
Through the first part of this project, it is proved, that the ALA in Chia seeds can be absorbed of the
human body, so that it can be used. Even, if we don’t know, if all of the ALA contained in the seeds can
be split out of the Lignin, it is sure, that the quantity is sufficient to show some beneficial effects to the
human body (which we can see in the second part of this project) such as memorizing abilities and
short-time muscle activity.
In conclusion, the regular intake of chia seeds is profitable for no-fish eaters because it increases the
level of essential omega-3 fatty acids.
8. Acknowledgement
We would like to show our gratitude to M. Wedel, vice director of our school who made this project
possible by granting us all the support the school could give us.
We also thank M. Castanet for accepting to be our mentor without knowing us, and for giving us
support whenever we needed it.
We would like to thank M. Stefanic for supporting us and answer all the questions we had and for
mostly taking the role of our mentor when our mentor couldn’t help us due to health reasons.
We are also very grateful to Christelle Bahlawane, microbiologist for lending us tools the school
doesn’t have and for her scientific advice although any errors are our own.
Our acknowledges also goes out to the language teachers, Tiffany and Chiara who accepted to proof
read our report, and to the 19 students from our school who accepted to take part in our study.
Our gratitude also goes to Soraya Bahlawane for taking over one of the 3 stations from the study
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9. References

Metabolism of linoleic acid by human gut bacteria: different routes for biosynthesis of
conjugated linoleic acid by Estelle Devillard, Freda McIntosh, Sylvia Ducan, John Wallace

https://www.vimeda.de/magazin/gesundheit/biomarker/alpha-linolensaeure (15.10.2016)

Chia seeds does not promote weight loss or alter disease risk factors in overweight adults by
David Nieman, Erin Cayea, Melanie Austin, Dru A. Henson, Steven R: McAnulty, Fuxia Jin

No positive influence of Ingesting Chia seed oil on Human Running Performance by David
Nieman, Nicholas Gillitt, Mary Pat Meaney , Dustin A. Dew

Raman Spectroscopic Barcode Use for Differensation of Vegetable Oils and Determination of
their Major Fatty Acid Composition by Serap Durakli Velioglu, Elif Ercioglu, H. Tumay Temiz,
Murat Velioglu, Ali Topcu, Ismail H. Boyaci

https://draxe.com/chia-seeds-benefits-side-effects/ (02.01.2017)

Chia seed supplementation and disease risk factors in overweight women: a metabolomics
investigation by Nieman DC1, Gillitt N, Jin F, Henson DA, Kennerly K, Shanely RA, Ore B, Su M,
Schwartz S.

https://pubchem.ncbi.nlm.nih.gov/compound/linolenic_acid (09.01.2017)

https://en.wikipedia.org/wiki/Alpha-Linolenic_acid (11.12.2016)

http://www.biothemen.de/Heilpflanzen/steckbrief/chia.html (04.12.2016)

http://www.spiegel.de/gesundheit/ernaehrung/chia-samen-im-check-gut-aber-nichteinzigartig-a-1056210.html (29.11.2016)

http://www.chia-samen.info/naehrwerte-kalorien-inhaltsstoffe.html (13.01.2017)

http://chia-samen-bio.de/naehrwerte-inhaltsstoffe/ (13.01.2017)

http://www.omega-3.ch/omega-3/omega-3-menude-116/das-kleine-abc-ber-omega-3menude-52.html (12.01.2017)

http://thepaleodiet.com/seed-fatty-acid-composition/ (05.01.2017)

https://www.zentrum-der-gesundheit.de/chia-samen.html (10.02.2017)

https://notfromapacketmix.com/2014/01/24/chia-seeds-and-fructose-malabsorptionfodmap/ (09.02.2017)
https://de.wikipedia.org/wiki/Α-Linolensäure (05.02.2017)

https://en.wikipedia.org/wiki/Arachidonic_acid (04.02.2017)

https://de.wikipedia.org/wiki/Eicosanoide (04.02.2017)

https://de.wikipedia.org/wiki/Omega-3-Fettsäuren#.CE.B1Linolens.C3.A4ure_.28ALA.29_aus_Pflanzen_gegen.C3.BCber_Eicosapentaens.C3.A4ure_.28E
PA.29_und_Docosahexaens.C3.A4ure_.28DHA.29_aus_Fisch.C3.B6l (04.02.2017)
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
https://de.wikipedia.org/wiki/Bauchspeicheldrüse (15.01.2017)

http://www.onmeda.de/anatomie/gallenblase-funktion-der-gallenblase-19969-3.html
(09.01.2017)

http://flexikon.doccheck.com/de/Pankreassekret (09.01.2017)

http://www.chemie.de/lexikon/Linolensäure.html (01.10.2016)
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